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WO2025255541A1 - Haploid inducer strawberry lines and methods of producing and using thereof - Google Patents

Haploid inducer strawberry lines and methods of producing and using thereof

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Publication number
WO2025255541A1
WO2025255541A1 PCT/US2025/032766 US2025032766W WO2025255541A1 WO 2025255541 A1 WO2025255541 A1 WO 2025255541A1 US 2025032766 W US2025032766 W US 2025032766W WO 2025255541 A1 WO2025255541 A1 WO 2025255541A1
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WIPO (PCT)
Prior art keywords
plant
strawberry
octoploid
facenh3
gene
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Pending
Application number
PCT/US2025/032766
Other languages
French (fr)
Inventor
Judson Arthur Ward
Franca Gabriela ROSSI
Lien Diana Bertier
Xingang Wang
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Ohalo Genetics Inc
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Ohalo Genetics Inc
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Publication of WO2025255541A1 publication Critical patent/WO2025255541A1/en
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Anticipated expiration legal-status Critical

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Classifications

    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • C12N9/14—Hydrolases (3)
    • C12N9/16—Hydrolases (3) acting on ester bonds (3.1)
    • C12N9/22—Ribonucleases [RNase]; Deoxyribonucleases [DNase]
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
    • A01H1/00—Processes for modifying genotypes ; Plants characterised by associated natural traits
    • A01H1/06—Processes for producing mutations, e.g. treatment with chemicals or with radiation
    • A01H1/08—Methods for producing changes in chromosome number
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
    • A01H5/00—Angiosperms, i.e. flowering plants, characterised by their plant parts; Angiosperms characterised otherwise than by their botanic taxonomy
    • A01H5/08—Fruits
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
    • A01H6/00—Angiosperms, i.e. flowering plants, characterised by their botanic taxonomy
    • A01H6/74—Rosaceae, e.g. strawberry, apple, almonds, pear, rose, blackberries or raspberries
    • A01H6/7409—Fragaria, i.e. strawberries
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/415—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from plants
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
    • C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
    • C12N15/82—Vectors or expression systems specially adapted for eukaryotic hosts for plant cells, e.g. plant artificial chromosomes (PACs)
    • C12N15/8241—Phenotypically and genetically modified plants via recombinant DNA technology
    • C12N15/8261—Phenotypically and genetically modified plants via recombinant DNA technology with agronomic (input) traits, e.g. crop yield
    • C12N15/8287—Phenotypically and genetically modified plants via recombinant DNA technology with agronomic (input) traits, e.g. crop yield for fertility modification, e.g. apomixis

Definitions

  • FIELD [0003] The present disclosure relates generally to strawberry breeding, and more specifically to generation of fully homozygous octoploid strawberry lines for producing true breeding F1 hybrid octoploid strawberry seed.
  • BACKGROUND [0004] Strawberries are a major agronomic fruit crop worldwide, with China, Mexico, and the U.S. being among the primary producers and consumers. According to recent data, 94% of U.S. households consume strawberries, with an annual per capita consumption of 4.85 pounds. California in particular produces 75% of the entire U.S. strawberry crop, with Florida being the next closest U.S. state in terms of production.
  • a strawberry plant such as a haploid-inducing strawberry plant, or a plant part thereof, comprising one or more genetic modifications resulting in decreased expression of one or more CENH3 genes.
  • the one or more genetic modifications comprise a modification of an enhancer of one or more of the CENH3 2 sf-6744554 197072001240 genes, a modification of a promoter of one or more of the CENH3 genes, a modification of a coding region of one or more of the CENH3 genes, a modification of an intron of one or more of the CENH3 genes, a modification of methylation status of one or more of the CENH3 genes, expression of a repressor protein that targets the DNA or an mRNA of one or more of the CENH3 genes, expression of an RNA interference construct that targets an mRNA of one or more of the CENH3 genes, or any combination thereof.
  • the one or more genetic modifications comprise a modification of an enhancer of one or more of the CENH3 genes, a modification of a promoter of one or more of the CENH3 genes, a modification of a coding region of one or more of the CENH3 genes, modification of an intron of one or more of the CENH3 genes, or any combination thereof relative to an unmodified strawberry plant of the same species or an unmodified control strawberry plant.
  • the one or more genetic modifications comprise a deletion, insertion, or one or more nucleotide changes in a coding region of one or more of the CENH3 genes.
  • the strawberry plant e.g., haploid-inducing strawberry plant
  • the strawberry plant has decreased expression of CENH3 proteins relative to a strawberry plant of the same species lacking the one or more genetic modifications.
  • the strawberry plant e.g., haploid-inducing strawberry plant
  • the strawberry plant t or plant part lacks detectable expression of CENH3 proteins.
  • the strawberry plant (e.g., haploid- inducing strawberry plant) is diploid, triploid, tetraploid, pentaploid, hexaploid, septaploid, octoploid, or nonaploid, or has a ploidy of 10x, 11x, 12x, 13x, 14x, 15x, 16x, 17x, 18x, 19x, or 20x.
  • the plant part is selected from the group consisting of pollen, an anther, a seed, an achene, a leaf, a flower, a fruit, and a stolon.
  • the plant part is pollen.
  • the plant part is a seed.
  • the strawberry plant e.g., haploid-inducing strawberry plant
  • the strawberry plant is diploid.
  • the strawberry plant e.g., haploid-inducing strawberry plant
  • the haploid- inducing strawberry plant is a plant of the species Fragaria vesca, Fragaria iinumae, Fragaria nipponica, Fragaria viridis, Fragaria ⁇ bifera, Fragaria bucharica, Fragaria chinensis, Fragaria daltoniana, Fragaria emeiensis, Fragaria hayatae, Fragaria iinumae, Fragaria mandshurica, Fragaria nilgerrensis, Fragaria nubicola, or Fragaria pentaphylla.
  • the strawberry plant (e.g., haploid-inducing strawberry plant) is a plant of the species Fragaria vesca. 3 sf-6744554 197072001240 [0010] In certain embodiments, the strawberry plant (e.g., haploid-inducing strawberry plant) is a Fragaria vesca plant of the subspecies Fragaria vesca ssp. vesca, Fragaria vesca sp. americana, or Fragaria vesca ssp. bracteata.
  • the one or more Fragaria vesca CENH3 (herein referred to as “FvCENH3”) genes comprise a polynucleotide sequence encoding an amino acid sequence having at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity the amino acid sequence of SEQ ID NO: 8.
  • the one or more FvCENH3 genes comprise a polynucleotide sequence having at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity to the polynucleotide sequence of SEQ ID NO: 1.
  • the strawberry plant (e.g., haploid-inducing strawberry plant) is a plant of the species Fragaria vesca and wherein the one or more CENH3 genes comprise FvCENH3.
  • the one or more genetic modifications comprise a modification of an enhancer of FvCENH3, a modification of a promoter of FvCENH3, a modification of a coding region of FvCENH3, modification of an intron of FvCENH3, or any combination thereof relative to a wild-type Fragaria vesca plant.
  • the strawberry plant (e.g., haploid-inducing strawberry plant) is octoploid.
  • the strawberry plant (e.g., haploid-inducing strawberry plant) is a plant of the species Fragaria x ananassa, Fragaria chiloensis, Fragaria virginiana, or Fragaria iturupensis.
  • the strawberry plant (e.g., haploid-inducing strawberry plant) is Fragaria chiloensis plant of the subspecies Fragaria chiloensis ssp. chiloensis, Fragaria chiloensis ssp. lucida, Fragaria chiloensis ssp. pacifica, or Fragaria chiloensis ssp.
  • the strawberry plant e.g., haploid-inducing strawberry plant
  • the strawberry plant is a plant of the species Fragaria x ananassa.
  • one or more of the Fragaria x ananassa CENH3 (herein referred to as “FaCENH3”) genes comprises a polynucleotide sequence encoding an amino acid sequence having at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity to an amino acid sequence selected from the group consisting of SEQ ID NOs: 9-14 and 53-56.
  • one or more of the FaCENH3 genes comprises a polynucleotide sequence having at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity to a polynucleotide sequence selected from the group consisting of SEQ ID NOs: 2-7 and 58-61.
  • the one or more FaCENH3 genes comprise: a 4 sf-6744554 197072001240 polynucleotide sequence encoding an amino acid sequence having at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity to the amino acid sequence of SEQ ID NO: 9 (Fragaria x ananassa CENH3-1a protein, hereinafter “FaCENH3-1a”); a polynucleotide sequence encoding an amino acid sequence having at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity to the amino acid sequence of SEQ ID NO: 10 (Fragaria x ananassa CENH3-1b protein, hereinafter “FaCENH3-1b”); a polynucleotide sequence encoding an amino acid sequence having at least 90%, at least 95%, at least 96%, at least 97%, at least
  • the one or more FaCENH3 genes comprise: a polynucleotide sequence encoding an amino acid sequence having at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity to the amino acid sequence of SEQ ID NO: 53 (Fragaria x ananassa CENH3-1a protein, hereinafter “FaCENH3-7a”); a polynucleotide sequence encoding an amino acid sequence having at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity to the amino acid sequence of SEQ ID NO: 54 (Fragaria x ananassa CENH3-1b protein, hereinafter “FaCENH3-7b”); a polynucleotide sequence encoding an amino acid sequence having at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity to
  • the one or more FaCENH3 genes comprise: a polynucleotide sequence having at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity to the polynucleotide sequence of SEQ ID NO: 2 (Fragaria x ananassa CENH3-1a gene, hereinafter “FaCENH3-1a”); a polynucleotide sequence having at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity to the polynucleotide sequence of SEQ ID NO: 3 (Fragaria x ananassa CENH3-1b gene, hereinafter “FaCENH3-1b”); a polynucleotide sequence having at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity to the polynucleotide sequence of SEQ ID NO:
  • the one or more FaCENH3 genes comprise: a polynucleotide sequence having at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity to the polynucleotide sequence of SEQ ID NO: 58 (Fragaria x ananassa CENH3-1a gene, hereinafter “FaCENH3-7a”); a polynucleotide sequence having at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity to the polynucleotide sequence of SEQ ID NO: 59 (Fragaria x ananassa CENH3-1b gene, hereinafter “FaCENH3-7b”); a polynucleotide sequence having at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity to the polynucleotide sequence of SEQ ID
  • the one or more FaCENH3 genes comprise FaCENH3-1a, FaCENH3-1b, FaCENH3-2a, FaCENH3-2b, FaCENH3-3, and FaCENH3-4.
  • the one or more genetic modifications comprise: a modification of an enhancer of FaCENH3-1a, a modification of a promoter of FaCENH3-1a, a modification of a coding region of FaCENH3-1a, a modification of an intron of FaCENH3-1a, or any combination thereof relative to an unmodified Fragaria x ananassa plant; a modification of an enhancer of FaCENH3-1b, a modification of a promoter of FaCENH3-1b, a modification of a coding region of FaCENH3-1b, a modification of an intron of FaCENH3-1b, or any combination thereof relative to an unmodified Fragaria x ananassa plant; a modification of an enhancer of FaCENH3-2a, a
  • the strawberry plant e.g., haploid-inducing strawberry plant
  • the one or more FaCENH3 genes comprise one, two, three, or all four of FaCENH3-7a, FaCENH3-7b, FaCENH3-7c, and FaCENH3-7d.
  • the one or more genetic modifications comprise: a modification of an enhancer of FaCENH3-7a, a modification of a promoter of FaCENH3-7a, a modification of a coding region of FaCENH3-7a, a modification of an intron of FaCENH3- 7a, or any combination thereof relative to an unmodified Fragaria x ananassa plant; a modification of an enhancer of FaCENH3-7b, a modification of a promoter of FaCENH3-7b, a modification of a coding region of FaCENH3-7b, a modification of an intron of FaCENH3- 7b, or any combination thereof relative to an unmodified Fragaria x ananassa plant; a 7 sf-6744554 197072001240 modification of an enhancer of FaCENH3-7c, a modification of a promoter of FaCENH3-7c, a modification of a coding region of FaCENH3-7c, a modification of an intron of
  • the decreased expression of the one or more CENH3 genes is achieved by gene disruption, gene knockout, gene knockdown, disruption of an endogenous amino acid sequence (including, for example, conserved amino acids or amino acid motifs), introduction of amino acids or amino acid motifs, gene silencing, RNA interference, induction of methylation, or any combination thereof.
  • the method comprises introducing one or more of the genetic modifications by mutagenesis, gene editing, transgenesis, or a combination thereof.
  • the method comprises introducing one or more of the genetic modifications by gene editing using a site-directed nuclease.
  • the site-directed nuclease is a CRISPR-associated (Cas) nuclease, a zinc finger nuclease (ZFN), a transcription activator-like effector nuclease (TALEN), or a megaTAL.
  • Cas CRISPR-associated nuclease
  • ZFN zinc finger nuclease
  • TALEN transcription activator-like effector nuclease
  • the method comprises contacting a plurality of cells of a parent strawberry plant with one or more Cas nucleases complexed with an RNA molecule comprising a polynucleotide sequence having at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% complementarity to 15, 16, 17, 18, 19, or 20 consecutive nucleotides of a protospacer sequence within one or more of the FvCENH3 and/or FaCENH3 genes.
  • the Cas nuclease is complexed with an RNA molecule comprising a polynucleotide sequence having at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity or complementarity to a polynucleotide sequence selected from the group consisting of SEQ ID NOs: 15-34.
  • the RNA molecule is a crRNA, a gRNA, or a pegRNA.
  • the plurality of cells is a plurality of protoplasts.
  • the method further comprises, subsequent to the contacting step, allowing the plurality of cells of the parent strawberry plant to form calli, plants, or a combination thereof, and identifying one or more calli or plants having the genetic modifications resulting in decreased expression of 8 sf-6744554 197072001240 one or more CENH3 genes.
  • the method comprises contacting a plurality of cells of a parent strawberry plant with one or more expression vectors together comprising an expression cassette for a Cas nuclease and an expression cassette for an RNA molecule having at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% complementarity to 15, 16, 17, 18, 19, or 20 consecutive nucleotides of one or more of the CENH3 genes.
  • step (a) comprises contracting a plurality of tetrahaploid reduced egg cells from the donor octoploid strawberry plant with a plurality of sperm cells of the haploid inducing strawberry plant.
  • step (a) comprises contacting pollen of the haploid-inducing strawberry plant with the stigma of a pistil of the donor octoploid strawberry plant comprising the reduced egg cell and allowing formation of a pollen tube and migration of the sperm cell to the reduced egg cell, thereby contacting the reduced egg cell with the sperm cell.
  • the method further comprises allowing the donor octoploid strawberry plant to form a plurality of seeds.
  • the method further comprises selecting one or more tetrahaploid seeds by 1) determining the ploidy of the embryo and endosperm of one or more seeds of the plurality of seeds, and 2) selecting one or more seeds having tetraploid embryo and an endosperm having a ploidy of greater than 8x.
  • provided herein is method of producing true homozygous octoploid strawberry seed, the method comprising (a) contacting an egg cell from a haploid- inducing strawberry plant described herein with a tetrahaploid reduced sperm cell from a donor octoploid strawberry plant, and (b) producing a doubled tetrahaploid cell from the egg cell, wherein the doubled tetrahaploid cell comprises the nuclear genome of the donor octoploid strawberry plant and the cytoplasmic genome of the haploid-inducing strawberry plant.
  • step (a) comprises contacting a plurality of egg cells from the haploid-inducing strawberry plants with a plurality of sperm cells from a donor octoploid strawberry plant.
  • the haploid-inducing strawberry plant and the donor octoploid strawberry plants are from different species of Fragaria.
  • the 9 sf-6744554 197072001240 haploid-inducing strawberry plant is diploid.
  • the haploid-inducing strawberry plant is a plant of the species Fragaria vesca.
  • the donor octoploid strawberry plant is a plant of the species Fragaria x ananassa.
  • step (a) comprises contacting the stigma of a pistil of the haploid-inducing strawberry plant with the pollen from the donor octoploid strawberry plant and allowing formation of a pollen tube and migration of the tetrahaploid reduced sperm cell to the egg cell, thereby contacting the egg cell with the tetrahaploid reduced sperm cell.
  • the method further comprises allowing the haploid-inducing strawberry plant to form a plurality of seeds.
  • a method of producing true homozygous octoploid strawberry plants comprising (a) contacting a plurality of tetrahaploid reduced egg cells from a maternal octoploid strawberry plant (e.g., an unedited maternal octoploid strawberry plant) with a plurality of sperm cells of an edited haploid-inducing octoploid strawberry plant described herein, (b) allowing the plurality of reduced egg cells to form a plurality of tetrahaploid seeds comprising the maternal genome of the octoploid strawberry plant (e.g., the unedited maternal genome) and lacking the edited, haploid- inducing paternal genome, (c) collecting the plurality of seeds and allowing them to germinate to form a plurality of plants, (d) determining the presence of one or more tetrahaploid plants in the plurality of plants, and (e) producing one or more doubled tetrahaploid plants (also referred
  • step (d) may include determining the presence of one or more spontaneously doubled tetrahaploids.
  • a method of producing true homozygous octoploid strawberry plants comprising (a) contacting a plurality of tetrahaploid reduced egg cells from an edited maternal haploid-inducing octoploid strawberry plant described herein with a plurality of sperm cells of an a paternal octoploid strawberry plant (e.g., an unedited octoploid strawberry plant), (b) allowing the plurality of reduced egg cells to form a plurality of tetrahaploid seeds comprising the paternal genome from the paternal octoploid strawberry plant (e.g., the unedited paternal genome) and lacking the edited, haploid-inducing maternal genome, (c) collecting the plurality of seeds and allowing them to germinate to form a plurality of plants, (d) determining the presence of one or more
  • step (d) may include determining the presence of one or more spontaneously doubled tetrahaploids.
  • a method of producing true homozygous octoploid strawberry plants comprising (a) contacting a plurality of tetrahaploid reduced egg cell from an edited haploid-inducing diploid strawberry plant described herein with a plurality of sperm cells of an octoploid strawberry plant (e.g., an unedited octoploid strawberry plant), (b) allowing the plurality of reduced egg cells to form a plurality of tetrahaploid seeds comprising the paternal genome of the octoploid strawberry plant and lacking the edited, haploid-inducing maternal genome of the diploid strawberry plant, (c) collecting the plurality of seeds and allowing them to germinate to form a plurality of plants, (d) determining the presence of one or more tetrahaploid plants in the plurality of plants, and (e) producing one or
  • step (d) may include determining the presence of one or more spontaneously doubled tetrahaploids.
  • the tetrahaploid seedling will comprise the unedited cytoplasmic genome of the diploid mother and the unedited octoploid genome of the father.
  • step (a) comprises contacting pollen of the haploid-inducing strawberry plant with the stigma of a pistil of the donor octoploid strawberry plant comprising the reduced egg cell and allowing formation of a pollen tube and migration of the pollen tube to the reduced egg cell, thereby contacting the reduced egg cell with the sperm cell.
  • step (d) comprises determining the presence of the maternal genome and the paternal genome in the one or more plants, wherein tetrahaploid plants within the plurality of plants are determined based on the absence of the maternal genome and the presence of the paternal genome.
  • step (a) comprises contacting pollen of the donor octoploid strawberry plant with the stigma of a pistil of the haploid-inducing strawberry plant comprising the reduced egg cell and allowing formation of a pollen tube and migration of the pollen tube to the reduced egg cell, thereby contacting the reduced egg cell with the sperm cell.
  • step (d) comprises determining the presence of the maternal genome and the 11 sf-6744554 197072001240 paternal genome in the one or more plants, wherein tetrahaploid plants within the plurality of plants are determined based on the presence of the maternal genome and the absence of the paternal genome.
  • step (d) further comprises determining the ploidy of the embryo from each of the one or more tetrahaploid plants and discarding any plants that (i) lacks a tetraploid embryo and/or (ii) comprises an embryo ploidy of greater than 8x.
  • the presence (or absence) of the maternal genome and/or the paternal genome, respectively, in the one or more plants is determined by the detection of one or more genetic markers which are present in the paternal genome and which are absent in the maternal genome. In certain embodiments, the presence (or absence) of the maternal genome and/or the paternal genome, respectively, in the one or more plants, is determined by the detection of one or more genetic markers which are present in the maternal genome and which are absent in the paternal genome.
  • step (d) comprises determining the presence of the maternal genome and the paternal genome in the one or more plants, wherein tetrahaploid plants within the plurality of plants are determined based on the absence of the maternal genome and the presence of the paternal genome. In other embodiments, step (d) comprises determining the presence of the maternal genome and the paternal genome in the one or more plants, wherein tetrahaploid plants within the plurality of plants are determined based on the presence of the maternal genome and the absence of the paternal genome.
  • step (d) further comprises determining the ploidy of the embryo from each of the one or more tetrahaploid plants and discarding any plants that (i) lacks a tetraploid embryo and/or (ii) comprises an embryo ploidy of greater than 8x.
  • the presence (or absence) of the maternal genome and/or the paternal genome, respectively, in the one or more plants is determined by the detection of one or more genetic markers which are present in the paternal genome and which are absent in the maternal genome.
  • step (e) comprises subjecting the selected tetrahaploid plant to a tetrahaploid doubling treatment.
  • the tetrahaploid doubling treatment comprises contacting a cell from the tetrahaploid plant with an anti-microtubule agent to form a doubled tetrahaploid cell (i.e., a true homozygous octoploid cell).
  • the anti-microtubule agent is an anti-mitotic herbicide.
  • the anti- microtubule agent comprises colchicine, oryzalin, nitrous oxide, trifluralin, or any combination thereof.
  • the doubled tetrahaploid cell is allowed to grow into a doubled tetrahaploid plant.
  • the doubled tetrahaploid plant i.e., the true homozygous octoploid plant
  • the doubled tetrahaploid plant is contacted with self-pollen and allowed to form seed, producing a plurality of true homozygous octoploid strawberry seed.
  • the doubled tetrahaploid plant i.e., the true homozygous octoploid plant
  • the donor octoploid strawberry plant is a plant of the species Fragaria x ananassa, Fragaria chiloensis, Fragaria virginiana, or Fragaria iturupensis, or a hybrid of any combination thereof.
  • the donor octoploid strawberry plant is Fragaria chiloensis plant of the subspecies Fragaria chiloensis ssp. chiloensis, Fragaria chiloensis ssp. lucida, Fragaria chiloensis ssp.
  • the donor octoploid strawberry plant is a plant of the species Fragaria x ananassa. In certain embodiments, the donor octoploid strawberry plant is a Fragaria x ananassa plant of the variety named “Camarosa.” In certain embodiments, the donor octoploid strawberry plant is a Fragaria x ananassa plant of the variety named “Chandler.” In certain embodiments, the donor octoploid strawberry plant is a Fragaria x ananassa plant of the variety named “Albion.”.
  • the method of producing a true homozygous octoploid strawberry plant or seed further comprises introducing one or more genetic modifications resulting in decreased expression of a gibberellin 20-oxidase (“ga20ox”) gene into the true homozygous octoploid strawberry plant or seed.
  • a gibberellin 20-oxidase (“ga20ox”) gene
  • a method of producing a runnerless true homozygous octoploid strawberry plant comprising (a) contacting a tetrahaploid reduced egg cell from a donor octoploid strawberry plant with the sperm cell of a haploid- inducing strawberry plant described herein, (b) producing a doubled tetrahaploid cell from the tetrahaploid reduced egg cell, and (c) introducing one or more genetic modifications resulting in decreased expression of a ga20ox gene into the true homozygous octoploid strawberry plant, wherein the runnerless true homozygous octoploid strawberry plant does not produce runners.
  • the true homozygous octoploid strawberry plant, or a progenitor thereof was subject to introduction of one or more of the genetic modifications by mutagenesis, gene editing, transgenesis, or a combination thereof followed by selection for a genetic modification that results in decreased expression of the one or more CENH3 genes.
  • the plant part is selected from the group consisting of pollen, an anther, a seed, an achene, a leaf, a flower, a fruit, and a stolon.
  • the cytoplasmic genome is from a plant of the species Fragaria vesca.
  • the nuclear genome is from a plant of the species Fragaria x ananassa.
  • the plant part is selected from the group consisting of pollen, an anther, a seed, an achene, a leaf, a flower, a fruit, and a stolon.
  • a true homozygous octoploid strawberry seed comprising an embryo having the nuclear genome of a first species of Fragaria and the cytoplasmic genome of a second plant species of Fragaria.
  • the first species of Fragaria is Fragaria x ananassa.
  • the second species of Fragaria is a diploid species of Fragaria.
  • the second species of Fragaria is Fragaria vesca.
  • the true homozygous octoploid strawberry seed comprises one or more genetic modifications resulting in decreased expression of a gibberellin ga20ox gene.
  • a true homozygous octoploid strawberry plant or plant part thereof comprising an embryo having the nuclear genome of a first species of Fragaria and the cytoplasmic genome of a second plant species of Fragaria.
  • the first species of Fragaria is Fragaria x ananassa.
  • the second species of Fragaria is a diploid species of Fragaria.
  • the second species of Fragaria is Fragaria vesca.
  • the true homozygous octoploid strawberry seed comprises one or more genetic modifications resulting in decreased expression of a gibberellin ga20ox gene.
  • the true homozygous 15 sf-6744554 197072001240 octoploid strawberry plant does not produce runners.
  • the true homozygous octoploid strawberry plant, or a progenitor thereof was subject to introduction of one or more of the genetic modifications by mutagenesis, gene editing, transgenesis, or a combination thereof followed by selection for a genetic modification that results in decreased expression of the one or more CENH3 genes.
  • the plant part is selected from the group consisting of pollen, an anther, a seed, an achene, a leaf, a flower, a fruit, and a stolon.
  • a method of producing uniform octoploid F1 hybrid strawberry seed comprising crossing two of the true homozygous octoploid strawberry plants described herein.
  • a uniform octoploid F1 hybrid strawberry seed produced according to said method is provided herein.
  • the uniform octoploid F1 hybrid strawberry seed comprises one or more genetic modifications resulting in decreased expression of a ga20ox gene.
  • a uniform octoploid F1 hybrid strawberry plant or a plant part thereof produced according to said method comprises one or more genetic modifications resulting in decreased expression of a ga20ox gene.
  • the uniform octoploid F1 hybrid strawberry plant does not produce runners.
  • the plant part is selected from the group consisting of pollen, an anther, a seed, an achene, a leaf, a flower, a fruit, and a stolon.
  • the expression vector or isolated DNA molecule comprises a DNA sequence having at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity or complementarity to 15, 16, 17, 18, 19, or 20 consecutive nucleotides of a polynucleotide having at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity to a polynucleotide sequence selected from the group consisting of SEQ ID NOs: 1-7 and 58-61. 16 sf-6744554 197072001240 [0039]
  • the expression vector or isolated DNA molecule comprises a DNA sequence encoding a non-coding RNA.
  • the non-coding RNA comprises a polynucleotide sequence having at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity or complementarity to 15, 16, 17, 18, 19, or 20 consecutive nucleotides of a polynucleotide having at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity to a polynucleotide sequence selected from the group consisting of SEQ ID NOs: 1-7 and 58-61.
  • the non-coding RNA is a crRNA, a gRNA, a pegRNA, a siRNA, a miRNA, or a dsRNA.
  • the expression vector or isolated DNA molecule comprises a DNA sequence encoding a site-directed nuclease.
  • the site-directed nuclease is a Cas nuclease, a TALEN, ZFN, or a mega-TAL.
  • the site- directed nuclease comprises an amino acid sequence that confers binding to a polynucleotide sequence having at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity or complementarity to 15, 16, 17, 18, 19, or 20 consecutive nucleotides of a polynucleotide sequence encoding an amino acid sequence having at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity to an amino acid sequence selected from the group consisting of SEQ ID NOs: 8-14 and 53-56.
  • the site-directed nuclease comprises an amino acid sequence that confers binding to a polynucleotide sequence having at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity or complementarity to 15, 16, 17, 18, 19, or 20 consecutive nucleotides of a polynucleotide having at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity to a polynucleotide sequence selected from the group consisting of SEQ ID NOs: 1-7 and 58-61.
  • a bacterial cell comprising an expression vector or isolated DNA molecule described herein.
  • the bacterial cell is an Agrobacterium cell.
  • a kit comprising an expression 17 sf-6744554 197072001240 vector or isolated DNA molecule or a bacterial cell described herein.
  • a genetically modified plant, plant part, plant cell, or seed comprising an expression vector or isolated DNA molecule described herein.
  • a genetically modified non-regenerable plant cell of a strawberry plant e.g., haploid-inducing strawberry plant
  • a genetically modified plant genome of the strawberry plant e.g., haploid-inducing strawberry plant
  • FIG. 1 depicts a schematic illustrating the production of homozygous octoploid strawberry lines by tetrahaploid induction of the maternal genome and then doubling the resulting tetrahaploid.
  • Step (A) exemplifies the starting plant material where chromosomes are indicated by gray and white patterned vertical bars, and the oval lines surrounding the vertical bars indicate exemplary nuclei. Ploidy is indicated at the top of the nuclei.
  • An edited cenh3 gene is indicated on the chromosomes by a white “X” mark.
  • step (B) the top row exemplifies reduced, recombined 1x cenh3-edited pollen derived from a cenh3-edited diploid Fragaria vesca strawberry plant; the middle row illustrates a reduced, recombined 4x unedited tetrahaploid egg cell derived from an unedited octoploid Fragaria x ananassa strawberry plant; and the bottom row exemplifies reduced, recombined 4x cenh3-edited pollen from a cenh3-edited octoploid Fragaria x ananassa strawberry plant.
  • the reduced, recombined 4x unedited tetrahaploid egg cell (middle row) is contacted by either (i) the reduced, recombined 1x cenh3-edited pollen (or sperm cell) (top row), (ii) the reduced, recombined 4x cenh3-edited pollen (or sperm cell), or (iii) both.
  • step (C) some amount of fertilizations cause the maternal tetrahaploid egg cell to undergo embryogenesis without retention of genetic material from the cenh3-edited pollen, as indicated by the straight-rightwards arrow.
  • the remaining eggs result in fertilizations by the haploid inducer, illustrated by: (i) the up-rightwards arrow pointing to an illustration of an exemplary 5x embryo with one set of paternal chromosomes from the cenh3-edited diploid male parent, and (ii) the down-rightwards arrow pointing to an illustration of an exemplary 18 sf-6744554 197072001240 8x embryo with 4 sets of paternal chromosomes from the cenh3-edited octoploid parent.
  • FIG. 2A depicts a schematic illustrating the production of homozygous octoploid strawberry lines by tetrahaploid induction of the paternal genome and then doubling the resulting tetrahaploid.
  • Step (A) exemplifies the starting plant material where chromosomes are indicated by gray and white patterned vertical bars, and the oval lines surrounding the vertical bars indicate exemplary nuclei. Ploidy is indicated at the top of the nuclei.
  • step (B) An edited cenh3 gene is indicated on the chromosomes by a white “X” mark. Pollen is indicated by a male symbol, and egg cells and megaspore mother cells are indicated by a female symbol.
  • step (B) the bottom row exemplifies reduced, recombined 4x pollen derived from an unedited octoploid Fragaria x ananassa strawberry plant, and the top row illustrates a reduced, recombined 4x tetrahaploid egg cell derived from a cenh3-edited octoploid Fragaria x ananassa strawberry plant.
  • the egg cell is a reduced, recombined 1x haploid egg cell derived from a cenh3-edited diploid Fragaria vesca strawberry plant (not illustrated).
  • the reduced, recombined 4x cenh3-edited tetrahaploid egg cell (illustrated in the top row) or the reduced, recombined 1x cenh3-edited haploid egg cell (not illustrated), is contacted by the reduced, recombined 4x unedited pollen (bottom row).
  • step (C) some of the fertilizations cause the maternal tetrahaploid egg cell (illustrated in the top row) or the reduced, recombined 1x cenh3-edited haploid egg cell (not illustrated), to undergo embryogenesis without retention of nuclear genetic material from the cenh3-edited egg cell, as indicated by the straight-rightwards arrow.
  • the remaining eggs result in fertilizations by the haploid inducer, illustrated by the down-rightwards arrow pointing to an illustration of an exemplary 8x embryo with four sets of both maternal and paternal chromosomes from octoploid parents.
  • the reduced, recombined egg cells is a 1x cenh3- edited haploid egg cell
  • fertilization by the haploid inducer results in 5x embryos (not illustrated).
  • Embryos are screened for presence of the maternal and/or paternal genome and have their ploidy confirmed before subjecting the tetrahaploid embryos to a doubling treatment, thereby producing true homozygous octoploid strawberry plants in step (D).
  • 2B depicts a schematic illustrating the production of homozygous octoploid strawberry lines by tetrahaploid induction of the paternal genome, with a cytoplasmic swap 19 sf-6744554 197072001240 resulting in a seed having the cytoplasmic genome of the maternal plant and the nuclear genome of the paternal plant.
  • Pollen of octoploid (8x) F. x ananassa is used to pollinate a haploid-inducing line of diploid (2x) F. vesca, resulting in tetrahaploid embryogenesis of an embryo retaining the nuclear tetrahaploid (4x) genome of F. x ananassa and the cytoplasmic genome of F. vesca.
  • FIGS. 3A-3B show the configuration of CENH3 loci in octoploid strawberry.
  • FIG. 3A shows the configuration of six homoeologous FaCENH3 loci in octoploid strawberry Fragaria x ananassa according to reference genome “Fragaria x ananassa Camarosa Genome v1.0.a2”.
  • FaCENH3-1a is illustrated as the left-most black bar on chromosome 7, subgenome 1
  • FaCENH3-1b is illustrated as the right-most on chromosome 7, subgenome 1
  • FaCENH3-2a is illustrated as the left-most black bar on chromosome 7, subgenome 2
  • FaCENH3-2b is illustrated as the right-most black bar on chromosome 7, subgenome 2
  • FaCENH3-3 is illustrated as the black bar on chromosome 7, subgenome 3
  • FaCENH3-4 is illustrated as the black bar on chromosome 7, subgenome 4.
  • FIG. 3B shows the configuration of six homoeologous FaCENH3 loci in octoploid strawberry Fragaria x ananassa according to reference genome “Fragaria x ananassa Royal Royce Genome v1.0.” The four homeologs are shown, with black bars indicating the locations of the CENH3 loci.
  • FIG. 4 shows the CENH3 gene in the diploid strawberry Fragaria vesca according to the reference genome “Fragaria vesca v_4.0”. The gene (referred to herein as (“FvCENH3”) is shown with a black bar indicating its location of chromosome 7.
  • FIGS. 5A-C show a protein sequence alignment of CENH3 orthologs from Fragaria vesca (FvCENH3) (SEQ ID NO: 8), Fragaria x ananassa Camarosa (FaCENH3- 1a, FaCENH3-1b, FaCENH3-2,a FaCENH3-2b, FaCENH3-3, FaCENH3-4) (SEQ ID NOs: 20 sf-6744554 197072001240 9-14) Fragaria chiloensis (FcCENH3-7-AV, FcCENH3-7-Bi, FcCENH3-7-B1, and FcCENH3-7-B2) (SEQ ID NOs: 36-39), Fragaria iinumae (FiCENH3) (SEQ ID NO: 40), Fragaria viridis (FvirCENH3) (SEQ ID NO: 41), Potentilla micrantha (PmCENH3), (SEQ ID NO: 42), Arabidopsis thal
  • FIG. 5A shows positions 1-42 of the sequence alignment.
  • FIG. 5B shows positions 43-108 of the sequence alignment.
  • FIG. 5C shows positions 109-178 of the sequence alignment.
  • FIGS. 5D-G show a protein sequence alignment of CENH3 orthologs from Fragaria vesca (FvCENH3) (SEQ ID NO: 8), Fragaria x ananassa Royal Royce (FaCENH3-7a, FaCENH3-7b, FaCENH3-7c, and FaCENH3-7d) (SEQ ID NOs: 53-56) Fragaria chiloensis (FcCENH3-7-AV, FcCENH3-7-Bi, FcCENH3-7-B1, and FcCENH3-7- B2) (SEQ ID NOs: 36-39), Fragaria iinumae (FiCENH3) (SEQ ID NO: 40), Fragaria viridis (FvirCENH3) (SEQ ID NO: 41), Potentilla micrantha
  • FIG. 5D shows positions 1-60 of the sequence alignment.
  • FIG. 5E shows positions 61- 120 of the sequence alignment.
  • FIG. 5F shows positions 121-180 of the sequence alignment.
  • FIG. 5G shows positions 181-189 of the sequence alignment.
  • FIG. 6A shows a phylogenetic tree of identified putative and characterized CENH3 orthologs as in FIGS. 5A-5C.
  • the phylogenetic tree is a UPGMA consensus tree.
  • the scale bar represents phylogenetic distance of 0.05 nucleotide substitutions per site. The tree is generated using software Geneious Prime 2023.2.1, with Genetic Distance Model: Jukes-Cantor.
  • FIG. 6A shows a phylogenetic tree of identified putative and characterized CENH3 orthologs as in FIGS. 5A-5C.
  • the phylogenetic tree is a UPGMA consensus tree.
  • the scale bar represents phylogenetic distance of 0.05 nucleotide substitutions per site. The tree
  • FIG. 6B shows a phylogenetic tree of identified putative and characterized CENH3 orthologs as in FIGS. 5D-5G.
  • the phylogenetic tree is a UPGMA consensus tree.
  • the scale bar represents phylogenetic distance of 0.05 nucleotide substitutions per site. The tree is generated using software Geneious Prime 2025.0.2, with Genetic Distance Model: Jukes-Cantor.
  • FIG. 7 shows a bar graph of editing efficiencies of guide RNAs targeting different protospacers in FaCENH3 in four different varieties of Fragaria x ananassa.
  • the x-axis 21 sf-6744554 197072001240 indicates the different target protospacer sites which were tested; the y-axis indicates the percent of protoplasts with successful editing. Different shades indicate different varieties of Fragaria x ananassa strawberry varieties tested. The sample size ranged from 3 replicates up to 9 replicates, with a median of 6 replicates across all data sets. The error bars indicate standard error of the mean. [0055] FIG.
  • FIG. 8A shows a schematic illustrating the editing windows associated with PRS878 for FvCENH3 (SEQ ID NO: 1), FaCENH3-1a (SEQ ID NO: 2), FaCENH3-1b (SEQ ID NO: 3), FaCENH3-2a (SEQ ID NO: 4), FaCENH3-2b (SEQ ID NO: 5), FaCENH3-3 (SEQ ID NO: 6), and FaCENH3-4 (SEQ ID NO: 7).
  • the protospacer sequence is shown in bold, underlined text. UTRs are denoted in light gray; coding sequences are denoted in dark gray; and introns are denoted as a line.
  • FIG. 8B shows a schematic illustrating the editing windows associated with PRS878 for FvCENH3 (SEQ ID NO: 1), FaCENH3-7a (SEQ ID NO: 58), FaCENH3-7b (SEQ ID NO: 59), FaCENH3-7c (SEQ ID NO: 60), and FaCENH3-7d (SEQ ID NO: 61).
  • the protospacer sequence is shown in bold, underlined text.
  • UTRs are denoted in light gray; coding sequences are denoted in dark gray; and introns are denoted as a line.
  • the nucleotides in bolded, underlined font show the (+) strand reverse complement of the PRS878 sequence (SEQ ID NO: 21). At the top of the figure, its general location is illustrated by a downwards- pointing black triangle over the third exon. [0057] FIG.
  • FIG. 9A shows a schematic illustrating the editing windows associated with PRS874 for FvCENH3 (SEQ ID NO: 1), FaCENH3-1a (SEQ ID NO: 2), FaCENH3-1b (SEQ ID NO: 3), FaCENH3-2a (SEQ ID NO: 4), FaCENH3-2b (SEQ ID NO: 5), FaCENH3-3 (SEQ ID NO: 6), and FaCENH3-4 (SEQ ID NO: 7).
  • the protospacer sequence is shown in bold, underlined text. UTRs are denoted in light gray; coding sequences are denoted in dark gray; and introns are denoted as a line.
  • FIG. 9B shows a schematic illustrating the editing windows associated with PRS874 for FvCENH3 (SEQ ID NO: 1), FaCENH3-7a (SEQ ID NO: 58), FaCENH3-7b (SEQ ID NO: 59), FaCENH3-7c (SEQ ID NO: 60), and FaCENH3-7d (SEQ ID NO: 61).
  • the protospacer sequence is shown in bold, underlined text.
  • FIG. 10A shows a schematic illustrating the editing windows associated with PRS876 for FvCENH3 (SEQ ID NO: 1), FaCENH3-1a (SEQ ID NO: 2), FaCENH3-1b (SEQ ID NO: 3), FaCENH3-2a (SEQ ID NO: 4), FaCENH3-2b (SEQ ID NO: 5), FaCENH3-3 (SEQ ID NO: 6), and FaCENH3-4 (SEQ ID NO: 7).
  • FIGS. 14A-14F depict a sequence alignment of FaCENH3-7c alleles in wild-type (N; SEQ ID NO: 103 and 104) and edited (E; SEQ ID NOs: 105-113) F. x ananassa Royal Royce plants with the translated protein sequence (SEQ ID NO: 114).
  • FIG. 14A shows the alignment at positions 1-108 of the wild-type sequence.
  • FIG. 14B shows the alignment at positions 109-216 of the wild-type sequence.
  • FIG. 14C shows the alignment at positions 217-270 of the wild-type sequence. [0065] FIGS.
  • FIGS. 23A-23B depict photographs of wild-type and cenh3 mutant strawberry lines.
  • FIG. 23A depicts cenh3-007 (E-PED524-4091; left) and wild-type (right) plants.
  • FIG. 23B depict two flowers and three stamens each from the wild-type (top) and cenh3-007 (E- PED524-4091; bottom) plants.
  • DETAILED DESCRIPTION [0074] The following description is presented to enable a person of ordinary skill in the art to make and use the various embodiments.
  • Pollen from the haploid inducer lines is used to fertilize or stimulate development of reduced eggs cells (4x) from elite octoploid (8x) strawberry varieties.
  • pollen from the haploid inducer lines induce the reduced egg cells to undergo embryogenesis without transfer or retention of genetic material from the paternal genome.
  • pollen from the haploid inducer lines fuses with the reduced egg cell to form a zygote wherein the maternal genome is thereafter eliminated from the zygote, generating a tetrahaploid (4x) embryo without transfer or retention of genetic material from the maternal genome.
  • the resulting haploid strawberry lines (4x) are doubled (for example, using colchicine) to achieve fully homozygous octoploid (8x) strawberry lines.
  • Embryogenesis without retention of the genomic material from the pollen occurs in some of the egg cells (FIG. 1, steps B).
  • the resulting tetrahaploid lines (4x) are doubled with colchicine (FIG. 1, step C) to obtain fully homozygous octoploid strawberry derived from the maternal genome (8x) (FIG. 1, step D).
  • Multiple fully homozygous octoploid strawberry lines are produced and then crossed according to standard methods in the art. The best crosses are chosen to produce genetically uniform F1 hybrid seed.
  • unmodified 4x pollen (FIG.
  • FIG. 2A bottom row
  • a modified cenh3-edited female wherein the egg cell is induced to undergo embryogenesis, resulting in a tetrahaploid (4x) comprising the paternal nuclear genome and lacking the cenh3-edited maternal genome (FIG. 2, middle row).
  • embryogenesis without retention of the genomic material from the egg occurs in some of the egg cells (FIG. 2, steps B).
  • the resulting haploid lines (4x) are doubled with colchicine (FIG. 2, step C) or double spontaneously to obtain fully homozygous octoploid strawberry lines derived from the paternal genome (8x) (FIG. 2, step D).
  • haploid-inducing strawberry plants, and plant parts thereof comprising one or more genetic modifications resulting in decreased expression (including, e.g., decreased activity) of one or more CENH3 genes.
  • Said haploid-inducing strawberry plant and plant parts may be crossed with octoploid strawberry plants to produce haploid (4x) strawberry plants, which may then be subject to genome doubling to produce true homozygous octoploid (8x) strawberry plants.
  • true homozygous octoploid strawberry plants, and parts thereof are true homozygous octoploid strawberry plants, and parts thereof.
  • kits useful in performing the methods described herein.
  • the true homozygous octoploid strawberry seed and plant lines of the present disclosure are homozygous within the same subgenome, but can remain heterozygous across the four different subgenomes, respective to each other. This stands in stark contrast to existing methods of producing inbred octoploid strawberry lines, which requires a prohibitive number of generations of selfing and may not achieve true homozygosity across all loci. Also described herein are true homozygous octoploid strawberry seeds, plants, and plant parts produced according to the methods described herein. Further provided herein are uniform octoploid F1 hybrid strawberry seed, plants, and plant parts thereof produced by crossing two of said true homozygous octoploid strawberry plants.
  • the term “plant” includes the whole plant or any parts or derivatives thereof, such as plant organs (e.g., harvested or non-harvested flowers, leaves, etc.), plant cells, plant protoplasts, plant cell or tissue cultures from which whole plants can be regenerated, regenerable or non-regenerable plant cells, plant calli, plant cell clumps, and plant cells that are intact in plants, or parts of plants, such as embryos, pollen, ovules, ovaries, reduced egg cells, pistil, stigma, seeds, achenes, fruits (e.g., aggregate fruits), flowers, leaves, seeds, runners, clonally propagated plants, roots, stems, cotyledons, hypocotyls, root tips (e.g., harvested tissues or organs) and the like.
  • plant organs e.g., harvested or non-harvested flowers, leaves, etc.
  • plant cells e.g., plant protoplasts, plant cell or tissue cultures from which whole plants can be regenerated, regenerable or
  • the term “strawberry plant” typically refers to any plant commonly understood by one of ordinary skill in the art to be a strawberry plant, and includes plants of the genera Fragaria.
  • the term “flower” or “strawberry flower” may refer to either the aggregate inflorescence of flowers or the individual flowers making up the aggregate.
  • the term “fruit” or “strawberry fruit” typically refers to the aggregate fruit of the strawberry, which is the fleshy structure that is typically used in food applications and considered by laypeople as the strawberry fruit.
  • seed or “strawberry seed” includes both true strawberry seed and strawberry achenes.
  • Achenes are the small seed-like structures found on the outer surface of the aggregate fruits of strawberries. Each achene arises from a single ovule of the aggregate inflorescence and, if the egg is fertilized, contains an endosperm and a most commonly a single seed. Each achene is therefore a true fruit which most commonly contains a seed.
  • non-regenerable generally refers to a plant part, a plant cell, a processed plant product, or a portion of any of the foregoing, that cannot be induced to form a whole plant or that cannot be induced to form a whole plant that is capable of sexual and/or asexual reproduction.
  • nucleus of a plant cell including, e.g., an egg cell, a sperm cell, or an embryo.
  • cytoplasmic genome typically refers to the genome present in the organelles present outside the nucleus of a plant cell, such as the mitochondrial genome or the chloroplast genome of a plant cell.
  • ploidy refers to the number of complete sets of chromosomes in a cell or organism. Ploidy may be annotated using “n” as the unit of complete sets of chromosomes. For example, a cell or organism with a single set of chromosomes may be referred to as “1x”, or the single set of chromosomes itself may be referred to as “1x”.
  • Polyploid may refer to organisms which are triploid (3x), tetraploid (4x), pentaploid (5x), hexaploid (6x), septaploid (or heptaploid, 7x), octoploid (8x), or higher ploidies (greater than 8x). 29 sf-6744554 197072001240 [0090] As used herein, “octoploid” (which may also be spelled “octaploid”) refers to a cell or organism with a ploidy of 8x. [0091] As used herein, “haploid” typically refers to a cell or organism with a ploidy half that of the parent organism.
  • haploid gametes typically refers to gamete cells with a ploidy half that of the parent organism.
  • the 4x haploid gametes may also be referred to as tetrahaploid gametes.
  • haploid-inducing strawberry plant typically refers to a strawberry plant that produces pollen that triggers the development of haploid embryos and seeds when crossed with another strawberry plant.
  • a “haploid-inducing strawberry plant” or “haploid inducer strawberry plant” may also refer to a strawberry plant that produces an egg whose genome is replaced by the genome of an unmodified strawberry plant upon fertilization by the unmodified plant and results in the development of haploid embryos and seeds.
  • haploid induction typically refers to the process of crossing pollen from a haploid- inducing plant with a non-haploid-inducing plant to generate haploid embryos and seeds.
  • two haploid sperm cells migrate down the pollen tube to the ovule.
  • One of the sperm cells fertilizes the central cell of the ovule to form the endosperm, while the other sperm cell fertilizes the haploid reduced egg cell to form an embryo having the ploidy of the parent plant.
  • haploid (4x) sperm and egg form an octoploid (8x) embryo in a normal fertilization process.
  • haploid induction the sperm cells from the haploid-inducing plant migrate down the pollen tube to the ovule, where one of the sperm cells fertilizes the central cell to form the endosperm.
  • embryogenesis is initiated, but the DNA from the sperm cell is not delivered to the egg cell or is otherwise eliminated, resulting in formation of a haploid embryo.
  • haploid embryo comprising the paternal nuclear 30 sf-6744554 197072001240 genome.
  • the haploid (4x) egg cell either (i) does not incorporate the DNA from the haploid sperm cell (4x), resulting in a haploid (4x) embryo comprising the maternal nuclear genome (4x), or (ii) does not retain the DNA from the haploid egg cell (4x), resulting in a haploid (4x) embryo comprising the paternal nuclear genome (4x).
  • the non-haploid- inducing plant that is pollinated by the haploid-inducing plant is typically referred to as the donor plant, however the resulting haploid embryos can contain chromosomes from either the donor plant or the haploid-inducing plant, and usually have half the ploidy of the donor plant (although spontaneous doubling is also sometimes observed).
  • the term “donor octoploid strawberry plant” typically refers to an octoploid strawberry plant wherein the nuclear genome undergoes the haploid induction process, resulting in haploid embryos each having one set of chromosomes from the donor octoploid strawberry plant, and lacking nuclear genome from the haploid-inducing strawberry plant.
  • the haploid- inducing plant may be a female, as in the case of female genome elimination and induction of the paternal genome; or the haploid-inducing plant may be a male, as in the case of maternal genome induction.
  • the haploid-inducing strawberry plant and the donor strawberry plant do not need to have the same ploidy.
  • allele refers to one of two or more alternative forms of a single gene or locus within the genome.
  • “monoallelic” typically describes the presence of a single allele at a given locus or set of loci within a cell or organism.
  • “monoallelic at over n% of the loci in the genome” typically refers to the percentage of total loci in the genome which are monoallelic.
  • “biallelic” typically describes the presence of two different alleles at a given locus or set of loci within a cell or organism.
  • “multiallelic” typically describes the presence of three or more alleles at a given locus or set of loci within a cell or organism.
  • “monoallelic” and “biallelic” typically refer to the allelic makeup of a set of chromosomes within the same subgenome.
  • an octoploid strawberry plant of the species Fragaria x ananassa which has four subgenomes, is monoallelic at all loci if the two chromosomes in a subgenome are monoallelic at all loci.
  • “monoallelic at over 31 sf-6744554 197072001240 n% of the loci in the genome” typically refers to the percentage of total loci in the entire genome that are monoallelic within each subgenome, but does not indicate that n% of the loci are monoallelic across all subgenomes.
  • haplotype refers to a distinct 1n set of chromosomes with a unique set of alleles.
  • each haplotype is distinct from other haplotypes in that it contains a set of alleles that confers a unique set of characteristics not conferred by other haplotypes.
  • “monoallelic plant” typically refers to a plant line containing a single haplotype within a subgenome
  • a “biallelic plant” typically refers to a plant line containing two haplotypes.
  • clonal bodies of DNA, clonal cells, or clonal organisms may not be completely identical.
  • “Clonal” may describe two sets of chromosomes that are not completely identical in sequence but that contain the same set of alleles.
  • “genetically uniform” describes a set of individual plants, plant parts (e.g., seeds), or plant cells whose genomes are clonal. For example, a population of strawberry F1 hybrid seed wherein at least 99% of the population is genetically uniform indicates that 99% of the seeds of the population are clonal across sub-genomes.
  • clonal may describe a body of DNA that is substantially identical to another body of DNA, however due to random errors in natural DNA replication, the clonal bodies of DNA, clonal plants, plant parts (e.g., seeds), or plant cells may not be completely identical. “Clonal” may also describe two genomes that are not completely identical in sequence but that contain the same set of alleles. [0097] As used herein, “homozygous” describes a cell or organism in which all sets of chromosomes within a genome or subgenome encode the same allele or set of alleles at a certain chromosomal locus, a set of chromosomal loci, or at all chromosomal loci.
  • homozygous plant typically refers to a monoallelic plant or plant line having allelic uniformity across all chromosomal loci.
  • homozygous typically refers to sets of chromosomes within the same subgenome having the same alleles or sets of alleles at one or more chromosomal loci
  • homozygous plant typically refers to a monoallelic plant or plant line having allelic uniformity across all chromosomal loci within each subgenome, but does not indicate that they have allelic uniformity across subgenomes.
  • an octoploid strawberry cell or plant having the same allele at a specific locus in two homologous chromosomes within in a subgenome may also be considered homozygous even though one or more alternative alleles exist in one or more different subgenomes.
  • heterozygous describes a cell or organism in which at least one set of chromosomes in one subgenome encodes an allele or set of alleles at a certain chromosomal locus or set of chromosomal loci that is distinct from those of the other sets of chromosomes within the cell or organism.
  • a strawberry cell or plant having allele a1 at locus A in one set of chromosomes in a subgenome and having allele a2 at locus A in a second set of chromosomes is heterozygous for alleles a1 and a2.
  • heterozygous plant typically refers to a biallelic plant or plant line.
  • heterozygous typically refers to two sets of chromosomes within the same subgenome having distinct alleles or sets of alleles at one or more chromosomal loci
  • heterozygous plant typically refers to a plant that is biallelic for one or more loci in at least one of its subgenomes.
  • crossing refers to the act of forming an embryo from gametes of two distinct plants or plant lines.
  • Crossing may refer to pollinating a plant or plant line using the pollen of a different plant or plant line, e.g., using the pollen of a haploid-inducing plant line to pollinate a donor plant line.
  • crossing may refer to successful fertilization of an egg by a sperm cell resulting in an embryo or may refer to induction of haploid or tetrahaploid embryo development without successful fertilization of the egg.
  • true homozygous plant line and “true homozygous seed” typically refer to a monoallelic plant line or seed, respectively, having allelic uniformity across all chromosomal loci within a subgenome.
  • hybrid describes a plant or a part thereof, such as a seed, comprising two haplotypes within a subgenome.
  • uniform F1 hybrid refers to the genetically uniform first filial generation of hybrid seeds or plants resulting from the cross of two fully homozygous parent plants each having a different haplotype.
  • uniform octoploid F1 hybrid strawberry seed refers to a genetically uniform population of seed generated from the cross of two true homozygous octoploids or octoploids derived from doubled tetrahaploid strawberry plants or plant lines wherein each parent may possess a different haplotype.
  • genetic modification refers to any sequence or portion thereof within a nucleic acid molecule that differs from the sequence of an ancestral nucleic acid molecule. For example, a seed that contains an inserted or deleted genomic sequence that is not present in one of its parent plants comprises a genetic modification. A genetic modification may be naturally occurring or introduced.
  • a genetic modification may be introduced via, for example: plant breeding to introduce a naturally-occurring genetic modification of one plant line into another plant line; transgenic methods; gene editing; chemical mutagenesis; and the like.
  • expression and “expression level” refer to the relative or absolute amount of a functional gene product present in a cell.
  • gene products include, but are not limited to, nucleic acids (e.g., RNA), post-transcriptionally modified nucleic acids (e.g., spliced RNA, poly-adenylated mRNA), proteins (e.g., enzymes, structural proteins, etc.), and post-translationally modified proteins (e.g., glycoproteins, lipoproteins, etc.).
  • the function of the gene product refers to the wild-type, unmodified, uninhibited function of the gene product.
  • decreased expression refers to a relative decrease in the amount of a functional gene product present in a cell.
  • the decreased expression may refer to a decrease in the total amount of a gene product present in a cell (e.g., a decrease in the amount of a protein) or to a decrease in the amount of functional gene products present in a cell (e.g., a decrease in the percentage of proteins with wild-type function) or to a decrease in the function of gene products present in a cell (e.g., a decrease in the activity of proteins as compared to proteins with wild-type function).
  • the decreased expression may be of a gene product encoded at a certain genomic locus, and may be relative to a control strawberry plant (e.g., a strawberry plant of the same species).
  • non-expression refers to the absence of a functional gene product present in a cell, or to an 34 sf-6744554 197072001240 expression level insufficient for detection of the gene product in the cell, or to an expression level insufficient to result in the function of the gene product within the cell, or to an activity level insufficient to result in the detectable activity of the gene product within the cell.
  • CENH3 gene typically refers to a gene encoding “centromere- specific histone 3 variant” (abbreviated as “CENH3”), or a gene having nucleotide or amino acid homology thereto.
  • Genes may include DNA sequences encoding a CENH3 protein or any gene product thereof, for example, nucleic acids (e.g., RNA), post-transcriptionally modified nucleic acids (e.g., spliced RNA, poly-adenylated mRNA), CENH3 protein, and post-translationally modified CENH3 protein. Genes also include any CENH3 gene orthologs, paralogs, and homeologs within a strawberry plant or cell. [0105] As used herein, “transgenesis” typically refers to the insertion of an exogenous genetic element into the genome of an organism.
  • any exogenous genetic element may be inserted via transgenesis, including, but not limited to, genes, protein coding sequences, non- protein coding sequences, regulatory sequences, spacer DNA, and the like.
  • gene editing typically refers to a type of genetic modification in which DNA is inserted, deleted or substituted in the genome of an organism using one or more natural or engineered nucleases. Gene editing may be carried out using site-specific nucleases, guided nucleases, or a combination thereof. The nuclease creates one or more site- specific breaks, such as double-strand breaks (DSBs) at target loci in the genome.
  • DSBs double-strand breaks
  • Each site- specific break may be repaired, for example via non-homologous end joining (NHEJ), resulting in a genetic modification in the genome at the target locus; or via homologous recombination of the target locus with a provided repair nucleic acid molecule comprising homology to the target genomic sequence and the desired genetic modification.
  • NHEJ non-homologous end joining
  • a haploid-inducing strawberry plant or a plant part thereof, that triggers the development of haploid embryos and seeds when crossed with another strawberry plant.
  • the haploid-inducing strawberry plant or part thereof comprises one or more genetic modifications resulting in decreased expression of one or more CENH3 genes, such as any of the genetic modifications described herein.
  • provided herein is pollen, seed, pistil, stigma or a stolon of a haploid-inducing strawberry plant. 35 sf-6744554 197072001240 [0108]
  • described herein is a haploid-inducing plant of the genus Potentilla, or a plant part thereof, that that triggers the development of haploid embryos and seeds when crossed with another strawberry plant.
  • the haploid- inducing plant of the genus Potentilla or part thereof comprises one or more genetic modifications resulting in decreased expression of one or more CENH3 genes, such as any of the genetic modifications described herein.
  • provided herein is pollen, seed, pistil, stigma or a stolon of a haploid-inducing plant of the genus Potentilla.
  • the haploid- inducing strawberry plant is a plant of the genus Potentilla.
  • the haploid- inducing strawberry plant is a plant of the species Potentilla micrantha.
  • haploid-inducing strawberry plants may be of any ploidy, for example, diploid (2x), triploid (3x), tetraploid (4x), pentaploid (5x), hexaploid (6x), septaploid (or heptaploid, 7x), octoploid (8x), or of a higher ploidy (e.g., greater than 8x, e.g., 9x, 10x, 11x, 12x, 13x, 14x, 15x, 16x, 17x, 18x, 19x, or 20x).
  • the haploid-inducing strawberry plants may be of any taxonomic distinction known to those of skill in the art as strawberry plants.
  • the haploid- inducing strawberry plant is a plant of the family Rosaceae.
  • the haploid- inducing strawberry plant is a plant of the genus Fragaria.
  • the haploid-inducing strawberry plant, or plant part thereof is diploid, for example, a diploid species of the genus Fragaria.
  • Diploid species of strawberry plants include, for example, Fragaria vesca, Fragaria iinumae, Fragaria nipponica, Fragaria viridis, Fragaria ⁇ bifera, Fragaria bucharica, Fragaria chinensis, Fragaria daltoniana, Fragaria emeiensis, Fragaria hayatae, Fragaria iinumae, Fragaria mandshurica, Fragaria viridis, Fragaria nilgerrensis, Fragaria nipponica, Fragaria nubicola, and Fragaria pentaphylla.
  • the haploid-inducing strawberry plant is a plant of the species Fragaria vesca.
  • the haploid-inducing strawberry plant may be of any subspecies of Fragaria vesca, for example, Fragaria vesca ssp. vesca, Fragaria vesca ssp. americana, or Fragaria vesca ssp. bracteate.
  • the haploid-inducing strawberry plant may be of any variety of Fragaria vesca, for example, Rügen, Alexandria, Hawaii 4, 36 sf-6744554 197072001240 Baron Soleraum, Weisse Solemacher, Golden Alexandria, Quarantaine de Prin, Blanc Amélioré, Illa Martin, or Kleinfreude.
  • the haploid-inducing strawberry plant, or plant part thereof is tetraploid, for example, a tetraploid species or hybrid of the genus Fragaria.
  • Tetraploid species of strawberries include, for example, Fragaria corymbosa, Fragaria gracilis, Fragaria ⁇ intermedia, Fragaria moupinensis, Fragaria orientalis, and Fragaria tibetica.
  • the haploid-inducing strawberry plant, or plant part thereof is hexaploid, for example, a hexaploid species or hybrid of the genus Fragaria.
  • Hexaploid species of strawberries include, for example, Fragaria moschata.
  • the haploid-inducing strawberry plant, or plant part thereof is octoploid, for example, an octoploid species or hybrid of the genus Fragaria.
  • the haploid-inducing strawberry plant may be of any subspecies of Fragaria chiloensis, for example, Fragaria chiloensis subsp. chiloensis, Fragaria chiloensis subsp. lucida, Fragaria chiloensis subsp. pacifica, and Fragaria chiloensis subsp. sandwicensis. [0117]
  • the haploid-inducing strawberry plant, or plant part thereof has a ploidy of 10x, for example, a species or hybrid of the genus Fragaria having a ploidy of 10x.
  • Strawberry species having a ploidy of 10x include, for example, Fragaria cascadensis and Fragaria iturupensis.
  • the haploid-inducing strawberry plant, or plant part thereof is a hybrid between any of the strawberry species described herein, or is a synthetic polyploid 37 sf-6744554 197072001240 derived from a diploid species.
  • the haploid inducing strawberry plant is a synthetic polyploid derived from any diploid species, hybrid or synthetic polyploid.
  • CENH3 Genes [0119]
  • the haploid-inducing strawberry plant, or plant part thereof comprises one or more genetic modifications resulting in decreased expression of one or more CENH3 genes.
  • the haploid-inducing strawberry plant, or plant part thereof comprises one or more genetic modifications resulting in a decreased amount of a functional gene product encoded by one or more CENH3 genes (e.g., a functional CENH3 protein).
  • the gene products encoded by the CENH3 genes may include, but are not limited to, nucleic acids (e.g., RNA), post-transcriptionally modified nucleic acids (e.g., spliced RNA, poly-adenylated mRNA), proteins (e.g., enzymes, structural proteins, etc.), and post-translationally modified proteins (e.g., glycoproteins, lipoproteins, etc.).
  • the one or more genetic modifications resulting in decreased expression of one or more CENH3 genes may include, but are not limited to, modification of an enhancer in one or more CENH3 genes, modification of a promoter of one or more CENH3 genes, modification of a coding region in one or more CENH3 genes, modification of an intron in one or more CENH3 genes, modification of methylation status of one or more CENH3 genes, expression of a repressor protein that targets the DNA or an mRNA of one or more CENH3 genes, and expression of an RNA interference construct that targets one or more mRNAs from one or more CENH3 genes.
  • the haploid-inducing strawberry plant, or plant part thereof may comprise one or more genetic modifications resulting in non-expression of one or more CENH3 genes. In some embodiments, the haploid-inducing strawberry plant, or plant part thereof, may comprise one or more genetic modifications resulting in decreased expression, non-expression, or a combination thereof of a plurality of CENH3 genes. In some embodiments, the haploid- inducing strawberry plant, or plant part thereof, may comprise one or more genetic 38 sf-6744554 197072001240 modifications resulting in decreased activity or function of one or more CENH3 proteins or one or more domains of a CENH3 protein.
  • the domain of the CENH3 protein comprise an N-terminal tail, a Histone Fold Domain (HSD), or a combination thereof.
  • HSD Histone Fold Domain
  • the structure of CENH3 proteins, including the N-terminal tail and Histone Fold Domain, have been described previously, for example, in Wang et al. 2019 (Centromere histone H3- and phospholipase-mediated haploid induction in plants. Plant Methods 15, 42).
  • the haploid-inducing strawberry plant, or plant part thereof comprises one or more genetic modifications resulting in decreased expression of one or more CENH3 genes.
  • the haploid-inducing strawberry plant, or plant part thereof comprises one or more genetic modifications resulting in decreased expression of two or more (e.g., two, three, four, or more) CENH3 genes.
  • a CENH3 gene is exemplified by a FvCENH3 gene from diploid strawberry Fragaria vesca, and specifically by the FvCENH3 nucleotide sequences, FvCENH3 protein sequences, and percent identities described herein.
  • FvCENH3 protein (SEQ ID NO: 8) and nucleotide (SEQ ID NO: 1) sequences are provided in Table 1.
  • FvCENH3 genes, and gene products thereof include nucleotide sequences having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity or complementarity to SEQ ID NO. 8, or a fragment thereof.
  • Gene products of FvCENH3 genes include FvCENH3 proteins having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity, or at least 60%, 65%, 70%, 75%, 80%, 85%, 95%, 96%, 97%, 98%, or 99% sequence similarity to the FvCENH3 protein of SEQ ID NO. 1, or a fragment thereof.
  • a CENH3 gene is further exemplified by a FaCENH3-1a gene from chromosome 7 (subgenome 1, Fvb7-1) of octoploid strawberry Fragaria x ananassa (e.g., variety Camarosa), and specifically by the FaCENH3-1a nucleotide sequences, FaCENH3-1a protein sequences, and percent identities described herein.
  • FaCENH3-1a protein (SEQ ID NO: 9) and FaCENH3-1a nucleotide (SEQ ID NO: 2) sequences are provided in Table 1. The locus of FaCENH3-1a in the Fragaria x ananassa genome is shown in FIG.
  • FaCENH3-1a genes, and gene products thereof include nucleotide sequences having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity or complementarity to the FaCENH3-1a gene nucleotide sequence of SEQ ID NO. 2, or a fragment thereof.
  • FaCENH3-1a genes also include FaCENH3-1a proteins having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity, or at least 60%, 65%, 70%, 75%, 80%, 85%, 95%, 96%, 97%, 98%, or 99% sequence similarity to the FaCENH3-1a protein of SEQ ID NO. 9, or a fragment thereof.
  • a CENH3 gene is further exemplified by a FaCENH3-1b gene from chromosome 7 (subgenome 1, Fvb7-1) of octoploid strawberry Fragaria x ananassa (e.g., variety Camarosa), and specifically by the FaCENH3-1b nucleotide sequences, FaCENH3-1b protein sequences, and percent identities described herein.
  • FaCENH3-1b protein (SEQ ID NO: 10) and FaCENH3-1b nucleotide (SEQ ID NO: 3) sequences are provided in Table 1. The locus of FaCENH3-1b in the Fragaria x ananassa genome is shown in FIG.
  • FaCENH3-1b genes, and gene products thereof include nucleotide sequences having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity or complementarity to the FaCENH3-1b gene nucleotide sequence of SEQ ID NO. 3, or a fragment thereof.
  • FaCENH3-1b genes also include FaCENH3-1b proteins having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity, or at least 60%, 65%, 70%, 75%, 80%, 85%, 95%, 96%, 97%, 98%, or 99% sequence similarity to the FaCENH3-1b protein of SEQ ID NO. 10, or a fragment thereof.
  • a CENH3 gene is further exemplified by a FaCENH3-2a gene from chromosome 7 (subgenome 2, Fvb7-2) of octoploid strawberry Fragaria x ananassa (e.g., variety Camarosa), and specifically by the FaCENH3-2a nucleotide sequences, FaCENH3-2a protein sequences, and percent identities described herein.
  • FaCENH3-2a protein SEQ ID NO: 11
  • FaCENH3-2a nucleotide SEQ ID NO: 4
  • 40 sf-6744554 197072001240 sequences are provided in Table 1.
  • the locus of FaCENH3-2a in the Fragaria x ananassa genome is shown in FIG.
  • FaCENH3-2a genes, and gene products thereof include nucleotide sequences having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity or complementarity to the FaCENH3-2a gene nucleotide sequence of SEQ ID NO. 4, or a fragment thereof.
  • FaCENH3-2a genes also include FaCENH3-2a proteins having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity, or at least 60%, 65%, 70%, 75%, 80%, 85%, 95%, 96%, 97%, 98%, or 99% sequence similarity to the FaCENH3-2a protein of SEQ ID NO. 11, or a fragment thereof.
  • a CENH3 gene is further exemplified by a FaCENH3-2b gene from chromosome 7 (subgenome 2, Fvb7-2) of octoploid strawberry Fragaria x ananassa (e.g., variety Camarosa), and specifically by the FaCENH3-2b nucleotide sequences, FaCENH3-2b protein sequences, and percent identities described herein.
  • FaCENH3-2b protein (SEQ ID NO: 12) and FaCENH3-2b nucleotide (SEQ ID NO: 5) sequences are provided in Table 1. The locus of FaCENH3-2b in the Fragaria x ananassa genome is shown in FIG.
  • FaCENH3-2b genes, and gene products thereof include nucleotide sequences having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity or complementarity to the FaCENH3-2b gene nucleotide sequence of SEQ ID NO. 5, or a fragment thereof.
  • FaCENH3-2b genes also include FaCENH3-2b proteins having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity, or at least 60%, 65%, 70%, 75%, 80%, 85%, 95%, 96%, 97%, 98%, or 99% sequence similarity to the FaCENH3-2b protein of SEQ ID NO. 12, or a fragment thereof.
  • a CENH3 gene is further exemplified by a FaCENH3-3 gene from chromosome 7 (subgenome 3, Fvb7-3) of octoploid strawberry Fragaria x ananassa (e.g., variety Camarosa), and specifically by the FaCENH3-3 nucleotide sequences, 41 sf-6744554 197072001240 FaCENH3-3 protein sequences, and percent identities described herein.
  • FaCENH3-3 protein (SEQ ID NO: 13) and FaCENH3-3 nucleotide (SEQ ID NO: 6) sequences are provided in Table 1. The locus of FaCENH3-3 in the Fragaria x ananassa genome is shown in FIG.
  • FaCENH3-3 genes, and gene products thereof include nucleotide sequences having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity or complementarity to the FaCENH3-3 gene nucleotide sequence of SEQ ID NO. 6, or a fragment thereof.
  • FaCENH3-3 genes also include FaCENH3-3 proteins having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity, or at least 60%, 65%, 70%, 75%, 80%, 85%, 95%, 96%, 97%, 98%, or 99% sequence similarity to the FaCENH3-3 protein of SEQ ID NO. 13, or a fragment thereof.
  • a CENH3-4 gene is further exemplified by a FaCENH3- 4 gene from chromosome 7 (subgenome 2, Fvb7-4) of octoploid strawberry Fragaria x ananassa (e.g., variety Camarosa), and specifically by the FaCENH3-4 nucleotide sequences, FaCENH3-4 protein sequences, and percent identities described herein.
  • FaCENH3-4 protein (SEQ ID NO: 14) and FaCENH3-4 nucleotide (SEQ ID NO: 7) sequences are provided in Table 1. The locus of FaCENH3-4 in the Fragaria x ananassa genome is shown in FIG.
  • FaCENH3-4 genes, and gene products thereof include nucleotide sequences having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity or complementarity to the FaCENH3-4 gene nucleotide sequence of SEQ ID NO. 7, or a fragment thereof.
  • FaCENH3-4 genes also include FaCENH3-4 proteins having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity, or at least 60%, 65%, 70%, 75%, 80%, 85%, 95%, 96%, 97%, 98%, or 99% sequence similarity to the FaCENH3-4 protein of SEQ ID NO. 14, or a fragment thereof.
  • a CENH3 gene is further exemplified by a FaCENH3-7a gene from chromosome 7 (subgenome A) of octoploid strawberry Fragaria x ananassa (e.g., variety Royal Royce), and specifically by the FaCENH3-7a nucleotide sequences, FaCENH3-7a protein sequences, and percent identities described herein.
  • FaCENH3-7a protein (SEQ ID NO: 53) and FaCENH3-7a nucleotide (SEQ ID NOs: 49 and 58) sequences are provided in Table 1.
  • FIG. 3B top chromosome, chr_7A
  • FIG. 5D-G a phylogenetic tree showing the relationship between FaCENH3-7a and CENH3 proteins from other Fragaria x ananassa and other plant species is shown in FIG. 5D-G.
  • FaCENH3-7a genes, and gene products thereof include nucleotide sequences having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity or complementarity to the FaCENH3-7a gene nucleotide sequence of SEQ ID NO. 49 or 58, or a fragment thereof.
  • FaCENH3-7a genes also include FaCENH3-7a proteins having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity, or at least 60%, 65%, 70%, 75%, 80%, 85%, 95%, 96%, 97%, 98%, or 99% sequence similarity to the FaCENH3-7a protein of SEQ ID NO. 53, or a fragment thereof.
  • a CENH3 gene is further exemplified by a FaCENH3-7b gene from chromosome 7 (subgenome B) of octoploid strawberry Fragaria x ananassa (e.g., variety Royal Royce), and specifically by the FaCENH3-7b nucleotide sequences, FaCENH3-7b protein sequences, and percent identities described herein.
  • FaCENH3-7b protein (SEQ ID NO: 54) and FaCENH3-7b nucleotide (SEQ ID NOs: 50 and 59) sequences are provided in Table 1. The locus of FaCENH3-7b in the Fragaria x ananassa genome is shown in FIG.
  • FaCENH3-7b genes, and gene products thereof include nucleotide sequences having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity or complementarity to the FaCENH3-7b gene nucleotide sequence of SEQ ID NO. 50 or 59, or a fragment thereof.
  • a CENH3 gene is further exemplified by a FaCENH3-7c gene from chromosome 7 (subgenome C) of octoploid strawberry Fragaria x ananassa (e.g., variety Royal Royce), and specifically by the FaCENH3-7c nucleotide sequences, FaCENH3-7c protein sequences, and percent identities described herein.
  • FaCENH3-7c protein (SEQ ID NO: 55) and FaCENH3-7c nucleotide (SEQ ID NOs: 51 and 60) sequences are provided in Table 1. The locus of FaCENH3-7c in the Fragaria x ananassa genome is shown in FIG.
  • FaCENH3-7c genes also include FaCENH3-7c proteins having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity, or at least 60%, 65%, 70%, 75%, 80%, 85%, 95%, 96%, 97%, 98%, or 99% sequence similarity to the FaCENH3-7c protein of SEQ ID NO. 55, or a fragment thereof.
  • a CENH3 gene is further exemplified by a FaCENH3-7d gene from chromosome 7 (subgenome D) of octoploid strawberry Fragaria x ananassa (e.g., variety Royal Royce), and specifically by the FaCENH3-7d nucleotide sequences, FaCENH3-7d protein sequences, and percent identities described herein.
  • FaCENH3-7d protein (SEQ ID NO: 56) and FaCENH3-7d nucleotide (SEQ ID NOs: 52 and 61) sequences are provided in Table 1. The locus of FaCENH3-7d in the Fragaria x ananassa genome is shown in FIG.
  • FaCENH3-7d genes, and gene products thereof include nucleotide sequences having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity or complementarity to the FaCENH3-7d gene nucleotide sequence of SEQ ID NO. 52 or 61, or a fragment thereof.
  • FaCENH3-7d genes also include 44 sf-6744554 197072001240 FaCENH3-7d proteins having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity, or at least 60%, 65%, 70%, 75%, 80%, 85%, 95%, 96%, 97%, 98%, or 99% sequence similarity to the FaCENH3-7d protein of SEQ ID NO. 56, or a fragment thereof.
  • CENH3 gene and protein sequences described herein are reference sequences and may be used by one of ordinary skill in the art to identify CENH3 in any species or variety of strawberry. For example, using methods that are routine in the art, one of ordinary skill in the art could utilize any of the CENH3 gene or protein sequences described herein to query a nucleic acid database or a genome sequence to identify CENH3 genes in the database or genome sequence.
  • Genetic Modifications [0133]
  • the haploid-inducing strawberry plant, or plant part thereof, comprises one or more genetic modifications. Genetic modifications may be generated by modification of any nucleic acid sequence or genetic element by insertion, deletion, or substitution of one or more nucleotides in a nucleic acid molecule.
  • Such modifications can be achieved by any of several well-known methods known in the art including, but not limited to, random mutagenesis, gene editing, insertion of a recombinant nucleic acid, crossing of an unmodified plant (e.g., strawberry plant) with a modified plant (e.g., strawberry plant) to introduce the modification of the modified plant into the unmodified plant, and the like.
  • a genetic modification may be naturally occurring or non-naturally occurring.
  • the genetic modifications described herein may be present in any known genetic element including, but not limited to, protein-coding sequences, non-protein-coding sequences, promoter regions, 5' untranslated leaders, genes, exons, introns, poly-A signal sequences, 3' untranslated regions, regions encoding small RNAs (such as microRNAs and small-interfering RNAs), and any other sequences that affect transcription or translation of one or more nucleic acid sequences.
  • genetic modifications may 45 sf-6744554 197072001240 include, but are not limited to, modifying or replacing nucleotide sequences of interest (such as a regulatory elements), gene disruption, gene knockout, gene knockdown, gene knock-in, gene silencing (including, e.g., by expressing an inverted repeat into a gene of interest), RNA interference (including, e.g., by insertion and/or expression of an RNA interference construct), modification of methylation status, modification of splicing sites, introducing alternate splicing sites, or any combination thereof.
  • nucleotide sequences of interest such as a regulatory elements
  • gene disruption such as a regulatory elements
  • gene knockout gene knockout
  • gene knock-in gene silencing
  • gene silencing including, e.g., by expressing an inverted repeat into a gene of interest
  • RNA interference including, e.g., by insertion and/or expression of an RNA interference construct
  • modification of methylation status modification of splicing
  • gene disruption refers to the alteration or insertion of a sequence into a gene or locus that results in decreased expression (including, e.g., decreased activity or non-expression) of a functional gene product (e.g., a functional protein gene product) of the gene.
  • a gene disruption may be achieved by introduction of a genetic modification in a protein-coding sequence, including, but not limited to, as a missense or nonsense mutation, or an insertion, deletion, or substitution.
  • a knockout is a genetic modification wherein a gene or gene product has been rendered completely inoperative.
  • a knockout of a gene product may be achieved by introduction of a genetic modification in a protein-coding sequence of a gene or any non- protein-coding or regulatory sequence described herein.
  • a knockdown is a genetic modification wherein a gene or gene product has been rendered partially inoperative.
  • a knockdown of a gene product may be achieved by introduction of a genetic modification in a protein-coding sequence of a gene or in a non-protein-coding or regulatory sequence, or insertion of a trans-acting element, such as a construct that expresses an inverted repeat of the gene product or a construct that expresses a DNA- or RNA-binding protein such as a transcriptional repressor which may include, for example, a deactivated targeted nuclease such as deactivated CRISPR-associated (Cas) nuclease (e.g., dCas9).
  • Cas CRISPR-associated nuclease
  • knock- in represents the replacement or insertion of a DNA sequence at a specific DNA locus in a cell.
  • Knock-ins may include, but are not limited to, specific insertion of a heterologous amino acid coding sequence in a coding region of a gene, an insertion of a transcriptional regulatory element in a genetic locus, or any of several methods of inserting a DNA sequence into a call that are known to one of ordinary skill in the art.
  • the haploid-inducing strawberry plant, or plant part thereof comprises one or more genetic modifications resulting in decreased expression decreased expression (including, e.g., decreased activity or non-expression) of a gene product of a genomic locus.
  • genetic modifications resulting in decreased expression (including, e.g., decreased activity or non-expression) of a gene product or locus may include, but are not limited to, modification of an enhancer, modification of a promoter, 46 sf-6744554 197072001240 modification of a 5’ untranslated leader, modification of a coding region, modification of a non-coding region, insertion and/or expression of an RNA interference construct that targets an mRNA, modification of a region encoding a small RNA, modification of methylation status of a genomic locus, expression of a repressor protein that targets a DNA or mRNA sequence, and any other sequences that affect transcription or translation of one or more nucleic acid sequences.
  • genetic modifications resulting in decreased expression (including, e.g., decreased activity or non-expression) of a gene product or locus may include, but are not limited to, modifying or replacing nucleotide sequences of interest (such as a regulatory elements), gene disruption, gene knockout, gene knockdown, gene knock-in, gene silencing (including, e.g., by inserting and/or expressing an inverted repeat into a gene of interest), RNA interference (including, e.g., by insertion and/or expression of an RNA interference construct), expression of a repressor protein (e.g., dCas9), modification of methylation status of gene loci, modification of splicing sites, introducing alternate splicing sites, or any combination thereof.
  • a repressor protein e.g., dCas9
  • the haploid-inducing strawberry plant comprises one or more genetic modifications resulting in decreased expression (e.g., decreased activity) of one or more CENH3 proteins described herein. In some embodiments, the haploid-inducing strawberry plant comprises one or more genetic modifications of one or more CENH3 genes.
  • the haploid-inducing strawberry comprises one or more genetic modifications resulting in decreased expression of one or more CENH3 proteins having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity, or at least 60%, 65%, 70%, 75%, 80%, 85%, 95%, 96%, 97%, 98%, or 99% sequence similarity to an amino acid sequence selected from the list consisting of SEQ ID NOs: 8-14 and 53-56, or fragments thereof.
  • the haploid-inducing strawberry plant comprises one or more genetic modifications of one or more CENH3 genes comprising a nucleotide sequence having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity or complementarity to a nucleotide sequence selected from the group consisting SEQ ID NOs: 1-7 and 58-61, or a fragment thereof.
  • the one or more genetic modifications comprise a modification of an enhancer of one or more of the CENH3 genes, a modification of a promoter of one or more of the CENH3 genes, a modification of a coding region of one or more of the CENH3 genes, a modification of an intron of one or more of the CENH3 genes, a 47 sf-6744554 197072001240 modification of methylation status of one or more of the CENH3 genes, expression of a repressor protein that targets the DNA or an mRNA of one or more of the CENH3 genes, expression of an RNA interference construct that targets an mRNA of one or more of the CENH3 genes, or any combination thereof.
  • the haploid-inducing strawberry plant has decreased expression of one or more CENH3 genes described herein relative to a strawberry plant (e.g., a control strawberry plant, e.g., a strawberry plant of the same species) lacking the one or more genetic modifications.
  • the haploid-inducing strawberry lacks detectable expression of one or more CENH3 gene products described herein relative to a strawberry plant (e.g., a control strawberry plant, e.g., a strawberry plant of the same species) lacking the one or more genetic modifications.
  • the haploid-inducing strawberry lacks detectable expression of any CENH3 gene products.
  • the haploid-inducing strawberry plant or plant part further comprises one or more naturally-occurring inactive alleles of one or more CENH3 genes.
  • the decreased expression (e.g., decreased activity) of one or more CENH3 proteins is decreased by at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90% , or about 100% as compared to a control plant, plant part, or plant cell lacking the one or more genetic modifications.
  • the decreased expression (e.g., decreased activity) of one or more CENH3 proteins is decreased by no more than about 10%, about 20%, about 30%, about 40%, about 50%, about 60%, about 70%, about 80%, about 90%, about 95%, or about 99% as compared to a control plant, plant part, or plant cell lacking the one or more genetic modifications.
  • the decreased expression (e.g., decreased activity) of one or more CENH3 proteins is decreased is decreased by between about 10% and about 100%, between about 20% and about 100%, between about 30% and about 100%, between about 40% and about 100%, between about 50% and about 100%, between about 60% and about 100%, between about 70% and about 100%, between about 80% and about 100%, between about 90% and about 100%, between about 10% and about 90%, between about 20% and about 90%, between about 30% and about 90%, between about 40% and about 90%, between about 50% and about 90%, between about 60% and about 90%, between about 70% and about 90%, between about 80% and about 90%, between about 10% and about 80%, between about 20% and about 80%, between about 30% and about 80%, between about 40% and about 80%, between about 50% and about 80%, between about 60% and 48 sf-6744554 197072001240 about 80%, between about 70% and about 80%, 10% and about 70%, between about 20% and about 70%, between about 30% and about 70%, between about 40% and about 70%, between about 50% and about 70%, between about 60% and 48 s
  • the haploid-inducing strawberry plant is a plant of the species Fragaria vesca comprising one or more genetic modifications resulting in decreased expression of one or more CENH3 genes. In certain embodiments, the haploid-inducing strawberry plant is a plant of the species Fragaria vesca comprising one or more genetic modifications resulting in decreased expression of an FvCENH3 gene. In certain embodiments, the haploid-inducing strawberry plant is a plant of the species Fragaria vesca comprising one or more genetic modifications resulting in decreased expression of an FvCENH3 protein.
  • the haploid-inducing strawberry plant is a plant of the species Fragaria vesca comprising one or more genetic modifications resulting in decreased expression of a CENH3 protein having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity, or at least 60%, 65%, 70%, 75%, 80%, 85%, 95%, 96%, 97%, 98%, or 99% sequence similarity to the FvCENH3 protein of SEQ ID NO. 8, or a fragment thereof.
  • the haploid-inducing strawberry plant is a plant of the species Fragaria vesca comprising one or more genetic modifications of an FvCENH3 gene or gene product.
  • the haploid-inducing strawberry plant is a plant of the species Fragaria vesca comprising one or more genetic modifications in a CENH3 gene comprising a nucleotide sequence having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity or complementarity to the FvCENH3 gene nucleotide sequence of SEQ ID NO.
  • the haploid-inducing strawberry plant is a plant of the species Fragaria vesca comprising genetic modifications in both alleles of an 49 sf-6744554 197072001240 FvCENH3 gene.
  • the one or more genetic modifications comprise a modification of an enhancer of FvCENH3, a modification of a promoter of FvCENH3, a modification of a coding region of FvCENH3, modification of an intron of FvCENH3, or any combination thereof relative to a control Fragaria vesca plant (e.g., a Fragaria vesca plant lacking one or more of the genetic modifications).
  • the haploid- inducing strawberry plant is a plant of the species Fragaria vesca and has decreased expression of FvCENH3 protein relative to a Fragaria vesca plant lacking the one or more genetic modifications. In certain embodiments, the haploid-inducing strawberry plant is a plant of the species Fragaria vesca and lacks detectable expression of FvCENH3 protein. [0140] In some embodiments, the haploid-inducing strawberry plant is a plant of the species Fragaria x ananassa comprising one or more genetic modifications resulting in decreased expression of one or more FaCENH3 genes.
  • the haploid- inducing strawberry plant is a plant of the species Fragaria x ananassa comprising one or more genetic modifications resulting in decreased expression of an FaCENH3-1a gene, an FaCENH3-1b gene, an FaCENH3-2a gene, an FaCENH3-2b gene, an FaCENH3-3 gene, an FaCENH3-4 gene, or any combination thereof.
  • the haploid-inducing strawberry plant is a plant of the species Fragaria x ananassa comprising one or more genetic modifications resulting in decreased expression of FaCENH3-1a gene, an FaCENH3- 1b gene, an FaCENH3-2a gene, an FaCENH3-2b gene, an FaCENH3-3 gene, and an FaCENH3-4 gene.
  • the haploid-inducing strawberry plant is a plant of the species Fragaria x ananassa comprising one or more genetic modifications resulting in decreased expression of an FaCENH3-1a protein, an FaCENH3-1b protein, an FaCENH3-2a protein, an FaCENH3-2b protein, an FaCENH3-3 protein, an FaCENH3-4 protein, or any combination thereof.
  • the haploid-inducing strawberry plant is a plant of the species Fragaria x ananassa comprising one or more genetic modifications resulting in decreased expression of an FaCENH3-1a protein, an FaCENH3-1b protein, an FaCENH3- 2a protein, an FaCENH3-2b protein, an FaCENH3-3 protein, and an FaCENH3-4 protein.
  • the haploid-inducing strawberry plant is a plant of the species Fragaria x ananassa comprising one or more genetic modifications resulting in decreased expression of one or more CENH3 proteins having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence 50 sf-6744554 197072001240 identity, or at least 60%, 65%, 70%, 75%, 80%, 85%, 95%, 96%, 97%, 98%, or 99% sequence similarity to an amino acid sequence selected from the list consisting of SEQ ID NOs: 9-14, or fragments thereof.
  • the haploid-inducing strawberry plant is a plant of the species Fragaria x ananassa comprising one or more genetic modifications resulting in decreased expression of 1) a CENH3 protein having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity, or at least 60%, 65%, 70%, 75%, 80%, 85%, 95%, 96%, 97%, 98%, or 99% sequence similarity to the amino acid sequence of SEQ ID NO: 9; 2) a CENH3 protein having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity, or at least 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%
  • the haploid-inducing strawberry plant is a plant of the species Fragaria x ananassa comprising one or more genetic modifications resulting in decreased expression of an FaCENH3-7a protein, an FaCENH3-7b protein, an FaCENH3-7c protein, an FaCENH3-7d protein, or any combination thereof.
  • the haploid-inducing strawberry plant is a plant of the species Fragaria x ananassa comprising one or more genetic modifications resulting in decreased expression of one or more CENH3 51 sf-6744554 197072001240 proteins having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity, or at least 60%, 65%, 70%, 75%, 80%, 85%, 95%, 96%, 97%, 98%, or 99% sequence similarity to an amino acid sequence selected from the list consisting of SEQ ID NOs: 53-56, or fragments thereof.
  • the haploid-inducing strawberry plant is a plant of the species Fragaria x ananassa comprising one or more genetic modifications resulting in decreased expression of 1) a CENH3 protein having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity, or at least 60%, 65%, 70%, 75%, 80%, 85%, 95%, 96%, 97%, 98%, or 99% sequence similarity to the amino acid sequence of SEQ ID NO: 53; 2) a CENH3 protein having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity, or at least 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%
  • the haploid-inducing strawberry plant is a plant of the species Fragaria x ananassa comprising one or more genetic modifications of an FaCENH3- 1a gene or gene product, an FaCENH3-1b gene or gene product, an FaCENH3-2a gene or gene product, an FaCENH3-2b gene or gene product, an FaCENH3-3 gene or gene product, an FaCENH3-4 gene or gene product, or any combination thereof.
  • the haploid-inducing strawberry plant is a plant of the species Fragaria x ananassa comprising one or more genetic modifications of an FaCENH3-1a gene or gene product, an FaCENH3- 1b gene or gene product, an FaCENH3-2a gene or gene product, an FaCENH3-2b gene or gene product, an FaCENH3-3 gene or gene product, and an FaCENH3-4 gene or gene product.
  • the haploid-inducing strawberry plant is a plant of the species Fragaria x ananassa comprising one or more genetic modifications in a CENH3 gene comprising a nucleotide sequence having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 52 sf-6744554 197072001240 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity or complementarity to a nucleotide sequence selected from the group consisting of SEQ ID NOs: 2-71, or a fragment thereof.
  • the haploid- inducing strawberry plant is a plant of the species Fragaria x ananassa comprising: 1) one or more genetic modifications in a CENH3 gene comprising a nucleotide sequence having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity or complementarity to the nucleotide sequence of SEQ ID NO: 2, or a fragment thereof; 2) one or more genetic modifications in a CENH3 gene comprising a nucleotide sequence having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity or complementarity to the nucleotide sequence of SEQ ID NO: 3, or a fragment thereof
  • the one or more genetic modifications comprise a modification of an enhancer of an FaCENH3 gene (e.g., an FaCENH3-1a gene, an FaCENH3-1b gene, an FaCENH3-2a gene, an FaCENH3-2b gene, an FaCENH3-3 gene, and/or an FaCENH3-4 gene), a modification of a promoter of an FaCENH3 gene (e.g., an FaCENH3-1a gene, an FaCENH3-1b gene, an FaCENH3-2a gene, an FaCENH3-2b gene, an FaCENH3-3 gene, and/or an FaCENH3-4 gene), a modification of a coding region of an FaCENH3 gene (e.g., an FaCENH3-1a gene, an FaCENH3-1b gene, an FaCENH3-2a gene, an FaCENH3-2b gene, an FaCENH3-3 gene, and/or an FaCENH3-4 gene), modification of an intron of an 53 sf-6744554 197072001240 FaCENH3 gene (
  • the haploid-inducing strawberry plant is a plant of the species Fragaria x ananassa comprising one or more genetic modifications of an FaCENH3- 7a gene or gene product, an FaCENH3-7b gene or gene product, an FaCENH3-7c gene or gene product, an FaCENH3-7d gene or gene product, or any combination thereof.
  • the haploid-inducing strawberry plant is a plant of the species Fragaria x ananassa comprising one or more genetic modifications in a CENH3 gene comprising a nucleotide sequence having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity or complementarity to a nucleotide sequence selected from the group consisting of SEQ ID NOs: 58-61, or a fragment thereof.
  • the haploid-inducing strawberry plant is a plant of the species Fragaria x ananassa comprising: 1) one or more genetic modifications in a CENH3 gene comprising a nucleotide sequence having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity or complementarity to the nucleotide sequence of SEQ ID NO: 58, or a fragment thereof; 2) one or more genetic modifications in a CENH3 gene comprising a nucleotide sequence having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity or complementarity to the nucleotide sequence of SEQ ID NO: 59, or
  • the one or more genetic modifications comprise a modification of an enhancer of an FaCENH3 gene (e.g., an FaCENH3-7a gene, an FaCENH3-7b gene, an 54 sf-6744554 197072001240 FaCENH3-7c gene, and/or an FaCENH3-7d gene), a modification of a promoter of an FaCENH3 gene (e.g., an FaCENH3-7a gene, an FaCENH3-7b gene, an FaCENH3-7c gene, and/or an FaCENH3-7d gene), a modification of a coding region of an FaCENH3 gene (e.g., an FaCENH3-7a gene, an FaCENH3-7b gene, an FaCENH3-7c gene, and/or an FaCENH3- 7d gene), modification of an intron of an FaCENH3 gene (e.g., an FaCENH3-7a gene, an FaCENH3-7b gene, an FaCENH3-7c gene, and/or an FaCENH3-
  • the one or more genetic modifications comprise one or more deletions, insertions, or nucleotide substitutions in a coding region of an FaCENH3 gene (e.g., an FaCENH3-7a gene, an FaCENH3-7b gene, an FaCENH3-7c gene, and/or an FaCENH3-7d gene).
  • the one or more genetic modifications in a CENH3 gene each independently comprise an insertion, a deletion, one or more nucleotide changes, or an inversion that results in decreased expression (e.g., decreased activity) of the CENH3 gene.
  • the insertion, the deletion, the one or more nucleotide changes, or the inversion eliminates expression (e.g., eliminates activity) of the CENH3 gene. In certain embodiments, the insertion, the deletion, the one or more nucleotide changes, or the inversion decreases, but does not eliminate, expression of the CENH3 gene.
  • the insertion, the deletion, the one or more nucleotide changes, or the inversion is positioned in the first 70%, the first 60%, the first 50%, the first 40%, the first 30%, the first 20%, or the first 10% of the nucleotides of the CENH3 gene (e.g., of a polynucleotide sequence selected from the group consisting of SEQ ID NOs: 1-7).
  • the insertion, the deletion, the one or more nucleotide changes, or the inversion is positioned in the first 100, the first 200, the first 300, the first 400, the first 500, the first 600, the first 700, the first 800, the first 900, the first 1000, the first 1250, the first 1500, the first 1750, the first 2000, the first 2500, or the first 3000 nucleotides of the CENH3 gene (e.g., of a polynucleotide sequence selected from the group consisting of SEQ ID NOs: 1-7).
  • the insertion, the deletion, the one or more nucleotide changes, or the inversion eliminates expression (e.g., eliminates activity) of the CENH3 gene.
  • the insertion, the deletion, the one or more nucleotide changes, or the inversion results in a premature stop codon present in the first 70%, the first 60%, the first 50%, the first 40%, the first 30%, the first 20%, or the first 10% of the nucleotides of a polynucleotide sequence selected from the 55 sf-6744554 197072001240 group consisting of SEQ ID NOs: 1-7 or of the nucleotides of the coding sequence of the CENH3 gene following the start codon in the 3’ direction, thereby eliminating expression (e.g., activity) of the CENH3 gene.
  • the insertion, the deletion, the one or more nucleotide changes, or the inversion results in a premature stop codon present in the first 100, the first 200, the first 300, the first 400, the first 500, the first 600, the first 700, the first 800, the first 900, the first 1000, the first 1250, the first 1500, the first 1750, the first 2000, the first 2500, or the first 3000 nucleotides of a polynucleotide sequence selected from the group consisting of SEQ ID NOs: 1-7 or of the coding sequence of the CENH3 gene following the start codon in the 3’ direction, thereby eliminating expression (e.g., activity) of the CENH3 gene.
  • the haploid-inducing strawberry plant is a plant of the species Fragaria x ananassa and has decreased expression of an FaCENH3-1a protein, an FaCENH3-1b protein, an FaCENH3-2a protein, an FaCENH3-2b protein, an FaCENH3-3 protein, an FaCENH3-4 protein, or any combination (e.g., two, three, four, five or all six) thereof relative to a control Fragaria x ananassa plant (e.g., a Fragaria ananassa plant lacking one or more or all of the genetic modifications).
  • a control Fragaria x ananassa plant e.g., a Fragaria ananassa plant lacking one or more or all of the genetic modifications.
  • the haploid- inducing strawberry plant is a plant of the species Fragaria x ananassa and lacks detectable expression of an FaCENH3-1a protein, an FaCENH3-1b protein, an FaCENH3-2a protein, an FaCENH3-2b protein, an FaCENH3-3 protein, an FaCENH3-4 protein, or any combination (e.g., two, three, four, five or all six) thereof.
  • the haploid-inducing strawberry plant, or plant part thereof may comprise one or more genetic modifications resulting in decreased expression of any combination of CENH3 genes described herein or known in the art.
  • the haploid-inducing strawberry plant, or plant part thereof may comprise one or more genetic modifications resulting in non-expression of any combination of CENH3 genes described here or known in the art.
  • the haploid-inducing strawberry plant, or plant part thereof may comprise one or more genetic modifications resulting in decreased expression, non- expression, or a combination thereof of any combination of CENH3 genes described here or known in the art.
  • the haploid-inducing strawberry plant or plant part lacks detectable expression of CENH3 proteins.
  • the haploid-inducing strawberry plant or plant part thereof has decreased expression (including decreased activity) of any of the CENH3 genes and/or 56 sf-6744554 197072001240 CENH3 proteins described herein relative to a control strawberry plant.
  • the haploid-inducing strawberry plant or plant part thereof has decreased expression (e.g., decreased activity) of one or more CENH3 genes and/or one or more CENH3 proteins, wherein the decreased expression (e.g., decreased activity) of one or more CENH3 proteins CENH3 genes and/or is decreased by at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90% , or about 100% as compared to a control plant, plant part, or plant cell lacking the one or more genetic modifications.
  • the haploid-inducing strawberry plant or plant part thereof has decreased expression (e.g., decreased activity) of one or more CENH3 genes and/or one or more CENH3 proteins, wherein the decreased expression (e.g., decreased activity) of CENH3 genes and/or one or more CENH3 proteins is decreased by no more than about 10%, about 20%, about 30%, about 40%, about 50%, about 60%, about 70%, about 80%, about 90%, about 95%, or about 99% as compared to a control plant, plant part, or plant cell lacking the one or more genetic modifications.
  • the haploid- inducing strawberry plant or plant part thereof has decreased expression (e.g., decreased activity) of one or more CENH3 genes and/or one or more CENH3 proteins, wherein the decreased expression (e.g., decreased activity) of the one or more CENH3 genes and/or one or more CENH3 proteins is decreased is decreased by between about 10% and about 100%, between about 20% and about 100%, between about 30% and about 100%, between about 40% and about 100%, between about 50% and about 100%, between about 60% and about 100%, between about 70% and about 100%, between about 80% and about 100%, between about 90% and about 100%, between about 10% and about 90%, between about 20% and about 90%, between about 30% and about 90%, between about 40% and about 90%, between about 50% and about 90%, between about 60% and about 90%, between about 70% and about 90%, between about 80% and about 90%, between about 10% and about 80%, between about 20% and about 80%, between about 30% and about 80%, between about 40% and about 80%, between about 50% and about 80%, between about 60% and about about 100%, between about
  • Control strawberry plants include, for example, a plant of the same species as the haploid-inducing strawberry plant that lacks one or more or all of the genetic modifications comprised by the haploid-inducing strawberry plant, a wild-type plant of the same species as the haploid-inducing strawberry plant, an unmodified plant of the same species as the haploid-inducing strawberry plant, or a null segregant strawberry plant, which underwent the same genetic manipulation, but is lacking one or more or all of the genetic modifications comprised by the haploid-inducing strawberry plant.
  • the haploid-inducing strawberry plant or plant part thereof has decreased expression of CENH3 proteins relative to a strawberry plant of the same species lacking the one or more genetic modifications.
  • the strawberry plants (e.g., haploid-inducing strawberry plants) described herein having one or more genetic modifications resulting in decreased expression of one or more CENH3 genes may have characteristics that differ from those of a strawberry plant lacking the one or more genetic modifications. Without wishing to be bound by theory, it is believed that decreased expression of one or more CENH3 genes may result in certain changes in mitotic and meiotic activity in plants.
  • the strawberry plants (e.g., haploid-inducing strawberry plants) described herein have decreased pollen germination (e.g., a decreased pollen germination rate) than a control strawberry plant lacking the one or more genetic modifications.
  • the strawberry plant has a pollen germination rate of less than 65%, 60%, 55%, 50%, 45%, 40%, 35%, 30%, 25%, 20%, 15%, 10%, or 5%.
  • the strawberry plants e.g., haploid-inducing strawberry plants described herein have decreased flower diameter as compared to a control strawberry plant lacking the one or more genetic modifications.
  • the strawberry plant has a flower diameter of less than 25, 24, 23, 22, 21, 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, or 10 mm.
  • the strawberry plant has a flower diameter of between 5 mm and 25, 24, 23, 22, 21, 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, or 10 mm.
  • the strawberry plants (e.g., haploid-inducing strawberry plants) described herein have decreased anther width as compared to a control strawberry plant lacking the one or more genetic modifications.
  • the strawberry plant has a anther width of less than 1.5, 1.4, 1.3, 1.2, 1.1, 1.0, 0.9, 0.8, 0.7, 0.6, or 0.5 mm.
  • the strawberry plant has a anther width of between 0.1 mm and 1.5, 1.4, 1.3, 1.2, 1.1, 1.0, 0.9, 0.8, 0.7, 0.6, or 0.5 mm.
  • the strawberry plants (e.g., haploid-inducing strawberry plants) described herein have decreased filament length as compared to a control strawberry plant lacking the one or more genetic modifications.
  • the strawberry plant has a filament length of less than 2.7, 2.6, 2.5, 2.4, 2.3, 2.2, 2.1, 2.0, 1.9, 1.8, 1.7, 1.6, 1.5, 1.4, 1.3, 1.2, 1.1, 1.0, 0.9, 0.8, 0.7, 0.6, or 0.5 mm.
  • the strawberry plant has a filament length of less between 0.1 mm and 2.7, 2.6, 2.5, 2.4, 2.3, 2.2, 2.1, 2.0, 1.9, 1.8, 1.7, 1.6, 1.5, 1.4, 1.3, 1.2, 1.1, 1.0, 0.9, 0.8, 0.7, 0.6, or 0.5 mm.
  • the strawberry plants (e.g., haploid-inducing strawberry plants) described herein have delayed microspore formation as compared to a control strawberry plant lacking the one or more genetic modifications.
  • the strawberry plants (e.g., haploid-inducing strawberry plants) described herein have larger bud size at the time of microspore formation as compared to a control strawberry plant lacking the one or more genetic modifications.
  • the strawberry plant has an average bud size at the time of microspore formation of greater than 4.5, 4.6, 4.7, 4.8, 4.9, 5.0, 5.1, 5.2, 5.35.4, 5.5, 5.6, 5.7, 5.8, 5.9, or 6.0 at the time of microspore formation.
  • the strawberry plant has an average bud size at the time of microspore formation of greater than 4.7 at the time of microspore formation.
  • the strawberry plant has an average bud size at the time of microspore formation of greater than 5.0 at the time of microspore formation.
  • the strawberry plants (e.g., haploid-inducing strawberry plants) described herein have buds with observable tetrads, e.g., as shown in FIGS. 16E and 16F.
  • the strawberry plants (e.g., haploid-inducing strawberry plants) described herein have observable tetrads in at least 10%, 20%, 30%, 40%, 50%, 60%, or 70% of buds.
  • the strawberry plants (e.g., haploid-inducing strawberry plants) described herein have fewer seeds per berry as compared to a control strawberry plant lacking the one or more genetic modifications.
  • the strawberry plant has fewer than 100, 90, 80, 70, 60, 50, 40, 30, 20, or 10 seeds per berry.
  • the strawberry plant has between 1 and 100, 90, 80, 70, 60, 50, 40, 30, 20, or 10 seeds per berry.
  • the plant part may be any plant part known in the art, including, but not limited to, a flower, a pistil, a leaf, a stem, a petiole, a cutting, a tissue, a seed coat, an ovule, pollen, a sperm cell, a root, a fruit (e.g., an aggregate fruit), a cotyledon, a hypocotyl, a protoplast, an embryo, an anther, a seed, an achene, a stolon (also known as a runner), a callus, a cell culture, or any portion thereof.
  • Plant parts can be obtained by lifting, cutting, snapping, grinding, or otherwise disassociating the plant part from the plant.
  • pollen of a haploid-inducing strawberry plant comprises one or more genetic modifications resulting in decreased expression of one or more CENH3 genes described herein. In certain embodiments, the pollen comprises one or more genetic modifications in one or more CENH3 genes described herein.
  • stolon of a haploid-inducing strawberry plant comprises one or more genetic modifications resulting in decreased expression of one or more CENH3 genes described herein. In certain embodiments, the stolon comprises one or more genetic modifications in one or more CENH3 genes described herein.
  • seed of a haploid-inducing strawberry plant comprises one or more genetic modifications resulting in decreased 60 sf-6744554 197072001240 expression of one or more CENH3 genes described herein. In certain embodiments, the seed comprises one or more genetic modifications in one or more CENH3 genes described herein.
  • an achene of a haploid-inducing strawberry plant comprises one or more genetic modifications resulting in decreased expression of one or more CENH3 genes described herein. In certain embodiments, the achene comprises one or more genetic modifications in one or more CENH3 genes described herein.
  • the pollen, stolon, seed, or achene of the haploid-inducing strawberry plant comprises one or more genetic modifications resulting in decreased expression of one or more CENH3 proteins described herein. In some embodiments, the pollen, stolon, seed, or achene of the haploid-inducing strawberry plant comprises one or more genetic modifications of one or more CENH3 genes.
  • the pollen, stolon, seed, or achene of the haploid-inducing strawberry plant comprises one or more genetic modifications resulting in decreased expression of one or more CENH3 proteins having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity, or at least 60%, 65%, 70%, 75%, 80%, 85%, 95%, 96%, 97%, 98%, or 99% sequence similarity to an amino acid sequence selected from the list consisting of SEQ ID NOs: 8-14 and 53-56, or fragments thereof.
  • the pollen, stolon, seed, or achene of the haploid-inducing strawberry plant comprises one or more genetic modifications of one or more CENH3 genes comprising a nucleotide sequence having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity or complementarity to a nucleotide sequence selected from the group consisting of SEQ ID NOs: 1-7 and 58-61, or a fragment thereof.
  • the one or more genetic modifications comprise a modification of an enhancer of one or more of the CENH3 genes, a modification of a promoter of one or more of the CENH3 genes, a modification of a coding region of one or more of the CENH3 genes, a modification of an intron of one or more of the CENH3 genes, a modification of methylation status of one or more of the CENH3 genes, expression of a repressor protein that targets the DNA or an mRNA of one or more of the CENH3 genes, expression of an RNA interference construct that targets an mRNA of one or more of the CENH3 genes, or any combination thereof.
  • the pollen, stolon, seed, or achene of the haploid-inducing strawberry 61 sf-6744554 197072001240 plant has decreased expression of one or more CENH3 genes described herein relative to a pollen, stolon, seed, or achene of a strawberry plant (e.g., a control strawberry plant, e.g., a strawberry plant of the same species) lacking the one or more genetic modifications.
  • the haploid-inducing strawberry lacks detectable expression of one or more CENH3 genes described herein relative to a pollen, stolon, seed, or achene of a strawberry plant (e.g., a control strawberry plant, e.g., a strawberry plant of the same species) lacking the one or more genetic modifications.
  • the pollen, stolon, seed, or achene of the haploid-inducing strawberry plant lacks detectable expression of any CENH3 genes.
  • the pollen, stolon, seed, or achene of the haploid- inducing strawberry plant further comprises one or more naturally occurring inactive alleles of one or more CENH3 genes.
  • the pollen, stolon, seed, or achene is from a haploid- inducing strawberry plant of the species Fragaria vesca and comprises one or more genetic modifications resulting in decreased expression of one or more CENH3 genes.
  • the pollen, stolon, seed, or achene is from a haploid-inducing strawberry plant of the species Fragaria vesca and comprises one or more genetic modifications resulting in decreased expression of an FvCENH3 gene.
  • the pollen, stolon, seed, or achene is from a haploid-inducing strawberry plant of the species Fragaria vesca and comprises one or more genetic modifications resulting in decreased expression of an FvCENH3 protein.
  • the pollen, stolon, seed, or achene is from a haploid-inducing strawberry plant of the species Fragaria vesca and comprises one or more genetic modifications resulting in decreased expression of a CENH3 protein having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity, or at least 60%, 65%, 70%, 75%, 80%, 85%, 95%, 96%, 97%, 98%, or 99% sequence similarity to the FvCENH3 protein of SEQ ID NO. 8, or a fragment thereof.
  • the pollen, stolon, seed, or achene is from a haploid- inducing strawberry plant of the species Fragaria vesca and comprises one or more genetic modifications of an FvCENH3 gene or gene product.
  • the pollen, stolon, seed, or achene is from a haploid-inducing strawberry plant of the species Fragaria vesca and comprises one or more genetic modifications in a FvCENH3 gene comprising a nucleotide sequence having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 62 sf-6744554 197072001240 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity or complementarity to the FvCENH3 gene nucleotide sequence of SEQ ID NO. 1, or a fragment thereof.
  • the pollen, stolon, seed, or achene is from a haploid- inducing strawberry plant of the species Fragaria vesca and comprises genetic modifications in both alleles of an FvCENH3 gene.
  • the one or more genetic modifications comprise a modification of an enhancer of FvCENH3, a modification of a promoter of FvCENH3, a modification of a coding region of FvCENH3, modification of an intron of FvCENH3, or any combination thereof relative to a wild- type or unmodified Fragaria vesca plant.
  • the pollen, stolon, seed, or achene is from a haploid-inducing strawberry plant of the species Fragaria vesca and has decreased expression of FvCENH3 protein relative to a pollen, stolon, seed, or achene from a control Fragaria vesca plant (e.g., a Fragaria vesca plant lacking one or more of the genetic modifications).
  • the pollen, stolon, seed, or achene of the haploid- inducing strawberry plant is a plant of the species Fragaria vesca and lacks detectable expression of FvCENH3 protein.
  • the pollen, stolon, seed, or achene is from a haploid- inducing strawberry plant of the species Fragaria x ananassa and comprises one or more genetic modifications resulting in decreased expression of one or more CENH3 genes.
  • the pollen, stolon, seed, or achene is from a haploid-inducing strawberry plant of the species Fragaria x ananassa and comprises one or more genetic modifications resulting in decreased expression of an FaCENH3-1a gene, an FaCENH3-1b gene, an FaCENH3-2a gene, an FaCENH3-2b gene, an FaCENH3-3 gene, an FaCENH3-4 gene, or any combination thereof.
  • the pollen, stolon, seed, or achene is from a haploid-inducing strawberry plant of the species Fragaria x ananassa and comprises one or more genetic modifications resulting in decreased expression of an FaCENH3-1a gene, an FaCENH3-1b gene, an FaCENH3-2a gene, an FaCENH3-2b gene, an FaCENH3-3 gene, and an FaCENH3-4 gene.
  • the pollen, stolon, seed, or achene is from a haploid- inducing strawberry plant of the species Fragaria x ananassa and comprises one or more genetic modifications resulting in decreased expression of an FaCENH3-1a protein, an FaCENH3-1b protein, an FaCENH3-2a protein, an FaCENH3-2b protein, an FaCENH3-3 protein, an FaCENH3-4 protein, or any combination thereof.
  • the 63 sf-6744554 197072001240 pollen, stolon, seed, or achene is from a haploid-inducing strawberry plant of the species Fragaria x ananassa and comprises one or more genetic modifications resulting in decreased expression of an FaCENH3-1a protein, an FaCENH3-1b protein, an FaCENH3-2a protein, an FaCENH3-2b protein, an FaCENH3-3 protein, and an FaCENH3-4 protein.
  • the pollen, stolon, seed, or achene is from a haploid-inducing strawberry plant of the species Fragaria x ananassa and comprises one or more genetic modifications resulting in decreased expression of one or more CENH3 proteins having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity, or at least 60%, 65%, 70%, 75%, 80%, 85%, 95%, 96%, 97%, 98%, or 99% sequence similarity to an amino acid sequence selected from the list consisting of SEQ ID NOs: 9-14, or fragments thereof.
  • the pollen, stolon, seed, or achene is from a haploid-inducing strawberry plant of the species Fragaria x ananassa and comprises one or more genetic modifications resulting in decreased expression of 1) a CENH3 protein having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity, or at least 60%, 65%, 70%, 75%, 80%, 85%, 95%, 96%, 97%, 98%, or 99% sequence similarity to the amino acid sequence of SEQ ID NO: 9; 2) a CENH3 protein having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity, or at least
  • the pollen, stolon, seed, or achene is from a haploid- inducing strawberry plant of the species Fragaria x ananassa and comprises one or more genetic modifications resulting in decreased expression of an FaCENH3-7a protein, an FaCENH3-7b protein, an FaCENH3-7c protein, an FaCENH3-7d protein, or any combination thereof.
  • the pollen, stolon, seed, or achene is from a haploid- inducing strawberry plant of the species Fragaria x ananassa and comprises one or more genetic modifications resulting in decreased expression of one or more CENH3 proteins having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity, or at least 60%, 65%, 70%, 75%, 80%, 85%, 95%, 96%, 97%, 98%, or 99% sequence similarity to an amino acid sequence selected from the list consisting of SEQ ID NOs: 53-56, or fragments thereof.
  • the pollen, stolon, seed, or achene is from a haploid-inducing strawberry plant of the species Fragaria x ananassa and comprises one or more genetic modifications resulting in decreased expression of 1) a CENH3 protein having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity, or at least 60%, 65%, 70%, 75%, 80%, 85%, 95%, 96%, 97%, 98%, or 99% sequence similarity to the amino acid sequence of SEQ ID NO: 53; 2) a CENH3 protein having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity, or at least
  • the pollen, stolon, seed, or achene is from a haploid- inducing strawberry plant of the species Fragaria x ananassa and comprises one or more 65 sf-6744554 197072001240 genetic modifications of an FaCENH3-1a gene, an FaCENH3-1b gene, an FaCENH3-2a gene, an FaCENH3-2b gene, an FaCENH3-3 gene, an FaCENH3-4 gene, or any combination thereof.
  • the pollen, stolon, seed, or achene is from a haploid-inducing strawberry plant of the species Fragaria x ananassa and comprises one or more genetic modifications of an FaCENH3-1a gene, an FaCENH3-1b gene, an FaCENH3-2a gene, an FaCENH3-2b gene, an FaCENH3-3 gene, and an FaCENH3-4 gene.
  • the pollen, stolon, seed, or achene is from a haploid-inducing strawberry plant of the species Fragaria x ananassa and comprises one or more genetic modifications in a CENH3 gene comprising a nucleotide sequence having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity or complementarity to a nucleotide sequence selected from the group consisting of SEQ ID NOs: 2-7, or a fragment thereof.
  • the pollen, stolon, seed, or achene is from a haploid-inducing strawberry plant of the species Fragaria x ananassa and comprises: 1) one or more genetic modifications in a CENH3 gene comprising a nucleotide sequence having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity or complementarity to the nucleotide sequence of SEQ ID NO: 2, or a fragment thereof; 2) one or more genetic modifications in a CENH3 gene comprising a nucleotide sequence having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity or complementarity to the nucle
  • the one or more genetic modifications comprise a modification of an enhancer of an FaCENH3 gene (e.g., an FaCENH3-1a gene, an FaCENH3-1b gene, an FaCENH3-2a gene, an FaCENH3-2b gene, an FaCENH3-3 gene, and/or an FaCENH3-4 gene), a modification of a promoter of an Facenh3 gene (e.g., an FaCENH3-1a gene, an FaCENH3-1b gene, an FaCENH3-2a gene, an FaCENH3-2b gene, an FaCENH3-3 gene, and/or an FaCENH3-4 gene), a modification of a coding region of an Facenh3 gene (e.g., an FaCENH3-1a gene, an FaCENH3-1b gene, an FaCENH3-2a gene, an FaCENH3-2b gene, an FaCENH3-3 gene, and/or an FaCENH3-4 gene), modification of an intron of an Facenh3 gene (e.g., an FaCENH3 gene (
  • the pollen, stolon, seed, or achene is from a haploid- inducing strawberry plant of the species Fragaria x ananassa and comprises one or more genetic modifications of an FaCENH3-7a gene, an FaCENH3-7b gene, an FaCENH3-7c gene, an FaCENH3-7d gene, or any combination thereof.
  • the pollen, stolon, seed, or achene is from a haploid-inducing strawberry plant of the species Fragaria x ananassa and comprises one or more genetic modifications in a CENH3 gene comprising a nucleotide sequence having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity or complementarity to a nucleotide sequence selected from the group consisting of SEQ ID NOs: 58-61, or a fragment thereof.
  • the pollen, stolon, seed, or achene is from a haploid-inducing strawberry plant of the species Fragaria x ananassa and comprises: 1) one or more genetic modifications in a CENH3 gene comprising a nucleotide sequence having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity or complementarity to the nucleotide sequence of SEQ ID NO: 58, or a fragment thereof; 2) one or more genetic modifications in a CENH3 gene comprising a nucleotide sequence having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity or complementarity to the
  • the one or more genetic modifications comprise a modification of an enhancer of an FaCENH3 gene (e.g., an FaCENH3-7a gene, an FaCENH3-7b gene, an FaCENH3-7c gene, and/or an FaCENH3-7d gene), a modification of a promoter of an FaCENH3 gene (e.g., an FaCENH3-7a gene, an FaCENH3-7b gene, an FaCENH3-7c gene, and/or an FaCENH3-7d gene), a modification of a coding region of an FaCENH3 gene (e.g., an FaCENH3-7a gene, an FaCENH3-7b gene, an FaCENH3-7c gene, and/or an FaCENH3-7d gene), modification of an intron of an FaCENH3 gene (e.g., an FaCENH3-7a gene, an FaCENH3-7b gene, an FaCENH3-7c gene, and/or an FaCENH3-7d gene), or any combination thereof relative to
  • the one or more genetic modifications comprise one or more deletions, insertions, or nucleotide substitutions in a coding region of an FaCENH3 gene (e.g., an FaCENH3-7a gene, an FaCENH3-7b gene, an FaCENH3-7c gene, and/or an FaCENH3-7d gene).
  • an FaCENH3 gene e.g., an FaCENH3-7a gene, an FaCENH3-7b gene, an FaCENH3-7c gene, and/or an FaCENH3-7d gene.
  • the pollen, stolon, seed, or achene is from a haploid- inducing strawberry plant of the species Fragaria x ananassa and has decreased expression of an FaCENH3-1a protein, an FaCENH3-1b protein, an FaCENH3-2a protein, an FaCENH3-2b protein, an FaCENH3-3 protein, an FaCENH3-4 protein, or any combination (e.g., two, three, four, five or all six) thereof relative to a pollen, stolon, seed, or achene of a control Fragaria x ananassa plant (e.g., a Fragaria ananassa plant lacking one or more or all of the genetic modifications).
  • a control Fragaria x ananassa plant e.g., a Fragaria ananassa plant lacking one or more or all of the genetic modifications.
  • the pollen, stolon, seed, or achene is from a haploid-inducing strawberry plant of the species Fragaria x ananassa and lacks detectable expression of an FaCENH3-1a protein, an FaCENH3-1b protein, an FaCENH3-2a protein, an FaCENH3-2b protein, an FaCENH3-3 protein, an FaCENH3-4 protein, or any combination (e.g., two, three, four, five or all six) thereof.
  • the pollen, stolon, seed, or achene of the haploid-inducing strawberry plant may comprise one or more genetic modifications resulting in decreased expression of any combination of CENH3 genes described herein or known in the art.
  • the pollen, stolon, seed, or achene of the haploid-inducing strawberry plant may comprise one or more genetic modifications resulting in non-expression of any combination of CENH3 genes described here or known in the art.
  • the pollen, stolon, seed, or achene of the haploid-inducing strawberry plant may comprise one or more genetic modifications resulting in decreased expression, non-expression, or a combination thereof of any combination of CENH3 genes described here or known in the art.
  • the pollen, stolon, seed, or achene of the haploid-inducing strawberry plant lacks detectable expression of CENH3 proteins.
  • the pollen, stolon, seed, or achene of the haploid-inducing strawberry plant has decreased expression of CENH3 proteins relative to a control strawberry plant.
  • Control strawberry plants include, for example, a plant of the same species as the haploid- inducing strawberry plant that lacks one or more or all of the genetic modifications comprised by the haploid-inducing strawberry plant, a wild-type plant of the same species as the haploid- inducing strawberry plant, an unmodified plant of the same species as the haploid-inducing strawberry plant, or a null segregant strawberry plant, which underwent the same genetic manipulation, but is lacking one or more or all of the genetic modifications comprised by the haploid-inducing strawberry plant.
  • the pollen, stolon, seed, or achene of the haploid-inducing strawberry plant has decreased expression of CENH3 proteins relative to a pollen, stolon, seed, or achene of a strawberry plant of the same species lacking the one or more genetic modifications.
  • Methods and Compositions for Producing Haploid-inducing Strawberry Plants [0174] In one aspect, described herein are methods of producing haploid-inducing strawberry plants. In some embodiments, the method comprises introducing one or more genetic modifications resulting in decreased expression of one or more CENH3 genes described herein into a strawberry plant.
  • the method further comprises introducing one or more naturally occurring inactive alleles of one or more CENH3 genes into a strawberry plant by crossing with a plant having the one or more naturally occurring inactive alleles of one or more CENH3 genes.
  • 69 sf-6744554 197072001240 Methods of Introducing Genetic Modifications [0175]
  • the methods of producing haploid-inducing strawberry plants described herein comprise introducing one or more genetic modifications into a strawberry plant.
  • the step of introducing one or more genetic modifications into a strawberry plant results in a decreased expression of one or more CENH3 genes.
  • the decreased expression is achieved by gene disruption (e.g., disruption of one or more CENH3 genes), gene knockout (e.g., knockout of one or more CENH3 genes), gene knockdown (e.g., knockdown of one or more CENH3 genes), gene silencing (e.g., silencing of one or more CENH3 genes), RNA interference (e.g., RNA interference of one or more CENH3 genes), induction of methylation (e.g., induction of methylation of one or more CENH3 genes), or any combination thereof.
  • the method comprises introducing one or more of the genetic modifications by gene editing using a site-directed nuclease.
  • the genetic modifications are introduced by gene editing. Any of several gene editing methods known in the art may be used to introduce the genetic modifications of the one or more CENH3 genes.
  • gene editing is performed with one or more natural or engineered site-directed nucleases including, but not limited to, RNA-guided nucleases, meganucleases, zinc finger nucleases (ZFNs), and transcription activator-like effector-based nucleases (TALENs).
  • gene editing is performed with RNA-guided nucleases including, but not limited to, Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) associated (Cas) nucleases.
  • CRISPR Clustered Regularly Interspaced Short Palindromic Repeats
  • gene editing may be performed using a deactivated Cas nuclease fused to an engineered reverse transcriptase in a process known as prime editing.
  • Methods of gene editing are numerous, well-known and routine in the art, and are described in US17/045747, US16/977020, and US16/961396, which are herein incorporated in their entirety.
  • An engineered nuclease may be a guided nuclease, which may function as a ribonucleoprotein (RNP) complex with a guide RNA.
  • RNP ribonucleoprotein
  • a guided nuclease may be selected from the group consisting of Cas1, Cas1B, Cas2, Cas3, Cas4, Cas5, Cash, Cas7, Cas8, Cas9 (also known as Csn1 and Csx12), Cas10, Cas12a, Csy1, Csy2, Csy3, Cse1, Cse2, Csc1, Csc2, Csa5, Csn2, Csm2, Csm3, Csm4, Csm5, Csm6, Cmr1, Cmr3, Cmr4, Cmr5, Cmr6, Csb1, Csb2, Csb3, Csx17, Csx14, Csx10, Csx16, CsaX, Csx3, 70 sf-6744554 197072001240 Csx1, Csx15, Csf1, Csf2, Csf3, Csf4, Cpf1, CasX, CasY, CasZ, MAD7, and homolog
  • the DNA construct or molecule encoding a guided nuclease may be delivered with or without a guide nucleic acid.
  • a guide nucleic acid molecule may be further provided to direct the guided nuclease to a target site in the genome of the plant via base-pairing or hybridization to cause a DSB or nick at or near the target site.
  • the guide nucleic acid may be transformed or introduced into a plant cell or tissue as a guide nucleic acid molecule, or as a recombinant DNA molecule, construct or vector comprising a transcribable DNA sequence encoding the guide nucleic acid operably linked to a promoter or plant-expressible promoter.
  • the promoter may be a constitutive promoter, a tissue-specific or tissue-preferred promoter, a developmental stage promoter, or an inducible promoter.
  • the guide nucleic acid comprises a first segment comprising a nucleotide sequence that is complementary to a sequence in a target nucleic acid (e.g., a target nucleic acid of a CENH3 gene described herein) and a second segment that interacts with a guided nuclease protein.
  • the first segment of a guide comprising a nucleotide sequence that is complementary to a sequence in a target nucleic acid corresponds to a CRISPR RNA (crRNA or crRNA repeat).
  • the second segment of a guide comprising a nucleic acid sequence that interacts with a guided nuclease protein corresponds to a trans-acting CRISPR RNA (tracrRNA).
  • the guide nucleic acid comprises two separate nucleic acid molecules (a polynucleotide that is complementary to a sequence in a target nucleic acid and a polynucleotide that interacts with a guided nuclease protein) that hybridize with one another.
  • the guide nucleic acid is a single polynucleotide (e.g., a gRNA).
  • the guide nucleic acid may comprise DNA, RNA or a combination of DNA and RNA.
  • a protospacer-adjacent motif may be present in the genome immediately adjacent and upstream to the 5' end of the genomic target site sequence complementary to the targeting sequence of the guide RNA, immediately downstream (3') to the sense (+) strand of the genomic target site (relative to the targeting sequence of the guide RNA) as known in the art. See, e.g., Wu, X. et al. 2014. "Target specificity of the CRISPR-Cas9 system," Quant 71 sf-6744554 197072001240 Biol. 2(2): 59-70.
  • the genomic PAM sequence on the sense (+) strand adjacent to the target site may comprise 5'-NGG-3'.
  • the corresponding sequence of the guide nucleic acid may generally not be complementary to the genomic PAM sequence.
  • the guide nucleic acid may typically be a non-coding RNA molecule that does not encode a protein.
  • the targeting sequence of the guide nucleic acid may be at least 10 nucleotides in length, such as 12-40 nucleotides, 12-30 nucleotides, 12-20 nucleotides, 12-35 nucleotides, 12-30 nucleotides, 15-30 nucleotides, 17-30 nucleotides, or 17-25 nucleotides in length, or about 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25 or more nucleotides in length.
  • the targeting sequence may be at least 95%, at least 96%, at least 97%, at least 99% or 100% identical or complementary to at least 10, at least 11, at least 12, at least 13, at least 14, at least 15, at least 16, at least 17, at least 18, at least 19, at least 20, at least 21, at least 22, at least 23, at least 24, at least 25, or more consecutive nucleotides of a DNA sequence at the genomic target site.
  • a guide nucleic acid may further comprise one or more other structural or scaffold sequence(s), which may bind or interact with an RNA-guided endonuclease. Such scaffold or structural sequences may further interact with other RNA molecules (e.g., tracrRNA).
  • a modified guided nuclease such as a Cas nickase
  • RT reverse transcriptase
  • pegRNA a non-coding RNA typically referred to as a pegRNA
  • a pegRNA comprises three components, from 5’ to 3’: 1) guide sequence complementary to the protospacer on non-PAM strand of the target site, 2) a template sequence comprising the desired nucleotide edit, and 3) a primer binding site complementary to the protospacer on the PAM strand of the target site.
  • Guide sequence- mediated binding of the nuclease-RT fusion to the target site results in a nick in the PAM strand of the target site.
  • the primer binding site of the pegRNA then binds to the nicked (PAM) strand of the target, which positions the template sequence 3’ of the PAM strand to serve as a template for reverse transcription.
  • An engineered nuclease may be a site-specific nuclease.
  • site-specific nucleases such as recombinases, zinc finger nucleases (ZFNs), meganucleases, and TALENs
  • ZFNs zinc finger nucleases
  • TALENs TALENs
  • non-nucleic acid-guided site-specific nucleases such as recombinases, zinc finger nucleases (ZFNs), meganucleases, and TALENs
  • ZFNs zinc finger nucleases
  • TALENs may be designed, engineered and constructed according to known methods to target and bind to a target site at or near the genomic locus of an endogenous gene of a plant to create a DSB or nick at such genomic locus to knockout or knockdown expression of the gene via repair of the DSB or nick, which may lead to the creation of a mutation or insertion of a sequence at the site of the DSB or nick, through cellular repair mechanisms, which may be guided by a donor template molecule.
  • a site-specific nuclease is a recombinase.
  • a recombinase may be a serine recombinase attached to a DNA recognition motif, a tyrosine recombinase attached to a DNA recognition motif, or other recombinase enzyme known in the art.
  • a recombinase or transposase may be a DNA transposase or recombinase attached or fused to a DNA binding domain.
  • Non-limiting examples of recombinases include a tyrosine recombinase attached, etc., to a DNA recognition motif provided herein is selected from the group consisting of a Cre recombinase, a Gin recombinase, a Flp recombinase, and a Tnp1 recombinase.
  • a Cre recombinase or a Gin recombinase provided herein is tethered to a zinc-finger DNA-binding domain, or a TALE DNA-binding domain, or a Cas9 nuclease.
  • a serine recombinase attached to a DNA recognition motif provided herein is selected from the group consisting of a PhiC31 integrase, an R4 integrase, and a TP-901 integrase.
  • a DNA transposase attached to a DNA binding domain provided herein is selected from the group consisting of a TALE-piggyBac and TALE-Mutator.
  • a site-specific nuclease may be a zinc finger nuclease (ZFN).
  • ZFNs are synthetic proteins consisting of an engineered zinc finger DNA-binding domain fused to a cleavage domain (or a cleavage half-domain), which may be derived from a restriction endonuclease (e.g., FokI).
  • the DNA binding domain may be canonical (C2H2) or non-canonical (e.g., C3H or C4).
  • the DNA-binding domain can comprise one or more zinc fingers (e.g., 2, 3, 4, 5, 6, 7, 8, 9 or more zinc fingers) depending on the target site. Multiple zinc fingers in a DNA- binding domain may be separated by linker sequence(s).
  • ZFNs can be designed to cleave almost any stretch of double-stranded DNA by modification of the zinc finger DNA-binding domain.
  • ZFNs form dimers from monomers composed of a non-specific DNA cleavage domain (e.g., derived from the FokI nuclease) fused to a DNA-binding domain comprising a zinc finger array engineered to bind a target site DNA sequence.
  • the DNA-binding domain of a ZFN may typically be composed of 3-4 (or more) zinc-fingers.
  • the amino acids at positions -1, +2, +3, and +6 relative to the start of the zinc finger alpha-helix, which contribute to site-specific binding to the target site, can be changed and customized to fit specific target sequences.
  • the other amino acids may form a consensus backbone to generate ZFNs with different sequence specificities.
  • ZFNs for targeting and binding to specific target sequences are known in the art. See, e.g., US Patent App. Nos. 2005/0064474, 2009/0117617, and 2012/0142062.
  • the FokI nuclease domain may require dimerization to cleave DNA and therefore two ZFNs with their C-terminal regions are needed to bind opposite DNA strands of the cleavage site (separated by 5-7 bp).
  • the ZFN monomer can cut the target site if the two- ZF-binding sites are palindromic.
  • a ZFN, as used herein, is broad and includes a monomeric ZFN that can cleave double stranded DNA without assistance from another ZFN.
  • ZFN may also be used to refer to one or both members of a pair of ZFNs that are engineered to work together to cleave DNA at the same site.
  • customized ZFNs can theoretically be constructed to target nearly any target sequence (e.g., at or near a gene in a plant genome).
  • Publicly available methods for engineering zinc finger domains include Context-dependent Assembly (CoDA), Oligomerized Pool Engineering (OPEN), and Modular Assembly.
  • a method and/or composition provided herein comprises one or more, two or more, three or more, four or more, or five or more ZFNs.
  • a ZFN provided herein is capable of generating a targeted DSB or nick.
  • a site-specific nuclease may be a TALEN enzyme.
  • TALENs are artificial restriction enzymes generated by fusing the transcription activator-like effector (TALE) DNA binding domain to a nuclease domain (e.g., FokI).
  • TALE transcription activator-like effector
  • FokI nuclease domain
  • TALENs are artificial restriction enzymes generated by fusing the transcription activator-like effector (TALE) DNA binding domain to a nuclease domain.
  • TALE transcription activator-like effector
  • the nuclease is selected from a group consisting of PvuII, MutH, TevI, FokI, AlwI, MlyI, SbfI, SdaI, StsI, CleDORF, Clo051, and Pept071.
  • the FokI monomers dimerize and cause a double- stranded DNA break at the target site.
  • TALEN as used herein, is broad and includes a monomeric TALEN that can cleave double stranded DNA without assistance from another TALEN.
  • TALE Transcription activator-like effectors
  • TALE can be engineered to bind practically any DNA sequence, such as at or near the genomic locus of a gene in a plant.
  • TALE has a central DNA-binding domain composed of 13-28 repeat monomers of 33-34 amino acids. The amino acids of each monomer are highly conserved, except for hypervariable amino acid residues at positions 12 and 13. The two variable amino acids are called repeat-variable diresidues (RVDs).
  • RVDs repeat-variable diresidues
  • the amino acid pairs NI, NG, HD, and NN of RVDs preferentially recognize adenine, thymine, cytosine, and guanine/adenine, respectively, and modulation of RVDs can recognize consecutive DNA bases.
  • This simple relationship between amino acid sequence and DNA recognition has allowed for the engineering of specific DNA binding domains by selecting a combination of repeat segments containing the appropriate RVDs.
  • 75 sf-6744554 197072001240 [0191]
  • variants of the FokI cleavage domain with mutations have been designed to improve cleavage specificity and cleavage activity.
  • the FokI domain functions as a dimer, requiring two constructs with unique DNA binding domains for sites in the target genome with proper orientation and spacing. Both the number of amino acid residues between the TALEN DNA binding domain and the FokI cleavage domain and the number of bases between the two individual TALEN binding sites are parameters for achieving high levels of activity. PvuII, MutH, and TevI cleavage domains are useful alternatives to FokI and FokI variants for use with TALEs. PvuII functions as a highly specific cleavage domain when coupled to a TALE (see Yank et al. 2013. PLoS One. 8: e82539).
  • MutH is capable of introducing strand-specific nicks in DNA (see Gabsalilow et al. 2013. Nucleic Acids Research. 41: e83). TevI introduces double-stranded breaks in DNA at targeted sites (see Beurdeley et al., 2013. Nature Communications. 4: 1762). [0192] The relationship between amino acid sequence and DNA recognition of the TALE binding domain allows for designable proteins. Software programs such as DNAWorks can be used to design TALE constructs. Other methods of designing TALE constructs are known to those of skill in the art.
  • a site-specific nuclease may be a meganuclease. Meganucleases, which are commonly identified in microbes, such as the LAGLIDADG family of homing endonucleases, are unique enzymes with high activity and long recognition sequences (>14 bp) resulting in site-specific digestion of target DNA.
  • a meganuclease may comprise a scaffold or base enzyme selected from the group consisting of I-CreI, I-CeuI, I-MsoI, I-SceI, I-AniI, and I-DmoI.
  • the engineering of meganucleases can be more challenging than ZFNs and TALENs because the DNA recognition and cleavage functions of meganucleases are intertwined in a single domain.
  • meganucleases have been combined with transcription activator-like (TAL) effectors to generate megaTALs, which fuse the DNA- binding region of the TAL to allow recognition of specific sequences to meganucleases to 76 sf-6744554 197072001240 increase the activity and specificity of cleavage (see, e.g., Boissel et al. "megaTALs: a rare- cleaving nuclease architecture for therapeutic genome engineering.” Nucleic acids research 42.4 (2014): 2591-2601).
  • TAL transcription activator-like
  • a meganuclease may be selected or engineered to bind to a genomic target sequence in a plant, such as at or near the genomic locus of a gene.
  • a meganuclease provided herein is capable of generating a targeted DSB.
  • gene editing comprises (a) inducing a DSB in the genome of a cell at a cleavage site (e.g., a cleavage site within a CENH3 gene) at or near a recognition site (e.g., a recognition site within a CENH3 gene) for a natural or engineered nuclease by expressing in the cell the natural or engineered nuclease recognizing said recognition site and inducing said DSB at the cleavage site; (b) introducing into the cell a repair nucleic acid molecule comprising an upstream flanking region having homology to the DNA region upstream of the preselected site (e.g., the recognition site within the CENH3 gene) and/or a downstream flanking DNA region having homology to the DNA region downstream of the preselected site (e.g., the recognition site within the CENH3 gene) for allowing homologous recombination between said flanking region or regions and said DNA region or regions flanking
  • gene editing comprises (a) inducing a DSB in the genome of a cell at a cleavage site (e.g., a cleavage site within a CENH3 gene) at or near a recognition site (e.g., a recognition site within a CENH3 gene) for a natural or engineered nuclease by introducing into the cell the natural or engineered nuclease recognizing said recognition site and inducing said DSB at the cleavage site; (b) introducing into the cell a repair nucleic acid molecule comprising an upstream flanking region having homology to the DNA region upstream of the preselected site (e.g., the recognition site within the CENH3 gene) and/or a downstream flanking DNA region having homology to the DNA region downstream of the preselected site (e.g., the recognition site within the CENH3 gene) for allowing homologous recombination between said flanking region or regions and said DNA region or regions flanking said preselected
  • a repair nucleic acid molecule is a single-stranded or double- stranded DNA molecule or RNA molecule that is used as a template for modification of the 77 sf-6744554 197072001240 genomic DNA at the preselected site in the vicinity of or at the cleavage site.
  • repair nucleic acid molecule is copied or integrated at the preselected site by homologous recombination between the flanking region(s) and the corresponding homology region(s) in the target genome flanking the preselected site, optionally in combination with non-homologous end- joining (NHEJ) at one of the two ends of the repair nucleic acid molecule (e.g., in case there is only one flanking region).
  • NHEJ non-homologous end- joining
  • the genetic modifications introduced by gene editing result in the decreased expression (including, e.g., decreased activity or non-expression) of one or more CENH3 genes.
  • the introduction of a DSB or nick may be used to introduce targeted genetic modifications in the genome of a plant.
  • genetic modifications such as deletions, insertions, inversions and/or substitutions may be introduced at a target site (e.g., a target site within a nucleotide of a CENH3 gene) via imperfect repair of the DSB or nick to produce a knock- out or knock-down of a gene.
  • a target site e.g., a target site within a nucleotide of a CENH3 gene
  • Such genetic modifications may be generated by imperfect repair of the targeted locus even without the use of a donor template molecule, and can result in decreased expression (including, e.g., decreased activity or non-expression) of an endogenous gene product.
  • genetic modifications may be produced by an indel (insertion or deletion of nucleotide bases in a target DNA sequence through NHEJ), or by specific removal of sequence that reduces or completely destroys the function of sequence at or near the targeting site.
  • a knockout of a CENH3 gene may be achieved by inducing a DSB or nick at or near the endogenous locus of the gene that results in non-expression of the gene product (e.g., a CENH3 protein), whereas a knockdown of a gene may be achieved in a similar manner by inducing a DSB or nick at or near the endogenous locus of the gene that is repaired imperfectly at a site that does not affect the coding sequence of the gene in a manner that would eliminate the function of the gene product (e.g., CENH3 protein).
  • the site of the DSB or nick within the endogenous locus may be in the upstream or 5' region of the CENH3 gene (e.g., a promoter and/or enhancer sequence) to affect or reduce its level of expression.
  • the CENH3 gene e.g., a promoter and/or enhancer sequence
  • targeted knockout or knockdown mutations of a CENH3 gene may be generated with a donor template molecule to direct a particular or desired mutation at 78 sf-6744554 197072001240 or near the target site via repair of the DSB or nick.
  • the donor template molecule may comprise a homologous sequence with or without an insertion sequence and comprising one or more mutations, such as one or more deletions, insertions, inversions and/or substitutions, relative to the targeted genomic sequence at or near the site of the DSB or nick.
  • targeted knockout mutations of a CENH3 gene may be achieved by substituting, inserting, deleting or inverting at least a portion of the CENH3 gene, including, but not limited to, by introducing a frame shift or premature stop codon into a protein coding sequence of the gene.
  • a deletion of a portion of a CENH3 gene may also be introduced by generating DSBs or nicks at two target sites and causing a deletion of the intervening target region flanked by the target sites.
  • the genetic modifications are introduced by transgenesis.
  • Transgenes may include, but are not limited to, one or more protein-coding sequences operably linked to a plant-expressible promoter, one or more transcribable DNA sequences encoding an RNA molecule operably linked to a plant-expressible promoter, a gene of interest, a marker gene, or any combination thereof.
  • transgenesis comprises (a) inducing a DSB in the genome of a cell at a cleavage site at or near a recognition site for a natural or engineered nuclease by expressing in the cell the natural or engineered nuclease recognizing said recognition site and inducing said DSB at the cleavage site; (b) introducing into the cell a repair nucleic acid molecule comprising an upstream flanking region having homology to the DNA region upstream of the preselected site, a downstream flanking DNA region having homology to the DNA region downstream of the preselected site, and a transgene region flanked by the upstream and downstream flanking DNA regions and comprising the transgene to be inserted at the preselected site; and (c) selecting a cell wherein said repair nucleic acid molecule has been used as a template for making a modification of said genome at said preselected site.
  • transgenesis comprises (a) inducing a DSB in the genome of a cell at a cleavage site at or near a recognition site for a natural or engineered nuclease by introducing into the cell the natural or engineered nuclease recognizing said recognition site and inducing said DSB at the cleavage site; (b) introducing into the cell a repair nucleic acid molecule comprising an upstream flanking region having homology to the DNA region upstream of the preselected site, a downstream flanking DNA region having homology to the DNA region downstream of the preselected site, and a transgene region flanked by the upstream and downstream flanking 79 sf-6744554 197072001240 DNA regions and comprising the transgene to be inserted at the preselected site; and (c) selecting a cell wherein said repair nucleic acid molecule has been used as a template for making a modification of said genome at said preselected site.
  • the recognition site is a site within a CENH3 gene. In other variations, the recognition site is within a neutral (e.g., non-coding) site within the genome.
  • the genetic modification comprises introducing proteins, nucleic acids, or a combination thereof into a plant cell. The introduction of the proteins, nucleic acids, or combination thereof into the plant cell may be achieved by any of several means known and routinely-used in the art. In some embodiments, the introduction of the proteins, nucleic acids, or combination thereof into the plant cell comprises isolating protoplasts, transfecting the protoplasts, encapsulating the protoplasts, and regenerating plants from the protoplasts.
  • the introduction of the proteins, nucleic acids, or combination thereof into the plant cell comprises biolistic transformation.
  • the introduction of the proteins, nucleic acids, or combination thereof into the plant cell comprises isolating immature plant embryos, bombarding the embryos with particles comprising nucleic acids, and regenerating plants from the immature embryos.
  • Numerous additional transformation methods may be used to introduce the proteins, nucleic acids, or combination thereof into a suitable plant or plant cell. Transformation methods include the use of liposomes, electroporation, chemicals that increase free DNA uptake, injection of the DNA directly into the plant (cell) such as microinjection, particle gun bombardment, transformation using viruses or pollen and microprojection.
  • Methods may be selected from the calcium/polyethylene glycol method for protoplasts (Krens et al. (1982) Nature 296: 72-74; Negrutiu et al. (1987) Plant. Mol. Biol. 8: 363-373); electroporation of protoplasts (Shillito et al. (1985) Bio/Technol. 3: 1099-1102); microinjection into plant material (Crossway et al. (1986) Mol. Gen. Genet. 202: 179-185); DNA or RNA-coated particle bombardment (Klein et al. (1987) Nature 327: 70) infection with (non-integrative) viruses and the like.
  • the method of producing the haploid- inducing strawberry plant comprises contacting a plurality of cells of a strawberry plant with one or more nucleic acid molecules, proteins, or a combination thereof into a plurality of cells of a strawberry plant.
  • the method comprises introducing one or more nucleic acid molecules, proteins, or a combination thereof into a plurality of cells of a 80 sf-6744554 197072001240 strawberry plant.
  • the plurality of cells is a plurality of protoplasts.
  • the method further comprises, subsequent to the contacting step: allowing the plurality of cells (e.g., protoplasts) of the parent strawberry plant to form calli, plants, or a combination thereof; determining the presence of one or more of the genetic modifications in the calli or plants; and identifying one or more calli or plants as having the genetic modifications resulting in decreased expression of one or more CENH3 genes.
  • Methods of detecting genetic modifications are known in the art and include, for example, PCR amplification, DNA sequencing, hybridization assays (e.g., in situ hybridization), chip- based assays, reporter assays, and the like. A method of detecting genetic modifications in strawberry plants is described in Example 2.
  • the method comprises contacting a plurality of cells of a parent strawberry plant with one or more expression vectors comprising an expression cassette for a site-directed nuclease (e.g., a Cas nuclease, a meganuclease, a TALEN, a ZFN, or a megaTAL).
  • a site-directed nuclease e.g., a Cas nuclease, a meganuclease, a TALEN, a ZFN, or a megaTAL.
  • the method comprises contacting a plurality of cells of a parent strawberry plant with one or more expression vectors comprising an expression cassette for a site-directed nuclease contacting a plurality of cells of a parent strawberry plant with one or more expression vectors together comprising an expression cassette for a Cas nuclease and an expression cassette for a guide RNA molecule.
  • the method of producing the haploid-inducing strawberry plant comprises contacting a plurality of cells (e.g., protoplasts) of a parent strawberry plant with one or more expression vectors together comprising an expression cassette for a Cas nuclease and an expression cassette for a guide RNA molecule having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity or complementarity to 15, 16, 17, 18, 19, 20, or more consecutive nucleotides of one or more of the CENH3 genes described herein.
  • the method of producing the haploid- inducing strawberry plant comprises contacting a plurality of cells (e.g., protoplasts) of a parent strawberry plant with one or more Cas nucleases, wherein each Cas nuclease is complexed with a guide RNA molecule having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity or complementarity to 15, 16, 17, 18, 19, 20, or more consecutive nucleotides of one or more of the CENH3 genes described herein.
  • the guide RNA molecule may be a crRNA, a gRNA, or a pegRNA.
  • the guide RNA molecule has at least 80%, at 81 sf-6744554 197072001240 least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity or complementarity to 15, 16, 17, 18, 19, 20, or more consecutive nucleotides of a nucleotide sequence encoding one or more CENH3 proteins having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity, or at least 60%, 65%, 70%, 75%, 80%, 85%, 95%, 96%, 97%, 98%, or 99% sequence similarity to an amino acid sequence selected from the group consisting of SEQ ID NOs: 8-14 and 53-56.
  • the guide RNA molecule has at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity or complementarity to 15, 16, 17, 18, 19, 20, or more consecutive nucleotides of a nucleotide sequence having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity or complementarity to a nucleotide sequence selected from the group consisting of SEQ ID NOs: 1-7 and 58-61.
  • the guide RNA molecule has at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity or complementarity to 15, 16, 17, 18, 19, 20, or more consecutive nucleotides of an enhancer of one or more CENH3 genes described herein, a promoter of one or more CENH3 genes described herein, a coding region of one or more CENH3 genes, or an intron of one or more of the CENH3 genes described herein.
  • the guide RNA has 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity or complementarity to a nucleotide sequence selected from the group consisting of SEQ ID NOs: 25-34. In certain embodiments, the guide RNA has 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity or complementarity to SEQ ID NO: 27, SEQ ID NO: 29, or SEQ ID NO: 31.
  • the method of producing the haploid-inducing strawberry plant comprises contacting a plurality of cells (e.g., protoplasts) of a parent strawberry plant of the species Fragaria vesca with one or more expression vectors together comprising an expression cassette for a Cas nuclease and an expression cassette for a guide RNA molecule having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity or complementarity to 15, 16, 17, 18, 19, 20, or more consecutive nucleotides of an FvCENH3 gene.
  • a plurality of cells e.g., protoplasts
  • a guide RNA molecule having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity or complementarity to 15, 16, 17, 18, 19, 20, or more consecutive nucleotides of an FvCENH3 gene.
  • the method of producing the haploid-inducing strawberry plant comprises contacting a plurality of cells (e.g., protoplasts) of a parent strawberry plant of the species Fragaria vesca with one or more 82 sf-6744554 197072001240 Cas nucleases, wherein each Cas nuclease is complexed with a guide RNA molecule having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity or complementarity to 15, 16, 17, 18, 19, 20, or more consecutive nucleotides of an FvCENH3 gene.
  • a guide RNA molecule having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity or complementarity to 15, 16, 17, 18, 19, 20, or more consecutive nucleotides of an FvCENH3 gene.
  • the guide RNA molecule may be a crRNA, a gRNA, or a pegRNA.
  • the guide RNA molecule has at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity or complementarity to 15, 16, 17, 18, 19, 20, or more consecutive nucleotides of a nucleotide sequence encoding a CENH3 protein having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity, or at least 60%, 65%, 70%, 75%, 80%, 85%, 95%, 96%, 97%, 98%, or 99% sequence similarity to the amino acid sequence of SEQ ID NO: 8.
  • the guide RNA molecule has at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity or complementarity to 15, 16, 17, 18, 19, 20, or more consecutive nucleotides of a nucleotide sequence having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity or complementarity to the nucleotide sequence of SEQ ID NO: 1.
  • the guide RNA molecule has at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity or complementarity to 15, 16, 17, 18, 19, 20, or more consecutive nucleotides of an enhancer of FvCENH3, a promoter of FvCENH3, a coding region of FvCENH3, or an intron of FvCENH3.
  • the method of producing the haploid-inducing strawberry plant comprises contacting a plurality of cells (e.g., protoplasts) of a parent strawberry plant of the species Fragaria vesca with one or more Cas nucleases, wherein each Cas nuclease is complexed with a guide RNA molecule having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity or complementarity to SEQ ID NO: 27, SEQ ID NO: 29, SEQ ID NO: 31, or a combination thereof.
  • a guide RNA molecule having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity or complementarity to SEQ ID NO: 27, SEQ ID NO: 29, SEQ ID NO: 31, or a combination thereof.
  • the method of producing the haploid-inducing strawberry plant comprises contacting a plurality of cells protoplasts of a parent strawberry plant of the species Fragaria vesca with a plurality of Cas nucleases, wherein a first portion of the plurality of the Cas nucleases are each complexed with a guide RNA molecule having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity or complementarity to SEQ ID NO: 27, wherein a second portion of 83 sf-6744554 197072001240 the plurality of the Cas nucleases are each complexed with a guide RNA molecule having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity or complementarity to SEQ ID NO: 29; and wherein a third portion of the plurality of the plurality
  • the method of producing the haploid-inducing strawberry plant comprises contacting a plurality of cells (e.g., protoplasts) of a parent strawberry plant of the species Fragaria x ananassa with one or more expression vectors together comprising an expression cassette for a Cas nuclease and an expression cassette for one or more guide RNA molecules having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity or complementarity to 15, 16, 17, 18, 19, 20, or more consecutive nucleotides of an FaCENH3-1a gene, an FaCENH3-1b gene, an FaCENH3-2a gene, an FaCENH3-2b gene, an FaCENH3-3 gene, an FaCENH3-4 gene, or any combination (e.g., two, three, four, five or all six) thereof.
  • a plurality of cells e.g., protoplasts
  • the method of producing the haploid-inducing strawberry plant comprises contacting a plurality of cells (e.g., protoplasts) of a parent strawberry plant of the species Fragaria x ananassa with one or more Cas nucleases, wherein each Cas nuclease is complexed with a guide RNA molecule having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity or complementarity to 15, 16, 17, 18, 19, 20, or more consecutive nucleotides of an FaCENH3-1a gene, an FaCENH3-1b gene, an FaCENH3-2a gene, an FaCENH3-2b gene, an FaCENH3-3 gene, an FaCENH3-4 gene, or any combination (e.g., two, three, four, five or all six) thereof.
  • a guide RNA molecule having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%
  • the guide RNA molecule may be a crRNA, a gRNA, or a pegRNA.
  • the guide RNA molecule has at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity or complementarity to 15, 16, 17, 18, 19, 20, or more consecutive nucleotides of a nucleotide sequence encoding a CENH3 protein having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity, or at least 60%, 65%, 70%, 75%, 80%, 85%, 95%, 96%, 97%, 98%, or 99% sequence similarity to an amino acid sequence selected from the group consisting of SEQ ID NOs: 9-14 and 53-56.
  • the guide RNA molecule has at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity or complementarity to 15, 16, 17, 84 sf-6744554 197072001240 18, 19, 20, or more consecutive nucleotides of a nucleotide sequence having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity or complementarity to a nucleotide sequence selected from the group consisting of SEQ ID NOs: 2-7 and 58-61.
  • the guide RNA molecule has at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity or complementarity to 15, 16, 17, 18, 19, 20, or more consecutive nucleotides of an enhancer, a promoter, a coding region, or an intron of an FaCENH3-1a gene, an FaCENH3-1b gene, an FaCENH3-2a gene, an FaCENH3-2b gene, an FaCENH3-3 gene, an FaCENH3-4 gene, or any combination (e.g., two, three, four, five or all six) thereof.
  • the method of producing the haploid-inducing strawberry plant comprises contacting a plurality of cells (e.g., protoplasts) of a parent strawberry plant of the species Fragaria x ananassa with one or more Cas nucleases, wherein each Cas nuclease is complexed with a guide RNA molecule having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity or complementarity to SEQ ID NO: 27, SEQ ID NO: 29, or SEQ ID NO: 31, or a combination thereof.
  • a guide RNA molecule having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity or complementarity to SEQ ID NO: 27, SEQ ID NO: 29, or SEQ ID NO: 31, or a combination thereof.
  • the method of producing the haploid-inducing strawberry plant comprises contacting a plurality of cells protoplasts of a parent strawberry plant of the species Fragaria x ananassa with a plurality of Cas nucleases, wherein a first portion of the plurality of the Cas nucleases are each complexed with a guide RNA molecule having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity or complementarity to SEQ ID NO: 27, and wherein a second portion of the plurality of the Cas nucleases are each complexed with a guide RNA molecule having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity or complementarity to SEQ ID NO: 29, and wherein a third portion of the plurality of the Cas nucleases are
  • the decreased expression of each of the one or more of the CENH3 genes is independently achieved by introducing an insertion, a deletion, one or more nucleotide changes, or an inversion into the CENH3 gene that that results in decreased expression of the CENH3 gene.
  • the insertion, the deletion, the one or 85 sf-6744554 197072001240 more nucleotide changes, or the inversion is introduced into the first 70%, the first 60%, the first 50%, the first 40%, the first 30%, the first 20%, or the first 10% of the nucleotides of the CENH3 gene (e.g., a sequence selected from the group consisting of SEQ ID NOs: 1-7).
  • the insertion, the deletion, the one or more nucleotide changes, or the inversion is introduced into the first 100, the first 200, the first 300, the first 400, the first 500, the first 600, the first 700, the first 800, the first 900, the first 1000, the first 1250, the first 1500, the first 1750, the first 2000, the first 2500, or the first 3000 nucleotides of the CENH3 gene (e.g., a sequence selected from the group consisting of SEQ ID NOs: 1-7).
  • the insertion, the deletion, the one or more nucleotide changes, or the inversion decreases, but does not eliminate, expression of the CENH3 gene.
  • the insertion, the deletion, the one or more nucleotide changes, or the inversion eliminates expression (e.g., eliminates activity) of the CENH3 gene.
  • the activity of the CENH3 gene is eliminated by a premature stop codon introduced into the first 70%, the first 60%, the first 50%, the first 40%, the first 30%, the first 20%, or the first 10% of a sequence selected from the group consisting of SEQ ID NOs: 1-7 or of the nucleotides of the coding sequence of the CENH3 gene following the start codon in the 3’ direction.
  • the activity of the CENH3 gene is eliminated by a premature stop codon introduced into the first 100, the first 200, the first 300, the first 400, the first 500, the first 600, the first 700, the first 800, the first 900, the first 1000, the first 1250, the first 1500, the first 1750, the first 2000, the first 2500, or the first 3000 nucleotides of a sequence selected from the group consisting of SEQ ID NOs: 1-7 or of the coding sequence of the CENH3 gene following the start codon in the 3’ direction.
  • compositions for Producing Haploid-inducing Strawberry Plants [0207]
  • genetically modified plants, plant parts, plant cells, and seeds comprising said expression vectors and isolated DNA molecules.
  • expression vectors for making a haploid- inducing strawberry plant described herein comprises one or more expression cassettes.
  • the expression cassettes may comprise, for example, a nucleic acid encoding site-directed nuclease operably linked to a promoter, or a nucleic acid encoding a non- coding RNA (e.g., a guide RNA, an siRNA, or the like) 86 sf-6744554 197072001240 operably linked to a promoter.
  • the expression vector comprises at least a first expression cassette comprising a nucleic acid encoding a guide RNA operably linked to a promoter.
  • the expression vector comprises one or more additional expression cassettes, for example, a second expression cassette comprising a nucleic acid encoding a Cas nuclease operably linked to a promoter, and/or one or more additional expression cassettes each comprising a nucleic acid encoding a guide RNA operably linked to a promoter.
  • the expression vector comprises a DNA sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity or complementarity to 15, 16, 17, 18, 19, 20, or more consecutive nucleotides of a nucleotide sequence encoding one or more CENH3 proteins having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity, or at least 60%, 65%, 70%, 75%, 80%, 85%, 95%, 96%, 97%, 98%, or 99% sequence similarity to an amino acid sequence selected from the group consisting SEQ ID NOs: 8-14 and 53-56.
  • the expression vector comprises a DNA sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity or complementarity to 15, 16, 17, 18, 19, 20, or more consecutive nucleotides of a nucleotide sequence having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity or complementarity to a nucleotide sequence selected from the group consisting of SEQ ID NOs: 1-7 and 58-61.
  • the expression vector comprises a DNA sequence encoding a site- directed nuclease.
  • the site-directed nuclease is a Cas nuclease, a TALEN, ZFN, or a mega-TAL.
  • an expression vector for making a haploid- inducing strawberry plant wherein the expression vector comprises at least a first expression cassette comprising a nucleic acid encoding a non-coding RNA molecule operably linked to a promoter.
  • the non-coding RNA is a crRNA, a gRNA, a pegRNA, a siRNA, a miRNA, or a dsRNA.
  • the non-coding RNA molecule (e.g., guide RNA) has at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity or complementarity to 15, 16, 17, 18, 19, 20, or more consecutive nucleotides of a nucleotide sequence encoding one or more CENH3 proteins having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity, 87 sf-6744554 197072001240 or at least 60%, 65%, 70%, 75%, 80%, 85%, 95%, 96%, 97%, 98%, or 99% sequence similarity to an amino acid sequence selected from the group consisting of SEQ ID NOs: 8- 14 and 53-56.
  • the non- coding RNA molecule (e.g., guide RNA) has at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity or complementarity to 15, 16, 17, 18, 19, 20, or more consecutive nucleotides of a nucleotide sequence having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity or complementarity to a nucleotide sequence selected from the group consisting of SEQ ID NOs: 1-7 and 58-61.
  • the non-coding RNA molecule (e.g., guide RNA) has at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity or complementarity to 15, 16, 17, 18, 19, 20, or more consecutive nucleotides of an enhancer of one or more CENH3 genes described herein, a promoter of one or more CENH3 genes described herein, a coding region of one or more CENH3 genes, or an intron of one or more of the CENH3 genes described herein.
  • guide RNA has at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity or complementarity to 15, 16, 17, 18, 19, 20, or more consecutive nucleotides of an enhancer of one or more CENH3 genes described herein, a promoter of one or more CENH3 genes described herein, a coding region of one or more CENH
  • the non-coding RNA molecule is a guide RNA having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity or complementarity to SEQ ID NO: 17, SEQ ID NO: 19, SEQ ID NO: 21, or any combination thereof.
  • the expression vector comprises a first expression cassette comprising a nucleic acid encoding a guide RNA having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity or complementarity to SEQ ID NO: 17, SEQ ID NO: 19, SEQ ID NO: 21, or any combination thereof.
  • the expression vector comprises a first expression cassette comprising a nucleic acid encoding a guide RNA having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity or complementarity to SEQ ID NO: 17, a second expression cassette comprising a nucleic acid encoding a guide RNA having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity or complementarity to SEQ ID NO: 19, and a third expression cassette comprising a nucleic acid encoding a guide RNA having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity or complementarity to SEQ ID NO: 21.
  • the expression vector further comprises a DNA sequence encoding a Cas nuclease operably linked to a promoter.
  • 88 sf-6744554 197072001240 [0210]
  • an expression vector comprising a DNA sequence encoding a site-directed nuclease operably linked to a promoter.
  • the site-directed nuclease is a Cas nuclease, a TALEN, ZFN, or a mega-TAL.
  • the expression vector comprises a DNA sequence encoding a TALEN, ZFN, or a mega-TAL operably linked to a promoter, wherein the TALEN, ZFN, or a mega-TAL comprises an amino acid sequence that confers binding to a polynucleotide sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity or complementarity to 15, 16, 17, 18, 19, 20, or more consecutive nucleotides of a nucleotide sequence encoding one or more CENH3 proteins having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity, or at least 60%, 65%, 70%, 75%, 80%, 85%, 95%, 96%, 97%, 98%, or 99%, or 100% sequence identity
  • the expression vector comprises a DNA sequence encoding a TALEN, ZFN, or a mega-TAL operably linked to a promoter, wherein the TALEN, ZFN, or a mega-TAL comprises an amino acid sequence that confers binding to a polynucleotide sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity or complementarity to 15, 16, 17, 18, 19, 20, or more consecutive nucleotides of a nucleotide sequence having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity or complementarity to a nucleotide sequence selected from the group consisting of SEQ ID NOs: 1-7 and 58-61.
  • an isolated DNA molecule for making a haploid-inducing strawberry plant wherein the isolated DNA molecule has at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity or complementarity to at least 15, 16, 17, 18, 19, 20, 50, 100, or more consecutive nucleotides of a nucleotide sequence encoding one or more CENH3 proteins having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity, or at least 60%, 65%, 70%, 75%, 80%, 85%, 95%, 96%, 97%, 98%, or 99% sequence similarity to an amino acid sequence selected from the group consisting of SEQ ID NOs: 8-14 and 53-56.
  • an isolated DNA molecule for making a haploid-inducing strawberry plant wherein the isolated DNA molecule has at least 80%, at least 85%, at least 89 sf-6744554 197072001240 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity or complementarity to at least 15, 16, 17, 18, 19, 20, 50, 100, or more consecutive nucleotides of a nucleotide sequence having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity or complementarity to a nucleotide sequence selected from the group consisting of SEQ ID NOs: 1-7 and 58-61.
  • the isolated DNA molecule is a synthetic oligo nucleotide.
  • an isolated nucleic acid molecule for making a haploid-inducing strawberry plant wherein the isolated nucleic acid molecule has at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity or complementarity to at least 15, 16, 17, 18, 19, 20, 50, 100, or more consecutive nucleotides of a nucleotide sequence encoding one or more CENH3 proteins having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity, or at least 60%, 65%, 70%, 75%, 80%, 85%, 95%, 96%, 97%, 98%, or 99% sequence similarity
  • an isolated nucleic acid molecule for making a haploid- inducing strawberry plant wherein the isolated RNA molecule has at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity or complementarity to at least 15, 16, 17, 18, 19, 20, 50, 100, or more consecutive nucleotides of a nucleotide sequence having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity or complementarity to a nucleotide sequence selected from the group consisting of SEQ ID NOs: 25-34.
  • the isolated nucleic acid molecule has at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity or complementarity to a nucleotide sequence selected from the group consisting of SEQ ID NOs: 15-24. In certain embodiments, the isolated nucleic acid molecule has at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity or complementarity to SEQ ID NO: 17, SEQ ID NO: 19, or SEQ ID NO: 21, or a combination thereof.
  • the isolated nucleic acid molecule comprises DNA, RNA, or both. In some embodiments, the isolated nucleic acid molecule is an isolated RNA molecule. In certain embodiments, the isolated RNA molecule is a guide RNA. In certain embodiments, the guide 90 sf-6744554 197072001240 RNA has at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity or complementarity to SEQ ID NO: 27, SEQ ID NO: 29, or SEQ ID NO: 31, or a combination thereof.
  • the isolated nucleic acid molecule further comprises a nucleic acid sequence for association with a CRISPR-associated (Cas) enzyme.
  • Cas CRISPR-associated enzyme.
  • the genetically modified plants, plant parts, plant cells, or seeds are genetically modified strawberry plants, strawberry plant parts (e.g., achenes, stolons, or any strawberry plant part described herein), strawberry plant cells, or strawberry seeds.
  • a bacterial cell comprising any of the expression vectors or isolated DNA molecules described herein.
  • the bacterial cell is an Agrobacterium cell.
  • kits for producing a haploid-inducing strawberry plant comprising one or more of the expression vectors, isolated DNA molecules, isolated RNA molecules, bacterial cells, or any combination thereof described herein. In some embodiments, the kit further comprises instructions for using the kit components to produce a haploid-inducing strawberry plant. [0216] In yet other aspects, provided herein is a kit for producing a true homozygous octoploid strawberry plant comprising one or more cells or plant parts of any of the haploid- inducing strawberry plants described herein.
  • the kit further comprises instructions for using the kit components to produce a true homozygous octoploid strawberry plant.
  • the kit further comprises a chromosome doubling agent described herein.
  • the kit further comprises colchicine, oryzalin, oxide, trifluralin, or any combination thereof.
  • methods of producing true homozygous octoploid strawberry plant comprising (a) generating a tetrahaploid strawberry cell or plant 91 sf-6744554 197072001240 through haploid induction, and (b) subjecting the tetrahaploid strawberry plant to genome doubling to produce a doubled tetrahaploid strawberry cell or plant.
  • a doubled tetrahaploid cell or plant may also be described as a homozygous octoploid cell or plant having the same haplotype for each of the chromosomes within each of the subgenomes of the plant.
  • the method comprises (a) contacting a tetrahaploid cell from a donor octoploid strawberry plant with pollen from a haploid-inducing strawberry plant described herein, and (b) producing a doubled tetrahaploid cell or plant from the tetrahaploid cell.
  • the method comprises (a) contacting a tetrahaploid cell from a donor octoploid strawberry plant with pollen from a haploid-inducing strawberry plant described herein, and (b) producing an octoploid cell or plant from the doubled tetrahaploid cell or plant.
  • the method comprises (a1) contacting a reduced egg cell from a donor octoploid strawberry plant with a sperm cell from a haploid-inducing strawberry plant described herein, (a2) allowing a tetrahaploid embryo to form and grow into a tetrahaploid strawberry plant, and (b) subjecting the tetrahaploid strawberry plant to genome doubling to produce a doubled tetrahaploid strawberry plant.
  • the method comprises (a1) contacting a tetraploid reduced egg cell (a tetrahaploid cell) from a donor octoploid strawberry plant with a sperm cell from a haploid- inducing strawberry plant described herein, (a2) allowing a tetrahaploid embryo to form and grow into a tetrahaploid strawberry plant, and (b) subjecting the tetrahaploid strawberry plant to genome doubling to produce an octoploid strawberry plant from the doubled tetrahaploid.
  • a tetraploid reduced egg cell a tetrahaploid cell
  • the methods of producing true homozygous octoploid strawberry plant, or plant part thereof comprise generating a tetrahaploid strawberry plant through haploid induction, or the process of crossing gametes from a haploid-inducing strawberry plant with a donor (non-haploid-inducing) strawberry plant to generate embryos and seeds with half the ploidy of the donor strawberry plant.
  • a donor non-haploid-inducing
  • two haploid sperm cells migrate down the pollen tube to the ovule.
  • One of the sperm cells fertilizes the central cell of the ovule to form the endosperm, while the other sperm cell fertilizes the haploid reduced egg cell to form an embryo having the ploidy of the parent plant.
  • the haploid (4x) sperm and egg form an octoploid (8x) embryo in a normal fertilization process.
  • a haploid induction cross beteen a non-haploid inducing plant 92 sf-6744554 197072001240 and a haploid-inducing plants with reduced expression of CENH3
  • embryogenesis is initiated in the egg cell with only the haploid genome of the male or female gamete, resulting in a haploid embryo.
  • the sperm cells from the haploid-inducing plant migrate down the pollen tube to the ovule, where one of the sperm cells fertilizes the central cell to form the endosperm.
  • the other sperm cell contacts the reduced egg cell, embryogenesis is initiated, but the DNA from the sperm cell is not delivered to or retained in the egg cell, resulting in formation of a haploid embryo having a nuclear genome derived from only the maternal plant, and not the paternal plant.
  • haploid induction using pollen from the haploid-inducing plant if both parent plants are octoploid, the haploid (4x) egg cell does not incorporate the DNA from the haploid sperm cell (4x), resulting in a haploid (4x) embryo, which may also be referred to herein as a tetrahaploid.
  • the sperm cells from the non-haploid-inducing plant migrate down the pollen tube to the ovule, where one of the sperm cells fertilizes the central cell to form the endosperm and the other sperm cell fertilizes the egg. Fertilization of the haploid-inducing egg cell with the non-haploid-inducing sperm cell results in retention of the paternal nuclear DNA present in the sperm cell, but elimination of the maternal nuclear DNA present in the egg cell.
  • the step of generating a tetrahaploid strawberry plant through haploid induction comprises contacting a tetrahaploid cell from a donor octoploid strawberry plant with pollen, or a part thereof, from a haploid-inducing strawberry plant described herein.
  • the tetrahaploid cell is a reduced egg cell. In certain embodiments, the tetrahaploid cell is a reduced egg cell and the part of the pollen from the haploid-inducing strawberry plant is a sperm cell.
  • the step of generating a tetrahaploid strawberry plant through haploid induction comprises contacting pollen of the haploid-inducing strawberry plant with the stigma of a pistil of the donor octoploid strawberry plant comprising the reduced egg cell, allowing formation of a pollen tube and migration of one or more (e.g., two) sperm cells of the pollen to the reduced egg 93 sf-6744554 197072001240 cell, thereby contacting the reduced egg cell with the sperm cells.
  • the step of generating a tetrahaploid strawberry plant through haploid induction comprises allowing the donor octoploid strawberry plant to form a plurality of seeds.
  • the plurality of seeds comprises one or more tetrahaploid seeds.
  • the method may further comprise a step of selecting one or more tetrahaploid seeds of the plurality of seeds by 1) determining the ploidy of the embryo and/or endosperm of one or more seeds of the plurality of seeds, and 2) selecting one or more seeds having a tetrahaploid embryo.
  • the step of selecting one or more tetrahaploid seeds of the plurality of seeds comprises 1) determining the ploidy of the embryo and/or endosperm of one or more seeds of the plurality of seeds, and 2) selecting one or more seeds having a tetraploid embryo and an endosperm having a ploidy of greater than 8x (e.g., a ploidy of 9x, 10x, 11x, or 12x).
  • the step of generating a tetrahaploid strawberry plant through haploid induction comprises contacting a tetrahaploid cell from a donor octoploid strawberry plant with an egg cell from a haploid-inducing strawberry plant described herein.
  • the tetrahaploid cell is a reduced sperm cell.
  • the step of generating a tetrahaploid strawberry plant through haploid induction comprises contacting the stigma of a pistil of the haploid-inducing strawberry plant with the pollen of the donor octoploid strawberry plant comprising the reduced egg cell, allowing formation of a pollen tube and migration of one or more (e.g., two) reduced sperm cells of the pollen to the egg cell, thereby contacting the egg cell with the reduced sperm cells.
  • the step of generating a tetrahaploid strawberry plant through haploid induction comprises allowing the haploid-inducing strawberry plant to form a plurality of seeds.
  • the plurality of seeds comprises one or more tetrahaploid seeds.
  • the method may further comprise a step of selecting one or more tetrahaploid seeds of the plurality of seeds by 1) determining the ploidy of the embryo and/or endosperm of one or more seeds of the plurality of seeds, and 2) selecting one or more seeds having a tetrahaploid embryo.
  • the step of selecting one or more tetrahaploid seeds of the plurality of seeds comprises 1) determining the ploidy of the embryo and/or endosperm of one or more seeds of the plurality of seeds, and 2) selecting one or more seeds having a tetraploid embryo and an endosperm having a ploidy of greater than 8x (e.g., a ploidy of 9x, 10x, 11x, or 12x).
  • the haploid-inducing strawberry plant and the donor octoploid strawberry plants are from different species of Fragaria.
  • the haploid- inducing strawberry plant is diploid.
  • the haploid-inducing strawberry 94 sf-6744554 197072001240 plant is a plant of the species Fragaria vesca.
  • the donor octoploid strawberry plant is a plant of the species Fragaria x ananassa.
  • the embryo of the true homozygous octoploid strawberry seed has a nuclear genome derived from only the donor octoploid strawberry plant and a cytoplasmic genome (e.g., mitochondrial and/or chloroplast genome) derived from the haploid-inducing strawberry plant.
  • the haploid-inducing strawberry plant and the donor strawberry plant may have the same ploidy or different ploidies.
  • a haploid-inducing strawberry plant may be diploid (2x) and produce haploid (1x) pollen or egg cells that can be crossed with an octoploid (8x) strawberry donor plant, resulting in tetraploid (4x) embryos, also referred to as tetrahaploid embryos.
  • the method of producing true homozygous octoploid strawberry plant, or plant part thereof comprises (a) contacting a tetrahaploid cell from a donor octoploid strawberry plant with pollen from a diploid, triploid, tetraploid, pentaploid, hexaploid, septaploid, or octoploid haploid-inducing strawberry plant described herein, and (b) producing an octoploid doubled tetrahaploid cell or plant from the tetrahaploid cell.
  • the method comprises (a1) contacting a reduced egg cell from a donor octoploid strawberry plant with a sperm cell from a diploid, triploid, tetraploid, pentaploid, hexaploid, septaploid, or octoploid haploid-inducing strawberry plant described herein, (a2) allowing a tetrahaploid embryo to form and grow into a tetrahaploid strawberry plant, and (b) subjecting the tetrahaploid strawberry plant to genome doubling to produce a doubled tetrahaploid strawberry plant.
  • the method comprises (a) contacting a tetrahaploid cell from a donor octoploid strawberry plant with pollen from a diploid, triploid, tetraploid, pentaploid, hexaploid, septaploid, or octoploid haploid-inducing strawberry plant described herein, and (b) producing an octoploid doubled tetrahaploid cell or plant from the tetrahaploid cell.
  • the method comprises (a1) contacting a tetrahaploid reduced egg cell from a donor octoploid strawberry plant with a sperm cell from a diploid, triploid, tetraploid, pentaploid, hexaploid, septaploid, or octoploid haploid-inducing strawberry plant described herein, (a2) allowing a tetrahaploid embryo to form and grow into a tetrahaploid strawberry plant, and (b) subjecting the tetrahaploid strawberry plant to genome doubling to produce an octoploid doubled tetrahaploid strawberry plant.
  • the method of producing true homozygous octoploid strawberry plant, or plant part thereof comprises (a) contacting a tetrahaploid cell from a donor octoploid strawberry plant with an egg cell from a diploid, triploid, tetraploid, pentaploid, hexaploid, septaploid, or octoploid haploid-inducing strawberry plant described herein, and (b) producing an octoploid doubled tetrahaploid cell or plant from the tetrahaploid cell.
  • the method comprises (a1) contacting a reduced sperm cell from a donor octoploid strawberry plant with an egg cell from a diploid, triploid, tetraploid, pentaploid, hexaploid, septaploid, or octoploid haploid-inducing strawberry plant described herein, (a2) allowing a tetrahaploid embryo to form and grow into a tetrahaploid strawberry plant, and (b) subjecting the tetrahaploid strawberry plant to genome doubling to produce a doubled tetrahaploid strawberry plant.
  • the method comprises (a) contacting a tetrahaploid cell from a donor octoploid strawberry plant with an egg cell from a diploid, triploid, tetraploid, pentaploid, hexaploid, septaploid, or octoploid haploid-inducing strawberry plant described herein, and (b) producing an octoploid doubled tetrahaploid cell or plant from the tetrahaploid cell.
  • the method comprises (a1) contacting a tetrahaploid reduced sperm cell from a donor octoploid strawberry plant with an egg cell from a diploid, triploid, tetraploid, pentaploid, hexaploid, septaploid, or octoploid haploid-inducing strawberry plant described herein, (a2) allowing a tetrahaploid embryo to form and grow into a tetrahaploid strawberry plant, and (b) subjecting the tetrahaploid strawberry plant to genome doubling to produce an octoploid doubled tetrahaploid strawberry plant.
  • the haploid-inducing strawberry plant and the donor octoploid strawberry plants are from different species of Fragaria.
  • the haploid- inducing strawberry plant is diploid.
  • the haploid-inducing strawberry plant is a plant of the species Fragaria vesca.
  • the donor octoploid strawberry plant is a plant of the species Fragaria x ananassa.
  • the embryo of the true homozygous octoploid strawberry seed has a nuclear genome derived from only the donor octoploid strawberry plant and a cytoplasmic genome (e.g., mitochondrial and/or chloroplast genome) derived from the haploid-inducing strawberry plant.
  • a cytoplasmic genome e.g., mitochondrial and/or chloroplast genome
  • the method of producing a true homozygous octoploid strawberry plant, or plant part thereof comprises (a) contacting a haploid cell from a donor octoploid strawberry plant with pollen from a diploid haploid-inducing strawberry plant described herein, and (b) producing an octoploid cell from a doubled tetrahaploid or plant 96 sf-6744554 197072001240 from the haploid cell.
  • the method comprises (a1) contacting a reduced egg cell from a donor octoploid strawberry plant with a sperm cell from a diploid haploid-inducing strawberry plant described herein, (a2) allowing a tetrahaploid embryo to form and grow into a tetrahaploid strawberry plant, and (b) subjecting the tetrahaploid strawberry plant to genome doubling to produce a doubled tetrahaploid strawberry plant.
  • the method comprises (a) contacting a tetrahaploid cell from a donor octoploid strawberry plant with pollen from a diploid haploid-inducing strawberry plant described herein, and (b) producing an octoploid doubled tetrahaploid cell or plant from the tetrahaploid cell.
  • the method comprises (a1) contacting a tetrahaploid reduced egg cell from a donor octoploid strawberry plant with a sperm cell from a diploid haploid-inducing strawberry plant described herein, (a2) allowing a tetrahaploid embryo to form and grow into a tetrahaploid strawberry plant, and (b) subjecting the tetrahaploid strawberry plant to genome doubling to produce an octoploid doubled tetrahaploid strawberry plant.
  • the diploid haploid-inducing strawberry plant is a plant of the species Fragaria vesca, Fragaria iinumae, Fragaria nipponica, Fragaria viridis, Fragaria ⁇ bifera, Fragaria bucharica, Fragaria chinensis, Fragaria daltoniana, Fragaria emeiensis, Fragaria hayatae, Fragaria iinumae, Fragaria mandshurica, Fragaria viridis, Fragaria nilgerrensis, Fragaria nipponica, Fragaria nubicola, or Fragaria pentaphylla.
  • the diploid haploid-inducing strawberry plant is a plant of the species Fragaria vesca.
  • the method of producing a true homozygous octoploid strawberry plant, or plant part thereof comprises (a) contacting a haploid cell from a donor octoploid strawberry plant with an egg cell from a diploid haploid-inducing strawberry plant described herein, and (b) producing an octoploid cell from a doubled tetrahaploid or plant from the haploid cell.
  • the method comprises (a1) contacting a reduced sperm cell from a donor octoploid strawberry plant with an egg cell from a diploid haploid-inducing strawberry plant described herein, (a2) allowing a tetrahaploid embryo to form and grow into a tetrahaploid strawberry plant, and (b) subjecting the tetrahaploid strawberry plant to genome doubling to produce a doubled tetrahaploid strawberry plant.
  • the method comprises (a) contacting a tetrahaploid cell from a donor octoploid strawberry plant with an egg cell from a diploid haploid-inducing strawberry plant described herein, and (b) producing an octoploid doubled tetrahaploid cell or plant from the tetrahaploid cell.
  • the method comprises (a1) contacting a 97 sf-6744554 197072001240 tetrahaploid reduced sperm cell from a donor octoploid strawberry plant with an egg cell from a diploid haploid-inducing strawberry plant described herein, (a2) allowing a tetrahaploid embryo to form and grow into a tetrahaploid strawberry plant, and (b) subjecting the tetrahaploid strawberry plant to genome doubling to produce an octoploid doubled tetrahaploid strawberry plant.
  • the diploid haploid-inducing strawberry plant is a plant of the species Fragaria vesca, Fragaria iinumae, Fragaria nipponica, Fragaria viridis, Fragaria ⁇ bifera, Fragaria bucharica, Fragaria chinensis, Fragaria daltoniana, Fragaria emeiensis, Fragaria hayatae, Fragaria iinumae, Fragaria mandshurica, Fragaria viridis, Fragaria nilgerrensis, Fragaria nipponica, Fragaria nubicola, or Fragaria pentaphylla.
  • the diploid haploid-inducing strawberry plant is a plant of the species Fragaria vesca.
  • the donor octoploid strawberry plant is a plant of the species Fragaria x ananassa.
  • the embryo of the true homozygous octoploid strawberry seed has a nuclear genome derived from only the donor octoploid strawberry plant (e.g., Fragaria x ananassa) and a cytoplasmic genome (e.g., mitochondrial and/or chloroplast genome) derived from the haploid-inducing strawberry plant (e.g., Fragaria vesca).
  • the method of producing true homozygous octoploid strawberry plant, or plant part thereof comprises (a) contacting a tetrahaploid cell from a donor octoploid strawberry plant with pollen from an octoploid haploid-inducing strawberry plant described herein, and (b) producing an octoploid doubled tetrahaploid cell or plant from the tetrahaploid cell.
  • the method comprises (a1) contacting a reduced egg cell from a donor octoploid strawberry plant with a sperm cell from an octoploid haploid- inducing strawberry plant described herein, (a2) allowing a tetrahaploid embryo to form and grow into a tetrahaploid strawberry plant, and (b) subjecting the tetrahaploid strawberry plant to genome doubling to produce a doubled tetrahaploid strawberry plant.
  • the method comprises (a) contacting a tetrahaploid cell from a donor octoploid strawberry plant with pollen from an octoploid haploid-inducing strawberry plant described herein, and (b) producing an octoploid doubled tetrahaploid cell or plant from the tetrahaploid cell.
  • the method comprises (a1) contacting a tetrahaploid reduced egg cell from a donor octoploid strawberry plant with a sperm cell from an octoploid haploid- inducing strawberry plant described herein, (a2) allowing a tetrahaploid embryo to form and grow into a tetrahaploid strawberry plant, and (b) subjecting the tetrahaploid strawberry plant to genome doubling to produce an octoploid doubled tetrahaploid strawberry plant.
  • the octoploid haploid-inducing strawberry plant is a plant of the species Fragaria x ananassa or Fragaria chiloensis. In certain embodiments, the octoploid haploid- inducing strawberry plant is a plant of the species Fragaria x ananassa.
  • the method of producing true homozygous octoploid strawberry plant, or plant part thereof comprises (a) contacting a tetrahaploid cell from a donor octoploid strawberry plant with an egg cell from an octoploid haploid-inducing strawberry plant described herein, and (b) producing an octoploid doubled tetrahaploid cell or plant from the tetrahaploid cell.
  • the method comprises (a1) contacting a reduced sperm cell from a donor octoploid strawberry plant with an egg cell from an octoploid haploid-inducing strawberry plant described herein, (a2) allowing a tetrahaploid embryo to form and grow into a tetrahaploid strawberry plant, and (b) subjecting the tetrahaploid strawberry plant to genome doubling to produce a doubled tetrahaploid strawberry plant.
  • the method comprises (a) contacting a tetrahaploid cell from a donor octoploid strawberry plant with an egg cell from an octoploid haploid- inducing strawberry plant described herein, and (b) producing an octoploid doubled tetrahaploid cell or plant from the tetrahaploid cell.
  • the method comprises (a1) contacting a tetrahaploid reduced sperm cell from a donor octoploid strawberry plant with an egg cell from an octoploid haploid-inducing strawberry plant described herein, (a2) allowing a tetrahaploid embryo to form and grow into a tetrahaploid strawberry plant, and (b) subjecting the tetrahaploid strawberry plant to genome doubling to produce an octoploid doubled tetrahaploid strawberry plant.
  • the octoploid haploid-inducing strawberry plant is a plant of the species Fragaria x ananassa or Fragaria chiloensis.
  • the octoploid haploid-inducing strawberry plant is a plant of the species Fragaria x ananassa. In some embodiments, the haploid-inducing strawberry plant and the donor octoploid strawberry plants are from different species of Fragaria. In some embodiments, the donor octoploid strawberry plant is a plant of the species Fragaria x ananassa.
  • the embryo of the true homozygous octoploid strawberry seed has a nuclear genome derived from only the donor octoploid strawberry plant (e.g., Fragaria x ananassa) and a cytoplasmic genome (e.g., mitochondrial and/or chloroplast genome) derived from the haploid-inducing strawberry plant.
  • a nuclear genome derived from only the donor octoploid strawberry plant e.g., Fragaria x ananassa
  • cytoplasmic genome e.g., mitochondrial and/or chloroplast genome
  • Haploid induction may comprise intraspecific haploid induction (e.g., using pollen of a haploid-inducing Fragaria x ananassa plant to fertilize a donor Fragaria x ananassa 99 sf-6744554 197072001240 plant) or interspecific haploid induction (e.g., using pollen of a haploid-inducing Fragaria vesca plant to fertilize a donor Fragaria x ananassa plant).
  • intraspecific haploid induction e.g., using pollen of a haploid-inducing Fragaria x ananassa plant to fertilize a donor Fragaria x ananassa plant
  • interspecific haploid induction e.g., using pollen of a haploid-inducing Fragaria vesca plant to fertilize a donor Fragaria x ananassa plant.
  • the method of producing true homozygous octoploid strawberry plant, or plant part thereof comprises (a) contacting a tetrahaploid cell from a donor octoploid strawberry plant with pollen from a haploid-inducing strawberry plant described herein, wherein the donor octoploid strawberry plant and the haploid-inducing strawberry plant are plants of different species; and (b) producing an octoploid doubled tetrahaploid cell or plant from the tetrahaploid cell.
  • the method comprises (a1) contacting a reduced egg cell from a donor octoploid strawberry plant with a sperm cell from a haploid-inducing strawberry plant described herein, wherein the donor octoploid strawberry plant and the haploid-inducing strawberry plant are plants of different species; (a2) allowing a tetrahaploid embryo to form and grow into a tetrahaploid strawberry plant; and (b) subjecting the tetrahaploid strawberry plant to genome doubling to produce a doubled tetrahaploid strawberry plant.
  • the method comprises (a) contacting a tetrahaploid cell from a donor octoploid strawberry plant with pollen from a haploid-inducing strawberry plant described herein, wherein the donor octoploid strawberry plant and the haploid- inducing strawberry plant are plants of different species; and (b) producing an octoploid doubled tetrahaploid cell or plant from the tetrahaploid cell.
  • the method comprises (a1) contacting a tetrahaploid reduced egg cell from a donor octoploid strawberry plant with a sperm cell from a haploid-inducing strawberry plant described herein, wherein the donor octoploid strawberry plant and the haploid-inducing strawberry plant are plants of different species; (a2) allowing a tetrahaploid embryo to form and grow into a tetrahaploid strawberry plant; and (b) subjecting the tetrahaploid strawberry plant to genome doubling to produce an octoploid doubled haploid strawberry plant.
  • the donor octoploid strawberry plant is a plant of the species Fragaria x ananassa
  • the haploid-inducing strawberry plant is a plant of the species Fragaria vesca.
  • the method of producing true homozygous octoploid strawberry plant, or plant part thereof comprises (a) contacting a tetrahaploid cell from a donor octoploid strawberry plant with an egg cell from a haploid-inducing strawberry plant described herein, wherein the donor octoploid strawberry plant and the haploid-inducing strawberry plant are plants of different species; and (b) producing an octoploid doubled tetrahaploid cell or plant from the tetrahaploid cell.
  • the method comprises (a1) contacting a reduced sperm cell from a donor octoploid strawberry plant with 100 sf-6744554 197072001240 an egg cell from a haploid-inducing strawberry plant described herein, wherein the donor octoploid strawberry plant and the haploid-inducing strawberry plant are plants of different species; (a2) allowing a tetrahaploid embryo to form and grow into a tetrahaploid strawberry plant; and (b) subjecting the tetrahaploid strawberry plant to genome doubling to produce a doubled tetrahaploid strawberry plant.
  • the method comprises (a) contacting a tetrahaploid cell from a donor octoploid strawberry plant with an egg cell from a haploid-inducing strawberry plant described herein, wherein the donor octoploid strawberry plant and the haploid-inducing strawberry plant are plants of different species; and (b) producing an octoploid doubled tetrahaploid cell or plant from the tetrahaploid cell.
  • the method comprises (a1) contacting a tetrahaploid reduced sperm cell from a donor octoploid strawberry plant with an egg cell from a haploid-inducing strawberry plant described herein, wherein the donor octoploid strawberry plant and the haploid-inducing strawberry plant are plants of different species; (a2) allowing a tetrahaploid embryo to form and grow into a tetrahaploid strawberry plant; and (b) subjecting the tetrahaploid strawberry plant to genome doubling to produce an octoploid doubled haploid strawberry plant.
  • the donor octoploid strawberry plant is a plant of the species Fragaria x ananassa
  • the haploid-inducing strawberry plant is a plant of the species Fragaria vesca.
  • the embryo of the true homozygous octoploid strawberry seed has a nuclear genome derived from Fragaria x ananassa and a cytoplasmic genome (e.g., mitochondrial and/or chloroplast genome) derived from Fragaria vesca.
  • the method of producing true homozygous octoploid strawberry plant, or plant part thereof comprises (a) contacting a tetrahaploid cell from a donor octoploid strawberry plant with pollen from a haploid-inducing strawberry plant described herein, wherein the donor octoploid strawberry plant and the haploid-inducing strawberry plant are plants of the same species; and (b) producing an octoploid doubled tetrahaploid cell or plant from the tetrahaploid cell.
  • the method comprises (a1) contacting a reduced egg cell from a donor octoploid strawberry plant with a sperm cell from a haploid-inducing strawberry plant described herein, wherein the donor octoploid strawberry plant and the haploid-inducing strawberry plant are plants of the same species; (a2) allowing a tetrahaploid embryo to form and grow into a tetrahaploid strawberry plant; and (b) subjecting the tetrahaploid strawberry plant to genome doubling to produce a doubled tetrahaploid strawberry plant.
  • the method comprises (a) contacting a tetrahaploid cell from a donor octoploid strawberry plant with pollen from a 101 sf-6744554 197072001240 haploid-inducing strawberry plant described herein, wherein the donor octoploid strawberry plant and the haploid-inducing strawberry plant are plants of the same species; and (b) producing an octoploid doubled tetrahaploid cell or plant from the tetrahaploid cell.
  • the method comprises (a1) contacting a tetrahaploid reduced egg cell from a donor octoploid strawberry plant with a sperm cell from a haploid-inducing strawberry plant described herein, wherein the donor octoploid strawberry plant and the haploid-inducing strawberry plant are plants of the same species; (a2) allowing a tetrahaploid embryo to form and grow into a tetrahaploid strawberry plant; and (b) subjecting the tetrahaploid strawberry plant to genome doubling to produce an octoploid doubled tetrahaploid strawberry plant.
  • both the donor octoploid strawberry plant and the haploid-inducing strawberry plant are plants of the species Fragaria x ananassa.
  • the method of producing true homozygous octoploid strawberry plant, or plant part thereof comprises (a) contacting a tetrahaploid cell from a donor octoploid strawberry plant with an egg cell from a haploid-inducing strawberry plant described herein, wherein the donor octoploid strawberry plant and the haploid-inducing strawberry plant are plants of the same species; and (b) producing an octoploid doubled tetrahaploid cell or plant from the tetrahaploid cell.
  • the method comprises (a1) contacting a reduced sperm cell from a donor octoploid strawberry plant with an egg cell from a haploid-inducing strawberry plant described herein, wherein the donor octoploid strawberry plant and the haploid-inducing strawberry plant are plants of the same species; (a2) allowing a tetrahaploid embryo to form and grow into a tetrahaploid strawberry plant; and (b) subjecting the tetrahaploid strawberry plant to genome doubling to produce a doubled tetrahaploid strawberry plant.
  • the method comprises (a) contacting a tetrahaploid cell from a donor octoploid strawberry plant with an egg cell from a haploid-inducing strawberry plant described herein, wherein the donor octoploid strawberry plant and the haploid-inducing strawberry plant are plants of the same species; and (b) producing an octoploid doubled tetrahaploid cell or plant from the tetrahaploid cell.
  • the method comprises (a1) contacting a tetrahaploid reduced sperm cell from a donor octoploid strawberry plant with an egg cell from a haploid-inducing strawberry plant described herein, wherein the donor octoploid strawberry plant and the haploid-inducing strawberry plant are plants of the same species; (a2) allowing a tetrahaploid embryo to form and grow into a tetrahaploid strawberry plant; and (b) subjecting the tetrahaploid strawberry plant to genome doubling to produce an octoploid doubled tetrahaploid strawberry plant.
  • both the donor octoploid strawberry plant and the haploid-inducing strawberry plant are plants of the species Fragaria x ananassa.
  • the step of generating a tetrahaploid strawberry plant through haploid induction comprises allowing the donor octoploid strawberry plant to form a plurality of seeds and subsequently allowing one or more seeds of the plurality to germinate and form one or more plants.
  • the step of generating a tetrahaploid strawberry plant through haploid induction comprises allowing the donor octoploid strawberry plant to form a plurality of seeds and subsequently allowing two or more seeds of the plurality to germinate and form a plurality of plants.
  • the step of generating a tetrahaploid strawberry plant through haploid induction comprises collecting one or more embryos from the plurality of seeds, contacting the one or more embryos with a nutrient medium suitable for inducing plant growth, and allowing the one or more embryos to form one or more plants.
  • the step of generating a tetrahaploid strawberry plant through haploid induction comprises collecting a plurality of embryos from the plurality of seeds, contacting the plurality of embryos with a nutrient medium suitable for inducing plant growth, and allowing the plurality of embryos to form a plurality of plants.
  • the method may further comprise a step of selecting one or more tetrahaploid plants by 1) determining the ploidy of one or more cells of one or more plants of the plurality of plants, and 2) selecting one or more plants having one or more tetrahaploid cells from the plurality of plants.
  • the step of selecting one or more tetrahaploid seeds of the plurality of seeds comprises 1) determining the ploidy of one or more cells of one or more plants of the plurality of plants, and 2) selecting one or more plants having one or more tetraploid cells from the plurality of plants.
  • determining the ploidy of one or more cells of the one or more plants comprises obtaining a tissue sample of each of the one or more plants and determining the ploidy of one or more cells of each tissue sample.
  • the step of generating a tetrahaploid strawberry plant through haploid induction comprises allowing the haploid-inducing strawberry plant to form a plurality of seeds and subsequently allowing one or more seeds of the plurality to germinate and form one or more plants. In certain embodiments, the step of generating a tetrahaploid strawberry plant through haploid induction comprises allowing the haploid-inducing strawberry plant to form a plurality of seeds and subsequently allowing two or more seeds of the plurality to germinate and form a plurality of plants.
  • the step of 103 sf-6744554 197072001240 generating a tetrahaploid strawberry plant through haploid induction comprises collecting one or more embryos from the plurality of seeds, contacting the one or more embryos with a nutrient medium suitable for inducing plant growth, and allowing the one or more embryos to form one or more plants.
  • the step of generating a tetrahaploid strawberry plant through haploid induction comprises collecting a plurality of embryos from the plurality of seeds, contacting the plurality of embryos with a nutrient medium suitable for inducing plant growth, and allowing the plurality of embryos to form a plurality of plants.
  • the method may further comprise a step of selecting one or more tetrahaploid plants by 1) determining the ploidy of one or more cells of one or more plants of the plurality of plants, and 2) selecting one or more plants having one or more tetrahaploid cells from the plurality of plants.
  • the step of selecting one or more tetrahaploid seeds of the plurality of seeds comprises 1) determining the ploidy of one or more cells of one or more plants of the plurality of plants, and 2) selecting one or more plants having one or more tetraploid cells from the plurality of plants.
  • determining the ploidy of one or more cells of the one or more plants comprises obtaining a tissue sample of each of the one or more plants and determining the ploidy of one or more cells of each tissue sample.
  • Methods of determining the ploidy of a cell are known in the art and are described, for example, in Example 3 of the present disclosure and Galbraith et al (Galbraith et al. 1983. “Rapid flow cytometric analysis of the cell cycle in intact plant tissues.” Science 220, no. 4601: 1049-1051) and reviewed in, for example, Ochatt et al (Ochatt et al. 2011.
  • the methods of producing true homozygous octoploid strawberry plants, or plant parts thereof comprise subjecting a tetrahaploid strawberry cell or plant to genome doubling to produce a doubled tetrahaploid strawberry cell or plant.
  • Genome doubling also known as chromosome doubling, refers to the process of disrupting mitosis in a parent cell or plant in order to generate daughter cells or plants having two clonal copies of the genome of the parent cell or plant.
  • haploid doubling mitosis is disrupted in a haploid parent cell or plant in order to generate doubled-haploid daughter cells or plants having two clonal copies of the haploid gene of the parent cell or plant. This results in a true homozygous daughter cell or plant which is monoallelic at all loci across its two haploid sets 104 sf-6744554 197072001240 of chromosomes.
  • the methods of producing true homozygous octoploid strawberry plants, or plant parts thereof comprise subjecting one or more tetrahaploid cells, tetrahaploid seeds, or tetrahaploid plants to a haploid doubling treatment.
  • the tetrahaploid cells, tetrahaploid seeds, or tetrahaploid plants may have been selected from a plurality of cells, seeds, or plants grown from an octoploid strawberry donor plant that has been contacted with pollen or a part thereof (e.g., sperm cells) from a haploid-inducing strawberry plant, as described herein.
  • subjecting one or more tetrahaploid cells, tetrahaploid seeds, or tetrahaploid plants to a haploid doubling treatment comprises contacting the one or more tetrahaploid cells, tetrahaploid seeds, or tetrahaploid plants with an agent that causes genome doubling or chromosome doubling, such as an antimitotic agent.
  • the antimitotic agent is an anti-microtubule agent. In some embodiments, the antimitotic agent comprises colchicine, oryzalin, oxide, trifluralin, or any combination thereof. In some embodiments, the antimitotic agent comprises colchicine. In some embodiments, the antimitotic agent is an antimitotic herbicide. Antimitotic herbicides include, for example, oryzalin, trifluralin, and amiprofos-methyl (APM).
  • APM amiprofos-methyl
  • the haploid doubling treatment comprises contacting the tetrahaploid cell, tetrahaploid seed, or tetrahaploid plant with an anti-microtubule agent to form a doubled tetrahaploid cell, a doubled tetrahaploid seed, or a doubled tetrahaploid plant.
  • the doubled tetrahaploid cell or the doubled tetrahaploid seed is allowed to grow into a doubled tetrahaploid plant.
  • the doubled tetrahaploid plant is allowed to form seed, thus producing the true homozygous octoploid strawberry seed.
  • Methods of chromosome doubling in plants are known in the art and reviewed, for example, in Hooghvorst et al. (2021. "Chromosome doubling methods in doubled haploid and haploid inducer-mediated genome-editing systems in major crops.” Plant Cell Reports 40.2: 255- 270.). Methods of chromosome doubling include, but are not limited to, treatment with mitotic inhibitors such as colchicine, oryzalin, trifluralin, or nitrous oxide. 105 sf-6744554 197072001240 [0238] In some embodiments, haploid doubling occurs spontaneously.
  • methods of producing true homozygous octoploid strawberry plant comprising (a) generating a tetrahaploid strawberry cell or plant through haploid induction, and (b) allowing the tetrahaploid strawberry plant to undergo spontaneous genome doubling to produce a doubled tetrahaploid strawberry cell or plant.
  • the method may further comprise a step of selecting one or more doubled tetrahaploid cells, seeds, or plants. From a plurality of tetrahaploid cells, seeds, or plants.
  • the method comprises selecting one or more doubled tetrahaploid seeds from a plurality of seeds by 1) determining the ploidy of the embryo of one or more seeds of the plurality of seeds, and 2) selecting one or more seeds having an octoploid embryo.
  • the step of selecting one or more tetrahaploid seeds of the plurality of seeds comprises 1) determining the ploidy of the embryo and endosperm of one or more seeds of the plurality of seeds, and 2) selecting one or more seeds having an octoploid embryo and an endosperm having a ploidy of greater than 8x (e.g., a ploidy of 9x, 10x, 11x, or 12x).
  • true homozygous octoploid strawberry plants are true homozygous octoploid strawberry plants, plant parts thereof, plant lines, and seeds produced by the methods described herein.
  • the true homozygous octoploid strawberry plant is a plant of the species Fragaria x ananassa.
  • the true homozygous octoploid strawberry plant, or part thereof comprises two clonal sets of chromosomes.
  • the true homozygous octoploid strawberry plant, or part thereof comprises two clonal sets of chromosomes, each clonal set comprising a haploid set of chromosomes from each of four subgenomes.
  • the true homozygous octoploid strawberry plant, plant part, plant line, or seed is monoallelic across a majority of loci in the genome.
  • the true homozygous octoploid strawberry plant, plant part, plant line, or seed is monoallelic at more than 50%, more than 60%, more than 70%, more than 80%, more than 85%, more than 90%, more than 91%, more than 92%, more than 93%, more than 94%, more than 95%, more than 96%, more than 97%, more than 98%, more than 99%, more than 99.9%, more than 99.99%, or 100% of the loci in the genome.
  • 106 sf-6744554 197072001240 [0241] Methods of measuring homozygosity and heterozygosity within genomes and subgenomes of strawberry plants are known in the art.
  • BWA Burrows-Wheller Alignment
  • Read pairs are retained for analysis if they uniquely map to a single chromosome within a single subgenome. Variants are called at specific loci and the rate of within-subgenome heterozygosity is calculated. [0242] In the parental octoploid lines the average rate of within-subgenome heterozygosity is typically calculated to be approximately 1.5%, while the rate of within- subgenome heterozygosity in doubled tetrahaploids is typically less than 0.05% using the same methods, showing that the doubled tetrahaploids are approximately 30 times more homozygous than the octoploid parents after a single generation.
  • the true homozygous octoploid strawberry plant, plant part, plant line, or seed has a rate of within-subgenome heterozygosity of less than 1.5%, less than 1.0%, less than 0.5%, less than 0.1%, less than 0.05%, less than 0.01%, or less than 0.005%.
  • a homozygous octoploid strawberry plant, plant part, plant line, or seed comprising at least one cell having a nuclear genome from a first species of Fragaria and a cytoplasmic genome from a second species of Fragaria.
  • the first and second species of Fragaria may each independently be any species of Fragaria known in the art or described herein.
  • the nuclear genome and the cytoplasmic genome are both from octoploid species of Fragaria.
  • the nuclear genome is from an octoploid species of Fragaria and the cytoplasmic genome is from a diploid species of Fragaria.
  • the nuclear genome is from a plant of the species Fragaria x anannasa.
  • the cytoplasmic genome is a from a plant of the species Fragaria vesca.
  • the nuclear genome is from a plant of the species Fragaria x anannasa and the cytoplasmic genome is a from a plant of the species Fragaria vesca.
  • a runnerless true homozygous octoploid strawberry plant comprises one or more genetic modifications resulting in decreased expression of a gibberellin 20-oxidase (ga20ox) gene.
  • the runnerless true homozygous octoploid strawberry plant does not produce runners or produces fewer runners as compared to a control plant that lacks the one or more genetic modifications resulting in decreased expression of the ga20ox gene.
  • a population of true homozygous octoploid strawberry seed is provided herein.
  • At least 50% of the true homozygous octoploid strawberry seed are genetically uniform.
  • at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.9%, or at least 99.99% of the true homozygous octoploid strawberry seed are genetically uniform.
  • Genetic uniformity may be determined by any means known in the art including, for example, DNA-hybridization based methods (e.g., SNP-chip, DNA sequencing methods (e.g., whole genome sequencing or genotyping by sequencing (GBS)), and the like.
  • Methods of Breeding and Producing Uniform Octoploid F1 Hybrid Strawberry Seed [0247]
  • described herein are methods of breeding and producing a uniform octoploid F1 hybrid strawberry seed comprising crossing two true homozygous octoploid strawberry plants.
  • the methods comprise producing one or more true homozygous octoploid strawberry plants using the methods described herein.
  • the methods comprise performing a plurality of crosses between pairs of true homozygous octoploid strawberry lines selected from a plurality of true homozygous octoploid strawberry lines, evaluating the traits of the F1 generation of plants, and selecting one or more of the pairs of true homozygous octoploid strawberry lines that, when crossed, resulted in desirable traits in the F1 generation.
  • the method further comprises crossing the one or more selected pairs of true homozygous octoploid strawberry lines to produce the uniform octoploid F1 hybrid strawberry seed.
  • the methods of breeding and producing a uniform octoploid F 1 hybrid strawberry seed comprise crossing two true homozygous octoploid strawberry plants.
  • crossing two true homozygous octoploid strawberry 108 sf-6744554 197072001240 plants comprises contacting pollen from a first true homozygous octoploid strawberry plant with the stigma of a pistil of a second true homozygous octoploid strawberry plant.
  • the uniform octoploid F1 hybrid strawberry seed is produced by crossing the two true homozygous octoploid strawberry plants and allowing seeds to form.
  • the methods of breeding and producing uniform octoploid F1 hybrid strawberry seed comprise maintaining one or more true homozygous octoploid strawberry lines, one or more F1 hybrid strawberry lines, or a combination thereof.
  • the inbred and/or hybrid octoploid strawberry lines are maintained via vegetative propagation, selfing, apomixis, or any combination thereof. Additional methods of maintaining strawberry lines are well-known in the art.
  • the methods of breeding and producing uniform octoploid F1 hybrid strawberry seed comprise maintaining an inventory of true homozygous octoploid strawberry lines from which haplotypes may be selected for rapid deterministic combination of the haplotypes.
  • a uniform F1 hybrid strawberry seed produced by the methods described herein.
  • provided herein is a population of uniform octoploid F1 hybrid strawberry seed.
  • At least 50% of the uniform octoploid F1 hybrid strawberry seed are genetically uniform.
  • at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.9%, or at least 99.99% of the uniform octoploid F1 hybrid strawberry seed are genetically uniform.
  • the methods of breeding and producing uniform octoploid F1 hybrid strawberry seed comprise obtaining a set of octoploid strawberry lines.
  • the set of octoploid strawberry lines is genetically diverse and comprises a large and diverse pool of haplotypes.
  • the methods of breeding and producing uniform octoploid F1 hybrid strawberry seed comprises obtaining a set of lines of the same or related species of octoploid strawberry plant (e.g., Fragaria x ananassa and/or Fragaria chiloensis).
  • the set of strawberry lines may be obtained from any source and by any methods known in the art.
  • the set of octoploid strawberry lines is 109 sf-6744554 197072001240 obtained from sources including, but not limited to, natural diversity, existing breeding programs, or any combination thereof.
  • the set of octoploid strawberry lines is a genetically diverse founder population of plants. These plants may be collected from existent germplasm sources such as wild progenitor species and landraces. In some instances, these plants may have high genetic load and may not have undergone the narrowing of genetic variability attributable to the elite selection practices imposed on modern cultivated materials. As such, many possess traits promoting their fitness within non-agrarian environments in the case of wild progenitors, or have been adapted to vastly differing cultivation practices and agricultural environments in the case of landraces.
  • these founding octoploid strawberry lines may contain suites of both desirable and undesirable agronomic characteristics that may be recombined, selected, and complemented to develop a commercializable product.
  • Heterotic Groups the methods of breeding and producing uniform octoploid F1 hybrid strawberry seed comprise organizing the set of octoploid strawberry lines obtained into a plurality of heterotic groups, wherein each heterotic group comprises a haplotype, and wherein the haplotypes are grouped based on observed or predicted heterotic performance when combined in the octoploid F1 hybrid strawberry plant.
  • the methods of breeding and producing uniform octoploid F1 hybrid strawberry seed comprise organizing the set of octoploid strawberry lines into three or more heterotic groups, four or more heterotic groups, five or more heterotic groups, six or more heterotic groups, seven or more heterotic groups, or eight or more heterotic groups.
  • the obtained set of octoploid strawberry lines may be organized by assigning their membership into complementary heterotic groups based upon heterotic patterns observed from preliminary estimation or prediction of their combining ability for traits and environments of interest.
  • the combining ability is an estimation of the value of an octoploid strawberry plant as a parent as inferred by progeny testing in an established factorial or hierarchical mating design.
  • the main goal is identifying subpopulations of the set of octoploid strawberry lines based on employment of a clustering procedure that maximizes some measure of interpopulation combining ability.
  • Exhaustive evaluation of all possible parental combinations for the final octoploid F1 hybrid strawberry is infeasible for all but a trivial number of potential parents (i.e. even a partial diallel ignoring reciprocal crosses scales at (n+3)!/(4!(n-1)! crosses per n parents).
  • Hierarchical mating schemes necessitate an understanding and judgment of which set of “testers” or analogous constructs should serve as a relevant and efficient basis or frame of reference for inferring combining ability.
  • the sample of testers selected invariably biases perceptions of existing heterotic patterns.
  • the relative importance of traits and environments of interest used to infer these combining abilities are dynamic and depend upon market trends.
  • predictive modeling is essential and the process of assigning and refining heterotic groups and testers to represent them is one of iterative improvement and refinement throughout repeated cycles of the breeding process. Nonetheless, once preliminary heterotic group membership is assigned, interpopulation improvement of the true homozygous octoploid strawberry lines and development of hybrid octoploid strawberry plants may proceed.
  • the methods of breeding and producing uniform octoploid F1 hybrid strawberry seed comprise evaluating one or more characteristics of an octoploid F1 hybrid plant grown from the uniform octoploid F1 hybrid strawberry seed.
  • Methods for evaluating strawberry plant characteristics are numerous and well-known in the art.
  • the one or more characteristics evaluated may include, but are not limited to, plant size, plant vigor, fruit yield, fruit color, fruit flavor, fruit sweetness, fruit aroma, abiotic stress resistance, disease resistance, pest resistance, and the like.
  • the strawberry plants to be evaluated may then be grown under different geographical, climatic, and soil conditions, and further selections can be made during, and at the end of, the growing season.
  • the method of breeding a uniform octoploid F1 hybrid strawberry seed comprises repeating the steps of the method using the one or more characteristics of one or more octoploid F1 hybrid strawberry plants grown from the uniform octoploid F1 hybrid strawberry seed evaluated to guide the breeding of candidate octoploid 111 sf-6744554 197072001240 strawberry lines, the selecting of candidate octoploid strawberry lines, or both.
  • the method comprises repeating the steps of the method two, three, four, five, six, seven, eight, nine, or ten times or more.
  • the repeating of the steps of the method iteratively informs a genome prediction model for improved prediction of heterotic performance.
  • the improved prediction of heterotic performance allows for rapid combination of haplotypes with strong heterotic performance and acceleration of breeding programs.
  • the genetically modified plant parts, genetically modified plant cells, and processed plant products provided herein are non- regenerable.
  • genetically modified strawberry plants and genetically modified strawberry plant parts are provided herein.
  • the genetically modified strawberry plants and strawberry plant parts may be grown from true homozygous octoploid strawberry seed or uniform octoploid F1 strawberry seed described herein.
  • the genetically modified strawberry plants and strawberry plant parts may be regenerated from a genetically modified cell of a haploid-inducing strawberry plant, a true homozygous octoploid strawberry plant, or a uniform octoploid F1 strawberry plant described herein.
  • the genetically modified strawberry plants and strawberry plant parts may be regenerated from a genetically modified cell of a haploid-inducing strawberry plant, wherein the genetically modified cell comprises a genetic modification resulting in decreased expression of one or more CENH3 genes described herein.
  • Genetically modified strawberry plants can be obtained from a genetically modified strawberry seed.
  • Genetically modified strawberry plant parts can be obtained by cutting, snapping, grinding or otherwise disassociating the part from the strawberry plant.
  • the strawberry plant part may be any strawberry plant part known in the art, including, but not limited to, a flower, a pistil, a leaf, a stem, a petiole, a cutting, a tissue, a seed coat, an ovule, pollen, a sperm cell, a root, a fruit (e.g., an aggregate fruit), a cotyledon, a hypocotyl, a protoplast, an embryo, an anther, a seed, an achene, a stolon (also 112 sf-6744554 197072001240 known as a runner) or any portion thereof.
  • a flower e.g., a pistil, a leaf, a stem, a petiole, a cutting, a tissue, a seed coat, an ovule, pollen, a sperm cell, a root, a fruit (e.g., an aggregate fruit), a cotyledon, a hypocoty
  • a genetically modified strawberry plant part provided herein is a non-regenerable portion of a genetically modified strawberry plant part.
  • a “non-regenerable” portion of a genetically modified strawberry plant part refers to a portion that cannot be induced to form a whole strawberry plant or that cannot be induced to form a whole strawberry plant (e.g., through in vitro culture) that is capable of sexual and/or asexual reproduction.
  • a non-regenerable portion of a genetically modified strawberry plant part may be a portion of a flower, a pistil, a leaf, a stem, a petiole, a cutting, a tissue, a seed coat, an ovule, pollen, a sperm cell, a root, a fruit (e.g., an aggregate fruit), a cotyledon, a hypocotyl, a protoplast, an embryo, an anther, an achene, a stolon (also known as a runner) or any portion thereof.
  • a non-regenerable or non-propagating strawberry plant cell is provided herein.
  • a “non-regenerable strawberry plant cell” is a cell from a strawberry plant which cannot be regenerated into a whole strawberry plant that is capable of sexual and/or asexual reproduction through in vitro culture.
  • the non-regenerable strawberry plant cell may be in a strawberry plant or strawberry plant part described herein.
  • the non-regenerable strawberry plant cell may be a cell in a stolon, an achene, the hull or pericarp of said achene, a seed, or in the seedcoat of said seed.
  • Mature strawberry plant organs, including a mature leaf, a mature stem, a mature root, or a mature stolon contain at least one non-regenerable cell.
  • the non-regenerable strawberry plant cell is a somatic cell.
  • the regenerable cells are derived from embryos, protoplasts, meristematic cells, callus, pollen, leaves, anthers, pistils, ovules, stems, petioles, roots, root tips, stolons (also known as runners), fruits (e.g., aggregate fruits), seeds, achenes, flowers, cotyledons, and/or hypocotyls of a genetically modified plant or a genetically modified plant part grown from a haploid-inducing strawberry plant embryo or seed, a true homozygous octoploid strawberry seed, or a uniform octoploid F1 strawberry seed described herein.
  • a processed strawberry plant product derived from a genetically modified strawberry plant, plant part, or plant cell described 113 sf-6744554 197072001240 herein.
  • the processed plant product contains sufficient nucleic acid (e.g., DNA or RNA) and/or protein material from the genetically modified strawberry plant, plant part, or plant cell to detect nucleic acid and/or protein sequences corresponding to the haplotypes of one or two true homozygous octoploid strawberry plants described herein, the haplotypes of a uniform octoploid F1 strawberry seed described herein, or both.
  • the processed strawberry plant product is non-regenerable, i.e., cannot be induced to form a whole strawberry plant or that cannot be induced to form a whole strawberry plant that is capable of sexual and/or asexual reproduction.
  • a processed strawberry plant product may be a seed, an achene, a fruit (e.g., an aggregate fruit), a root, a vegetable, or any plant part described herein, and may be blended as a commodity or other product which moves through commerce and is derived from a genetically modified strawberry plant or a genetically modified plant part.
  • the commodity or other product can be tracked through commerce by detecting nucleic acid and/or protein sequences of the genetically modified plant or plant part from which they were obtained.
  • the processed strawberry plant product comprises a detectable amount of nucleotide and/or protein sequences corresponding to the haplotypes of one or two true homozygous octoploid strawberry plants described herein, the haplotypes of a uniform octoploid F1 strawberry seed described herein, or both.
  • the commodity or other product is produced in or maintained in the genetically modified strawberry plant or plant part from which the commodity or other product has been obtained.
  • Such commodities or other products of commerce include, but are not limited to, plant parts, fruit, biomass, oil, meal, food starch, syrup, sugar, animal feed, flour, flakes, bran, lint, hull, processed seed, seed, seedless fruit, puree, juice, juice concentrate, pulp, pomace, preserve, or sauce.
  • the processed plant product may be a food product that is processed by any means known in the art, e.g., canned, steamed, boiled, fried, dried, blanched, juiced, pureed, and/or frozen etc.
  • a genetically modified strawberry plant, strawberry plant part, or strawberry plant cell comprising one or more genetic modifications resulting in decreased expression of one or more CENH3 proteins described herein.
  • genetically modified strawberry plant, strawberry plant part, or strawberry plant cell comprises one or more genetic modifications of one or more CENH3 genes.
  • genetically modified strawberry plant, strawberry plant part, or 114 sf-6744554 197072001240 strawberry plant cell comprises one or more genetic modifications resulting in decreased expression of one or more CENH3 proteins having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity, or at least 60%, 65%, 70%, 75%, 80%, 85%, 95%, 96%, 97%, 98%, or 99% sequence similarity to an amino acid sequence selected from the list consisting of SEQ ID NOs: 8-14 and 53-56, or fragments thereof.
  • the genetically modified strawberry plant, strawberry plant part, or strawberry plant cell comprises one or more genetic modifications of one or more CENH3 genes comprising a nucleotide sequence having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity or complementarity to a nucleotide sequence selected from the group consisting of SEQ ID NOs: 1-7 and 58-61, or a fragment thereof.
  • the one or more genetic modifications comprise a modification of an enhancer of one or more of the CENH3 genes, a modification of a promoter of one or more of the CENH3 genes, a modification of a coding region of one or more of the CENH3 genes, a modification of an intron of one or more of the CENH3 genes, a modification of methylation status of one or more of the CENH3 genes, expression of a repressor protein that targets the DNA or an mRNA of one or more of the CENH3 genes, expression of an RNA interference construct that targets an mRNA of one or more of the CENH3 genes, or any combination thereof.
  • genetically modified strawberry plant, strawberry plant part, or strawberry plant cell has decreased expression of one or more CENH3 genes described herein relative to a strawberry plant (e.g., a control strawberry plant, e.g., a strawberry plant of the same species) lacking the one or more genetic modifications.
  • genetically modified strawberry plant, strawberry plant part, or strawberry plant cell lacks detectable expression of one or more CENH3 genes described herein relative to a strawberry plant (e.g., a control strawberry plant, e.g., a strawberry plant of the same species) lacking the one or more genetic modifications.
  • genetically modified strawberry plant, strawberry plant part, or strawberry plant cell lacks detectable expression of any CENH3 genes.
  • genetically modified strawberry plant, strawberry plant part, or strawberry plant cell further comprises one or more naturally occurring inactive alleles of one or more CENH3 genes.
  • the genetically modified strawberry plant, strawberry plant part, or strawberry plant cell is a plant or from a plant of the species Fragaria vesca and 115 sf-6744554 197072001240 comprises one or more genetic modifications resulting in decreased expression of one or more CENH3 genes.
  • the genetically modified strawberry plant, strawberry plant part, or strawberry plant cell is a plant or from a plant of the species Fragaria vesca and comprises one or more genetic modifications resulting in decreased expression of an FvCENH3 gene.
  • the genetically modified strawberry plant, strawberry plant part, or strawberry plant cell is a plant or from a plant of the species Fragaria vesca and comprises one or more genetic modifications resulting in decreased expression of an FvCENH3 protein.
  • the genetically modified strawberry plant, strawberry plant part, or strawberry plant cell is a plant or from a plant of the species Fragaria vesca and comprises one or more genetic modifications resulting in decreased expression of a CENH3 protein having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity, or at least 60%, 65%, 70%, 75%, 80%, 85%, 95%, 96%, 97%, 98%, or 99% sequence similarity to the FvCENH3 protein of SEQ ID NO.
  • the genetically modified strawberry plant, strawberry plant part, or strawberry plant cell is a plant or from a plant of the species Fragaria vesca and comprises one or more genetic modifications of an FvCENH3 gene or gene product.
  • the genetically modified strawberry plant, strawberry plant part, or strawberry plant cell is a plant or from a plant of the species Fragaria vesca and comprises one or more genetic modifications in a FvCENH3 gene comprising a nucleotide sequence having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity or complementarity to the FvCENH3 gene nucleotide sequence of SEQ ID NO. 1, or a fragment thereof.
  • the genetically modified strawberry plant, strawberry plant part, or strawberry plant cell is a plant or from a plant of the species Fragaria vesca and comprises genetic modifications in both alleles of an FvCENH3 gene.
  • the one or more genetic modifications comprise a modification of an enhancer of FvCENH3, a modification of a promoter of FvCENH3, a modification of a coding region of FvCENH3, modification of an intron of FvCENH3, or any combination thereof relative to a control Fragaria vesca plant (e.g., a Fragaria vesca plant lacking one or more of the genetic modifications).
  • the genetically modified strawberry plant, strawberry plant part, or strawberry plant cell is a plant or from a plant of the species Fragaria vesca and has decreased 116 sf-6744554 197072001240 expression of FvCENH3 protein relative to a control Fragaria vesca plant (e.g., a Fragaria vesca plant lacking one or more of the genetic modifications).
  • the genetically modified strawberry plant, strawberry plant part, or strawberry plant cell is a plant or from a plant of the species Fragaria vesca and lacks detectable expression of FvCENH3 protein.
  • the genetically modified strawberry plant, strawberry plant part, or strawberry plant cell is a plant or from a plant of the species Fragaria x ananassa and comprises one or more genetic modifications resulting in decreased expression of one or more FaCENH3 genes.
  • the genetically modified strawberry plant, strawberry plant part, or strawberry plant cell is a plant or from a plant of the species Fragaria x ananassa and comprises one or more genetic modifications resulting in decreased expression of an FaCENH3-1a gene, an FaCENH3-1b gene, an FaCENH3-2a gene, an FaCENH3-2b gene, an FaCENH3-3 gene, an FaCENH3-4 gene, or any combination thereof.
  • the genetically modified strawberry plant, strawberry plant part, or strawberry plant cell is a plant or from a plant of the species Fragaria x ananassa and comprises one or more genetic modifications resulting in decreased expression of an FaCENH3-1a gene, an FaCENH3-1b gene, an FaCENH3-2a gene, an FaCENH3-2b gene, an FaCENH3-3 gene, and an FaCENH3-4 gene.
  • the genetically modified strawberry plant, strawberry plant part, or strawberry plant cell is a plant or from a plant of the species Fragaria x ananassa and comprises one or more genetic modifications resulting in decreased expression of an FaCENH3-1a protein, an FaCENH3-1b protein, an FaCENH3-2a protein, an FaCENH3-2b protein, an FaCENH3-3 protein, an FaCENH3-4 protein, or any combination thereof.
  • the genetically modified strawberry plant, strawberry plant part, or strawberry plant cell is a plant or from a plant of the species Fragaria x ananassa and comprises one or more genetic modifications resulting in decreased expression of an FaCENH3-1a protein, an FaCENH3-1b protein, an FaCENH3-2a protein, an FaCENH3-2b protein, an FaCENH3-3 protein, and an FaCENH3-4 protein.
  • the genetically modified strawberry plant, strawberry plant part, or strawberry plant cell is a plant or from a plant of the species Fragaria x ananassa and comprises one or more genetic modifications resulting in decreased expression of one or more CENH3 proteins having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 117 sf-6744554 197072001240 95%, 96%, 97%, 98%, 99%, or 100% sequence identity, or at least 60%, 65%, 70%, 75%, 80%, 85%, 95%, 96%, 97%, 98%, or 99% sequence similarity to an amino acid sequence selected from the list consisting of SEQ ID NOs: 9-14, or fragments thereof.
  • the genetically modified strawberry plant, strawberry plant part, or strawberry plant cell is a plant or from a plant of the species Fragaria x ananassa and comprises one or more genetic modifications resulting in decreased expression of 1) a CENH3 protein having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity, or at least 60%, 65%, 70%, 75%, 80%, 85%, 95%, 96%, 97%, 98%, or 99% sequence similarity to the amino acid sequence of SEQ ID NO: 9; 2) a CENH3 protein having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity, or at least 60%, 65%, 70%
  • the genetically modified strawberry plant, strawberry plant part, or strawberry plant cell is a plant or from a plant of the species Fragaria x ananassa and comprises one or more genetic modifications resulting in decreased expression of an FaCENH3-7a protein, an FaCENH3-7b protein, an FaCENH3-7c protein, an FaCENH3-7d protein, or any combination thereof.
  • the genetically modified 118 sf-6744554 197072001240 strawberry plant, strawberry plant part, or strawberry plant cell is a plant or from a plant of the species Fragaria x ananassa and comprises one or more genetic modifications resulting in decreased expression of one or more CENH3 proteins having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity, or at least 60%, 65%, 70%, 75%, 80%, 85%, 95%, 96%, 97%, 98%, or 99% sequence similarity to an amino acid sequence selected from the list consisting of SEQ ID NOs: 53-56, or fragments thereof.
  • the genetically modified strawberry plant, strawberry plant part, or strawberry plant cell is a plant or from a plant of the species Fragaria x ananassa and comprises one or more genetic modifications resulting in decreased expression of 1) a CENH3 protein having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity, or at least 60%, 65%, 70%, 75%, 80%, 85%, 95%, 96%, 97%, 98%, or 99% sequence similarity to the amino acid sequence of SEQ ID NO: 53; 2) a CENH3 protein having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity, or at least 60%, 65%, 70%
  • the genetically modified strawberry plant, strawberry plant part, or strawberry plant cell is a plant or from a plant of the species Fragaria x ananassa and comprises one or more genetic modifications of an FaCENH3-1a gene or gene product, an FaCENH3-1b gene or gene product, an FaCENH3-2a gene or gene product, an FaCENH3-2b gene or gene product, an FaCENH3-3 gene or gene product, an FaCENH3-4 gene or gene product, or any combination thereof.
  • the genetically modified strawberry plant, strawberry plant part, or strawberry plant cell is a plant or from a plant of the species Fragaria x ananassa and comprises one or more genetic modifications of an FaCENH3-1a gene or gene product, an FaCENH3-1b gene or gene product, an FaCENH3-2a 119 sf-6744554 197072001240 gene or gene product, an FaCENH3-2b gene or gene product, an FaCENH3-3 gene or gene product, and an FaCENH3-4 gene or gene product.
  • the genetically modified strawberry plant, strawberry plant part, or strawberry plant cell is a plant or from a plant of the species Fragaria x ananassa and comprises one or more genetic modifications in a CENH3 gene comprising a nucleotide sequence having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity or complementarity to a nucleotide sequence selected from the group consisting of SEQ ID NOs: 2-7, or a fragment thereof.
  • the genetically modified strawberry plant, strawberry plant part, or strawberry plant cell is a plant or from a plant of the species Fragaria x ananassa and comprises: 1) one or more genetic modifications in a CENH3 gene comprising a nucleotide sequence having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity or complementarity to the nucleotide sequence of SEQ ID NO: 2, or a fragment thereof; 2) one or more genetic modifications in a CENH3 gene comprising a nucleotide sequence having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity or complementarity to the nucleotide sequence of SEQ ID NO:
  • the one or more genetic modifications comprise a modification of an enhancer of an FaCENH3 gene (e.g., an FaCENH3-1a gene, an FacCENH3-1b gene, an 120 sf-6744554 197072001240 FaCENH3-2a gene, an FaCENH3-2b gene, an FaCENH3-3 gene, and/or an FaCENH3-4 gene), a modification of a promoter of an FaCENH3 gene (e.g., an FaCENH3-1a gene, an FacCENH3-1b gene, an FaCENH3-2a gene, an FaCENH3-2b gene, an FaCENH3-3 gene, and/or an FaCENH3-4 gene), a modification of a coding region of an FaCENH3 gene (e.g., an FaCENH3-1a gene, an FacCENH3-1b gene, an FaCENH3-2a gene, an FaCENH3-2b gene, an FaCENH3-3 gene, and/or an FaCENH3-4 gene), modification of an intron of an FaCENH
  • the genetically modified strawberry plant, strawberry plant part, or strawberry plant cell is a plant or from a plant of the species Fragaria x ananassa and comprises one or more genetic modifications of an FaCENH3-7a gene or gene product, an FaCENH3-7b gene or gene product, an FaCENH3-7c gene or gene product, an FaCENH3-7d gene or gene product, or any combination thereof.
  • the genetically modified strawberry plant, strawberry plant part, or strawberry plant cell is a plant or from a plant of the species Fragaria x ananassa and comprises one or more genetic modifications in a CENH3 gene comprising a nucleotide sequence having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity or complementarity to a nucleotide sequence selected from the group consisting of SEQ ID NOs: 58-61, or a fragment thereof.
  • the genetically modified strawberry plant, strawberry plant part, or strawberry plant cell is a plant or from a plant of the species Fragaria x ananassa and comprises: 1) one or more genetic modifications in a CENH3 gene comprising a nucleotide sequence having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity or complementarity to the nucleotide sequence of SEQ ID NO: 58, or a fragment thereof; 2) one or more genetic modifications in a CENH3 gene comprising a nucleotide sequence having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity or complementarity to the nucleotide sequence
  • the one or more genetic modifications comprise a modification of an enhancer of an FaCENH3 gene (e.g., an FaCENH3-7a gene, an FaCENH3-7b gene, an FaCENH3-7c gene, and/or an FaCENH3-7d gene), a modification of a promoter of an FaCENH3 gene (e.g., an FaCENH3-7a gene, an FaCENH3-7b gene, an FaCENH3-7c gene, and/or an FaCENH3-7d gene), a modification of a coding region of an FaCENH3 gene (e.g., an FaCENH3-7a gene, an FaCENH3-7b gene, an FaCENH3-7c gene, and/or an FaCENH3-7d gene), modification of an intron of an FaCENH3 gene (e.g., an FaCENH3-7a gene, an FaCENH3-7b gene, an FaCENH3-7c gene, and/or an FaCENH3-7d gene), or any combination
  • the one or more genetic modifications comprise one or more deletions, insertions, or nucleotide substitutions in a coding region of an FaCENH3 gene (e.g., an FaCENH3-7a gene, an FaCENH3-7b gene, an FaCENH3-7c gene, and/or an FaCENH3-7d gene).
  • an FaCENH3 gene e.g., an FaCENH3-7a gene, an FaCENH3-7b gene, an FaCENH3-7c gene, and/or an FaCENH3-7d gene.
  • the genetically modified strawberry plant, strawberry plant part, or strawberry plant cell is a plant or from a plant of the species Fragaria x ananassa and has decreased expression of an FaCENH3-1a protein, an FaCENH3-1b protein, an FaCENH3-2a protein, an FaCENH3-2b protein, an FaCENH3-3 protein, an FaCENH3-4 protein, or any combination (e.g., one, two, three, four, five or all six) thereof relative to a control Fragaria x ananassa plant (e.g., a Fragaria ananassa plant lacking one or more or all of the genetic modifications).
  • a control Fragaria x ananassa plant e.g., a Fragaria ananassa plant lacking one or more or all of the genetic modifications.
  • the genetically modified strawberry plant, strawberry plant part, or strawberry plant cell is a plant or from a plant of the species Fragaria x ananassa and lacks detectable expression of FaCENH3-1a protein, FaCENH3-1b protein, FaCENH3-2a protein, FaCENH3-2b protein, FaCENH3-3 protein, FaCENH3-4 protein, or any combination (e.g., two, three, four, five or all six) thereof.
  • a processed strawberry plant product derived from any of the foregoing embodiments of genetically modified strawberry plants, plant parts, or plant cells, wherein the processed strawberry plant product comprises a detectable amount of nucleotide and/or protein sequences corresponding to the haplotypes of one or two true homozygous octoploid strawberry plants described herein, the haplotypes of a uniform octoploid F1 strawberry seed described herein, or both of the genetically modified plant, plant part, or plant cell.
  • the product is selected from the group consisting of plant biomass, oil, meal, food starch, syrup, animal feed, flour, flakes, bran, lint, hulls, processed seed, puree, juice, juice concentrate, pulp, pomace, preserve, or sauce.
  • the processed plant product is non-regenerable. Table 1. Nucleotide and amino acid sequences of CENH3 genes and gene products from Fragaria vesca and Fragaria x ananassa, and associated target protospacer site sequences and guide RNA sequences.
  • the one or more genetic modifications comprise a modification of an enhancer of one or more of the CENH3 genes, a modification of a promoter of one or more of the CENH3 genes, a modification of a coding region of one or more of the CENH3 genes, a modification of an intron of one or more of the CENH3 genes, a modification of methylation status of one or more of the CENH3 genes, expression of a repressor protein that targets the DNA or an mRNA of one or more of the CENH3 genes, expression of an RNA interference construct that targets an mRNA of one or more of the CENH3 genes, or any combination thereof. 5.
  • the strawberry plant is a plant of the species Fragaria vesca, Fragaria iinumae, Fragaria nipponica, Fragaria viridis, Fragaria ⁇ bifera, Fragaria bucharica, Fragaria chinensis, Fragaria daltoniana, Fragaria emeiensis, Fragaria hayatae, Fragaria iinumae, Fragaria mandshurica, Fragaria viridis, Fragaria nilgerrensis, Fragaria nipponica, Fragaria nubicola, or Fragaria pentaphylla.
  • the one or more genetic modifications comprise a modification of an enhancer of FvCENH3, a modification of a promoter of FvCENH3, a modification of a coding region of FvCENH3, modification of an intron of FvCENH3, or any combination thereof relative to a wild-type Fragaria vesca plant.
  • a polynucleotide sequence encoding an amino acid sequence having at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity to the amino acid sequence of SEQ ID NO: 53; b. a polynucleotide sequence encoding an amino acid sequence having at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity to the amino acid sequence of SEQ ID NO: 54; c.
  • a polynucleotide sequence encoding an amino acid sequence having at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity to the amino acid sequence of SEQ ID NO: 55; and/or d. a polynucleotide sequence encoding an amino acid sequence having at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity to the amino acid sequence of SEQ ID NO: 56.
  • the strawberry plant, or plant part thereof, of embodiment 15-23 wherein the strawberry plant is a plant of the species Fragaria x ananassa and the one or more CENH3 genes comprise one, two, three, four, five, or all six of FaCENH3-7a, FaCENH3-7b, FaCENH3-7c, and FaCENH3-7d.
  • the one or more genetic modifications comprise: a.
  • a modification of an enhancer of FaCENH3-7a a modification of a promoter of FaCENH3-7a, a modification of a coding region of FaCENH3-7a, a modification of an intron of FaCENH3-7a, or any combination thereof relative to an unmodified Fragaria x ananassa plant
  • 33. The strawberry plant, or plant part thereof, of any one of embodiments 1-32, wherein the plant part is pollen.
  • 34. The strawberry plant, or plant part thereof, of any one of embodiments 1-33, wherein the plant part is a seed. 35.
  • haploid-inducing strawberry plant, or plant part thereof, of embodiment 35 or 36 wherein one or more of the CENH3 genes comprises a polynucleotide sequence having at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity to a polynucleotide sequence selected from the group consisting of SEQ ID NOs: 1- 7 and 58-61. 38.
  • pacifica or Fragaria chiloensis subsp. sandwicensis.
  • a polynucleotide sequence encoding an amino acid sequence having at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity to the amino acid sequence of SEQ ID NO: 9;
  • a polynucleotide sequence encoding an amino acid sequence having at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity to the amino acid sequence of SEQ ID NO: 10;
  • a polynucleotide sequence encoding an amino acid sequence having at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity to the amino acid sequence of SEQ ID NO: 11; 175 sf-6744554 197072001240 d.
  • a polynucleotide sequence encoding an amino acid sequence having at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity to the amino acid sequence of SEQ ID NO: 12; e.
  • a polynucleotide sequence encoding an amino acid sequence having at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity to the amino acid sequence of SEQ ID NO: 13; and/or f. a polynucleotide sequence encoding an amino acid sequence having at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity to the amino acid sequence of SEQ ID NO: 14.
  • a polynucleotide sequence encoding an amino acid sequence having at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity to the amino acid sequence of SEQ ID NO: 55; and/or d. a polynucleotide sequence encoding an amino acid sequence having at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity to the amino acid sequence of SEQ ID NO: 56.
  • the haploid-inducing strawberry plant, or plant part thereof, of embodiment 51-61 wherein the haploid-inducing strawberry plant is a plant of the species Fragaria x ananassa and the one or more CENH3 genes comprise one, two, three, four, five, or all six of FaCENH3-1a, FaCENH3-1b, FaCENH3-2a, FaCENH3-2b, FaCENH3-3, and FaCENH3-4.
  • the one or more CENH3 genes comprise FaCENH3-1a, FaCENH3-1b, FaCENH3- 2a, FaCENH3-2b, FaCENH3-3, and FaCENH3-4.
  • a modification of an enhancer of FaCENH3-1b a modification of a promoter of FaCENH3-1b, a modification of a coding region of FaCENH3-1b, a modification of an intron of FaCENH3-1b, or any combination thereof relative to an unmodified Fragaria x ananassa plant
  • a modification of an enhancer of FaCENH3-2b a modification of a promoter of FaCENH3-2b, a modification of a coding region of FaCENH3-2b, a modification of an intron of FaCENH3-2b, or any combination thereof relative to an unmodified Fragaria x ananassa plant
  • a modification of an enhancer of FaCENH3-3 a modification of a promoter of FaCENH3-3, a modification of a coding region of FaCENH3-3, a modification of an intron of FaCENH3-3, or any combination thereof relative to an unmodified Fragaria x ananassa plant
  • f a modification of an enhancer of FaCENH3-3, a modification of a promoter of FaCENH3-3, a modification of a coding region of FaCENH3-3, a modification of an intron of FaCENH3-3, or any combination thereof relative to an unmodified Fragaria x ananassa plant
  • a modification of an enhancer of FaCENH3-4 a modification of a promoter of FaCENH3-4, a modification of a coding region of FaCENH3-4, a modification of an intron of FaCENH3-3, or any combination thereof relative to an unmodified Fragaria x ananassa plant.
  • the haploid-inducing strawberry plant, or plant part thereof, of embodiment 65 wherein the one or more genetic modifications comprise: a. a modification of an enhancer of FaCENH3-7a, a modification of a promoter of FaCENH3-7a, a modification of a coding region of FaCENH3-7a, a 178 sf-6744554 197072001240 modification of an intron of FaCENH3-7a, or any combination thereof relative to an unmodified Fragaria x ananassa plant; b.
  • a modification of an enhancer of FaCENH3-7b a modification of a promoter of FaCENH3-7b, a modification of a coding region of FaCENH3-7b, a modification of an intron of FaCENH3-7b, or any combination thereof relative to an unmodified Fragaria x ananassa plant
  • c a modification of an enhancer of FaCENH3-7c, a modification of a promoter of FaCENH3-7c, a modification of a coding region of FaCENH3-7c, a modification of an intron of FaCENH3-7c, or any combination thereof relative to an unmodified Fragaria x ananassa plant
  • d a modification of an enhancer of FaCENH3-7b, a modification of a promoter of FaCENH3-7b, a modification of a coding region of FaCENH3-7b, a modification of an intron of FaCENH3-7c, or any combination thereof relative to an unmodified Fragaria
  • a modification of an enhancer of FaCENH3-7d a modification of a promoter of FaCENH3-7d, a modification of a coding region of FaCENH3-7d, a modification of an intron of FaCENH3-7d, or any combination thereof relative to an unmodified Fragaria x ananassa plant.
  • 67. The haploid-inducing strawberry plant, or plant part thereof, of any one of embodiments 35-66, wherein the expression or activity of one or more CENH3 genes is decreased by no more than about 50%, about 60%, about 70%, about 80%, about 90%, about 95%, or about 99% as compared to a control plant, plant part, or plant cell lacking the one or more genetic modifications.
  • 71. The haploid-inducing strawberry plant of any one of embodiments 35-67, wherein the haploid-inducing strawberry plant has a larger bud size at which microspores form as compared to a control strawberry plant lacking the one or more genetic modifications. 179 sf-6744554 197072001240 72.
  • 76. A method of producing the strawberry plant of any one of embodiments 1-34 or the haploid-inducing strawberry plant of any one of embodiments 35-75.
  • 77. The method embodiment 76, wherein the decreased expression of the one or more CENH3 genes is achieved by gene disruption, gene knockout, gene knockdown, gene silencing, RNA interference, induction of methylation, or any combination thereof. 78.
  • the method of embodiment 76 or 78 comprising introducing one or more of the genetic modifications by mutagenesis, gene editing, transgenesis, or a combination thereof into a strawberry plant, plant part, or plant cell to produce the strawberry plant or the haploid- inducing strawberry plant.
  • the method of any one of embodiments 76-78 further comprising selecting a strawberry plant, plant part, or plant cell having decreased expression of the one or more CENH3 genes to produce the strawberry plant or the haploid-inducing strawberry plant.
  • 80. The method of embodiment 76-79 comprising introducing one or more of the genetic modifications by gene editing using a site-directed nuclease. 81.
  • the site-directed nuclease is a CRISPR- associated (Cas) nuclease, a zinc finger nuclease (ZFN), a transcription activator-like effector nuclease (TALEN), or a megaTAL. 82.
  • Cas CRISPR-associated nuclease
  • ZFN zinc finger nuclease
  • TALEN transcription activator-like effector nuclease
  • any one of embodiments 76-81 comprising contacting a plurality of cells of a parent strawberry plant with one or more expression vectors together comprising an expression cassette for a Cas nuclease and an expression cassette for an RNA molecule having at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% complementarity to 15, 16, 17, 18, 19, or 20 consecutive nucleotides of one or more of the CENH3 genes. 180 sf-6744554 197072001240 83.
  • any one of embodiments 76-81 comprising contacting a plurality of cells of a parent strawberry plant with one or more Cas nucleases, wherein each Cas nuclease is complexed with a RNA molecule having at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% complementarity to 15, 16, 17, 18, 19, or 20 consecutive nucleotides of one or more of the CENH3 genes.
  • RNA molecule is a crRNA, a gRNA, or a pegRNA.
  • RNA molecule is complementary to one or more nucleotide sequences consisting of SEQ ID NOs: 15-24 or comprises a nucleotide sequence selected from the group consisting of SEQ ID NOs: 25-34.
  • RNA molecule is complementary to one or more nucleotide sequences consisting of SEQ ID NOs: 17, 19, and 21, or comprises a nucleotide sequence selected from the group consisting of SEQ ID NOs: 27, 29, or 31.
  • the method any one of embodiments 82-86, wherein the plurality of cells is a plurality of protoplasts. 88.
  • a method of producing true homozygous octoploid strawberry seed comprising (a) contacting a tetrahaploid reduced egg cell from a donor octoploid strawberry plant with a sperm cell of the haploid-inducing strawberry plant of any one of embodiments 35-75, and (b) producing a doubled tetrahaploid cell from the tetrahaploid reduced egg cell.
  • step (a) comprises contacting pollen of the haploid-inducing strawberry plant with the stigma of a pistil of the donor octoploid strawberry plant comprising the reduced egg cell and allowing formation of a pollen tube and migration of the pollen tube to the reduced egg cell, thereby contacting the reduced egg cell with the sperm cell.
  • step (a) comprises contacting pollen of the haploid-inducing strawberry plant with the stigma of a pistil of the donor octoploid strawberry plant comprising the reduced egg cell and allowing formation of a pollen tube and migration of the pollen tube to the reduced egg cell, thereby contacting the reduced egg cell with the sperm cell.
  • the method of embodiment 91 further comprising selecting one or more tetrahaploid seeds by 1) determining the ploidy of the embryo and endosperm of one or more seeds of the 181 sf-6744554 197072001240 plurality of seeds, and 2) selecting one or more seeds having tetraploid embryo and an endosperm having a ploidy of greater than 8x. 93.
  • a method of producing true homozygous octoploid strawberry seed comprising (a) contacting an egg cell from the haploid-inducing strawberry plant of any one of embodiments 35-75 with a tetrahaploid reduced sperm cell from a donor octoploid strawberry plant, and (b) producing a doubled tetrahaploid cell from the egg cell, wherein the doubled tetrahaploid cell comprises the nuclear genome of the donor octoploid strawberry plant and the cytoplasmic genome of the haploid-inducing strawberry plant.
  • 94 The method of embodiment 93, wherein the haploid-inducing strawberry plant and the donor octoploid strawberry plants are from different species of Fragaria. 95.
  • the method of embodiment 94, wherein the haploid-inducing strawberry plant is diploid.
  • the method of embodiment 95, wherein the haploid-inducing strawberry plant is a plant of the species Fragaria vesca.
  • the method of embodiment 95, wherein the haploid-inducing strawberry plant is a plant of the species Fragaria x ananassa.
  • the method of any one of embodiments 93-97, wherein the donor octoploid strawberry plant is a plant of the species Fragaria x ananassa. 99.
  • step (a) comprises contacting the stigma of a pistil of the haploid-inducing strawberry plant with the pollen from the donor octoploid strawberry plant and allowing formation of a pollen tube and migration of the pollen tube to the egg cell, thereby contacting the egg cell with the tetrahaploid reduced sperm cell.
  • step (a) comprises contacting the stigma of a pistil of the haploid-inducing strawberry plant with the pollen from the donor octoploid strawberry plant and allowing formation of a pollen tube and migration of the pollen tube to the egg cell, thereby contacting the egg cell with the tetrahaploid reduced sperm cell.
  • step (a) comprises contacting the stigma of a pistil of the haploid-inducing strawberry plant with the pollen from the donor octoploid strawberry plant and allowing formation of a pollen tube and migration of the pollen tube to the egg cell, thereby contacting the egg cell with the tetrahaploid reduced sperm cell.
  • the method of embodiment 91, 92, or 99 further comprising collecting a plurality of embryos from the plurality of seeds, contacting the plurality of embryos with a nutrient medium suitable for inducing seedling growth, and allowing them to form a plurality of seedlings.
  • 103 The method of embodiment 101 or 102, further comprising selecting one or more tetrahaploid seedlings by 1) determining the ploidy of the cells of one or more seedlings of 182 sf-6744554 197072001240 the plurality of seedlings, and 2) selecting one or more tetrahaploid seedlings from the plurality of seedling. 104.
  • determining the ploidy of the cells of the one or more seedlings comprises obtaining a tissue sample of one or more seedlings and determining the ploidy of the cells of the tissue sample.
  • step (b) comprises subjecting the selected tetrahaploid cell, tetrahaploid seed, or tetrahaploid seedling to a tetrahaploid doubling treatment.
  • the tetrahaploid doubling treatment comprises contacting the tetrahaploid cell, tetrahaploid seed, or tetrahaploid seedling with an anti-microtubule agent to form a doubled tetrahaploid cell, a doubled tetrahaploid seed, or a doubled tetrahaploid seedling.
  • the anti-microtubule agent is an anti-mitotic herbicide.
  • the anti-microtubule agent comprises colchicine, oryzalin, oxide, trifluralin, or any combination thereof.
  • any one of embodiments 106-108 wherein the doubled tetrahaploid cell or the doubled tetrahaploid seed is allowed to grow into a doubled tetrahaploid seedling.
  • 110. The method of any one of embodiments 106-109, wherein the doubled tetrahaploid seedling is allowed to form seed, thus producing the true homozygous octoploid strawberry seed.
  • 111. The method of any one of embodiments 89-110, wherein the donor octoploid strawberry plant is a plant of the species Fragaria x ananassa, Fragaria chiloensis, Fragaria virginiana, or Fragaria iturupensis, or a hybrid of any combination thereof. 112.
  • the method of embodiment 111, wherein the donor octoploid strawberry plant is Fragaria chiloensis plant of the subspecies Fragaria chiloensis subsp. chiloensis, Fragaria chiloensis subsp. lucida, Fragaria chiloensis subsp. pacifica, or Fragaria chiloensis subsp. sandwicensis. 113.
  • the method of embodiment 111, wherein the donor octoploid strawberry plant is a plant of the species Fragaria x ananassa. 114.
  • the donor octoploid strawberry plant is a Fragaria x ananassa plant of the variety Camarosa or Albion.
  • the method further comprises introducing one or more genetic modifications resulting in decreased expression of 183 sf-6744554 197072001240 a gibberellin 20-oxidase (GA20ox) gene into the true homozygous octoploid strawberry plant or seed.
  • GA20ox gibberellin 20-oxidase
  • a method of producing a runnerless true homozygous octoploid strawberry plant comprising the steps of the method of any one of embodiments 89-115 and further comprising introducing one or more genetic modifications resulting in decreased expression of a gibberellin 20-oxidase (GA20ox) gene into the true homozygous octoploid strawberry plant, wherein the runnerless true homozygous octoploid strawberry plant does not produce runners.
  • GA20ox gibberellin 20-oxidase
  • the true homozygous octoploid strawberry seed of embodiment 117 wherein the true homozygous octoploid strawberry seed comprises one or more genetic modifications resulting in decreased expression of a gibberellin 20-oxidase (GA20ox) gene.
  • GA20ox gibberellin 20-oxidase
  • the true homozygous octoploid strawberry plant of embodiment 120 wherein the true homozygous octoploid strawberry plant does not produce runners.
  • the true homozygous octoploid strawberry seed of embodiment 130 wherein the different varieties of strawberry are different species of strawberry.
  • GA20ox gibberellin 20-oxidase
  • a true homozygous octoploid strawberry seed comprising an embryo having the nuclear genome of a first variety of strawberry and the cytoplasmic genome of a variety of strawberry.
  • 140. The true homozygous octoploid strawberry seed of embodiment 139, wherein the first and second varieties of strawberry are different species of strawberry. 141.
  • the true homozygous octoploid strawberry seed of embodiment 139 wherein the first and second varieties of strawberry are two different varieties of Fragaria x ananassa. 142.
  • a true homozygous octoploid strawberry seed comprising an embryo having the nuclear genome of a first species of Fragaria and the cytoplasmic genome of a second plant species of Fragaria. 143.
  • GA20ox gibberellin 20-oxidase
  • the true homozygous octoploid strawberry plant of embodiment 150, wherein the first species of Fragaria is Fragaria x ananassa. 186 sf-6744554 197072001240 152.
  • GA20ox gibberellin 20-oxidase
  • a method of producing uniform octoploid F1 hybrid strawberry seed comprising crossing two true homozygous octoploid strawberry plants of any one of embodiments 119- 122, 130-138, and 147-156.
  • the uniform octoploid F1 hybrid strawberry plant of embodiment 160, wherein the uniform octoploid F 1 hybrid strawberry plant comprises one or more genetic modifications resulting in decreased expression of a gibberellin 20-oxidase (GA20ox) gene. 162.
  • the expression vector or isolated DNA molecule of embodiment 164 comprising a DNA sequence having at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity or complementarity to 15, 16, 17, 18, 19, or 20 consecutive nucleotides of a polynucleotide sequence encoding an amino acid sequence having at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity to an amino acid sequence selected from the group consisting of SEQ ID NOs: 8-14 and 53-56. 166.
  • the expression vector or isolated DNA molecule of embodiment 164 or 165 comprising a DNA sequence having at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity or complementarity to 15, 16, 17, 18, 19, or 20 consecutive nucleotides of a polynucleotide having at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity to an polynucleotide sequence selected from the group consisting of SEQ ID NOs: 1-7 and 58-61. 167.
  • the expression vector or isolated DNA molecule of any one of embodiments 164- 166 comprising a DNA sequence encoding a non-coding RNA. 168.
  • non-coding RNA comprises a polynucleotide sequence having at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity or complementarity to 15, 16, 17, 18, 19, or 20 consecutive nucleotides of a polynucleotide sequence encoding an amino acid sequence having at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity to an amino acid sequence selected from the group consisting of SEQ ID NOs: 8-14 and 53-56. 169.
  • the expression vector or isolated DNA molecule of embodiment 167-169 wherein the RNA molecule is complementary to one or more nucleotide sequences consisting of SEQ ID NOs: 15-24 or comprises a nucleotide sequence selected from the group consisting of SEQ ID NOs: 25-34. 171.
  • a bacterial cell comprising the expression vector or isolated DNA molecule of any one of embodiments 164-176. 178.
  • a genetically modified plant, plant part, plant cell, or seed comprising the expression vector or isolated DNA molecule of any one of embodiments 164-176.
  • 180. A kit comprising the expression vector or isolated DNA molecule of 164-176 or the bacterial cell of embodiment 177 or 178. 181.
  • Example 1 General Methods Plants [0275] An octoploid Fragaria x ananassa ‘Chandler' (PED524) and a diploid Fragaria vesca Hawaii 4 (PED505) and Alexandria (PED503) were initiated in vitro by meristem extraction using hormone free 1 ⁇ 2 MS media (Murashige & Skoog (1962) “A revised medium for rapid growth and bio assays with tobacco tissue cultures” Physiologia Plantarum, 15(3): 473-497). The hormone free 1 ⁇ 2 MS media was supplemented with 30 g/L sucrose and plants were then grown under 16 hour day / 8 hour night lighting regime with cool white fluorescent lighting. After establishment, plants were micropropagated on the same medium.
  • hormone free 1 ⁇ 2 MS media was supplemented with 30 g/L sucrose and plants were then grown under 16 hour day / 8 hour night lighting regime with cool white fluorescent lighting. After establishment, plants were micropropagated on the same medium.
  • RNPs were prepared by standard methods known in the art. In short, 2 ⁇ L of NEB buffer 2.1 (10x stock) was placed into a 1.5 mL microcentrifuge tube with 10-40 ⁇ g of crRNA (CRISPR RNA) and an equal mass of Cas nuclease. The final volume was adjusted to 20 ⁇ L using nuclease free water.
  • Protoplasts Approximately 0.3 g of leaves from 3 to 4 week old explants were removed under aseptic conditions and sliced into thin sections 0.5 mm to 1 mm in width. Protoplasts were prepared from the sliced leaf tissue essentially as described in Barceló et al. (Barceló et al.
  • RNPs were added to 100 ⁇ L of isolated protoplast solution (approximately 2 x 105 protoplasts per mL) in 2 mL microfuge tubes.
  • An equal volume of PEG solution (40% PEG 4000 (wt/vol), 0.2 M mannitol, and 100 mM CaCl 2 ) was immediately mixed with the protoplasts by shaking gently and thoroughly.
  • the amount of plasmid DNA was determined experimentally between 5-80 ⁇ g.
  • Transfection mixtures were incubated at room temperature for 2-40 minutes, as determined experimentally, and then diluted with 440 ⁇ L W5 solution (2 mM MES (pH 5.7), 154 mM NaCl, 5 mM glucose, 125 mM CaCl 2 , and 5 mM KCl) at room temperature, and the transfection process terminated by gentle shaking.
  • Protoplasts were centrifuged at 100 x g for 1 minute at room temperature.
  • the protoplasts were gently resuspended with 200 ⁇ L WI solution (4 mM MES (pH 5.7), 0.5 M mannitol, and 20 mM KCl) and incubated in the dark for 20-25 hours at room temperature.
  • Example 2 Identification of target sites and guide RNA design Identification of cenh3 orthologs 191 sf-6744554 197072001240 [0281]
  • Candidate CENH3 orthologs from strawberry were identified by comparing CENH3 protein sequences from Zea mays (ZmCENH3) (SEQ ID NO: 44) and Arabidopsis thaliana (AtCENH3) (SEQ ID NO: 35) using Protein Blast, tblastn, and Clustal Omega workflows.
  • FvCENH3 (SEQ ID NO: 8) from diploid strawberry Fragaria vesca and six FaCENH3 homeologs (SEQ ID NOs: 9-14) from octoploid strawberry Fragaria x ananassa (Gene IDs based on Fragaria x ananassa Camarosa Genome v1.0.a2: www[dot]rosaceae[dot]org/Analysis/9642085) were identified as putative orthologs of ZmCENH3 (SEQ ID NO: 44) and AtCENH3 (SEQ ID NO: 35).
  • the configuration of the CENH3 loci in Fragaria x ananassa is illustrated in FIG. 3.
  • FIG. 4 The configuration of the CENH3 loci in Fragaria x ananassa is illustrated in FIG. 4.
  • Additional CENH3 orthologs were identified from Potentilla micrantha CENH3 (SEQ ID NO: 42) and closely related Fragaria species, Fragaria chiloensis CENH3-AV (SEQ ID NO: 36), Fragaria chiloensis CENH3-B1 (SEQ ID NO: 37), Fragaria chiloensis CENH3-B2 (SEQ ID NO: 38), Fragaria chiloensis CENH3-Bi (SEQ ID NO: 39), Fragaria iinumae CENH3 (SEQ ID NO: 40), and Fragaria viridis CENH3 (SEQ ID NO: 41) were identified.
  • FIG. 5A-5C The protein sequence alignment and a phylogenetic tree of CENH3 orthologs from Fragaria species, Zea mays, Arabidopsis thaliana are shown in FIG. 5A-5C.
  • the phylogenetic tree is a UPGMA consensus tree.
  • the scale bar represents phylogenetic distance of 0.05 nucleotide substitutions per site.
  • the tree is generated using software Geneious Prime 2023.2.1, with Genetic Distance Model: Jukes-Cantor.
  • Guide RNA design [0285] Protospacer sequences targeting FvCENH3 (SEQ ID NO: 1) or FaCENH3 homeologs (SEQ ID NO: 2-7) in Table 2 were selected.
  • RNAs were synthesized based on protospacer sequence and scaffold sequence and were screened in vivo.
  • Table 2 Protospacer sequences 192 sf-6744554 197072001240 Editing in protoplasts
  • RNPs with the synthesized guide RNAs and a Cas nuclease were prepared as in Example 1. Protoplasts were generated, transfected with the RNPs, and regenerated as described in Example 1. Sequence-based confirmation of editing [0287] Primers were designed to amplify each candidate edited cenh3 region based on the reference genome, and one or more long range direct PCRs were performed using crude lysate from transfected protoplasts. Alternatively, DNA was extracted from callus, leaf or other plant material.
  • PCR products were pooled by transfection sample and a seqWell library preparation performed to generate an Illumina library. Samples were loaded onto an Illumina iSeq and sequenced with a Paired End 150 nt sequencing kit. Sequences were analyzed by aligning fastq files to reference sequences, and mutations adjacent to target PAM sites for each targeted cenh3 gene were tabulated relative to a control. Editing efficiency was calculated based on the frequency of observed mutations as illustrated in FIG. 7, and used to calculate how many plants should be screened to identify the cenh3 gene knockouts required to confer a haploid induction phenotype.
  • Example 3 Haploid inducer system for strawberry Creation of haploid inducer lines
  • An octoploid Fragaria x ananassa (PED524) and a diploid Fragaria vesca (PED505) were edited with guide RNAs targeting PRS874 (SEQ ID NO. 27), PRS876 193 sf-6744554 197072001240 (SEQ ID NO: 29), and PRS878 (SEQ ID NO. 31) which were selected based on their unexpectedly high editing efficiency.
  • non- functional cenh3 alleles are denoted by a 0 and functional CENH3 alleles are denoted by a 1.
  • Each allele is given using the same terminology as FIG. 3 (-1a, -1b, -2a, -2b, -3 and 4), wherein “m” designates the maternal copy of that respective allele and “p” designates the paternal copy of that respective allele.
  • Additional Fragaria x ananassa plants are recovered, such that enough plants are recovered to cover all possible combinations of functional and edited, non-functional alleles (Table 3A provides an exemplary list and does not provide all possible combinations, all of which are recovered and within the scope of this example).
  • Table 3A Fragaria x ananassa haploid inducer lines. 194 sf-6744554 197072001240 [0290]
  • Table 3B provides the Fragaria vesca plants designated PED505-Fv-Inducer-01 through PED505-Fv-Inducer-03 which bear the indicated combinations of edited, nonfunctional cenh3 alleles, and functional CENH3 alleles, respectively. Edited, non- functional cenh3 alleles are denoted by a 0, a weak allele (resulting in reduced, but not eliminated, expression of CENH3) is denoted by a 1, and a functional CENH3 alleles are 195 sf-6744554 197072001240 denoted by a 2.
  • Table 3B Fragaria vesca haploid inducer lines [0291] After in vitro propagation, the populations of plants indicated the groups in Table 4 (below) are grown to maturity. Pollen is harvested from mature flowers of the plants from Groups A-i, C, E-i and G and used to pollinate the emasculated flowers of the plants from Groups B-i, D and F-i. All combinations for haploid induction of the maternal and paternal genome are tested including those described in Table 5 (below). For the unedited groups C and D, other strawberry varieties may be used, both cultivated and wild.
  • Table 4 Exemplary groups of strawberry plants for haploid induction 196 sf-6744554 197072001240
  • Table 5 Exemplary crosses for haploid induction [0292] Upon maturation of the berries all seeds (achenes) of the unmodified plants are collected, surface disinfected and placed into hormone free 1 ⁇ 2 MS basal salts for in vitro germination. Germinated seeds are grown in vitro until the second true leaf forms, at which point a leaf is harvested for crude lysate DNA extraction. Potential tetraploids are first screened by one of two molecular marker assays depending on the ploidy level of the inducer.
  • a presence-absence marker between Fragaria vesca and Fragaria x ananassa is used for the screening.
  • the marker is uniquely present in Fragaria vesca but not in any octoploid Fragaria x ananassa. F1 plants with the presence of this marker will be hybrids and F1 plants with the absence of this marker will be candidate 197 sf-6744554 197072001240 tetraploids.
  • a drop-off assay detecting CENH3 mutation at the editing site will be used for screening.
  • F1 plants with edited CENH3 alleles at the editing site will be hybrids and F1 plants with all WT CENH3 at the editing site will be candidate induced tetraploids or spontaneously doubled tetrahaploids. Confirmation of tetraploids is through flow cytometry performed essentially as described in Galbraith et al. (1983. “Rapid flow cytometric analysis of the cell cycle in intact plant tissues” Science 220(4601): 1049-1051). In short, intact nuclei are extracted into Galbraith buffer, then filtered and stained with propidium iodide. DNA content of nuclei is determined by applying the samples to a BD AccuriTM C6 Flow Cytometer.
  • Tetrahaploids are identified based on relative DNA content of nuclei between samples and internal controls, i.e. samples with approximately half the DNA content of unmodified octoploid Fragaria x ananassa controls are classified as tetrahaploid.
  • the tetrahaploid induction rate varies from among the ten populations of plants recovered, as determined by the number of haploid seedlings observed (including spontaneously doubled tetrahaploid) divided by the total number of germinated seedlings that develop.
  • Tetrahaploids are propagated in vitro on 1 ⁇ 2 MS medium with 0.5 mg/L 6- benzylaminopurine (BAP) and are subsequently doubled to produce fully homozygous octoploid plants with treatment of 0.1 to 0.3% colchicine solution in vitro for 4 to 24 hours.
  • Successful doubling of tetrahaploids is initially identified by plant morphology (larger and thicker leaves) and is then confirmed by flow cytometry using the same methods as above.
  • Resulting fully homozygous octoploid plants have approximately twice the DNA content of the unmodified tetrahaploid plants and are similar in DNA content to unmodified octoploid parents.
  • Editing occurs at any stage in the process including in the parent lines or in the tetrahaploid lines.
  • an octoploid Fragaria x ananassa (PED524) is edited to disrupt expression of FaGa20ox4-1 (SEQ. ID 46) and FaGa20ox4-3 (SEQ. ID 47), while simultaneously being edited for one or more FvCenh3 gene as described above.
  • a tetrahaploid (which has already undergone cenh3-induced haploid induction) is edited to disrupt expression of FaGa20ox4-1 (SEQ. ID 46) and FaGa20ox4-3 (SEQ. ID 47).
  • Example 4 Evaluation of CENH3-Edited Strawberry Lines Methods Sequence-based confirmation of editing [0298] Primers were designed to amplify each candidate edited cenh3 region based on the Fragaria x ananassa Camarosa or Royal Royce reference genome, and one or more long range direct PCRs were performed using crude lysate front transfected protoplasts. Alternatively, DNA was extracted from callus, leaf, or other plant material. PCR products were pooled either by transfection sample or plant material, and a seqWell library preparation was performed to generate Illumina libraries. Samples were sequenced on either an Illumina iSeq or MiSeq i100 using either a paired end 150 nt or paired end 300nt sequencing kit.
  • protoplast size was measured. High- resolution microscopic images of protoplasts were annotated and used to train a custom object detection model capable of identifying viable protoplasts and generating bounding boxes around each cell. Protoplasts without bounding boxes are non-viable. The bounding box dimensions were then used to determine cell size (FIG. 11A). Isolated protoplasts were encapsulated and maintained under sterile conditions, following methods described in Example 1.Six encapsulated lens were made for each cenh3 mutant line. The number of microcallus 0.2–0.4 mm in size per encapsulated lens was recorded 21 days after encapsulation. (FIG. 11B).
  • RNA quality and quantity were assessed and high-quality RNA samples were used to construct sequencing libraries for transcriptome analysis.
  • Three fully expanded primary or secondary flowers with bright-yellow anthers were destructively sampled from three plants.
  • the flower was categorized as either perfect (male and female reproductive structures), staminate (male only) or pistillate (female only).
  • Number of flowers per truss, , petal color, filament length, anther width and flower diameter were evaluated. Calipers were used to measure filament length, anther width, and flower diameter in mm.
  • Pollen germination was evaluated by removing three bright-yellow anthers from each of the three flowers. Anthers were placed in a 24-well plate and incubated at room temperature for an hour. After, the plate was gently tapped to release the pollen.
  • pollen germination media (20 mg/L boric acid, 100 g/L sucrose, pH 5.7) was added to each well and the plate was incubated at room temperature in the dark for four hours. Pollen germination was evaluated by counting the number of pollen grains with pollen tubes greater than the length of pollen grains. Either 50 or 100 pollen grains were observed, depending on the abundance of pollen grains. [0306] Flowers that were not destructively harvested were emasculated 3–4 days prior to opening. Then, 3–4 days after emasculation, receptive, glossy stigmas were pollinated using dried pollen collected from bright-yellow anthers of a wild type F. ⁇ ananassa (variety “Albion”).
  • Seeds were sanitized and germinated in- vitro by cutting the seed in half using a size 10 scalpel blade, placing the cut seed on 201 sf-6744554 197072001240 germination media (2.2 g/L MS basal salts with vitamins, 10 g/L sucrose, 3 g/L agar, pH 5.7), and incubating the seed at temperature between 20–25°C, a 16-hour photoperiod with a photon flux density of 40 ⁇ mol m -2 s -1 , and a relative humidity of 60–75%.
  • 201 sf-6744554 197072001240 germination media 2.2 g/L MS basal salts with vitamins, 10 g/L sucrose, 3 g/L agar, pH 5.7
  • Microspore development microscopy [0307] Strawberry buds at a variety of sizes were picked off the plant and measured. Six cenh3 mutant lines were analyzed (3 - 4 buds selected per plant) and 2 WT lines (3 buds/plant). Buds were then fixed in farmers Fixative 1:3 acetic acid:95% EtOH) overnight at 4C. Buds were washed in 70% EtOH and stored in fresh 70% EtOH at -20C. Each bud was then dissected to expose the anthers. Two anthers were extracted from each bud and slides were prepped for microscopy by squashing the tissue between a cover slip and microscope slide with a drop of acetocarmine solution. Slides were heated at 95C for 5 min and then visualized on a light microscope.
  • Haploid induction rate Ploidy was determined by sampling cotyledon tissue, approximately 0.2 cm 2 , from germinated seedlings 7-21 days after germination by flow cytometry. Tissue from F. vesca ‘Hawaii 4’ (2n), interspecific hybrid (5n), F. x ananassa ‘Chandler’ (8n) were collected as controls. Tissue was added to a 96-well plate with 30 ⁇ l of nuclei extraction buffer (Cystain kit). The tissue was finely cut using tattoo gun for 40-60 seconds. The instrument was rinsed thoroughly between samples. After, 170 ul of nuclei extraction buffer was added to each sample.
  • the “A” detector channel was used to determine the mean phycoerythrin fluorescent protein-chromophore (PE) labeled molecules bound to the nuclei.
  • PE phycoerythrin fluorescent protein-chromophore
  • the mean PE-A values of the peak(s) were recorded. If a pooled sample had two or more peaks, each individual was re-sampled individually by dispensing the remaining 150 ul of the sample stored in the refrigerator through the Celltric filters, removing the filters, adding 500 ml of the extraction buffer and incubating for 30-60 202 sf-6744554 197072001240 minutes at room temperature The number of aneuploids and tetrahaploids was recorded. The frequency of aneuploids was calculated as the number of aneuploid individuals divided by the total number of germinated seedlings.
  • the haploid induction rate was calculated as the number of tetrahaploids divided by the total number of germinated seedlings.
  • Results Identification of cenh3 mutant lines [0309] The regeneration rate, the number of calli with shoots of the total number of calli of F. ⁇ ananassa (variety “Chandler”) was 38.0% (Table 6). A total of 26 edited shoots of Chandler were identified from callus with shoots derived from protoplasts, respectively. The editing rate was, the number of calli with shoots that were edited of the total number of calli with shoots, 10.2%. The F. ⁇ ananassa cenh3 mutant lines had between 1–6 alleles of the total 8 alleles edited. The editing profiles of the F.
  • ⁇ ananassa cenh3 mutant lines is shown in Table 7, and edited FaCENH3 allele sequences for a subset of lines are shown in FIGS. 12A- 12F (FaCENH3-7a), FIGS. 13A-13C (FaCENH3-7b), FIGS. 14A-14F (FaCENH3-7c), and FIGS. 15A-15C (FaCENH3-7d).
  • Table 6 The number of calli, number of calli with shoots, regeneration rate (percentage of callus with shoots by the total callus) edited shoots, and editing rate (percentage of edited callus with shoots by the total callus with shoots) of F.
  • row 2 under Subgenome A “['0', '9']” indicates that the line has an unedited cenh3 allele on one 203 sf-6744554 197072001240 homologous chromosome of Subgenome A and a 9-nt deletion on the other homologous chromosome of Subgenome A.
  • Cell density, size and division evaluation [0310] Fifteen cenh3 mutant lines were evaluated for cell density (cells/g of tissue), size, and growth. Differences were observed between cenh3 mutant lines and wild type.
  • cenh3 mutant lines cenh3-13, cenh3-06, cenh3-13 withlower cell density compared to wild type.
  • cenh3 mutant lines cenh3-001 and cenh3-13, had significantly larger cells compared to wild type. These lines differed in cell growth (microcallus/ encapsulated lens.
  • cenh3 mutant lines cenh3-001, cenh3-005, cenh3-009, and cenh3-12 had fewer microcallus compared to wild type while on cenh3 mutant line, cenh3-010, had more microcallus compared to wild type. (Table 8).
  • Table 8 Table 8.
  • FIGS. 16B-16D Photographs of an exemplary squashed anther with microspheres is shown at various magnifications in FIGS. 16B-16D.
  • cenh3 mutant plant E-PED524-4145, cenh3-006
  • tetrads were observed in approximately two thirds of the buds. Tetrads are a normal stage prior to formation of microspores.
  • Exemplary images of microspores observed in cenh3-006 are shown in FIGS. 16E and 16F.
  • the PE-values were approximately 94,000 and 140,000. There were pooled samples from a F. x ananassa ‘Chandler’ cenh3-007 x F. x ananassa ‘Albion’ cross where a peak was observed between 44,000 – 94,000, between 94,000 – 140,000 or greater than 140,000 PA-E value. When the samples were analyzed individually seedlings were confirmed to have a PA-E value between 44,000 – 94,000 were characterized as a tetrahaploid. Individual seedlings confirmed to have a PA-E value between 94,000- 140,000 or greater than 140,000 were characterized as aneuploid. The haploid induction and aneuploid rate for F.
  • x ananassa ‘Chandler’ cenh3-006 crossed with F. x ananassa ‘Albion’ 208 sf-6744554 197072001240 is shown in Table 11.
  • the Cenh3-006 parent plant that resulted in a tetrahaploid seedling is the same plant that was observed to have tetrads in about two thirds of the buds, as described above and exemplified in FIGS. 16E-16F.
  • Table 11 Number of seedlings evaluated for ploidy from crosses with F. x ananassa ‘Chandler’ wild type or cenh3 mutant lines as the female and F. x ananassa ‘Albion’ as the male. 209 sf-6744554

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Abstract

The present disclosure relates to systems and methods for improved breeding of elite octoploid strawberry varieties. Described herein are haploid-inducing strawberry plants, plant parts, and plant lines comprising one or more genetic modifications resulting in decreased expression of one or more CENH3 (centromere-specific histone 3 variant) genes. Also described herein are methods of using thereof to generate true homozygous octoploid strawberry lines, as well as true breeding Fl hybrid octoploid strawberry seeds produced by crossing two true homozygous octoploid strawberry plants. Further described herein are methods and compositions for producing haploid-inducing strawberry plants, including methods for introducing genetic modifications resulting in decreased expression of one or more CENH3 genes.

Description

197072001240 HAPLOID INDUCER STRAWBERRY LINES AND METHODS OF PRODUCING AND USING THEREOF CROSS-REFERENCE TO RELATED APPLICATIONS [0001] This application claims the benefit of priority to U.S. Provisional Application No. 63/657,605, filed on June 7, 2024, which is incorporated by reference herein in its entirety. REFERENCE TO AN ELECTRONIC SEQUENCE LISTING [0002] The content of the electronic sequence listing (197072001240seqlisting.xml; Size: 184,457 bytes; and Date of Creation: June 6, 2025) is herein incorporated by reference in its entirety. FIELD [0003] The present disclosure relates generally to strawberry breeding, and more specifically to generation of fully homozygous octoploid strawberry lines for producing true breeding F1 hybrid octoploid strawberry seed. BACKGROUND [0004] Strawberries are a major agronomic fruit crop worldwide, with China, Mexico, and the U.S. being among the primary producers and consumers. According to recent data, 94% of U.S. households consume strawberries, with an annual per capita consumption of 4.85 pounds. California in particular produces 75% of the entire U.S. strawberry crop, with Florida being the next closest U.S. state in terms of production. [0005] Due to the complex genetics of the modern strawberry (e.g., having four distinct subgenomes, a tendency to outcross, and suffering from inbreeding depression), achieving fully homozygous inbred strawberry lines to cross for the production of true-breeding F1 hybrid strawberry seed has remained an elusive goal. Aside from inbreeding, anther culture has also been attempted in strawberry, but remains low efficiency with very few examples of limited success. (E.g., Xuan et al. 2012. Haploid plant production through anther culture in day-neutral strawberry (Fragaria x ananassa Duch) CV. Albion. J. ISSAAS Vol. 18, No. 1:173-184). Rather, current breeding practice involves recurrent selection and the clonal, vegetative propagation of strawberry plants through a resource intensive process, during which the plants are particularly susceptible to biotic and abiotic stressors. 1 sf-6744554 197072001240 [0006] Therefore, there is a need in the art for improved methods of producing fully homozygous strawberry lines that can be used for generating true-breeding F1 hybrid strawberry seed. Previous attempts, however, have failed to succeed in generating true- breeding populations that also exhibit commercially acceptable vigor and quality. (E.g., Dale et al. 2017. Breeding F1 hybrid day-neutral strawberries in eastern North America. ACTA HORTIC. 1156, 47-52). Another main issue with conversion of octoploid strawberry from a vegetatively propagated plant to a true seed plant is that breeding progress using current methodologies continues to provide consistent yield improvement, and allocation of resources toward inbreeding plants with traditional breeding would require many years and significant resources to develop populations and parents of sufficient homozygosity to make commercially relevant F1 hybrids. Therefore, most breeders have continued to use recurrent selection and vegetative propagation. Those results may also indicate limitations on the extent of homozygosity that can be achieved by selfing octoploid strawberry. Instead, it would be desirable to have a haploid inducing strawberry variety that could generate haploid progeny, which could then be doubled to achieve true homozygosity within each of the four subgenomes simultaneously. BRIEF SUMMARY [0007] For a variety of reasons, it would be desirable to have a system for producing fully homozygous strawberry lines to cross for the production of uniform (true-breeding) F1 hybrid strawberry seed. To our knowledge, the methods described herein provide the first such system enabled through use of a haploid inducer line in strawberry. [0008] In some aspects, provided herein is a strawberry plant, such as a haploid-inducing strawberry plant, or a plant part thereof, comprising one or more genetic modifications resulting in decreased expression of one or more CENH3 genes. In some embodiments, one or more of the CENH3 genes comprise a polynucleotide sequence encoding an amino acid sequence having at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity to an amino acid sequence selected from the group consisting of SEQ ID NOs: 8-14 and 53-56. In some embodiments, one or more of the CENH3 genes comprises a polynucleotide sequence having at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity to a polynucleotide sequence selected from the group consisting of SEQ ID NOs: 1-7 and 58-61. In some embodiments, the one or more genetic modifications comprise a modification of an enhancer of one or more of the CENH3 2 sf-6744554 197072001240 genes, a modification of a promoter of one or more of the CENH3 genes, a modification of a coding region of one or more of the CENH3 genes, a modification of an intron of one or more of the CENH3 genes, a modification of methylation status of one or more of the CENH3 genes, expression of a repressor protein that targets the DNA or an mRNA of one or more of the CENH3 genes, expression of an RNA interference construct that targets an mRNA of one or more of the CENH3 genes, or any combination thereof. In certain embodiments, the one or more genetic modifications comprise a modification of an enhancer of one or more of the CENH3 genes, a modification of a promoter of one or more of the CENH3 genes, a modification of a coding region of one or more of the CENH3 genes, modification of an intron of one or more of the CENH3 genes, or any combination thereof relative to an unmodified strawberry plant of the same species or an unmodified control strawberry plant. In some embodiments, the one or more genetic modifications comprise a deletion, insertion, or one or more nucleotide changes in a coding region of one or more of the CENH3 genes. In some embodiments, the strawberry plant (e.g., haploid-inducing strawberry plant) or plant part has decreased expression of CENH3 proteins relative to a strawberry plant of the same species lacking the one or more genetic modifications. In certain embodiments, the strawberry plant (e.g., haploid-inducing strawberry plant) t or plant part lacks detectable expression of CENH3 proteins. In some embodiments, the strawberry plant (e.g., haploid- inducing strawberry plant) is diploid, triploid, tetraploid, pentaploid, hexaploid, septaploid, octoploid, or nonaploid, or has a ploidy of 10x, 11x, 12x, 13x, 14x, 15x, 16x, 17x, 18x, 19x, or 20x. In some embodiments, the plant part is selected from the group consisting of pollen, an anther, a seed, an achene, a leaf, a flower, a fruit, and a stolon. In certain embodiments, the plant part is pollen. In certain embodiments, the plant part is a seed. [0009] In some embodiments, the strawberry plant (e.g., haploid-inducing strawberry plant) is diploid. In certain embodiments, the strawberry plant (e.g., haploid-inducing strawberry plant) is a member of the genus Fragaria. In certain embodiments, the haploid- inducing strawberry plant is a plant of the species Fragaria vesca, Fragaria iinumae, Fragaria nipponica, Fragaria viridis, Fragaria × bifera, Fragaria bucharica, Fragaria chinensis, Fragaria daltoniana, Fragaria emeiensis, Fragaria hayatae, Fragaria iinumae, Fragaria mandshurica, Fragaria nilgerrensis, Fragaria nubicola, or Fragaria pentaphylla. In certain embodiments, the strawberry plant (e.g., haploid-inducing strawberry plant) is a plant of the species Fragaria vesca. 3 sf-6744554 197072001240 [0010] In certain embodiments, the strawberry plant (e.g., haploid-inducing strawberry plant) is a Fragaria vesca plant of the subspecies Fragaria vesca ssp. vesca, Fragaria vesca ssp. americana, or Fragaria vesca ssp. bracteata. In some embodiments the one or more Fragaria vesca CENH3 (herein referred to as “FvCENH3”) genes comprise a polynucleotide sequence encoding an amino acid sequence having at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity the amino acid sequence of SEQ ID NO: 8. In some embodiments, the one or more FvCENH3 genes comprise a polynucleotide sequence having at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity to the polynucleotide sequence of SEQ ID NO: 1. In some embodiments, the strawberry plant (e.g., haploid-inducing strawberry plant) is a plant of the species Fragaria vesca and wherein the one or more CENH3 genes comprise FvCENH3. In certain embodiments, the one or more genetic modifications comprise a modification of an enhancer of FvCENH3, a modification of a promoter of FvCENH3, a modification of a coding region of FvCENH3, modification of an intron of FvCENH3, or any combination thereof relative to a wild-type Fragaria vesca plant. [0011] In some embodiments, the strawberry plant (e.g., haploid-inducing strawberry plant) is octoploid. In some embodiments, the strawberry plant (e.g., haploid-inducing strawberry plant) is a plant of the species Fragaria x ananassa, Fragaria chiloensis, Fragaria virginiana, or Fragaria iturupensis. In some embodiments, the strawberry plant (e.g., haploid-inducing strawberry plant) is Fragaria chiloensis plant of the subspecies Fragaria chiloensis ssp. chiloensis, Fragaria chiloensis ssp. lucida, Fragaria chiloensis ssp. pacifica, or Fragaria chiloensis ssp. sandwicensis. [0012] In some embodiments, the strawberry plant (e.g., haploid-inducing strawberry plant) is a plant of the species Fragaria x ananassa. In some embodiments, one or more of the Fragaria x ananassa CENH3 (herein referred to as “FaCENH3”) genes comprises a polynucleotide sequence encoding an amino acid sequence having at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity to an amino acid sequence selected from the group consisting of SEQ ID NOs: 9-14 and 53-56. In some embodiments, one or more of the FaCENH3 genes comprises a polynucleotide sequence having at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity to a polynucleotide sequence selected from the group consisting of SEQ ID NOs: 2-7 and 58-61. In certain embodiments, the one or more FaCENH3 genes comprise: a 4 sf-6744554 197072001240 polynucleotide sequence encoding an amino acid sequence having at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity to the amino acid sequence of SEQ ID NO: 9 (Fragaria x ananassa CENH3-1a protein, hereinafter “FaCENH3-1a”); a polynucleotide sequence encoding an amino acid sequence having at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity to the amino acid sequence of SEQ ID NO: 10 (Fragaria x ananassa CENH3-1b protein, hereinafter “FaCENH3-1b”); a polynucleotide sequence encoding an amino acid sequence having at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity to the amino acid sequence of SEQ ID NO: 11 (Fragaria x ananassa CENH3- 2a protein, hereinafter “FaCENH3-2a”); a polynucleotide sequence encoding an amino acid sequence having at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity to the amino acid sequence of SEQ ID NO: 12 (Fragaria x ananassa CENH3-2b protein, hereinafter “FaCENH3-2b”); a polynucleotide sequence encoding an amino acid sequence having at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity to the amino acid sequence of SEQ ID NO: 13 (Fragaria x ananassa CENH3-3 protein, hereinafter “FaCENH3-3”); and/or a polynucleotide sequence encoding an amino acid sequence having at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity to the amino acid sequence of SEQ ID NO: 14 (Fragaria x ananassa CENH3-4 protein, hereinafter “FaCENH3-4”). In certain embodiments, the one or more FaCENH3 genes comprise: a polynucleotide sequence encoding an amino acid sequence having at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity to the amino acid sequence of SEQ ID NO: 53 (Fragaria x ananassa CENH3-1a protein, hereinafter “FaCENH3-7a”); a polynucleotide sequence encoding an amino acid sequence having at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity to the amino acid sequence of SEQ ID NO: 54 (Fragaria x ananassa CENH3-1b protein, hereinafter “FaCENH3-7b”); a polynucleotide sequence encoding an amino acid sequence having at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity to the amino acid sequence of SEQ ID NO: 55 (Fragaria x ananassa CENH3-2a protein, hereinafter “FaCENH3-7c”); and/or a polynucleotide sequence encoding an amino acid sequence having at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity to the amino acid sequence of SEQ ID NO: 56 (Fragaria x ananassa CENH3-2b protein, hereinafter “FaCENH3-7d”). 5 sf-6744554 197072001240 [0013] In certain embodiments, the one or more FaCENH3 genes comprise: a polynucleotide sequence having at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity to the polynucleotide sequence of SEQ ID NO: 2 (Fragaria x ananassa CENH3-1a gene, hereinafter “FaCENH3-1a”); a polynucleotide sequence having at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity to the polynucleotide sequence of SEQ ID NO: 3 (Fragaria x ananassa CENH3-1b gene, hereinafter “FaCENH3-1b”); a polynucleotide sequence having at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity to the polynucleotide sequence of SEQ ID NO: 4 (Fragaria x ananassa CENH3-2a gene, hereinafter “FaCENH3-2a”); a polynucleotide sequence having at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity to the polynucleotide sequence of SEQ ID NO: 5 (Fragaria x ananassa CENH3-2b gene, hereinafter “FaCENH3-2b”); a polynucleotide sequence having at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity to the polynucleotide sequence of SEQ ID NO: 6 (Fragaria x ananassa CENH3-3 gene, hereinafter “FaCENH3- 3”); and/or a polynucleotide sequence having at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity to the polynucleotide sequence of SEQ ID NO: 7 (Fragaria x ananassa CENH3-4 gene, hereinafter “FaCENH3-4”). In certain embodiments, the one or more FaCENH3 genes comprise: a polynucleotide sequence having at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity to the polynucleotide sequence of SEQ ID NO: 58 (Fragaria x ananassa CENH3-1a gene, hereinafter “FaCENH3-7a”); a polynucleotide sequence having at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity to the polynucleotide sequence of SEQ ID NO: 59 (Fragaria x ananassa CENH3-1b gene, hereinafter “FaCENH3-7b”); a polynucleotide sequence having at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity to the polynucleotide sequence of SEQ ID NO: 60 (Fragaria x ananassa CENH3-2a gene, hereinafter “FaCENH3-7c”); and/or a polynucleotide sequence having at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity to the polynucleotide sequence of SEQ ID NO: 61 (Fragaria x ananassa CENH3-2b gene, hereinafter “FaCENH3-7d”) [0014] In some embodiments, the strawberry plant (e.g., haploid-inducing strawberry plant) is a plant of the species Fragaria x ananassa and the one or more FaCENH3 genes 6 sf-6744554 197072001240 comprise one, two, three, four, five, or all six of FaCENH3-1a, FaCENH3-1b, FaCENH3-2a, FaCENH3-2b, FaCENH3-3, and FaCENH3-4. In certain embodiments, the one or more FaCENH3 genes comprise FaCENH3-1a, FaCENH3-1b, FaCENH3-2a, FaCENH3-2b, FaCENH3-3, and FaCENH3-4. In some embodiments, the one or more genetic modifications comprise: a modification of an enhancer of FaCENH3-1a, a modification of a promoter of FaCENH3-1a, a modification of a coding region of FaCENH3-1a, a modification of an intron of FaCENH3-1a, or any combination thereof relative to an unmodified Fragaria x ananassa plant; a modification of an enhancer of FaCENH3-1b, a modification of a promoter of FaCENH3-1b, a modification of a coding region of FaCENH3-1b, a modification of an intron of FaCENH3-1b, or any combination thereof relative to an unmodified Fragaria x ananassa plant; a modification of an enhancer of FaCENH3-2a, a modification of a promoter of FaCENH3-2a, a modification of a coding region of FaCENH3-2a, a modification of an intron of FaCENH3-2a, or any combination thereof relative to an unmodified Fragaria x ananassa plant; a modification of an enhancer of FaCENH3-2b, a modification of a promoter of FaCENH3-2b, a modification of a coding region of FaCENH3-2b, a modification of an intron of FaCENH3-2b, or any combination thereof relative to an unmodified Fragaria x ananassa plant; a modification of an enhancer of FaCENH3-3, a modification of a promoter of FaCENH3-3, a modification of a coding region of FaCENH3-3, a modification of an intron of FaCENH3-3, or any combination thereof relative to an unmodified Fragaria x ananassa plant; and/or a modification of an enhancer of FaCENH3-4, a modification of a promoter of FaCENH3-4, a modification of a coding region of FaCENH3-4, a modification of an intron of FaCENH3-4, or any combination thereof relative to an unmodified Fragaria x ananassa plant. [0015] In some embodiments, the strawberry plant (e.g., haploid-inducing strawberry plant) is a plant of the species Fragaria x ananassa and the one or more FaCENH3 genes comprise one, two, three, or all four of FaCENH3-7a, FaCENH3-7b, FaCENH3-7c, and FaCENH3-7d. In some embodiments, the one or more genetic modifications comprise: a modification of an enhancer of FaCENH3-7a, a modification of a promoter of FaCENH3-7a, a modification of a coding region of FaCENH3-7a, a modification of an intron of FaCENH3- 7a, or any combination thereof relative to an unmodified Fragaria x ananassa plant; a modification of an enhancer of FaCENH3-7b, a modification of a promoter of FaCENH3-7b, a modification of a coding region of FaCENH3-7b, a modification of an intron of FaCENH3- 7b, or any combination thereof relative to an unmodified Fragaria x ananassa plant; a 7 sf-6744554 197072001240 modification of an enhancer of FaCENH3-7c, a modification of a promoter of FaCENH3-7c, a modification of a coding region of FaCENH3-7c, a modification of an intron of FaCENH3- 7c, or any combination thereof relative to an unmodified Fragaria x ananassa plant; and/or a modification of an enhancer of FaCENH3-7d, a modification of a promoter of FaCENH3-7d, a modification of a coding region of FaCENH3-7d, a modification of an intron of FaCENH3- 7d, or any combination thereof relative to an unmodified Fragaria x ananassa plant. [0016] In some aspects, provided is a method of producing the strawberry plants (e.g., haploid-inducing strawberry plants) described herein. In some embodiments, the decreased expression of the one or more CENH3 genes is achieved by gene disruption, gene knockout, gene knockdown, disruption of an endogenous amino acid sequence (including, for example, conserved amino acids or amino acid motifs), introduction of amino acids or amino acid motifs, gene silencing, RNA interference, induction of methylation, or any combination thereof. In some embodiments, the method comprises introducing one or more of the genetic modifications by mutagenesis, gene editing, transgenesis, or a combination thereof. In some embodiments, the method comprises introducing one or more of the genetic modifications by gene editing using a site-directed nuclease. In certain embodiments, the site-directed nuclease is a CRISPR-associated (Cas) nuclease, a zinc finger nuclease (ZFN), a transcription activator-like effector nuclease (TALEN), or a megaTAL. [0017] In some embodiments, the method comprises contacting a plurality of cells of a parent strawberry plant with one or more Cas nucleases complexed with an RNA molecule comprising a polynucleotide sequence having at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% complementarity to 15, 16, 17, 18, 19, or 20 consecutive nucleotides of a protospacer sequence within one or more of the FvCENH3 and/or FaCENH3 genes. In some embodiments, the Cas nuclease is complexed with an RNA molecule comprising a polynucleotide sequence having at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity or complementarity to a polynucleotide sequence selected from the group consisting of SEQ ID NOs: 15-34. In certain embodiments, the RNA molecule is a crRNA, a gRNA, or a pegRNA. In certain embodiments, the plurality of cells is a plurality of protoplasts. In some embodiments, the method further comprises, subsequent to the contacting step, allowing the plurality of cells of the parent strawberry plant to form calli, plants, or a combination thereof, and identifying one or more calli or plants having the genetic modifications resulting in decreased expression of 8 sf-6744554 197072001240 one or more CENH3 genes. In some embodiments, the method comprises contacting a plurality of cells of a parent strawberry plant with one or more expression vectors together comprising an expression cassette for a Cas nuclease and an expression cassette for an RNA molecule having at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% complementarity to 15, 16, 17, 18, 19, or 20 consecutive nucleotides of one or more of the CENH3 genes. [0018] In some aspects, provided herein is a method of producing true homozygous octoploid strawberry seed, the method comprising (a) contacting a tetrahaploid reduced egg cell from a donor octoploid strawberry plant with a sperm cell of a haploid-inducing strawberry plant described herein, and (b) producing a doubled tetrahaploid cell from the tetrahaploid reduced egg cell. In some embodiments, step (a) comprises contracting a plurality of tetrahaploid reduced egg cells from the donor octoploid strawberry plant with a plurality of sperm cells of the haploid inducing strawberry plant. In some embodiments, step (a) comprises contacting pollen of the haploid-inducing strawberry plant with the stigma of a pistil of the donor octoploid strawberry plant comprising the reduced egg cell and allowing formation of a pollen tube and migration of the sperm cell to the reduced egg cell, thereby contacting the reduced egg cell with the sperm cell. In some embodiments, the method further comprises allowing the donor octoploid strawberry plant to form a plurality of seeds. In certain embodiments, the method further comprises selecting one or more tetrahaploid seeds by 1) determining the ploidy of the embryo and endosperm of one or more seeds of the plurality of seeds, and 2) selecting one or more seeds having tetraploid embryo and an endosperm having a ploidy of greater than 8x. [0019] In some aspects, provided herein is method of producing true homozygous octoploid strawberry seed, the method comprising (a) contacting an egg cell from a haploid- inducing strawberry plant described herein with a tetrahaploid reduced sperm cell from a donor octoploid strawberry plant, and (b) producing a doubled tetrahaploid cell from the egg cell, wherein the doubled tetrahaploid cell comprises the nuclear genome of the donor octoploid strawberry plant and the cytoplasmic genome of the haploid-inducing strawberry plant. In some embodiments, step (a) comprises contacting a plurality of egg cells from the haploid-inducing strawberry plants with a plurality of sperm cells from a donor octoploid strawberry plant. In some embodiments, the haploid-inducing strawberry plant and the donor octoploid strawberry plants are from different species of Fragaria. In some embodiments, the 9 sf-6744554 197072001240 haploid-inducing strawberry plant is diploid. In certain embodiments, the haploid-inducing strawberry plant is a plant of the species Fragaria vesca. In some embodiments, the donor octoploid strawberry plant is a plant of the species Fragaria x ananassa. In some embodiments, step (a) comprises contacting the stigma of a pistil of the haploid-inducing strawberry plant with the pollen from the donor octoploid strawberry plant and allowing formation of a pollen tube and migration of the tetrahaploid reduced sperm cell to the egg cell, thereby contacting the egg cell with the tetrahaploid reduced sperm cell. In some embodiments, the method further comprises allowing the haploid-inducing strawberry plant to form a plurality of seeds. [0020] In some aspects, provided herein is a method of producing true homozygous octoploid strawberry plants, the method comprising (a) contacting a plurality of tetrahaploid reduced egg cells from a maternal octoploid strawberry plant (e.g., an unedited maternal octoploid strawberry plant) with a plurality of sperm cells of an edited haploid-inducing octoploid strawberry plant described herein, (b) allowing the plurality of reduced egg cells to form a plurality of tetrahaploid seeds comprising the maternal genome of the octoploid strawberry plant (e.g., the unedited maternal genome) and lacking the edited, haploid- inducing paternal genome, (c) collecting the plurality of seeds and allowing them to germinate to form a plurality of plants, (d) determining the presence of one or more tetrahaploid plants in the plurality of plants, and (e) producing one or more doubled tetrahaploid plants (also referred to herein as a “true homozygous octoploid”) from the one or more tetrahaploid plants. Additionally, step (d) may include determining the presence of one or more spontaneously doubled tetrahaploids. [0021] In some aspects, provided herein is a method of producing true homozygous octoploid strawberry plants, the method comprising (a) contacting a plurality of tetrahaploid reduced egg cells from an edited maternal haploid-inducing octoploid strawberry plant described herein with a plurality of sperm cells of an a paternal octoploid strawberry plant (e.g., an unedited octoploid strawberry plant), (b) allowing the plurality of reduced egg cells to form a plurality of tetrahaploid seeds comprising the paternal genome from the paternal octoploid strawberry plant (e.g., the unedited paternal genome) and lacking the edited, haploid-inducing maternal genome, (c) collecting the plurality of seeds and allowing them to germinate to form a plurality of plants, (d) determining the presence of one or more tetrahaploid plants in the plurality of plants, and (e) producing one or more doubled 10 sf-6744554 197072001240 tetrahaploid plants (also referred to herein as a “true homozygous octoploid”) from the one or more tetrahaploid plants. Additionally, step (d) may include determining the presence of one or more spontaneously doubled tetrahaploids. [0022] In some aspects, provided herein is a method of producing true homozygous octoploid strawberry plants, the method comprising (a) contacting a plurality of tetrahaploid reduced egg cell from an edited haploid-inducing diploid strawberry plant described herein with a plurality of sperm cells of an octoploid strawberry plant (e.g., an unedited octoploid strawberry plant), (b) allowing the plurality of reduced egg cells to form a plurality of tetrahaploid seeds comprising the paternal genome of the octoploid strawberry plant and lacking the edited, haploid-inducing maternal genome of the diploid strawberry plant, (c) collecting the plurality of seeds and allowing them to germinate to form a plurality of plants, (d) determining the presence of one or more tetrahaploid plants in the plurality of plants, and (e) producing one or more doubled tetrahaploid plants (also referred to herein as a “true homozygous octoploid”) from the one or more tetrahaploid plants. Additionally, step (d) may include determining the presence of one or more spontaneously doubled tetrahaploids. In certain such embodiments, the tetrahaploid seedling will comprise the unedited cytoplasmic genome of the diploid mother and the unedited octoploid genome of the father. [0023] In some embodiments of the method of producing a true homozygous octoploid strawberry plant, step (a) comprises contacting pollen of the haploid-inducing strawberry plant with the stigma of a pistil of the donor octoploid strawberry plant comprising the reduced egg cell and allowing formation of a pollen tube and migration of the pollen tube to the reduced egg cell, thereby contacting the reduced egg cell with the sperm cell. In certain embodiments, step (d) comprises determining the presence of the maternal genome and the paternal genome in the one or more plants, wherein tetrahaploid plants within the plurality of plants are determined based on the absence of the maternal genome and the presence of the paternal genome. [0024] In other embodiments of the method of producing a true homozygous octoploid strawberry plant, step (a) comprises contacting pollen of the donor octoploid strawberry plant with the stigma of a pistil of the haploid-inducing strawberry plant comprising the reduced egg cell and allowing formation of a pollen tube and migration of the pollen tube to the reduced egg cell, thereby contacting the reduced egg cell with the sperm cell. In certain embodiments, step (d) comprises determining the presence of the maternal genome and the 11 sf-6744554 197072001240 paternal genome in the one or more plants, wherein tetrahaploid plants within the plurality of plants are determined based on the presence of the maternal genome and the absence of the paternal genome. [0025] In certain embodiments of the method of producing a true homozygous octoploid strawberry plant, step (d) further comprises determining the ploidy of the embryo from each of the one or more tetrahaploid plants and discarding any plants that (i) lacks a tetraploid embryo and/or (ii) comprises an embryo ploidy of greater than 8x. In certain embodiments, the presence (or absence) of the maternal genome and/or the paternal genome, respectively, in the one or more plants, is determined by the detection of one or more genetic markers which are present in the paternal genome and which are absent in the maternal genome. In certain embodiments, the presence (or absence) of the maternal genome and/or the paternal genome, respectively, in the one or more plants, is determined by the detection of one or more genetic markers which are present in the maternal genome and which are absent in the paternal genome. [0026] In some embodiments of the method of producing a true homozygous octoploid strawberry plant, step (d) comprises determining the presence of the maternal genome and the paternal genome in the one or more plants, wherein tetrahaploid plants within the plurality of plants are determined based on the absence of the maternal genome and the presence of the paternal genome. In other embodiments, step (d) comprises determining the presence of the maternal genome and the paternal genome in the one or more plants, wherein tetrahaploid plants within the plurality of plants are determined based on the presence of the maternal genome and the absence of the paternal genome. In certain embodiments, step (d) further comprises determining the ploidy of the embryo from each of the one or more tetrahaploid plants and discarding any plants that (i) lacks a tetraploid embryo and/or (ii) comprises an embryo ploidy of greater than 8x. In certain embodiments, the presence (or absence) of the maternal genome and/or the paternal genome, respectively, in the one or more plants, is determined by the detection of one or more genetic markers which are present in the paternal genome and which are absent in the maternal genome. In certain embodiments, the presence (or absence) of the maternal genome and/or the paternal genome, respectively, in the one or more plants, is determined by the detection of one or more genetic markers which are present in the maternal genome and which are absent in the paternal genome. 12 sf-6744554 197072001240 [0027] In some embodiments of the method of producing a true homozygous octoploid strawberry plant, step (e) comprises subjecting the selected tetrahaploid plant to a tetrahaploid doubling treatment. In certain embodiments, the tetrahaploid doubling treatment comprises contacting a cell from the tetrahaploid plant with an anti-microtubule agent to form a doubled tetrahaploid cell (i.e., a true homozygous octoploid cell). In certain embodiments, the anti-microtubule agent is an anti-mitotic herbicide. In certain embodiments, the anti- microtubule agent comprises colchicine, oryzalin, nitrous oxide, trifluralin, or any combination thereof. In some embodiments of the method of producing a true homozygous octoploid strawberry plant, the doubled tetrahaploid cell is allowed to grow into a doubled tetrahaploid plant. In some embodiments, the doubled tetrahaploid plant (i.e., the true homozygous octoploid plant) is contacted with self-pollen and allowed to form seed, producing a plurality of true homozygous octoploid strawberry seed. In some embodiments, the doubled tetrahaploid plant (i.e., the true homozygous octoploid plant) is contacted with pollen from a different doubled tetrahaploid plant and allowed to form seed, producing a plurality of true breeding F1 hybrid octoploid strawberry seed. [0028] In some embodiments of the method of producing a true homozygous octoploid strawberry plant or seed, the donor octoploid strawberry plant is a plant of the species Fragaria x ananassa, Fragaria chiloensis, Fragaria virginiana, or Fragaria iturupensis, or a hybrid of any combination thereof. In some embodiments, the donor octoploid strawberry plant is Fragaria chiloensis plant of the subspecies Fragaria chiloensis ssp. chiloensis, Fragaria chiloensis ssp. lucida, Fragaria chiloensis ssp. pacifica, or Fragaria chiloensis ssp. sandwicensis. In some embodiments, the donor octoploid strawberry plant is a plant of the species Fragaria x ananassa. In certain embodiments, the donor octoploid strawberry plant is a Fragaria x ananassa plant of the variety named “Camarosa.” In certain embodiments, the donor octoploid strawberry plant is a Fragaria x ananassa plant of the variety named “Chandler.” In certain embodiments, the donor octoploid strawberry plant is a Fragaria x ananassa plant of the variety named “Albion.”. [0029] In some embodiments, the method of producing a true homozygous octoploid strawberry plant or seed further comprises introducing one or more genetic modifications resulting in decreased expression of a gibberellin 20-oxidase (“ga20ox”) gene into the true homozygous octoploid strawberry plant or seed. 13 sf-6744554 197072001240 [0030] In some aspects, provided herein is a method of producing a runnerless true homozygous octoploid strawberry plant, the method comprising (a) contacting a tetrahaploid reduced egg cell from a donor octoploid strawberry plant with the sperm cell of a haploid- inducing strawberry plant described herein, (b) producing a doubled tetrahaploid cell from the tetrahaploid reduced egg cell, and (c) introducing one or more genetic modifications resulting in decreased expression of a ga20ox gene into the true homozygous octoploid strawberry plant, wherein the runnerless true homozygous octoploid strawberry plant does not produce runners. [0031] In some aspects, provided herein is true homozygous octoploid strawberry seed produced according to the methods described herein. In certain embodiments, the true homozygous octoploid strawberry seed comprises one or more genetic modifications resulting in decreased expression of a ga20ox gene. [0032] In some aspects, provided herein is a true homozygous octoploid strawberry plant or a plant part thereof produced according to the methods described herein. In certain embodiments, the true homozygous octoploid strawberry plant comprises one or more genetic modifications resulting in decreased expression of a ga20ox gene. In certain embodiments, the true homozygous octoploid strawberry plant does not produce runners. In some embodiments, the true homozygous octoploid strawberry plant, or a progenitor thereof, was subject to introduction of one or more of the genetic modifications by mutagenesis, gene editing, transgenesis, or a combination thereof followed by selection for a genetic modification that results in decreased expression of the one or more CENH3 genes. In some embodiments, the plant part is selected from the group consisting of pollen, an anther, a seed, an achene, a leaf, a flower, a fruit, and a stolon. [0033] In some aspects, provided herein is a true homozygous octoploid strawberry seed produced according to a method described herein, comprising an embryo having the nuclear genome of the donor octoploid strawberry plant and the cytoplasmic genome of the haploid- inducing strawberry plant, wherein the nuclear genome and the cytoplasmic genome are from plants of different species of Fragaria. In some embodiments, the cytoplasmic genome is from a plant of the species Fragaria vesca. In some embodiments, the nuclear genome is from a plant of the species Fragaria x ananassa. In some embodiments, the true homozygous octoploid strawberry seed comprises one or more genetic modifications resulting in decreased expression of a ga20ox gene. 14 sf-6744554 197072001240 [0034] In some aspects, provided herein is a true homozygous octoploid strawberry plant or a plant part thereof produced according to the method described herein, comprising at least one somatic cell having the nuclear genome of the donor octoploid strawberry plant and the cytoplasmic genome of the haploid-inducing strawberry plant, wherein the nuclear genome and the cytoplasmic genome are from plants of different species of Fragaria. In some embodiments, the cytoplasmic genome is from a plant of the species Fragaria vesca. In some embodiments, the nuclear genome is from a plant of the species Fragaria x ananassa. In some embodiments, the true homozygous octoploid strawberry plant comprises one or more genetic modifications resulting in decreased expression of a ga20ox gene. In certain embodiments, the true homozygous octoploid strawberry plant does not produce runners. In some embodiments, the true homozygous octoploid strawberry plant, or a progenitor thereof, was subject to introduction of one or more of the genetic modifications by mutagenesis, gene editing, transgenesis, or a combination thereof followed by selection for a genetic modification that results in decreased expression of the one or more CENH3 genes. In some embodiments, the plant part is selected from the group consisting of pollen, an anther, a seed, an achene, a leaf, a flower, a fruit, and a stolon. [0035] In some aspects, provided herein is a true homozygous octoploid strawberry seed comprising an embryo having the nuclear genome of a first species of Fragaria and the cytoplasmic genome of a second plant species of Fragaria. In some embodiments, the first species of Fragaria is Fragaria x ananassa. In some embodiments, the second species of Fragaria is a diploid species of Fragaria. In certain embodiments, the second species of Fragaria is Fragaria vesca. In some embodiments, the true homozygous octoploid strawberry seed comprises one or more genetic modifications resulting in decreased expression of a gibberellin ga20ox gene. [0036] In some aspects, provided herein is a true homozygous octoploid strawberry plant or plant part thereof comprising an embryo having the nuclear genome of a first species of Fragaria and the cytoplasmic genome of a second plant species of Fragaria. In some embodiments, the first species of Fragaria is Fragaria x ananassa. In some embodiments, the second species of Fragaria is a diploid species of Fragaria. In certain embodiments, the second species of Fragaria is Fragaria vesca. In some embodiments, the true homozygous octoploid strawberry seed comprises one or more genetic modifications resulting in decreased expression of a gibberellin ga20ox gene. In certain embodiments, the true homozygous 15 sf-6744554 197072001240 octoploid strawberry plant does not produce runners. In some embodiments, the true homozygous octoploid strawberry plant, or a progenitor thereof, was subject to introduction of one or more of the genetic modifications by mutagenesis, gene editing, transgenesis, or a combination thereof followed by selection for a genetic modification that results in decreased expression of the one or more CENH3 genes. In some embodiments, the plant part is selected from the group consisting of pollen, an anther, a seed, an achene, a leaf, a flower, a fruit, and a stolon. [0037] In some aspects, provided herein is a method of producing uniform octoploid F1 hybrid strawberry seed comprising crossing two of the true homozygous octoploid strawberry plants described herein. In some aspects, provided herein is a uniform octoploid F1 hybrid strawberry seed produced according to said method. In certain embodiments, the uniform octoploid F1 hybrid strawberry seed comprises one or more genetic modifications resulting in decreased expression of a ga20ox gene. In some aspects, provided herein is a uniform octoploid F1 hybrid strawberry plant or a plant part thereof produced according to said method. In certain embodiments, the uniform octoploid F1 hybrid strawberry plant comprises one or more genetic modifications resulting in decreased expression of a ga20ox gene. In certain embodiments, the uniform octoploid F1 hybrid strawberry plant does not produce runners. In some embodiments, the plant part is selected from the group consisting of pollen, an anther, a seed, an achene, a leaf, a flower, a fruit, and a stolon. [0038] In some aspects, an expression vector or isolated DNA molecule for making a strawberry plant (e.g., haploid-inducing strawberry plant), or a plant part thereof, is described herein. In some embodiments, the expression vector or isolated DNA molecule comprises a DNA sequence having at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity or complementarity to 15, 16, 17, 18, 19, or 20 consecutive nucleotides of a polynucleotide sequence encoding an amino acid sequence having at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity to an amino acid sequence selected from the group consisting of SEQ ID NOs: 8-14 and 53-56. In some embodiments, the expression vector or isolated DNA molecule comprises a DNA sequence having at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity or complementarity to 15, 16, 17, 18, 19, or 20 consecutive nucleotides of a polynucleotide having at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity to a polynucleotide sequence selected from the group consisting of SEQ ID NOs: 1-7 and 58-61. 16 sf-6744554 197072001240 [0039] In some embodiments, the expression vector or isolated DNA molecule comprises a DNA sequence encoding a non-coding RNA. In some embodiments, the non-coding RNA comprises a polynucleotide sequence having at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity or complementarity to 15, 16, 17, 18, 19, or 20 consecutive nucleotides of a polynucleotide sequence encoding an amino acid sequence having at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity to an amino acid sequence selected from the group consisting of SEQ ID NOs: 8-14 and 53-56. In some embodiments, the non-coding RNA comprises a polynucleotide sequence having at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity or complementarity to 15, 16, 17, 18, 19, or 20 consecutive nucleotides of a polynucleotide having at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity to a polynucleotide sequence selected from the group consisting of SEQ ID NOs: 1-7 and 58-61. In some embodiments, the non-coding RNA is a crRNA, a gRNA, a pegRNA, a siRNA, a miRNA, or a dsRNA. [0040] In some embodiments, the expression vector or isolated DNA molecule comprises a DNA sequence encoding a site-directed nuclease. In some embodiments, the site-directed nuclease is a Cas nuclease, a TALEN, ZFN, or a mega-TAL. In some embodiments, the site- directed nuclease comprises an amino acid sequence that confers binding to a polynucleotide sequence having at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity or complementarity to 15, 16, 17, 18, 19, or 20 consecutive nucleotides of a polynucleotide sequence encoding an amino acid sequence having at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity to an amino acid sequence selected from the group consisting of SEQ ID NOs: 8-14 and 53-56. In some embodiments, the site-directed nuclease comprises an amino acid sequence that confers binding to a polynucleotide sequence having at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity or complementarity to 15, 16, 17, 18, 19, or 20 consecutive nucleotides of a polynucleotide having at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity to a polynucleotide sequence selected from the group consisting of SEQ ID NOs: 1-7 and 58-61. [0041] In some aspects, provided herein is a bacterial cell comprising an expression vector or isolated DNA molecule described herein. In some embodiments, the bacterial cell is an Agrobacterium cell. In some aspects, provided herein is a kit comprising an expression 17 sf-6744554 197072001240 vector or isolated DNA molecule or a bacterial cell described herein. In some aspects, provided herein is a genetically modified plant, plant part, plant cell, or seed comprising an expression vector or isolated DNA molecule described herein. [0042] In some aspects, provided herein is a genetically modified non-regenerable plant cell of a strawberry plant (e.g., haploid-inducing strawberry plant) described herein. [0043] In some aspects, provided herein is a genetically modified plant genome of the strawberry plant (e.g., haploid-inducing strawberry plant) described herein. DESCRIPTION OF THE FIGURES [0044] The present application can be understood by reference to the following description taken in conjunction with the accompanying figures. [0045] FIG. 1 depicts a schematic illustrating the production of homozygous octoploid strawberry lines by tetrahaploid induction of the maternal genome and then doubling the resulting tetrahaploid. Step (A) exemplifies the starting plant material where chromosomes are indicated by gray and white patterned vertical bars, and the oval lines surrounding the vertical bars indicate exemplary nuclei. Ploidy is indicated at the top of the nuclei. An edited cenh3 gene is indicated on the chromosomes by a white “X” mark. Pollen is indicated by a male symbol, and egg cells and megaspore mother cells are indicated by a female symbol. In step (B), the top row exemplifies reduced, recombined 1x cenh3-edited pollen derived from a cenh3-edited diploid Fragaria vesca strawberry plant; the middle row illustrates a reduced, recombined 4x unedited tetrahaploid egg cell derived from an unedited octoploid Fragaria x ananassa strawberry plant; and the bottom row exemplifies reduced, recombined 4x cenh3-edited pollen from a cenh3-edited octoploid Fragaria x ananassa strawberry plant. The reduced, recombined 4x unedited tetrahaploid egg cell (middle row) is contacted by either (i) the reduced, recombined 1x cenh3-edited pollen (or sperm cell) (top row), (ii) the reduced, recombined 4x cenh3-edited pollen (or sperm cell), or (iii) both. In step (C), some amount of fertilizations cause the maternal tetrahaploid egg cell to undergo embryogenesis without retention of genetic material from the cenh3-edited pollen, as indicated by the straight-rightwards arrow. The remaining eggs result in fertilizations by the haploid inducer, illustrated by: (i) the up-rightwards arrow pointing to an illustration of an exemplary 5x embryo with one set of paternal chromosomes from the cenh3-edited diploid male parent, and (ii) the down-rightwards arrow pointing to an illustration of an exemplary 18 sf-6744554 197072001240 8x embryo with 4 sets of paternal chromosomes from the cenh3-edited octoploid parent. Embryos are screened for presence of the maternal and/or paternal genome and have their ploidy confirmed before subjecting the tetrahaploid embryos to a doubling treatment, thereby producing true homozygous octoploid strawberry plants in step (D). [0046] FIG. 2A depicts a schematic illustrating the production of homozygous octoploid strawberry lines by tetrahaploid induction of the paternal genome and then doubling the resulting tetrahaploid. Step (A) exemplifies the starting plant material where chromosomes are indicated by gray and white patterned vertical bars, and the oval lines surrounding the vertical bars indicate exemplary nuclei. Ploidy is indicated at the top of the nuclei. An edited cenh3 gene is indicated on the chromosomes by a white “X” mark. Pollen is indicated by a male symbol, and egg cells and megaspore mother cells are indicated by a female symbol. In step (B), the bottom row exemplifies reduced, recombined 4x pollen derived from an unedited octoploid Fragaria x ananassa strawberry plant, and the top row illustrates a reduced, recombined 4x tetrahaploid egg cell derived from a cenh3-edited octoploid Fragaria x ananassa strawberry plant. In other embodiments, the egg cell is a reduced, recombined 1x haploid egg cell derived from a cenh3-edited diploid Fragaria vesca strawberry plant (not illustrated). The reduced, recombined 4x cenh3-edited tetrahaploid egg cell (illustrated in the top row) or the reduced, recombined 1x cenh3-edited haploid egg cell (not illustrated), is contacted by the reduced, recombined 4x unedited pollen (bottom row). In step (C), some of the fertilizations cause the maternal tetrahaploid egg cell (illustrated in the top row) or the reduced, recombined 1x cenh3-edited haploid egg cell (not illustrated), to undergo embryogenesis without retention of nuclear genetic material from the cenh3-edited egg cell, as indicated by the straight-rightwards arrow. The remaining eggs result in fertilizations by the haploid inducer, illustrated by the down-rightwards arrow pointing to an illustration of an exemplary 8x embryo with four sets of both maternal and paternal chromosomes from octoploid parents. In embodiments where the reduced, recombined egg cells is a 1x cenh3- edited haploid egg cell, fertilization by the haploid inducer results in 5x embryos (not illustrated). Embryos are screened for presence of the maternal and/or paternal genome and have their ploidy confirmed before subjecting the tetrahaploid embryos to a doubling treatment, thereby producing true homozygous octoploid strawberry plants in step (D). [0047] FIG. 2B depicts a schematic illustrating the production of homozygous octoploid strawberry lines by tetrahaploid induction of the paternal genome, with a cytoplasmic swap 19 sf-6744554 197072001240 resulting in a seed having the cytoplasmic genome of the maternal plant and the nuclear genome of the paternal plant. Pollen of octoploid (8x) F. x ananassa is used to pollinate a haploid-inducing line of diploid (2x) F. vesca, resulting in tetrahaploid embryogenesis of an embryo retaining the nuclear tetrahaploid (4x) genome of F. x ananassa and the cytoplasmic genome of F. vesca. Doubling of the resulting tetrahaploid plant produces the homozygous octoploid strawberry line, which is homozygous across the four subgenomes for the unedited 8x F. x ananassa paternal nuclear genome, and also has the cytoplasmic genome of the F. vesca maternal plant. [0048] FIGS. 3A-3B show the configuration of CENH3 loci in octoploid strawberry. FIG. 3A shows the configuration of six homoeologous FaCENH3 loci in octoploid strawberry Fragaria x ananassa according to reference genome “Fragaria x ananassa Camarosa Genome v1.0.a2”. The six homeologs are shown, with black bars indicating the locations of the CENH3 loci. As described further herein, “FaCENH3-1a” is illustrated as the left-most black bar on chromosome 7, subgenome 1; “FaCENH3-1b” is illustrated as the right-most on chromosome 7, subgenome 1; “FaCENH3-2a” is illustrated as the left-most black bar on chromosome 7, subgenome 2; “FaCENH3-2b” is illustrated as the right-most black bar on chromosome 7, subgenome 2; “FaCENH3-3” is illustrated as the black bar on chromosome 7, subgenome 3; “FaCENH3-4” is illustrated as the black bar on chromosome 7, subgenome 4. FIG. 3B shows the configuration of six homoeologous FaCENH3 loci in octoploid strawberry Fragaria x ananassa according to reference genome “Fragaria x ananassa Royal Royce Genome v1.0.” The four homeologs are shown, with black bars indicating the locations of the CENH3 loci. As described further herein, “FaCENH3-7a” is illustrated as a black bar on chromosome 7, subgenome A; “FaCENH3-7b” is illustrated as a black bar on chromosome 7, subgenome B; “FaCENH3-7c” is illustrated as a black bar on chromosome 7, subgenome C; and “FaCENH3-7d” is illustrated as a black bar on chromosome 7, subgenome D. [0049] FIG. 4 shows the CENH3 gene in the diploid strawberry Fragaria vesca according to the reference genome “Fragaria vesca v_4.0”. The gene (referred to herein as (“FvCENH3”) is shown with a black bar indicating its location of chromosome 7. [0050] FIGS. 5A-C show a protein sequence alignment of CENH3 orthologs from Fragaria vesca (FvCENH3) (SEQ ID NO: 8), Fragaria x ananassa Camarosa (FaCENH3- 1a, FaCENH3-1b, FaCENH3-2,a FaCENH3-2b, FaCENH3-3, FaCENH3-4) (SEQ ID NOs: 20 sf-6744554 197072001240 9-14) Fragaria chiloensis (FcCENH3-7-AV, FcCENH3-7-Bi, FcCENH3-7-B1, and FcCENH3-7-B2) (SEQ ID NOs: 36-39), Fragaria iinumae (FiCENH3) (SEQ ID NO: 40), Fragaria viridis (FvirCENH3) (SEQ ID NO: 41), Potentilla micrantha (PmCENH3), (SEQ ID NO: 42), Arabidopsis thaliana (AtCENH3) (SEQ ID NO: 35), Solanum lycopersicum (SlCENH3) (SEQ ID NO: 43), Zea mays (ZmCENH3) (SEQ ID NO: 44), and Consensus (SEQ ID NO: 48). FIG. 5A shows positions 1-42 of the sequence alignment. FIG. 5B shows positions 43-108 of the sequence alignment. FIG. 5C shows positions 109-178 of the sequence alignment. [0051] FIGS. 5D-G show a protein sequence alignment of CENH3 orthologs from Fragaria vesca (FvCENH3) (SEQ ID NO: 8), Fragaria x ananassa Royal Royce (FaCENH3-7a, FaCENH3-7b, FaCENH3-7c, and FaCENH3-7d) (SEQ ID NOs: 53-56) Fragaria chiloensis (FcCENH3-7-AV, FcCENH3-7-Bi, FcCENH3-7-B1, and FcCENH3-7- B2) (SEQ ID NOs: 36-39), Fragaria iinumae (FiCENH3) (SEQ ID NO: 40), Fragaria viridis (FvirCENH3) (SEQ ID NO: 41), Potentilla micrantha (PmCENH3), (SEQ ID NO: 42), Arabidopsis thaliana (AtCENH3) (SEQ ID NO: 35), Solanum lycopersicum (SlCENH3) (SEQ ID NO: 43), Zea mays (ZmCENH3) (SEQ ID NO: 44), and Consensus (SEQ ID NO: 57). FIG. 5D shows positions 1-60 of the sequence alignment. FIG. 5E shows positions 61- 120 of the sequence alignment. FIG. 5F shows positions 121-180 of the sequence alignment. FIG. 5G shows positions 181-189 of the sequence alignment. [0052] FIG. 6A shows a phylogenetic tree of identified putative and characterized CENH3 orthologs as in FIGS. 5A-5C. The phylogenetic tree is a UPGMA consensus tree. The scale bar represents phylogenetic distance of 0.05 nucleotide substitutions per site. The tree is generated using software Geneious Prime 2023.2.1, with Genetic Distance Model: Jukes-Cantor. [0053] FIG. 6B shows a phylogenetic tree of identified putative and characterized CENH3 orthologs as in FIGS. 5D-5G. The phylogenetic tree is a UPGMA consensus tree. The scale bar represents phylogenetic distance of 0.05 nucleotide substitutions per site. The tree is generated using software Geneious Prime 2025.0.2, with Genetic Distance Model: Jukes-Cantor. [0054] FIG. 7 shows a bar graph of editing efficiencies of guide RNAs targeting different protospacers in FaCENH3 in four different varieties of Fragaria x ananassa. The x-axis 21 sf-6744554 197072001240 indicates the different target protospacer sites which were tested; the y-axis indicates the percent of protoplasts with successful editing. Different shades indicate different varieties of Fragaria x ananassa strawberry varieties tested. The sample size ranged from 3 replicates up to 9 replicates, with a median of 6 replicates across all data sets. The error bars indicate standard error of the mean. [0055] FIG. 8A shows a schematic illustrating the editing windows associated with PRS878 for FvCENH3 (SEQ ID NO: 1), FaCENH3-1a (SEQ ID NO: 2), FaCENH3-1b (SEQ ID NO: 3), FaCENH3-2a (SEQ ID NO: 4), FaCENH3-2b (SEQ ID NO: 5), FaCENH3-3 (SEQ ID NO: 6), and FaCENH3-4 (SEQ ID NO: 7). The protospacer sequence is shown in bold, underlined text. UTRs are denoted in light gray; coding sequences are denoted in dark gray; and introns are denoted as a line. The nucleotides in bolded, underlined font show the (+) strand reverse complement of the PRS878 sequence (SEQ ID NO: 21). At the top of the figure, its general location is illustrated by a downwards-pointing black triangle over the third exon. [0056] FIG. 8B shows a schematic illustrating the editing windows associated with PRS878 for FvCENH3 (SEQ ID NO: 1), FaCENH3-7a (SEQ ID NO: 58), FaCENH3-7b (SEQ ID NO: 59), FaCENH3-7c (SEQ ID NO: 60), and FaCENH3-7d (SEQ ID NO: 61). The protospacer sequence is shown in bold, underlined text. UTRs are denoted in light gray; coding sequences are denoted in dark gray; and introns are denoted as a line. The nucleotides in bolded, underlined font show the (+) strand reverse complement of the PRS878 sequence (SEQ ID NO: 21). At the top of the figure, its general location is illustrated by a downwards- pointing black triangle over the third exon. [0057] FIG. 9A shows a schematic illustrating the editing windows associated with PRS874 for FvCENH3 (SEQ ID NO: 1), FaCENH3-1a (SEQ ID NO: 2), FaCENH3-1b (SEQ ID NO: 3), FaCENH3-2a (SEQ ID NO: 4), FaCENH3-2b (SEQ ID NO: 5), FaCENH3-3 (SEQ ID NO: 6), and FaCENH3-4 (SEQ ID NO: 7). The protospacer sequence is shown in bold, underlined text. UTRs are denoted in light gray; coding sequences are denoted in dark gray; and introns are denoted as a line. The general location of the sequence of PRS874 (SEQ ID NO: 17) is denoted by a downwards-pointing black triangle over the third exon. 22 sf-6744554 197072001240 [0058] FIG. 9B shows a schematic illustrating the editing windows associated with PRS874 for FvCENH3 (SEQ ID NO: 1), FaCENH3-7a (SEQ ID NO: 58), FaCENH3-7b (SEQ ID NO: 59), FaCENH3-7c (SEQ ID NO: 60), and FaCENH3-7d (SEQ ID NO: 61). The protospacer sequence is shown in bold, underlined text. UTRs are denoted in light gray; coding sequences are denoted in dark gray; and introns are denoted as a line. The general location of the sequence of PRS874 (SEQ ID NO: 17) is denoted by a downwards-pointing black triangle over the third exon. [0059] FIG. 10A shows a schematic illustrating the editing windows associated with PRS876 for FvCENH3 (SEQ ID NO: 1), FaCENH3-1a (SEQ ID NO: 2), FaCENH3-1b (SEQ ID NO: 3), FaCENH3-2a (SEQ ID NO: 4), FaCENH3-2b (SEQ ID NO: 5), FaCENH3-3 (SEQ ID NO: 6), and FaCENH3-4 (SEQ ID NO: 7). The protospacer sequence is shown in bold, underlined text. UTRs are denoted in light gray; coding sequences are denoted in dark gray; and introns are denoted as a line. Target site PRS876 (SEQ ID NO: 19) is illustrated by a downwards-pointing black triangle over the third exon. [0060] FIG. 10B shows a schematic illustrating the editing windows associated with PRS876 for FvCENH3 (SEQ ID NO: 1), FaCENH3-7a (SEQ ID NO: 58), FaCENH3-7b (SEQ ID NO: 59), FaCENH3-7c (SEQ ID NO: 60), and FaCENH3-7d (SEQ ID NO: 61). The protospacer sequence is shown in bold, underlined text. UTRs are denoted in light gray; coding sequences are denoted in dark gray; and introns are denoted as a line. Target site PRS876 (SEQ ID NO: 19) is illustrated by a downwards-pointing black triangle over the third exon. [0061] FIGS. 11A-11B depict the size and growth of an exemplary sample of protoplasts derived from a cenh3 mutant strawberry line. FIG. 11A shows the cell size of the protoplasts in the exemplary sample, with bounding boxes around the protoplasts for comparison of protoplast size. FIG. 11B depicts microcalli grown from protoplasts in the exemplary sample, 0.1 – 0.3 mm in size 14 days after encapsulation of the protoplasts. [0062] FIGS. 12A-12F depict a sequence alignment of FaCENH3-7a alleles in wild-type (N; SEQ ID NO: 77) and edited (E; SEQ ID NOs: 78-88) F. x ananassa Royal Royce plants with the translated protein sequence (SEQ ID NO: 89). FIG. 12A shows the alignment at positions 1-108 of the wild-type sequence. FIG. 12B shows the alignment at positions 109- 23 sf-6744554 197072001240 216 of the wild-type sequence. FIG. 12C shows the alignment at positions 217-270 of the wild-type sequence. [0063] FIGS. 13A-13C depict a sequence alignment of FaCENH3-7b alleles in wild-type (N; SEQ ID NO: 90) and edited (E; SEQ ID NOs: 91-101) F. x ananassa Royal Royce plants with the translated protein sequence (SEQ ID NO: 102). FIG. 13A shows the alignment at positions 1-108 of the wild-type sequence. FIG. 13B shows the alignment at positions 109- 216 of the wild-type sequence. FIG. 13C shows the alignment at positions 217-270 of the wild-type sequence. [0064] FIGS. 14A-14F depict a sequence alignment of FaCENH3-7c alleles in wild-type (N; SEQ ID NO: 103 and 104) and edited (E; SEQ ID NOs: 105-113) F. x ananassa Royal Royce plants with the translated protein sequence (SEQ ID NO: 114). FIG. 14A shows the alignment at positions 1-108 of the wild-type sequence. FIG. 14B shows the alignment at positions 109-216 of the wild-type sequence. FIG. 14C shows the alignment at positions 217-270 of the wild-type sequence. [0065] FIGS. 15A-15C depict a sequence alignment of FaCENH3-7d alleles in wild-type (N; SEQ ID NO: 115) and edited (E; SEQ ID NOs: 116-126) F. x ananassa Royal Royce plants with the translated protein sequence (SEQ ID NO: 127). FIG. 15A shows the alignment at positions 1-108 of the wild-type sequence. FIG. 15B shows the alignment at positions 109-216 of the wild-type sequence. FIG. 15C shows the alignment at positions 217-270 of the wild-type sequence. [0066] FIG. 16A-16F depict the results of microscopy to evaluate microspore formation in wild-type and cenh3 mutant strawberry plants. FIG. 16A shows a bar graph of the average size at which microspores developed. FIGS. 16B shows a micrograph of an exemplary smashed anther with microspores emerging. FIG. 16C shows a micrograph of the same exemplary anther at a higher magnification. FIG. 16D shows a micrograph of the same exemplary anther at yet higher magnification. FIG. 16E shows an exemplary micrograph from cenh3 mutant line E-PED524-4145 (cenh3-006), showing tetrads. FIG. 16F shows a second exemplary micrograph from cenh3 mutant line E-PED524-4145 (cenh3-006), showing tetrads. [0067] FIGS. 17A-17B depict photographs of wild-type and cenh3 mutant strawberry lines. FIG. 17A depicts cenh3-001 (E-PED524-4004; left) and wild-type (right) plants. FIG. 24 sf-6744554 197072001240 17B depict two flowers and three stamens each from the wild-type (top) and cenh3-001 (E- PED524-4004; bottom) plants. [0068] FIGS. 18A-18B depict photographs of wild-type and cenh3 mutant strawberry lines. FIG. 18A depicts cenh3-002 (E-PED524-3916; left) and wild-type (right) plants. FIG. 18B depict two flowers and three stamens each from the wild-type (top) and cenh3-002 (E- PED524-3916; bottom) plants. [0069] FIGS. 19A-19B depict photographs of wild-type and cenh3 mutant strawberry lines. FIG. 19A depicts cenh3-003 (E-PED524-4113; left) and wild-type (right) plants. FIG. 19B depict two flowers and three stamens each from the wild-type (top) and cenh3-003 (E- PED524-4113; bottom) plants. [0070] FIGS. 20A-20B depict photographs of wild-type and cenh3 mutant strawberry lines. FIG. 20A depicts cenh3-004 (E-PED524-4115; left) and wild-type (right) plants. FIG. 20B depict two flowers and three stamens each from the wild-type (top) and cenh3-004 (E- PED524-4115; bottom) plants. [0071] FIGS. 21A-21B depict photographs of wild-type and cenh3 mutant strawberry lines. FIG. 21A depicts cenh3-005 (E-PED524-4135; left) and wild-type (right) plants. FIG. 21B depict two flowers and three stamens each from the wild-type (top) and cenh3-005 (E- PED524-4135; bottom) plants. [0072] FIGS. 22A-22B depict photographs of wild-type and cenh3 mutant strawberry lines. FIG. 22A depicts cenh3-006 (E-PED524-4145; left) and wild-type (right) plants. FIG. 22B depict two flowers and three stamens each from the wild-type (top) and cenh3-006 (E- PED524-4145; bottom) plants. [0073] FIGS. 23A-23B depict photographs of wild-type and cenh3 mutant strawberry lines. FIG. 23A depicts cenh3-007 (E-PED524-4091; left) and wild-type (right) plants. FIG. 23B depict two flowers and three stamens each from the wild-type (top) and cenh3-007 (E- PED524-4091; bottom) plants. DETAILED DESCRIPTION [0074] The following description is presented to enable a person of ordinary skill in the art to make and use the various embodiments. Descriptions of specific devices, techniques, 25 sf-6744554 197072001240 and applications are provided only as examples. Various modifications to the examples described herein will be readily apparent to those of ordinary skill in the art, and the general principles defined herein may be applied to other examples and applications without departing from the spirit and scope of the various embodiments. Thus, the various embodiments are not intended to be limited to the examples described herein and shown, but are to be accorded the scope consistent with the claims. Overview [0075] The methods described herein first involve the de novo creation of strawberry haploid inducer lines. Pollen from the haploid inducer lines is used to fertilize reduced egg cells (4x) from elite octoploid (8x) strawberry varieties. Pollen from the haploid inducer lines is used to fertilize or stimulate development of reduced eggs cells (4x) from elite octoploid (8x) strawberry varieties. In some embodiments, pollen from the haploid inducer lines induce the reduced egg cells to undergo embryogenesis without transfer or retention of genetic material from the paternal genome. In some embodiments, pollen from the haploid inducer lines fuses with the reduced egg cell to form a zygote wherein the maternal genome is thereafter eliminated from the zygote, generating a tetrahaploid (4x) embryo without transfer or retention of genetic material from the maternal genome. The resulting haploid strawberry lines (4x) are doubled (for example, using colchicine) to achieve fully homozygous octoploid (8x) strawberry lines. Fully homozygous octoploid (8x) strawberry lines are crossed to determine optimal crosses for the production of genetically uniform (true-breeding) F1 hybrid seed. An exemplary such method is depicted in FIG. 1. [0076] Another advantage of the methods described herein is that the resulting F1 hybrid seed retains no foreign DNA and no targeted genetic alterations. First, de novo creation of strawberry haploid inducer lines is achieved by disrupting expression of CENH3 (centromere-specific histone 3 variant) ortholog genes in strawberry. In some embodiments, this involves the identification and targeting of CENH3 ortholog genes via nuclease-guided deletions. In some embodiments, this is done in a diploid line such as Fragaria vesca (FIG. 1, top row). In other embodiments, this is done in an octoploid Fragaria x ananassa (FIG. 1, bottom row). In the latter embodiment, edits are made to CENH3 orthologs from subgenome 1 (“sg1”), sg2 and sg3, and sg4. Next, pollen is collected from cenh3-edited strawberry lines (referred to herein as “cenh3-edited pollen” or just “cenh3-pollen”) and used to induce embryogenesis in reduced tetrahaploid egg cells (4x) from elite octoploid strawberry varieties 26 sf-6744554 197072001240 (8x) (FIG. 1, middle row). Embryogenesis without retention of the genomic material from the pollen occurs in some of the egg cells (FIG. 1, steps B). The resulting tetrahaploid lines (4x) are doubled with colchicine (FIG. 1, step C) to obtain fully homozygous octoploid strawberry derived from the maternal genome (8x) (FIG. 1, step D). Multiple fully homozygous octoploid strawberry lines are produced and then crossed according to standard methods in the art. The best crosses are chosen to produce genetically uniform F1 hybrid seed. [0077] In other embodiments, unmodified 4x pollen (FIG. 2A, bottom row) is used to pollinate a modified cenh3-edited female, wherein the egg cell is induced to undergo embryogenesis, resulting in a tetrahaploid (4x) comprising the paternal nuclear genome and lacking the cenh3-edited maternal genome (FIG. 2, middle row). In other words, embryogenesis without retention of the genomic material from the egg occurs in some of the egg cells (FIG. 2, steps B). The resulting haploid lines (4x) are doubled with colchicine (FIG. 2, step C) or double spontaneously to obtain fully homozygous octoploid strawberry lines derived from the paternal genome (8x) (FIG. 2, step D). Multiple fully homozygous octoploid strawberry lines are produced and then crossed according to standard methods in the art. The best crosses are chosen to produce genetically uniform F1 hybrid seed. [0078] In one aspect, provided herein are haploid-inducing strawberry plants, and plant parts thereof, comprising one or more genetic modifications resulting in decreased expression (including, e.g., decreased activity) of one or more CENH3 genes. Said haploid-inducing strawberry plant and plant parts may be crossed with octoploid strawberry plants to produce haploid (4x) strawberry plants, which may then be subject to genome doubling to produce true homozygous octoploid (8x) strawberry plants. Accordingly, also provided herein are true homozygous octoploid strawberry plants, and parts thereof. [0079] In another aspect, provided herein are methods of producing a haploid-inducing strawberry plants by decreasing the expression of one or more CENH3 genes in a strawberry plant. The methods may comprise introduction of one or more genetic modifications into a strawberry plant that result in decreased expression (including, e.g., decreased activity or non-expression) of one or more CENH3 genes, thereby producing the haploid-inducing strawberry plant. Further provided herein are expression vectors, isolated DNA molecules, bacterial cells, and kits useful in performing the methods described herein. 27 sf-6744554 197072001240 [0080] In yet another aspect, provided herein are methods of producing true homozygous octoploid strawberry seed and plant lines, the methods comprising crossing a haploid- inducing strawberry plant with an octoploid strawberry plant to generate a tetrahaploid (4x) cell or plant, and subjecting the tetrahaploid (4x) cell or plant to genome doubling to produce an octoploid (8x) plant having two clonal sets of 4x chromosomes. The true homozygous octoploid strawberry seed and plant lines of the present disclosure have the advantage of homozygosity within each subgenome, while only requiring two plant generations to produce. To clarify, the true homozygous octoploid strawberry seed and plant lines of the present disclosure are homozygous within the same subgenome, but can remain heterozygous across the four different subgenomes, respective to each other. This stands in stark contrast to existing methods of producing inbred octoploid strawberry lines, which requires a prohibitive number of generations of selfing and may not achieve true homozygosity across all loci. Also described herein are true homozygous octoploid strawberry seeds, plants, and plant parts produced according to the methods described herein. Further provided herein are uniform octoploid F1 hybrid strawberry seed, plants, and plant parts thereof produced by crossing two of said true homozygous octoploid strawberry plants. Definitions [0081] As used herein, the term “plant” includes the whole plant or any parts or derivatives thereof, such as plant organs (e.g., harvested or non-harvested flowers, leaves, etc.), plant cells, plant protoplasts, plant cell or tissue cultures from which whole plants can be regenerated, regenerable or non-regenerable plant cells, plant calli, plant cell clumps, and plant cells that are intact in plants, or parts of plants, such as embryos, pollen, ovules, ovaries, reduced egg cells, pistil, stigma, seeds, achenes, fruits (e.g., aggregate fruits), flowers, leaves, seeds, runners, clonally propagated plants, roots, stems, cotyledons, hypocotyls, root tips (e.g., harvested tissues or organs) and the like. Any developmental stage is also included, such as seedlings, immature and mature, and the like. [0082] As used herein, the term “strawberry plant” typically refers to any plant commonly understood by one of ordinary skill in the art to be a strawberry plant, and includes plants of the genera Fragaria. [0083] As used herein, the term “flower” or “strawberry flower” may refer to either the aggregate inflorescence of flowers or the individual flowers making up the aggregate. As 28 sf-6744554 197072001240 used herein, the term “fruit” or “strawberry fruit” typically refers to the aggregate fruit of the strawberry, which is the fleshy structure that is typically used in food applications and considered by laypeople as the strawberry fruit. As used herein, the term “seed” or “strawberry seed” includes both true strawberry seed and strawberry achenes. Achenes are the small seed-like structures found on the outer surface of the aggregate fruits of strawberries. Each achene arises from a single ovule of the aggregate inflorescence and, if the egg is fertilized, contains an endosperm and a most commonly a single seed. Each achene is therefore a true fruit which most commonly contains a seed. [0084] As used herein, the term “non-regenerable” generally refers to a plant part, a plant cell, a processed plant product, or a portion of any of the foregoing, that cannot be induced to form a whole plant or that cannot be induced to form a whole plant that is capable of sexual and/or asexual reproduction. [0085] As used herein, the term “nuclear genome” refers to the genome present in the nucleus of a plant cell, including, e.g., an egg cell, a sperm cell, or an embryo. [0086] As used herein, the term “cytoplasmic genome” typically refers to the genome present in the organelles present outside the nucleus of a plant cell, such as the mitochondrial genome or the chloroplast genome of a plant cell. [0087] As used herein, “ploidy” refers to the number of complete sets of chromosomes in a cell or organism. Ploidy may be annotated using “n” as the unit of complete sets of chromosomes. For example, a cell or organism with a single set of chromosomes may be referred to as “1x”, or the single set of chromosomes itself may be referred to as “1x”. A diploid cell or organism with two sets of chromosomes may be referred to as “2x”; a triploid cell or organism with three sets of chromosomes may be referred to as “3x”; and so on. [0088] As used herein, “diploid” refers to a cell or organism with a ploidy of 2x. [0089] As used herein, “polyploid” refers to a cell or organism with a ploidy of greater than 2x. “Polyploid” may refer to organisms which are triploid (3x), tetraploid (4x), pentaploid (5x), hexaploid (6x), septaploid (or heptaploid, 7x), octoploid (8x), or higher ploidies (greater than 8x). 29 sf-6744554 197072001240 [0090] As used herein, “octoploid” (which may also be spelled “octaploid”) refers to a cell or organism with a ploidy of 8x. [0091] As used herein, “haploid” typically refers to a cell or organism with a ploidy half that of the parent organism. As used herein, “haploid gametes” typically refers to gamete cells with a ploidy half that of the parent organism. For example, in a diploid strawberry plant (2n=2x=14), meiosis in germline cells results in 1n haploid gametes (e.g., pollen and egg cells) where 1n=1x=7. In another example, in an octoploid strawberry plant (2n=8x=56), meiosis in germline cells results in 1n haploid gametes (e.g., pollen and egg cells) where 1n=4x=28. In the latter example, the 4x haploid gametes may also be referred to as tetrahaploid gametes. [0092] As used herein, “haploid-inducing strawberry plant,” also referred to as a “haploid inducer strawberry plant,” typically refers to a strawberry plant that produces pollen that triggers the development of haploid embryos and seeds when crossed with another strawberry plant. As used herein, a “haploid-inducing strawberry plant” or “haploid inducer strawberry plant” may also refer to a strawberry plant that produces an egg whose genome is replaced by the genome of an unmodified strawberry plant upon fertilization by the unmodified plant and results in the development of haploid embryos and seeds. As used herein, “haploid induction” typically refers to the process of crossing pollen from a haploid- inducing plant with a non-haploid-inducing plant to generate haploid embryos and seeds. During a normal cross with two non-haploid-inducing plant plants, two haploid sperm cells migrate down the pollen tube to the ovule. One of the sperm cells fertilizes the central cell of the ovule to form the endosperm, while the other sperm cell fertilizes the haploid reduced egg cell to form an embryo having the ploidy of the parent plant. For example, if both parent plants are octoploid, the haploid (4x) sperm and egg form an octoploid (8x) embryo in a normal fertilization process. In haploid induction, the sperm cells from the haploid-inducing plant migrate down the pollen tube to the ovule, where one of the sperm cells fertilizes the central cell to form the endosperm. However, in some cases, when the other sperm cell contacts the reduced egg cell, embryogenesis is initiated, but the DNA from the sperm cell is not delivered to the egg cell or is otherwise eliminated, resulting in formation of a haploid embryo. In other cases, when the other sperm cell contacts the reduced egg cell, embryogenesis is initiated, but the maternal nuclear genome is not retained or otherwise eliminated, resulting in formation of a haploid embryo comprising the paternal nuclear 30 sf-6744554 197072001240 genome. For example, in haploid induction, if both parent plants are octoploid, the haploid (4x) egg cell either (i) does not incorporate the DNA from the haploid sperm cell (4x), resulting in a haploid (4x) embryo comprising the maternal nuclear genome (4x), or (ii) does not retain the DNA from the haploid egg cell (4x), resulting in a haploid (4x) embryo comprising the paternal nuclear genome (4x). In a haploid induction cross, the non-haploid- inducing plant that is pollinated by the haploid-inducing plant is typically referred to as the donor plant, however the resulting haploid embryos can contain chromosomes from either the donor plant or the haploid-inducing plant, and usually have half the ploidy of the donor plant (although spontaneous doubling is also sometimes observed). Accordingly, as used herein, the term “donor octoploid strawberry plant” typically refers to an octoploid strawberry plant wherein the nuclear genome undergoes the haploid induction process, resulting in haploid embryos each having one set of chromosomes from the donor octoploid strawberry plant, and lacking nuclear genome from the haploid-inducing strawberry plant. The haploid- inducing plant may be a female, as in the case of female genome elimination and induction of the paternal genome; or the haploid-inducing plant may be a male, as in the case of maternal genome induction. The haploid-inducing strawberry plant and the donor strawberry plant do not need to have the same ploidy. For example, a haploid-inducing strawberry plant may be diploid (2n=2x=14) and produce haploid (1n=1x=7) pollen that can be crossed with an octoploid (2n=8x=56) strawberry plant, resulting in tetraploid (1n=4x=28) haploid embryos, also referred to as tetrahaploid embryos. [0093] As used herein, “allele” refers to one of two or more alternative forms of a single gene or locus within the genome. As used herein, “monoallelic” typically describes the presence of a single allele at a given locus or set of loci within a cell or organism. As used herein, “monoallelic at over n% of the loci in the genome” typically refers to the percentage of total loci in the genome which are monoallelic. As used herein, “biallelic” typically describes the presence of two different alleles at a given locus or set of loci within a cell or organism. As used herein, “multiallelic” typically describes the presence of three or more alleles at a given locus or set of loci within a cell or organism. In allopolyploid strawberry cells or plants containing two or more subgenomes, “monoallelic” and “biallelic” typically refer to the allelic makeup of a set of chromosomes within the same subgenome. For example, an octoploid strawberry plant of the species Fragaria x ananassa, which has four subgenomes, is monoallelic at all loci if the two chromosomes in a subgenome are monoallelic at all loci. Accordingly, in the case of Fragaria x ananassa, “monoallelic at over 31 sf-6744554 197072001240 n% of the loci in the genome” typically refers to the percentage of total loci in the entire genome that are monoallelic within each subgenome, but does not indicate that n% of the loci are monoallelic across all subgenomes. [0094] As used herein, “haplotype” refers to a distinct 1n set of chromosomes with a unique set of alleles. For example, a tetraploid gamete or haploid plant where 1n = 4x = 28 would comprise as many as 4 unique haplotypes (1 per subgenome) for each of the basic seven chromosomes of strawberry. As used herein, each haplotype is distinct from other haplotypes in that it contains a set of alleles that confers a unique set of characteristics not conferred by other haplotypes. As used herein, “monoallelic plant” typically refers to a plant line containing a single haplotype within a subgenome, and a “biallelic plant” typically refers to a plant line containing two haplotypes. [0095] As used herein, “clonal” describes a body of DNA that is substantially identical to another body of DNA; or a set of cells or organisms that comprise such DNA. For example, genome doubling (e.g., by colchicine treatment) of a tetrahaploid strawberry plant (1n=4x=28) results in an octoploid plant having two clonal (substantially identical) sets of chromosomes for each of the four subgenomes. Due to random errors in natural DNA replication, clonal bodies of DNA, clonal cells, or clonal organisms may not be completely identical. “Clonal” may describe two sets of chromosomes that are not completely identical in sequence but that contain the same set of alleles. [0096] As used herein, “genetically uniform” describes a set of individual plants, plant parts (e.g., seeds), or plant cells whose genomes are clonal. For example, a population of strawberry F1 hybrid seed wherein at least 99% of the population is genetically uniform indicates that 99% of the seeds of the population are clonal across sub-genomes. As used herein, “clonal” may describe a body of DNA that is substantially identical to another body of DNA, however due to random errors in natural DNA replication, the clonal bodies of DNA, clonal plants, plant parts (e.g., seeds), or plant cells may not be completely identical. “Clonal” may also describe two genomes that are not completely identical in sequence but that contain the same set of alleles. [0097] As used herein, “homozygous” describes a cell or organism in which all sets of chromosomes within a genome or subgenome encode the same allele or set of alleles at a certain chromosomal locus, a set of chromosomal loci, or at all chromosomal loci. As used 32 sf-6744554 197072001240 herein, “homozygous plant” typically refers to a monoallelic plant or plant line having allelic uniformity across all chromosomal loci. In allopolyploid strawberry cells or plants containing two or more subgenomes, “homozygous” typically refers to sets of chromosomes within the same subgenome having the same alleles or sets of alleles at one or more chromosomal loci, and “homozygous plant” typically refers to a monoallelic plant or plant line having allelic uniformity across all chromosomal loci within each subgenome, but does not indicate that they have allelic uniformity across subgenomes. For example, an octoploid strawberry cell or plant having the same allele at a specific locus in two homologous chromosomes within in a subgenome may also be considered homozygous even though one or more alternative alleles exist in one or more different subgenomes. [0098] As used herein, “heterozygous” describes a cell or organism in which at least one set of chromosomes in one subgenome encodes an allele or set of alleles at a certain chromosomal locus or set of chromosomal loci that is distinct from those of the other sets of chromosomes within the cell or organism. For example, a strawberry cell or plant having allele a1 at locus A in one set of chromosomes in a subgenome and having allele a2 at locus A in a second set of chromosomes is heterozygous for alleles a1 and a2. As used herein, “heterozygous plant” typically refers to a biallelic plant or plant line. In polyploid strawberry cells or plants containing two or more subgenomes, “heterozygous” typically refers to two sets of chromosomes within the same subgenome having distinct alleles or sets of alleles at one or more chromosomal loci, and “heterozygous plant” typically refers to a plant that is biallelic for one or more loci in at least one of its subgenomes. [0099] As used herein, “crossing” refers to the act of forming an embryo from gametes of two distinct plants or plant lines. Crossing may refer to pollinating a plant or plant line using the pollen of a different plant or plant line, e.g., using the pollen of a haploid-inducing plant line to pollinate a donor plant line. In some cases “crossing” may refer to successful fertilization of an egg by a sperm cell resulting in an embryo or may refer to induction of haploid or tetrahaploid embryo development without successful fertilization of the egg. [0100] As used herein, “true homozygous plant line” and “true homozygous seed” typically refer to a monoallelic plant line or seed, respectively, having allelic uniformity across all chromosomal loci within a subgenome. As used herein, “true homozygous octoploid strawberry seed” typically refers to octoploid strawberry seed having allelic uniformity across all chromosomal loci within a subgenome. 33 sf-6744554 197072001240 [0101] As used herein, “hybrid” describes a plant or a part thereof, such as a seed, comprising two haplotypes within a subgenome. As used herein, “uniform F1 hybrid” refers to the genetically uniform first filial generation of hybrid seeds or plants resulting from the cross of two fully homozygous parent plants each having a different haplotype. As used herein, “uniform octoploid F1 hybrid strawberry seed” refers to a genetically uniform population of seed generated from the cross of two true homozygous octoploids or octoploids derived from doubled tetrahaploid strawberry plants or plant lines wherein each parent may possess a different haplotype. [0102] As used herein, “genetic modification” refers to any sequence or portion thereof within a nucleic acid molecule that differs from the sequence of an ancestral nucleic acid molecule. For example, a seed that contains an inserted or deleted genomic sequence that is not present in one of its parent plants comprises a genetic modification. A genetic modification may be naturally occurring or introduced. A genetic modification may be introduced via, for example: plant breeding to introduce a naturally-occurring genetic modification of one plant line into another plant line; transgenic methods; gene editing; chemical mutagenesis; and the like. [0103] As used herein, “expression” and “expression level” refer to the relative or absolute amount of a functional gene product present in a cell. As used herein, “gene products” include, but are not limited to, nucleic acids (e.g., RNA), post-transcriptionally modified nucleic acids (e.g., spliced RNA, poly-adenylated mRNA), proteins (e.g., enzymes, structural proteins, etc.), and post-translationally modified proteins (e.g., glycoproteins, lipoproteins, etc.). The function of the gene product refers to the wild-type, unmodified, uninhibited function of the gene product. As used herein, “decreased expression” refers to a relative decrease in the amount of a functional gene product present in a cell. The decreased expression may refer to a decrease in the total amount of a gene product present in a cell (e.g., a decrease in the amount of a protein) or to a decrease in the amount of functional gene products present in a cell (e.g., a decrease in the percentage of proteins with wild-type function) or to a decrease in the function of gene products present in a cell (e.g., a decrease in the activity of proteins as compared to proteins with wild-type function). The decreased expression may be of a gene product encoded at a certain genomic locus, and may be relative to a control strawberry plant (e.g., a strawberry plant of the same species). As used herein, “non-expression” refers to the absence of a functional gene product present in a cell, or to an 34 sf-6744554 197072001240 expression level insufficient for detection of the gene product in the cell, or to an expression level insufficient to result in the function of the gene product within the cell, or to an activity level insufficient to result in the detectable activity of the gene product within the cell. [0104] As used herein, “CENH3 gene” typically refers to a gene encoding “centromere- specific histone 3 variant” (abbreviated as “CENH3”), or a gene having nucleotide or amino acid homology thereto. Genes may include DNA sequences encoding a CENH3 protein or any gene product thereof, for example, nucleic acids (e.g., RNA), post-transcriptionally modified nucleic acids (e.g., spliced RNA, poly-adenylated mRNA), CENH3 protein, and post-translationally modified CENH3 protein. Genes also include any CENH3 gene orthologs, paralogs, and homeologs within a strawberry plant or cell. [0105] As used herein, “transgenesis” typically refers to the insertion of an exogenous genetic element into the genome of an organism. Any exogenous genetic element may be inserted via transgenesis, including, but not limited to, genes, protein coding sequences, non- protein coding sequences, regulatory sequences, spacer DNA, and the like. [0106] As used herein, “gene editing” typically refers to a type of genetic modification in which DNA is inserted, deleted or substituted in the genome of an organism using one or more natural or engineered nucleases. Gene editing may be carried out using site-specific nucleases, guided nucleases, or a combination thereof. The nuclease creates one or more site- specific breaks, such as double-strand breaks (DSBs) at target loci in the genome. Each site- specific break may be repaired, for example via non-homologous end joining (NHEJ), resulting in a genetic modification in the genome at the target locus; or via homologous recombination of the target locus with a provided repair nucleic acid molecule comprising homology to the target genomic sequence and the desired genetic modification. Haploid-inducing Strawberry Plants and Parts Thereof [0107] In one aspect, described herein is a haploid-inducing strawberry plant, or a plant part thereof, that triggers the development of haploid embryos and seeds when crossed with another strawberry plant. In some embodiments, the haploid-inducing strawberry plant or part thereof comprises one or more genetic modifications resulting in decreased expression of one or more CENH3 genes, such as any of the genetic modifications described herein. In some embodiments, provided herein is pollen, seed, pistil, stigma or a stolon of a haploid-inducing strawberry plant. 35 sf-6744554 197072001240 [0108] In another aspect, described herein is a haploid-inducing plant of the genus Potentilla, or a plant part thereof, that that triggers the development of haploid embryos and seeds when crossed with another strawberry plant. In some embodiments, the haploid- inducing plant of the genus Potentilla or part thereof comprises one or more genetic modifications resulting in decreased expression of one or more CENH3 genes, such as any of the genetic modifications described herein. In some embodiments, provided herein is pollen, seed, pistil, stigma or a stolon of a haploid-inducing plant of the genus Potentilla. In some embodiments, the haploid- inducing strawberry plant is a plant of the genus Potentilla. In certain embodiments, the haploid- inducing strawberry plant is a plant of the species Potentilla micrantha. Ploidy and Species of Strawberry Plant [0109] In one aspect, described herein is a haploid-inducing strawberry plants. The haploid- inducing strawberry plants may be of any ploidy, for example, diploid (2x), triploid (3x), tetraploid (4x), pentaploid (5x), hexaploid (6x), septaploid (or heptaploid, 7x), octoploid (8x), or of a higher ploidy (e.g., greater than 8x, e.g., 9x, 10x, 11x, 12x, 13x, 14x, 15x, 16x, 17x, 18x, 19x, or 20x). The haploid-inducing strawberry plants may be of any taxonomic distinction known to those of skill in the art as strawberry plants. In some embodiments, the haploid- inducing strawberry plant is a plant of the family Rosaceae. In some embodiments, the haploid- inducing strawberry plant is a plant of the genus Fragaria. [0110] In some embodiments, the haploid-inducing strawberry plant, or plant part thereof, is diploid, for example, a diploid species of the genus Fragaria. Diploid species of strawberry plants include, for example, Fragaria vesca, Fragaria iinumae, Fragaria nipponica, Fragaria viridis, Fragaria × bifera, Fragaria bucharica, Fragaria chinensis, Fragaria daltoniana, Fragaria emeiensis, Fragaria hayatae, Fragaria iinumae, Fragaria mandshurica, Fragaria viridis, Fragaria nilgerrensis, Fragaria nipponica, Fragaria nubicola, and Fragaria pentaphylla. [0111] In some embodiments, the haploid-inducing strawberry plant, or plant part thereof, is a plant of the species Fragaria vesca. The haploid-inducing strawberry plant may be of any subspecies of Fragaria vesca, for example, Fragaria vesca ssp. vesca, Fragaria vesca ssp. americana, or Fragaria vesca ssp. bracteate. The haploid-inducing strawberry plant may be of any variety of Fragaria vesca, for example, Rügen, Alexandria, Hawaii 4, 36 sf-6744554 197072001240 Baron Solemacher, Weisse Solemacher, Golden Alexandria, Quarantaine de Prin, Blanc Amélioré, Illa Martin, or Gartenfreude. [0112] In some embodiments, the haploid-inducing strawberry plant, or plant part thereof, is tetraploid, for example, a tetraploid species or hybrid of the genus Fragaria. Tetraploid species of strawberries include, for example, Fragaria corymbosa, Fragaria gracilis, Fragaria × intermedia, Fragaria moupinensis, Fragaria orientalis, and Fragaria tibetica. [0113] In some embodiments, the haploid-inducing strawberry plant, or plant part thereof, is hexaploid, for example, a hexaploid species or hybrid of the genus Fragaria. Hexaploid species of strawberries include, for example, Fragaria moschata. [0114] In some embodiments, the haploid-inducing strawberry plant, or plant part thereof, is octoploid, for example, an octoploid species or hybrid of the genus Fragaria. Octoploid species of strawberries include, for example, Fragaria x ananassa, Fragaria chiloensis, Fragaria virginiana, and Fragaria iturupensis. [0115] In some embodiments, the haploid-inducing strawberry plant, or plant part thereof, is a plant of the species Fragaria x ananassa. The haploid-inducing strawberry plant may be of any of Fragaria x ananassa, for example, Camarosa, Sweet Charlie, Chandler or Seascape. [0116] In some embodiments, the haploid-inducing strawberry plant, or plant part thereof, is a plant of the species Fragaria chiloensis. The haploid-inducing strawberry plant may be of any subspecies of Fragaria chiloensis, for example, Fragaria chiloensis subsp. chiloensis, Fragaria chiloensis subsp. lucida, Fragaria chiloensis subsp. pacifica, and Fragaria chiloensis subsp. sandwicensis. [0117] In some embodiments, the haploid-inducing strawberry plant, or plant part thereof, has a ploidy of 10x, for example, a species or hybrid of the genus Fragaria having a ploidy of 10x. Strawberry species having a ploidy of 10x include, for example, Fragaria cascadensis and Fragaria iturupensis. [0118] In some embodiments the haploid-inducing strawberry plant, or plant part thereof, is a hybrid between any of the strawberry species described herein, or is a synthetic polyploid 37 sf-6744554 197072001240 derived from a diploid species. In some embodiments, the haploid inducing strawberry plant is a synthetic polyploid derived from any diploid species, hybrid or synthetic polyploid. CENH3 Genes [0119] In some embodiments, the haploid-inducing strawberry plant, or plant part thereof, comprises one or more genetic modifications resulting in decreased expression of one or more CENH3 genes. In some variations, the haploid-inducing strawberry plant, or plant part thereof, comprises one or more genetic modifications resulting in a decreased amount of a functional gene product encoded by one or more CENH3 genes (e.g., a functional CENH3 protein). The gene products encoded by the CENH3 genes may include, but are not limited to, nucleic acids (e.g., RNA), post-transcriptionally modified nucleic acids (e.g., spliced RNA, poly-adenylated mRNA), proteins (e.g., enzymes, structural proteins, etc.), and post-translationally modified proteins (e.g., glycoproteins, lipoproteins, etc.). The function of the gene product at the CENH3 gene refers to the wild-type, unmodified function of the gene product (e.g., wild-type, unmodified function of the CENH3 protein). The decreased expression of a CENH3 gene may refer to a decrease in the total amount of a gene product encoded by a CENH3 gene present in a cell (e.g., a decrease in the amount of total CENH3 protein) or to a decrease in the amount of a functional gene product encoded by a CENH3 gene present in a cell (e.g., a decrease in the percentage of CENH3 proteins with wild-type function). In some embodiments, the one or more genetic modifications resulting in decreased expression of one or more CENH3 genes may include, but are not limited to, modification of an enhancer in one or more CENH3 genes, modification of a promoter of one or more CENH3 genes, modification of a coding region in one or more CENH3 genes, modification of an intron in one or more CENH3 genes, modification of methylation status of one or more CENH3 genes, expression of a repressor protein that targets the DNA or an mRNA of one or more CENH3 genes, and expression of an RNA interference construct that targets one or more mRNAs from one or more CENH3 genes. In some embodiments, the haploid-inducing strawberry plant, or plant part thereof, may comprise one or more genetic modifications resulting in non-expression of one or more CENH3 genes. In some embodiments, the haploid-inducing strawberry plant, or plant part thereof, may comprise one or more genetic modifications resulting in decreased expression, non-expression, or a combination thereof of a plurality of CENH3 genes. In some embodiments, the haploid- inducing strawberry plant, or plant part thereof, may comprise one or more genetic 38 sf-6744554 197072001240 modifications resulting in decreased activity or function of one or more CENH3 proteins or one or more domains of a CENH3 protein. In some variations, the domain of the CENH3 protein comprise an N-terminal tail, a Histone Fold Domain (HSD), or a combination thereof. The structure of CENH3 proteins, including the N-terminal tail and Histone Fold Domain, have been described previously, for example, in Wang et al. 2019 (Centromere histone H3- and phospholipase-mediated haploid induction in plants. Plant Methods 15, 42). [0120] In some embodiments, the haploid-inducing strawberry plant, or plant part thereof, comprises one or more genetic modifications resulting in decreased expression of one or more CENH3 genes. In other embodiments, the haploid-inducing strawberry plant, or plant part thereof, comprises one or more genetic modifications resulting in decreased expression of two or more (e.g., two, three, four, or more) CENH3 genes. [0121] By way of example only, a CENH3 gene is exemplified by a FvCENH3 gene from diploid strawberry Fragaria vesca, and specifically by the FvCENH3 nucleotide sequences, FvCENH3 protein sequences, and percent identities described herein. FvCENH3 protein (SEQ ID NO: 8) and nucleotide (SEQ ID NO: 1) sequences are provided in Table 1. The locus of FvCENH3 on chromosome 7 in the Fragaria vesca genome is shown in FIG. 4, and a phylogenetic tree showing the relationship between FvCENH3 and CENH3 proteins from other plant species is shown in FIG. 5. FvCENH3 genes, and gene products thereof, include nucleotide sequences having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity or complementarity to SEQ ID NO. 8, or a fragment thereof. Gene products of FvCENH3 genes include FvCENH3 proteins having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity, or at least 60%, 65%, 70%, 75%, 80%, 85%, 95%, 96%, 97%, 98%, or 99% sequence similarity to the FvCENH3 protein of SEQ ID NO. 1, or a fragment thereof. [0122] By way of example only, a CENH3 gene is further exemplified by a FaCENH3-1a gene from chromosome 7 (subgenome 1, Fvb7-1) of octoploid strawberry Fragaria x ananassa (e.g., variety Camarosa), and specifically by the FaCENH3-1a nucleotide sequences, FaCENH3-1a protein sequences, and percent identities described herein. FaCENH3-1a protein (SEQ ID NO: 9) and FaCENH3-1a nucleotide (SEQ ID NO: 2) sequences are provided in Table 1. The locus of FaCENH3-1a in the Fragaria x ananassa genome is shown in FIG. 3A (top chromosome, Fvb7-1), and a phylogenetic tree showing 39 sf-6744554 197072001240 the relationship between FaCENH3-1a and CENH3 from other Fragaria x ananassa and other plant species is shown in FIG. 5A-C. FaCENH3-1a genes, and gene products thereof, include nucleotide sequences having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity or complementarity to the FaCENH3-1a gene nucleotide sequence of SEQ ID NO. 2, or a fragment thereof. Gene products of FaCENH3-1a genes also include FaCENH3-1a proteins having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity, or at least 60%, 65%, 70%, 75%, 80%, 85%, 95%, 96%, 97%, 98%, or 99% sequence similarity to the FaCENH3-1a protein of SEQ ID NO. 9, or a fragment thereof. [0123] By way of example only, a CENH3 gene is further exemplified by a FaCENH3-1b gene from chromosome 7 (subgenome 1, Fvb7-1) of octoploid strawberry Fragaria x ananassa (e.g., variety Camarosa), and specifically by the FaCENH3-1b nucleotide sequences, FaCENH3-1b protein sequences, and percent identities described herein. FaCENH3-1b protein (SEQ ID NO: 10) and FaCENH3-1b nucleotide (SEQ ID NO: 3) sequences are provided in Table 1. The locus of FaCENH3-1b in the Fragaria x ananassa genome is shown in FIG. 3A (top chromosome, Fvb7-1), and a phylogenetic tree showing the relationship between FaCENH3-1b and CENH3 from other Fragaria x ananassa and other plant species is shown in FIG. 5A-C. FaCENH3-1b genes, and gene products thereof, include nucleotide sequences having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity or complementarity to the FaCENH3-1b gene nucleotide sequence of SEQ ID NO. 3, or a fragment thereof. Gene products of FaCENH3-1b genes also include FaCENH3-1b proteins having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity, or at least 60%, 65%, 70%, 75%, 80%, 85%, 95%, 96%, 97%, 98%, or 99% sequence similarity to the FaCENH3-1b protein of SEQ ID NO. 10, or a fragment thereof. [0124] By way of example only, a CENH3 gene is further exemplified by a FaCENH3-2a gene from chromosome 7 (subgenome 2, Fvb7-2) of octoploid strawberry Fragaria x ananassa (e.g., variety Camarosa), and specifically by the FaCENH3-2a nucleotide sequences, FaCENH3-2a protein sequences, and percent identities described herein. FaCENH3-2a protein (SEQ ID NO: 11) and FaCENH3-2a nucleotide (SEQ ID NO: 4) 40 sf-6744554 197072001240 sequences are provided in Table 1. The locus of FaCENH3-2a in the Fragaria x ananassa genome is shown in FIG. 3A (second chromosome from the top, Fvb7-2), and a phylogenetic tree showing the relationship between FaCENH3-2a and CENH3 from other Fragaria x ananassa and other plant species is shown in FIG. 5A-C. FaCENH3-2a genes, and gene products thereof, include nucleotide sequences having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity or complementarity to the FaCENH3-2a gene nucleotide sequence of SEQ ID NO. 4, or a fragment thereof. Gene products of FaCENH3-2a genes also include FaCENH3-2a proteins having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity, or at least 60%, 65%, 70%, 75%, 80%, 85%, 95%, 96%, 97%, 98%, or 99% sequence similarity to the FaCENH3-2a protein of SEQ ID NO. 11, or a fragment thereof. [0125] By way of example only, a CENH3 gene is further exemplified by a FaCENH3-2b gene from chromosome 7 (subgenome 2, Fvb7-2) of octoploid strawberry Fragaria x ananassa (e.g., variety Camarosa), and specifically by the FaCENH3-2b nucleotide sequences, FaCENH3-2b protein sequences, and percent identities described herein. FaCENH3-2b protein (SEQ ID NO: 12) and FaCENH3-2b nucleotide (SEQ ID NO: 5) sequences are provided in Table 1. The locus of FaCENH3-2b in the Fragaria x ananassa genome is shown in FIG. 3A (second chromosome from the top, Fvb7-2), and a phylogenetic tree showing the relationship between FaCENH3-2b and CENH3 from other Fragaria x ananassa and other plant species is shown in FIG. 5A-C. FaCENH3-2b genes, and gene products thereof, include nucleotide sequences having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity or complementarity to the FaCENH3-2b gene nucleotide sequence of SEQ ID NO. 5, or a fragment thereof. Gene products of FaCENH3-2b genes also include FaCENH3-2b proteins having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity, or at least 60%, 65%, 70%, 75%, 80%, 85%, 95%, 96%, 97%, 98%, or 99% sequence similarity to the FaCENH3-2b protein of SEQ ID NO. 12, or a fragment thereof. [0126] By way of example only, a CENH3 gene is further exemplified by a FaCENH3-3 gene from chromosome 7 (subgenome 3, Fvb7-3) of octoploid strawberry Fragaria x ananassa (e.g., variety Camarosa), and specifically by the FaCENH3-3 nucleotide sequences, 41 sf-6744554 197072001240 FaCENH3-3 protein sequences, and percent identities described herein. FaCENH3-3 protein (SEQ ID NO: 13) and FaCENH3-3 nucleotide (SEQ ID NO: 6) sequences are provided in Table 1. The locus of FaCENH3-3 in the Fragaria x ananassa genome is shown in FIG. 3A (third chromosome from the top, Fvb7-3), and a phylogenetic tree showing the relationship between FaCENH3-3 and CENH3 from other Fragaria x ananassa and other plant species is shown in FIG. 5A-C. FaCENH3-3 genes, and gene products thereof, include nucleotide sequences having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity or complementarity to the FaCENH3-3 gene nucleotide sequence of SEQ ID NO. 6, or a fragment thereof. Gene products of FaCENH3-3 genes also include FaCENH3-3 proteins having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity, or at least 60%, 65%, 70%, 75%, 80%, 85%, 95%, 96%, 97%, 98%, or 99% sequence similarity to the FaCENH3-3 protein of SEQ ID NO. 13, or a fragment thereof. [0127] By way of example only, a CENH3-4 gene is further exemplified by a FaCENH3- 4 gene from chromosome 7 (subgenome 2, Fvb7-4) of octoploid strawberry Fragaria x ananassa (e.g., variety Camarosa), and specifically by the FaCENH3-4 nucleotide sequences, FaCENH3-4 protein sequences, and percent identities described herein. FaCENH3-4 protein (SEQ ID NO: 14) and FaCENH3-4 nucleotide (SEQ ID NO: 7) sequences are provided in Table 1. The locus of FaCENH3-4 in the Fragaria x ananassa genome is shown in FIG. 3A (bottom chromosome, Fvb7-4), and a phylogenetic tree showing the relationship between FaCENH3-4 and CENH3 from other Fragaria x ananassa and other plant species is shown in FIG. 5A-C. FaCENH3-4 genes, and gene products thereof, include nucleotide sequences having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity or complementarity to the FaCENH3-4 gene nucleotide sequence of SEQ ID NO. 7, or a fragment thereof. Gene products of FaCENH3-4 genes also include FaCENH3-4 proteins having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity, or at least 60%, 65%, 70%, 75%, 80%, 85%, 95%, 96%, 97%, 98%, or 99% sequence similarity to the FaCENH3-4 protein of SEQ ID NO. 14, or a fragment thereof. 42 sf-6744554 197072001240 [0128] By way of example only, a CENH3 gene is further exemplified by a FaCENH3-7a gene from chromosome 7 (subgenome A) of octoploid strawberry Fragaria x ananassa (e.g., variety Royal Royce), and specifically by the FaCENH3-7a nucleotide sequences, FaCENH3-7a protein sequences, and percent identities described herein. FaCENH3-7a protein (SEQ ID NO: 53) and FaCENH3-7a nucleotide (SEQ ID NOs: 49 and 58) sequences are provided in Table 1. The locus of FaCENH3-7a in the Fragaria x ananassa genome is shown in FIG. 3B (top chromosome, chr_7A), and a phylogenetic tree showing the relationship between FaCENH3-7a and CENH3 proteins from other Fragaria x ananassa and other plant species is shown in FIG. 5D-G. FaCENH3-7a genes, and gene products thereof, include nucleotide sequences having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity or complementarity to the FaCENH3-7a gene nucleotide sequence of SEQ ID NO. 49 or 58, or a fragment thereof. Gene products of FaCENH3-7a genes also include FaCENH3-7a proteins having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity, or at least 60%, 65%, 70%, 75%, 80%, 85%, 95%, 96%, 97%, 98%, or 99% sequence similarity to the FaCENH3-7a protein of SEQ ID NO. 53, or a fragment thereof. [0129] By way of example only, a CENH3 gene is further exemplified by a FaCENH3-7b gene from chromosome 7 (subgenome B) of octoploid strawberry Fragaria x ananassa (e.g., variety Royal Royce), and specifically by the FaCENH3-7b nucleotide sequences, FaCENH3-7b protein sequences, and percent identities described herein. FaCENH3-7b protein (SEQ ID NO: 54) and FaCENH3-7b nucleotide (SEQ ID NOs: 50 and 59) sequences are provided in Table 1. The locus of FaCENH3-7b in the Fragaria x ananassa genome is shown in FIG. 3B (top chromosome, chr_7B), and a phylogenetic tree showing the relationship between FaCENH3-7b and CENH3 proteins from other Fragaria x ananassa and other plant species is shown in FIG. 5D-G. FaCENH3-7b genes, and gene products thereof, include nucleotide sequences having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity or complementarity to the FaCENH3-7b gene nucleotide sequence of SEQ ID NO. 50 or 59, or a fragment thereof. Gene products of FaCENH3-7b genes also include FaCENH3-7b proteins having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity, 43 sf-6744554 197072001240 or at least 60%, 65%, 70%, 75%, 80%, 85%, 95%, 96%, 97%, 98%, or 99% sequence similarity to the FaCENH3-7b protein of SEQ ID NO. 54, or a fragment thereof. [0130] By way of example only, a CENH3 gene is further exemplified by a FaCENH3-7c gene from chromosome 7 (subgenome C) of octoploid strawberry Fragaria x ananassa (e.g., variety Royal Royce), and specifically by the FaCENH3-7c nucleotide sequences, FaCENH3-7c protein sequences, and percent identities described herein. FaCENH3-7c protein (SEQ ID NO: 55) and FaCENH3-7c nucleotide (SEQ ID NOs: 51 and 60) sequences are provided in Table 1. The locus of FaCENH3-7c in the Fragaria x ananassa genome is shown in FIG. 3B (top chromosome, chr_7C), and a phylogenetic tree showing the relationship between FaCENH3-7c and CENH3 proteins from other Fragaria x ananassa and other plant species is shown in FIG. 5D-G. FaCENH3-7c genes, and gene products thereof, include nucleotide sequences having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity or complementarity to the FaCENH3-7c gene nucleotide sequence of SEQ ID NO. 51 or 60, or a fragment thereof. Gene products of FaCENH3-7c genes also include FaCENH3-7c proteins having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity, or at least 60%, 65%, 70%, 75%, 80%, 85%, 95%, 96%, 97%, 98%, or 99% sequence similarity to the FaCENH3-7c protein of SEQ ID NO. 55, or a fragment thereof. [0131] By way of example only, a CENH3 gene is further exemplified by a FaCENH3-7d gene from chromosome 7 (subgenome D) of octoploid strawberry Fragaria x ananassa (e.g., variety Royal Royce), and specifically by the FaCENH3-7d nucleotide sequences, FaCENH3-7d protein sequences, and percent identities described herein. FaCENH3-7d protein (SEQ ID NO: 56) and FaCENH3-7d nucleotide (SEQ ID NOs: 52 and 61) sequences are provided in Table 1. The locus of FaCENH3-7d in the Fragaria x ananassa genome is shown in FIG. 3B (top chromosome, chr_7D), and a phylogenetic tree showing the relationship between FaCENH3-7d and CENH3 proteins from other Fragaria x ananassa and other plant species is shown in FIG. 5D-G. FaCENH3-7d genes, and gene products thereof, include nucleotide sequences having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity or complementarity to the FaCENH3-7d gene nucleotide sequence of SEQ ID NO. 52 or 61, or a fragment thereof. Gene products of FaCENH3-7d genes also include 44 sf-6744554 197072001240 FaCENH3-7d proteins having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity, or at least 60%, 65%, 70%, 75%, 80%, 85%, 95%, 96%, 97%, 98%, or 99% sequence similarity to the FaCENH3-7d protein of SEQ ID NO. 56, or a fragment thereof. [0132] The CENH3 gene and protein sequences described herein are reference sequences and may be used by one of ordinary skill in the art to identify CENH3 in any species or variety of strawberry. For example, using methods that are routine in the art, one of ordinary skill in the art could utilize any of the CENH3 gene or protein sequences described herein to query a nucleic acid database or a genome sequence to identify CENH3 genes in the database or genome sequence. Genetic Modifications [0133] In some embodiments, the haploid-inducing strawberry plant, or plant part thereof, comprises one or more genetic modifications. Genetic modifications may be generated by modification of any nucleic acid sequence or genetic element by insertion, deletion, or substitution of one or more nucleotides in a nucleic acid molecule. This can occur by a replacement of at least one nucleotide, a deletion of at least one nucleotide, an insertion of at least one nucleotide, a chemical alteration of at least one nucleotide, or a combination thereof as long as the result is a detectable (e.g., by PCR, DNA sequencing, chromatography, etc.) change of nucleotide sequence compared to the sequence of the nucleic acid molecule prior to modification. Such modifications can be achieved by any of several well-known methods known in the art including, but not limited to, random mutagenesis, gene editing, insertion of a recombinant nucleic acid, crossing of an unmodified plant (e.g., strawberry plant) with a modified plant (e.g., strawberry plant) to introduce the modification of the modified plant into the unmodified plant, and the like. A genetic modification may be naturally occurring or non-naturally occurring. [0134] The genetic modifications described herein may be present in any known genetic element including, but not limited to, protein-coding sequences, non-protein-coding sequences, promoter regions, 5' untranslated leaders, genes, exons, introns, poly-A signal sequences, 3' untranslated regions, regions encoding small RNAs (such as microRNAs and small-interfering RNAs), and any other sequences that affect transcription or translation of one or more nucleic acid sequences. In some embodiments, genetic modifications may 45 sf-6744554 197072001240 include, but are not limited to, modifying or replacing nucleotide sequences of interest (such as a regulatory elements), gene disruption, gene knockout, gene knockdown, gene knock-in, gene silencing (including, e.g., by expressing an inverted repeat into a gene of interest), RNA interference (including, e.g., by insertion and/or expression of an RNA interference construct), modification of methylation status, modification of splicing sites, introducing alternate splicing sites, or any combination thereof. As used herein, gene disruption refers to the alteration or insertion of a sequence into a gene or locus that results in decreased expression (including, e.g., decreased activity or non-expression) of a functional gene product (e.g., a functional protein gene product) of the gene. A gene disruption may be achieved by introduction of a genetic modification in a protein-coding sequence, including, but not limited to, as a missense or nonsense mutation, or an insertion, deletion, or substitution. As used herein, a knockout is a genetic modification wherein a gene or gene product has been rendered completely inoperative. A knockout of a gene product may be achieved by introduction of a genetic modification in a protein-coding sequence of a gene or any non- protein-coding or regulatory sequence described herein. As used herein, a knockdown is a genetic modification wherein a gene or gene product has been rendered partially inoperative. A knockdown of a gene product may be achieved by introduction of a genetic modification in a protein-coding sequence of a gene or in a non-protein-coding or regulatory sequence, or insertion of a trans-acting element, such as a construct that expresses an inverted repeat of the gene product or a construct that expresses a DNA- or RNA-binding protein such as a transcriptional repressor which may include, for example, a deactivated targeted nuclease such as deactivated CRISPR-associated (Cas) nuclease (e.g., dCas9). As used herein, knock- in represents the replacement or insertion of a DNA sequence at a specific DNA locus in a cell. Knock-ins may include, but are not limited to, specific insertion of a heterologous amino acid coding sequence in a coding region of a gene, an insertion of a transcriptional regulatory element in a genetic locus, or any of several methods of inserting a DNA sequence into a call that are known to one of ordinary skill in the art. [0135] In certain embodiments, the haploid-inducing strawberry plant, or plant part thereof, comprises one or more genetic modifications resulting in decreased expression decreased expression (including, e.g., decreased activity or non-expression) of a gene product of a genomic locus. In some embodiments, genetic modifications resulting in decreased expression (including, e.g., decreased activity or non-expression) of a gene product or locus may include, but are not limited to, modification of an enhancer, modification of a promoter, 46 sf-6744554 197072001240 modification of a 5’ untranslated leader, modification of a coding region, modification of a non-coding region, insertion and/or expression of an RNA interference construct that targets an mRNA, modification of a region encoding a small RNA, modification of methylation status of a genomic locus, expression of a repressor protein that targets a DNA or mRNA sequence, and any other sequences that affect transcription or translation of one or more nucleic acid sequences. In some embodiments, genetic modifications resulting in decreased expression (including, e.g., decreased activity or non-expression) of a gene product or locus may include, but are not limited to, modifying or replacing nucleotide sequences of interest (such as a regulatory elements), gene disruption, gene knockout, gene knockdown, gene knock-in, gene silencing (including, e.g., by inserting and/or expressing an inverted repeat into a gene of interest), RNA interference (including, e.g., by insertion and/or expression of an RNA interference construct), expression of a repressor protein (e.g., dCas9), modification of methylation status of gene loci, modification of splicing sites, introducing alternate splicing sites, or any combination thereof. [0136] In some embodiments, the haploid-inducing strawberry plant comprises one or more genetic modifications resulting in decreased expression (e.g., decreased activity) of one or more CENH3 proteins described herein. In some embodiments, the haploid-inducing strawberry plant comprises one or more genetic modifications of one or more CENH3 genes. In some embodiments, the haploid-inducing strawberry comprises one or more genetic modifications resulting in decreased expression of one or more CENH3 proteins having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity, or at least 60%, 65%, 70%, 75%, 80%, 85%, 95%, 96%, 97%, 98%, or 99% sequence similarity to an amino acid sequence selected from the list consisting of SEQ ID NOs: 8-14 and 53-56, or fragments thereof. In some embodiments, the haploid-inducing strawberry plant comprises one or more genetic modifications of one or more CENH3 genes comprising a nucleotide sequence having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity or complementarity to a nucleotide sequence selected from the group consisting SEQ ID NOs: 1-7 and 58-61, or a fragment thereof. In certain embodiments, the one or more genetic modifications comprise a modification of an enhancer of one or more of the CENH3 genes, a modification of a promoter of one or more of the CENH3 genes, a modification of a coding region of one or more of the CENH3 genes, a modification of an intron of one or more of the CENH3 genes, a 47 sf-6744554 197072001240 modification of methylation status of one or more of the CENH3 genes, expression of a repressor protein that targets the DNA or an mRNA of one or more of the CENH3 genes, expression of an RNA interference construct that targets an mRNA of one or more of the CENH3 genes, or any combination thereof. In some embodiments, the haploid-inducing strawberry plant has decreased expression of one or more CENH3 genes described herein relative to a strawberry plant (e.g., a control strawberry plant, e.g., a strawberry plant of the same species) lacking the one or more genetic modifications. In certain embodiments, the haploid-inducing strawberry lacks detectable expression of one or more CENH3 gene products described herein relative to a strawberry plant (e.g., a control strawberry plant, e.g., a strawberry plant of the same species) lacking the one or more genetic modifications. In certain embodiments, the haploid-inducing strawberry lacks detectable expression of any CENH3 gene products. In certain embodiments, the haploid-inducing strawberry plant or plant part further comprises one or more naturally-occurring inactive alleles of one or more CENH3 genes. [0137] In some embodiments, the decreased expression (e.g., decreased activity) of one or more CENH3 proteins is decreased by at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90% , or about 100% as compared to a control plant, plant part, or plant cell lacking the one or more genetic modifications. In some embodiments, the decreased expression (e.g., decreased activity) of one or more CENH3 proteins is decreased by no more than about 10%, about 20%, about 30%, about 40%, about 50%, about 60%, about 70%, about 80%, about 90%, about 95%, or about 99% as compared to a control plant, plant part, or plant cell lacking the one or more genetic modifications. In some embodiments, the decreased expression (e.g., decreased activity) of one or more CENH3 proteins is decreased is decreased by between about 10% and about 100%, between about 20% and about 100%, between about 30% and about 100%, between about 40% and about 100%, between about 50% and about 100%, between about 60% and about 100%, between about 70% and about 100%, between about 80% and about 100%, between about 90% and about 100%, between about 10% and about 90%, between about 20% and about 90%, between about 30% and about 90%, between about 40% and about 90%, between about 50% and about 90%, between about 60% and about 90%, between about 70% and about 90%, between about 80% and about 90%, between about 10% and about 80%, between about 20% and about 80%, between about 30% and about 80%, between about 40% and about 80%, between about 50% and about 80%, between about 60% and 48 sf-6744554 197072001240 about 80%, between about 70% and about 80%, 10% and about 70%, between about 20% and about 70%, between about 30% and about 70%, between about 40% and about 70%, between about 50% and about 70%, between about 60% and about 70%, between about 10% and about 60%, between about 20% and about 60%, between about 30% and about 60%, between about 40% and about 60%, between about 50% and about 60%, between about 10% and about 50%, between about 20% and about 50%, between about 30% and about 50%, between about 40% and about 50%, between about 10% and about 40%, between about 20% and about 40%, between about 30% and about 40%, between about 10% and about 30%, between about 20% and about 30%, or about 10% and 20% as compared to a control plant, plant part, or plant cell lacking the one or more genetic modifications. [0138] In some embodiments, the haploid-inducing strawberry plant is a plant of the species Fragaria vesca comprising one or more genetic modifications resulting in decreased expression of one or more CENH3 genes. In certain embodiments, the haploid-inducing strawberry plant is a plant of the species Fragaria vesca comprising one or more genetic modifications resulting in decreased expression of an FvCENH3 gene. In certain embodiments, the haploid-inducing strawberry plant is a plant of the species Fragaria vesca comprising one or more genetic modifications resulting in decreased expression of an FvCENH3 protein. In certain embodiments, the haploid-inducing strawberry plant is a plant of the species Fragaria vesca comprising one or more genetic modifications resulting in decreased expression of a CENH3 protein having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity, or at least 60%, 65%, 70%, 75%, 80%, 85%, 95%, 96%, 97%, 98%, or 99% sequence similarity to the FvCENH3 protein of SEQ ID NO. 8, or a fragment thereof. [0139] In some embodiments, the haploid-inducing strawberry plant is a plant of the species Fragaria vesca comprising one or more genetic modifications of an FvCENH3 gene or gene product. In certain embodiments, the haploid-inducing strawberry plant is a plant of the species Fragaria vesca comprising one or more genetic modifications in a CENH3 gene comprising a nucleotide sequence having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity or complementarity to the FvCENH3 gene nucleotide sequence of SEQ ID NO. 1, or a fragment thereof. In certain embodiments, the haploid-inducing strawberry plant is a plant of the species Fragaria vesca comprising genetic modifications in both alleles of an 49 sf-6744554 197072001240 FvCENH3 gene. In some variations, the one or more genetic modifications comprise a modification of an enhancer of FvCENH3, a modification of a promoter of FvCENH3, a modification of a coding region of FvCENH3, modification of an intron of FvCENH3, or any combination thereof relative to a control Fragaria vesca plant (e.g., a Fragaria vesca plant lacking one or more of the genetic modifications). In some embodiments, the haploid- inducing strawberry plant is a plant of the species Fragaria vesca and has decreased expression of FvCENH3 protein relative to a Fragaria vesca plant lacking the one or more genetic modifications. In certain embodiments, the haploid-inducing strawberry plant is a plant of the species Fragaria vesca and lacks detectable expression of FvCENH3 protein. [0140] In some embodiments, the haploid-inducing strawberry plant is a plant of the species Fragaria x ananassa comprising one or more genetic modifications resulting in decreased expression of one or more FaCENH3 genes. In certain embodiments, the haploid- inducing strawberry plant is a plant of the species Fragaria x ananassa comprising one or more genetic modifications resulting in decreased expression of an FaCENH3-1a gene, an FaCENH3-1b gene, an FaCENH3-2a gene, an FaCENH3-2b gene, an FaCENH3-3 gene, an FaCENH3-4 gene, or any combination thereof. In certain embodiments, the haploid-inducing strawberry plant is a plant of the species Fragaria x ananassa comprising one or more genetic modifications resulting in decreased expression of FaCENH3-1a gene, an FaCENH3- 1b gene, an FaCENH3-2a gene, an FaCENH3-2b gene, an FaCENH3-3 gene, and an FaCENH3-4 gene. [0141] In some embodiments, the haploid-inducing strawberry plant is a plant of the species Fragaria x ananassa comprising one or more genetic modifications resulting in decreased expression of an FaCENH3-1a protein, an FaCENH3-1b protein, an FaCENH3-2a protein, an FaCENH3-2b protein, an FaCENH3-3 protein, an FaCENH3-4 protein, or any combination thereof. In certain embodiments, the haploid-inducing strawberry plant is a plant of the species Fragaria x ananassa comprising one or more genetic modifications resulting in decreased expression of an FaCENH3-1a protein, an FaCENH3-1b protein, an FaCENH3- 2a protein, an FaCENH3-2b protein, an FaCENH3-3 protein, and an FaCENH3-4 protein. In certain embodiments, the haploid-inducing strawberry plant is a plant of the species Fragaria x ananassa comprising one or more genetic modifications resulting in decreased expression of one or more CENH3 proteins having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence 50 sf-6744554 197072001240 identity, or at least 60%, 65%, 70%, 75%, 80%, 85%, 95%, 96%, 97%, 98%, or 99% sequence similarity to an amino acid sequence selected from the list consisting of SEQ ID NOs: 9-14, or fragments thereof. In certain embodiments, the haploid-inducing strawberry plant is a plant of the species Fragaria x ananassa comprising one or more genetic modifications resulting in decreased expression of 1) a CENH3 protein having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity, or at least 60%, 65%, 70%, 75%, 80%, 85%, 95%, 96%, 97%, 98%, or 99% sequence similarity to the amino acid sequence of SEQ ID NO: 9; 2) a CENH3 protein having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity, or at least 60%, 65%, 70%, 75%, 80%, 85%, 95%, 96%, 97%, 98%, or 99% sequence similarity to the amino acid sequence of SEQ ID NO: 10; 3) a CENH3 protein having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity, or at least 60%, 65%, 70%, 75%, 80%, 85%, 95%, 96%, 97%, 98%, or 99% sequence similarity to the amino acid sequence of SEQ ID NO: 11; 4) a CENH3 protein having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity, or at least 60%, 65%, 70%, 75%, 80%, 85%, 95%, 96%, 97%, 98%, or 99% sequence similarity to the amino acid sequence of SEQ ID NO: 12; 5) a CENH3 protein having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity, or at least 60%, 65%, 70%, 75%, 80%, 85%, 95%, 96%, 97%, 98%, or 99% sequence similarity to the amino acid sequence of SEQ ID NO: 13; and/or 5) a CENH3 protein having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity, or at least 60%, 65%, 70%, 75%, 80%, 85%, 95%, 96%, 97%, 98%, or 99% sequence similarity to the amino acid sequence of SEQ ID NO: 14. [0142] In some embodiments, the haploid-inducing strawberry plant is a plant of the species Fragaria x ananassa comprising one or more genetic modifications resulting in decreased expression of an FaCENH3-7a protein, an FaCENH3-7b protein, an FaCENH3-7c protein, an FaCENH3-7d protein, or any combination thereof. In certain embodiments, the haploid-inducing strawberry plant is a plant of the species Fragaria x ananassa comprising one or more genetic modifications resulting in decreased expression of one or more CENH3 51 sf-6744554 197072001240 proteins having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity, or at least 60%, 65%, 70%, 75%, 80%, 85%, 95%, 96%, 97%, 98%, or 99% sequence similarity to an amino acid sequence selected from the list consisting of SEQ ID NOs: 53-56, or fragments thereof. In certain embodiments, the haploid-inducing strawberry plant is a plant of the species Fragaria x ananassa comprising one or more genetic modifications resulting in decreased expression of 1) a CENH3 protein having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity, or at least 60%, 65%, 70%, 75%, 80%, 85%, 95%, 96%, 97%, 98%, or 99% sequence similarity to the amino acid sequence of SEQ ID NO: 53; 2) a CENH3 protein having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity, or at least 60%, 65%, 70%, 75%, 80%, 85%, 95%, 96%, 97%, 98%, or 99% sequence similarity to the amino acid sequence of SEQ ID NO: 54; 3) a CENH3 protein having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity, or at least 60%, 65%, 70%, 75%, 80%, 85%, 95%, 96%, 97%, 98%, or 99% sequence similarity to the amino acid sequence of SEQ ID NO: 55; and/or 4) a CENH3 protein having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity, or at least 60%, 65%, 70%, 75%, 80%, 85%, 95%, 96%, 97%, 98%, or 99% sequence similarity to the amino acid sequence of SEQ ID NO: 56. [0143] In some embodiments, the haploid-inducing strawberry plant is a plant of the species Fragaria x ananassa comprising one or more genetic modifications of an FaCENH3- 1a gene or gene product, an FaCENH3-1b gene or gene product, an FaCENH3-2a gene or gene product, an FaCENH3-2b gene or gene product, an FaCENH3-3 gene or gene product, an FaCENH3-4 gene or gene product, or any combination thereof. In some embodiments, the haploid-inducing strawberry plant is a plant of the species Fragaria x ananassa comprising one or more genetic modifications of an FaCENH3-1a gene or gene product, an FaCENH3- 1b gene or gene product, an FaCENH3-2a gene or gene product, an FaCENH3-2b gene or gene product, an FaCENH3-3 gene or gene product, and an FaCENH3-4 gene or gene product. In certain embodiments, the haploid-inducing strawberry plant is a plant of the species Fragaria x ananassa comprising one or more genetic modifications in a CENH3 gene comprising a nucleotide sequence having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 52 sf-6744554 197072001240 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity or complementarity to a nucleotide sequence selected from the group consisting of SEQ ID NOs: 2-71, or a fragment thereof. In certain embodiments, the haploid- inducing strawberry plant is a plant of the species Fragaria x ananassa comprising: 1) one or more genetic modifications in a CENH3 gene comprising a nucleotide sequence having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity or complementarity to the nucleotide sequence of SEQ ID NO: 2, or a fragment thereof; 2) one or more genetic modifications in a CENH3 gene comprising a nucleotide sequence having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity or complementarity to the nucleotide sequence of SEQ ID NO: 3, or a fragment thereof; 3) one or more genetic modifications in a CENH3 gene comprising a nucleotide sequence having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity or complementarity to the nucleotide sequence of SEQ ID NO: 4, or a fragment thereof; 4) one or more genetic modifications in a CENH3 gene comprising a nucleotide sequence having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity or complementarity to the nucleotide sequence of SEQ ID NO: 5, or a fragment thereof; 5) one or more genetic modifications in a CENH3 gene comprising a nucleotide sequence having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity or complementarity to the nucleotide sequence of SEQ ID NO: 6, or a fragment thereof; and/or 6) one or more genetic modifications in a CENH3 gene comprising a nucleotide sequence having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity or complementarity to the nucleotide sequence of SEQ ID NO: 7, or a fragment thereof. In some variations, the one or more genetic modifications comprise a modification of an enhancer of an FaCENH3 gene (e.g., an FaCENH3-1a gene, an FaCENH3-1b gene, an FaCENH3-2a gene, an FaCENH3-2b gene, an FaCENH3-3 gene, and/or an FaCENH3-4 gene), a modification of a promoter of an FaCENH3 gene (e.g., an FaCENH3-1a gene, an FaCENH3-1b gene, an FaCENH3-2a gene, an FaCENH3-2b gene, an FaCENH3-3 gene, and/or an FaCENH3-4 gene), a modification of a coding region of an FaCENH3 gene (e.g., an FaCENH3-1a gene, an FaCENH3-1b gene, an FaCENH3-2a gene, an FaCENH3-2b gene, an FaCENH3-3 gene, and/or an FaCENH3-4 gene), modification of an intron of an 53 sf-6744554 197072001240 FaCENH3 gene (e.g., an FaCENH3-1a gene, an FaCENH3-1b gene, an FaCENH3-2a gene, an FaCENH3-2b gene, an FaCENH3-3 gene, and/or an FaCENH3-4 gene), or any combination thereof relative to a control Fragaria x ananassa plant (e.g., a wild-type or unmodified Fragaria x ananassa plant lacking one or more or all of the genetic modifications). [0144] In some embodiments, the haploid-inducing strawberry plant is a plant of the species Fragaria x ananassa comprising one or more genetic modifications of an FaCENH3- 7a gene or gene product, an FaCENH3-7b gene or gene product, an FaCENH3-7c gene or gene product, an FaCENH3-7d gene or gene product, or any combination thereof. In certain embodiments, the haploid-inducing strawberry plant is a plant of the species Fragaria x ananassa comprising one or more genetic modifications in a CENH3 gene comprising a nucleotide sequence having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity or complementarity to a nucleotide sequence selected from the group consisting of SEQ ID NOs: 58-61, or a fragment thereof. In certain embodiments, the haploid-inducing strawberry plant is a plant of the species Fragaria x ananassa comprising: 1) one or more genetic modifications in a CENH3 gene comprising a nucleotide sequence having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity or complementarity to the nucleotide sequence of SEQ ID NO: 58, or a fragment thereof; 2) one or more genetic modifications in a CENH3 gene comprising a nucleotide sequence having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity or complementarity to the nucleotide sequence of SEQ ID NO: 59, or a fragment thereof; 3) one or more genetic modifications in a CENH3 gene comprising a nucleotide sequence having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity or complementarity to the nucleotide sequence of SEQ ID NO: 60, or a fragment thereof; and/or 4) one or more genetic modifications in a CENH3 gene comprising a nucleotide sequence having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity or complementarity to the nucleotide sequence of SEQ ID NO: 61, or a fragment thereof. In some variations, the one or more genetic modifications comprise a modification of an enhancer of an FaCENH3 gene (e.g., an FaCENH3-7a gene, an FaCENH3-7b gene, an 54 sf-6744554 197072001240 FaCENH3-7c gene, and/or an FaCENH3-7d gene), a modification of a promoter of an FaCENH3 gene (e.g., an FaCENH3-7a gene, an FaCENH3-7b gene, an FaCENH3-7c gene, and/or an FaCENH3-7d gene), a modification of a coding region of an FaCENH3 gene (e.g., an FaCENH3-7a gene, an FaCENH3-7b gene, an FaCENH3-7c gene, and/or an FaCENH3- 7d gene), modification of an intron of an FaCENH3 gene (e.g., an FaCENH3-7a gene, an FaCENH3-7b gene, an FaCENH3-7c gene, and/or an FaCENH3-7d gene), or any combination thereof relative to a control Fragaria x ananassa plant (e.g., a wild-type or unmodified Fragaria x ananassa plant lacking one or more or all of the genetic modifications). In some embodiments, the one or more genetic modifications comprise one or more deletions, insertions, or nucleotide substitutions in a coding region of an FaCENH3 gene (e.g., an FaCENH3-7a gene, an FaCENH3-7b gene, an FaCENH3-7c gene, and/or an FaCENH3-7d gene). [0145] In some embodiments, the one or more genetic modifications in a CENH3 gene each independently comprise an insertion, a deletion, one or more nucleotide changes, or an inversion that results in decreased expression (e.g., decreased activity) of the CENH3 gene. In certain embodiments, the insertion, the deletion, the one or more nucleotide changes, or the inversion eliminates expression (e.g., eliminates activity) of the CENH3 gene. In certain embodiments, the insertion, the deletion, the one or more nucleotide changes, or the inversion decreases, but does not eliminate, expression of the CENH3 gene. In some variations, the insertion, the deletion, the one or more nucleotide changes, or the inversion is positioned in the first 70%, the first 60%, the first 50%, the first 40%, the first 30%, the first 20%, or the first 10% of the nucleotides of the CENH3 gene (e.g., of a polynucleotide sequence selected from the group consisting of SEQ ID NOs: 1-7). In certain variations, the insertion, the deletion, the one or more nucleotide changes, or the inversion is positioned in the first 100, the first 200, the first 300, the first 400, the first 500, the first 600, the first 700, the first 800, the first 900, the first 1000, the first 1250, the first 1500, the first 1750, the first 2000, the first 2500, or the first 3000 nucleotides of the CENH3 gene (e.g., of a polynucleotide sequence selected from the group consisting of SEQ ID NOs: 1-7). In some embodiments, the insertion, the deletion, the one or more nucleotide changes, or the inversion eliminates expression (e.g., eliminates activity) of the CENH3 gene. In some variations, the insertion, the deletion, the one or more nucleotide changes, or the inversion results in a premature stop codon present in the first 70%, the first 60%, the first 50%, the first 40%, the first 30%, the first 20%, or the first 10% of the nucleotides of a polynucleotide sequence selected from the 55 sf-6744554 197072001240 group consisting of SEQ ID NOs: 1-7 or of the nucleotides of the coding sequence of the CENH3 gene following the start codon in the 3’ direction, thereby eliminating expression (e.g., activity) of the CENH3 gene. In some variations, the insertion, the deletion, the one or more nucleotide changes, or the inversion results in a premature stop codon present in the first 100, the first 200, the first 300, the first 400, the first 500, the first 600, the first 700, the first 800, the first 900, the first 1000, the first 1250, the first 1500, the first 1750, the first 2000, the first 2500, or the first 3000 nucleotides of a polynucleotide sequence selected from the group consisting of SEQ ID NOs: 1-7 or of the coding sequence of the CENH3 gene following the start codon in the 3’ direction, thereby eliminating expression (e.g., activity) of the CENH3 gene. [0146] In some embodiments, the haploid-inducing strawberry plant is a plant of the species Fragaria x ananassa and has decreased expression of an FaCENH3-1a protein, an FaCENH3-1b protein, an FaCENH3-2a protein, an FaCENH3-2b protein, an FaCENH3-3 protein, an FaCENH3-4 protein, or any combination (e.g., two, three, four, five or all six) thereof relative to a control Fragaria x ananassa plant (e.g., a Fragaria ananassa plant lacking one or more or all of the genetic modifications). In certain embodiments, the haploid- inducing strawberry plant is a plant of the species Fragaria x ananassa and lacks detectable expression of an FaCENH3-1a protein, an FaCENH3-1b protein, an FaCENH3-2a protein, an FaCENH3-2b protein, an FaCENH3-3 protein, an FaCENH3-4 protein, or any combination (e.g., two, three, four, five or all six) thereof. [0147] The haploid-inducing strawberry plant, or plant part thereof, may comprise one or more genetic modifications resulting in decreased expression of any combination of CENH3 genes described herein or known in the art. In some embodiments, the haploid-inducing strawberry plant, or plant part thereof, may comprise one or more genetic modifications resulting in non-expression of any combination of CENH3 genes described here or known in the art. In further embodiments, the haploid-inducing strawberry plant, or plant part thereof, may comprise one or more genetic modifications resulting in decreased expression, non- expression, or a combination thereof of any combination of CENH3 genes described here or known in the art. In certain embodiments, the haploid-inducing strawberry plant or plant part lacks detectable expression of CENH3 proteins. [0148] In some embodiments, the haploid-inducing strawberry plant or plant part thereof has decreased expression (including decreased activity) of any of the CENH3 genes and/or 56 sf-6744554 197072001240 CENH3 proteins described herein relative to a control strawberry plant. For example, in some embodiments, the haploid-inducing strawberry plant or plant part thereof has decreased expression (e.g., decreased activity) of one or more CENH3 genes and/or one or more CENH3 proteins, wherein the decreased expression (e.g., decreased activity) of one or more CENH3 proteins CENH3 genes and/or is decreased by at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90% , or about 100% as compared to a control plant, plant part, or plant cell lacking the one or more genetic modifications. In some embodiments, the haploid-inducing strawberry plant or plant part thereof has decreased expression (e.g., decreased activity) of one or more CENH3 genes and/or one or more CENH3 proteins, wherein the decreased expression (e.g., decreased activity) of CENH3 genes and/or one or more CENH3 proteins is decreased by no more than about 10%, about 20%, about 30%, about 40%, about 50%, about 60%, about 70%, about 80%, about 90%, about 95%, or about 99% as compared to a control plant, plant part, or plant cell lacking the one or more genetic modifications. In some embodiments, the haploid- inducing strawberry plant or plant part thereof has decreased expression (e.g., decreased activity) of one or more CENH3 genes and/or one or more CENH3 proteins, wherein the decreased expression (e.g., decreased activity) of the one or more CENH3 genes and/or one or more CENH3 proteins is decreased is decreased by between about 10% and about 100%, between about 20% and about 100%, between about 30% and about 100%, between about 40% and about 100%, between about 50% and about 100%, between about 60% and about 100%, between about 70% and about 100%, between about 80% and about 100%, between about 90% and about 100%, between about 10% and about 90%, between about 20% and about 90%, between about 30% and about 90%, between about 40% and about 90%, between about 50% and about 90%, between about 60% and about 90%, between about 70% and about 90%, between about 80% and about 90%, between about 10% and about 80%, between about 20% and about 80%, between about 30% and about 80%, between about 40% and about 80%, between about 50% and about 80%, between about 60% and about 80%, between about 70% and about 80%, 10% and about 70%, between about 20% and about 70%, between about 30% and about 70%, between about 40% and about 70%, between about 50% and about 70%, between about 60% and about 70%, between about 10% and about 60%, between about 20% and about 60%, between about 30% and about 60%, between about 40% and about 60%, between about 50% and about 60%, between about 10% and about 50%, between about 20% and about 50%, between about 30% and about 50%, between about 40% and about 50%, between about 10% and about 40%, between about 20% and about 40%, between 57 sf-6744554 197072001240 about 30% and about 40%, between about 10% and about 30%, between about 20% and about 30%, or about 10% and 20% as compared to a control plant, plant part, or plant cell lacking the one or more genetic modifications. Control strawberry plants include, for example, a plant of the same species as the haploid-inducing strawberry plant that lacks one or more or all of the genetic modifications comprised by the haploid-inducing strawberry plant, a wild-type plant of the same species as the haploid-inducing strawberry plant, an unmodified plant of the same species as the haploid-inducing strawberry plant, or a null segregant strawberry plant, which underwent the same genetic manipulation, but is lacking one or more or all of the genetic modifications comprised by the haploid-inducing strawberry plant. In certain embodiments, the haploid-inducing strawberry plant or plant part thereof has decreased expression of CENH3 proteins relative to a strawberry plant of the same species lacking the one or more genetic modifications. Characteristics of Strawberry Plants [0149] The strawberry plants (e.g., haploid-inducing strawberry plants) described herein having one or more genetic modifications resulting in decreased expression of one or more CENH3 genes may have characteristics that differ from those of a strawberry plant lacking the one or more genetic modifications. Without wishing to be bound by theory, it is believed that decreased expression of one or more CENH3 genes may result in certain changes in mitotic and meiotic activity in plants. [0150] In some embodiments, the strawberry plants (e.g., haploid-inducing strawberry plants) described herein have decreased pollen germination (e.g., a decreased pollen germination rate) than a control strawberry plant lacking the one or more genetic modifications. In some embodiments, the strawberry plant has a pollen germination rate of less than 65%, 60%, 55%, 50%, 45%, 40%, 35%, 30%, 25%, 20%, 15%, 10%, or 5%. [0151] In some embodiments, the strawberry plants (e.g., haploid-inducing strawberry plants) described herein have decreased flower diameter as compared to a control strawberry plant lacking the one or more genetic modifications. In some embodiments, the strawberry plant has a flower diameter of less than 25, 24, 23, 22, 21, 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, or 10 mm. In some embodiments, the strawberry plant has a flower diameter of between 5 mm and 25, 24, 23, 22, 21, 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, or 10 mm. 58 sf-6744554 197072001240 [0152] In some embodiments, the strawberry plants (e.g., haploid-inducing strawberry plants) described herein have decreased anther width as compared to a control strawberry plant lacking the one or more genetic modifications. In some embodiments, the strawberry plant has a anther width of less than 1.5, 1.4, 1.3, 1.2, 1.1, 1.0, 0.9, 0.8, 0.7, 0.6, or 0.5 mm. In some embodiments, the strawberry plant has a anther width of between 0.1 mm and 1.5, 1.4, 1.3, 1.2, 1.1, 1.0, 0.9, 0.8, 0.7, 0.6, or 0.5 mm. [0153] In some embodiments, the strawberry plants (e.g., haploid-inducing strawberry plants) described herein have decreased filament length as compared to a control strawberry plant lacking the one or more genetic modifications. In some embodiments, the strawberry plant has a filament length of less than 2.7, 2.6, 2.5, 2.4, 2.3, 2.2, 2.1, 2.0, 1.9, 1.8, 1.7, 1.6, 1.5, 1.4, 1.3, 1.2, 1.1, 1.0, 0.9, 0.8, 0.7, 0.6, or 0.5 mm. In some embodiments, the strawberry plant has a filament length of less between 0.1 mm and 2.7, 2.6, 2.5, 2.4, 2.3, 2.2, 2.1, 2.0, 1.9, 1.8, 1.7, 1.6, 1.5, 1.4, 1.3, 1.2, 1.1, 1.0, 0.9, 0.8, 0.7, 0.6, or 0.5 mm. [0154] In some embodiments, the strawberry plants (e.g., haploid-inducing strawberry plants) described herein have delayed microspore formation as compared to a control strawberry plant lacking the one or more genetic modifications. In some embodiments, the strawberry plants (e.g., haploid-inducing strawberry plants) described herein have larger bud size at the time of microspore formation as compared to a control strawberry plant lacking the one or more genetic modifications. In some embodiments, the strawberry plant has an average bud size at the time of microspore formation of greater than 4.5, 4.6, 4.7, 4.8, 4.9, 5.0, 5.1, 5.2, 5.35.4, 5.5, 5.6, 5.7, 5.8, 5.9, or 6.0 at the time of microspore formation. In certain embodiments, the strawberry plant has an average bud size at the time of microspore formation of greater than 4.7 at the time of microspore formation. In certain embodiments, the strawberry plant has an average bud size at the time of microspore formation of greater than 5.0 at the time of microspore formation. [0155] In some embodiments, the strawberry plants (e.g., haploid-inducing strawberry plants) described herein have buds with observable tetrads, e.g., as shown in FIGS. 16E and 16F. In some embodiments, the strawberry plants (e.g., haploid-inducing strawberry plants) described herein have observable tetrads in at least 10%, 20%, 30%, 40%, 50%, 60%, or 70% of buds. 59 sf-6744554 197072001240 [0156] In some embodiments, the strawberry plants (e.g., haploid-inducing strawberry plants) described herein have fewer seeds per berry as compared to a control strawberry plant lacking the one or more genetic modifications. In some embodiments, the strawberry plant has fewer than 100, 90, 80, 70, 60, 50, 40, 30, 20, or 10 seeds per berry. In some embodiments, the strawberry plant has between 1 and 100, 90, 80, 70, 60, 50, 40, 30, 20, or 10 seeds per berry. [0157] In some embodiments, the strawberry plants (e.g., haploid-inducing strawberry plants) described herein have reduced cell growth as compared to a control strawberry plant lacking the one or more genetic modifications, for example, as measured by the number of microcalli regenerated from protoplasts per area of growth medium, or per lens as measured using microscopy. Plant Parts of Haploid-inducing Strawberry Plants [0158] Also provided herein are plant parts of haploid-inducing strawberry plants. The plant part may be any plant part known in the art, including, but not limited to, a flower, a pistil, a leaf, a stem, a petiole, a cutting, a tissue, a seed coat, an ovule, pollen, a sperm cell, a root, a fruit (e.g., an aggregate fruit), a cotyledon, a hypocotyl, a protoplast, an embryo, an anther, a seed, an achene, a stolon (also known as a runner), a callus, a cell culture, or any portion thereof. Plant parts can be obtained by lifting, cutting, snapping, grinding, or otherwise disassociating the plant part from the plant. [0159] Accordingly, provided herein is pollen of a haploid-inducing strawberry plant. In some embodiments, the pollen comprises one or more genetic modifications resulting in decreased expression of one or more CENH3 genes described herein. In certain embodiments, the pollen comprises one or more genetic modifications in one or more CENH3 genes described herein. [0160] Also provided herein is stolon of a haploid-inducing strawberry plant. In some embodiments, the stolon comprises one or more genetic modifications resulting in decreased expression of one or more CENH3 genes described herein. In certain embodiments, the stolon comprises one or more genetic modifications in one or more CENH3 genes described herein. [0161] Also provided herein is seed of a haploid-inducing strawberry plant. In some embodiments, the seed comprises one or more genetic modifications resulting in decreased 60 sf-6744554 197072001240 expression of one or more CENH3 genes described herein. In certain embodiments, the seed comprises one or more genetic modifications in one or more CENH3 genes described herein. [0162] Also provided herein is an achene of a haploid-inducing strawberry plant. In some embodiments, the achene comprises one or more genetic modifications resulting in decreased expression of one or more CENH3 genes described herein. In certain embodiments, the achene comprises one or more genetic modifications in one or more CENH3 genes described herein. [0163] In some embodiments, the pollen, stolon, seed, or achene of the haploid-inducing strawberry plant comprises one or more genetic modifications resulting in decreased expression of one or more CENH3 proteins described herein. In some embodiments, the pollen, stolon, seed, or achene of the haploid-inducing strawberry plant comprises one or more genetic modifications of one or more CENH3 genes. In some embodiments, the pollen, stolon, seed, or achene of the haploid-inducing strawberry plant comprises one or more genetic modifications resulting in decreased expression of one or more CENH3 proteins having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity, or at least 60%, 65%, 70%, 75%, 80%, 85%, 95%, 96%, 97%, 98%, or 99% sequence similarity to an amino acid sequence selected from the list consisting of SEQ ID NOs: 8-14 and 53-56, or fragments thereof. In some embodiments, the pollen, stolon, seed, or achene of the haploid-inducing strawberry plant comprises one or more genetic modifications of one or more CENH3 genes comprising a nucleotide sequence having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity or complementarity to a nucleotide sequence selected from the group consisting of SEQ ID NOs: 1-7 and 58-61, or a fragment thereof. In certain embodiments, the one or more genetic modifications comprise a modification of an enhancer of one or more of the CENH3 genes, a modification of a promoter of one or more of the CENH3 genes, a modification of a coding region of one or more of the CENH3 genes, a modification of an intron of one or more of the CENH3 genes, a modification of methylation status of one or more of the CENH3 genes, expression of a repressor protein that targets the DNA or an mRNA of one or more of the CENH3 genes, expression of an RNA interference construct that targets an mRNA of one or more of the CENH3 genes, or any combination thereof. In some embodiments, the pollen, stolon, seed, or achene of the haploid-inducing strawberry 61 sf-6744554 197072001240 plant has decreased expression of one or more CENH3 genes described herein relative to a pollen, stolon, seed, or achene of a strawberry plant (e.g., a control strawberry plant, e.g., a strawberry plant of the same species) lacking the one or more genetic modifications. In certain embodiments, the haploid-inducing strawberry lacks detectable expression of one or more CENH3 genes described herein relative to a pollen, stolon, seed, or achene of a strawberry plant (e.g., a control strawberry plant, e.g., a strawberry plant of the same species) lacking the one or more genetic modifications. In certain embodiments, the pollen, stolon, seed, or achene of the haploid-inducing strawberry plant lacks detectable expression of any CENH3 genes. In certain embodiments, the pollen, stolon, seed, or achene of the haploid- inducing strawberry plant further comprises one or more naturally occurring inactive alleles of one or more CENH3 genes. [0164] In some embodiments, the pollen, stolon, seed, or achene is from a haploid- inducing strawberry plant of the species Fragaria vesca and comprises one or more genetic modifications resulting in decreased expression of one or more CENH3 genes. In certain embodiments, the pollen, stolon, seed, or achene is from a haploid-inducing strawberry plant of the species Fragaria vesca and comprises one or more genetic modifications resulting in decreased expression of an FvCENH3 gene. In certain embodiments, the pollen, stolon, seed, or achene is from a haploid-inducing strawberry plant of the species Fragaria vesca and comprises one or more genetic modifications resulting in decreased expression of an FvCENH3 protein. In certain embodiments, the pollen, stolon, seed, or achene is from a haploid-inducing strawberry plant of the species Fragaria vesca and comprises one or more genetic modifications resulting in decreased expression of a CENH3 protein having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity, or at least 60%, 65%, 70%, 75%, 80%, 85%, 95%, 96%, 97%, 98%, or 99% sequence similarity to the FvCENH3 protein of SEQ ID NO. 8, or a fragment thereof. [0165] In some embodiments, the pollen, stolon, seed, or achene is from a haploid- inducing strawberry plant of the species Fragaria vesca and comprises one or more genetic modifications of an FvCENH3 gene or gene product. In certain embodiments, the pollen, stolon, seed, or achene is from a haploid-inducing strawberry plant of the species Fragaria vesca and comprises one or more genetic modifications in a FvCENH3 gene comprising a nucleotide sequence having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 62 sf-6744554 197072001240 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity or complementarity to the FvCENH3 gene nucleotide sequence of SEQ ID NO. 1, or a fragment thereof. In certain embodiments, the pollen, stolon, seed, or achene is from a haploid- inducing strawberry plant of the species Fragaria vesca and comprises genetic modifications in both alleles of an FvCENH3 gene. In some variations, the one or more genetic modifications comprise a modification of an enhancer of FvCENH3, a modification of a promoter of FvCENH3, a modification of a coding region of FvCENH3, modification of an intron of FvCENH3, or any combination thereof relative to a wild- type or unmodified Fragaria vesca plant. In some embodiments, the pollen, stolon, seed, or achene is from a haploid-inducing strawberry plant of the species Fragaria vesca and has decreased expression of FvCENH3 protein relative to a pollen, stolon, seed, or achene from a control Fragaria vesca plant (e.g., a Fragaria vesca plant lacking one or more of the genetic modifications). In certain embodiments, the pollen, stolon, seed, or achene of the haploid- inducing strawberry plant is a plant of the species Fragaria vesca and lacks detectable expression of FvCENH3 protein. [0166] In some embodiments, the pollen, stolon, seed, or achene is from a haploid- inducing strawberry plant of the species Fragaria x ananassa and comprises one or more genetic modifications resulting in decreased expression of one or more CENH3 genes. In certain embodiments, the pollen, stolon, seed, or achene is from a haploid-inducing strawberry plant of the species Fragaria x ananassa and comprises one or more genetic modifications resulting in decreased expression of an FaCENH3-1a gene, an FaCENH3-1b gene, an FaCENH3-2a gene, an FaCENH3-2b gene, an FaCENH3-3 gene, an FaCENH3-4 gene, or any combination thereof. In certain embodiments, the pollen, stolon, seed, or achene is from a haploid-inducing strawberry plant of the species Fragaria x ananassa and comprises one or more genetic modifications resulting in decreased expression of an FaCENH3-1a gene, an FaCENH3-1b gene, an FaCENH3-2a gene, an FaCENH3-2b gene, an FaCENH3-3 gene, and an FaCENH3-4 gene. [0167] In some embodiments, the pollen, stolon, seed, or achene is from a haploid- inducing strawberry plant of the species Fragaria x ananassa and comprises one or more genetic modifications resulting in decreased expression of an FaCENH3-1a protein, an FaCENH3-1b protein, an FaCENH3-2a protein, an FaCENH3-2b protein, an FaCENH3-3 protein, an FaCENH3-4 protein, or any combination thereof. In certain embodiments, the 63 sf-6744554 197072001240 pollen, stolon, seed, or achene is from a haploid-inducing strawberry plant of the species Fragaria x ananassa and comprises one or more genetic modifications resulting in decreased expression of an FaCENH3-1a protein, an FaCENH3-1b protein, an FaCENH3-2a protein, an FaCENH3-2b protein, an FaCENH3-3 protein, and an FaCENH3-4 protein. In certain embodiments, the pollen, stolon, seed, or achene is from a haploid-inducing strawberry plant of the species Fragaria x ananassa and comprises one or more genetic modifications resulting in decreased expression of one or more CENH3 proteins having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity, or at least 60%, 65%, 70%, 75%, 80%, 85%, 95%, 96%, 97%, 98%, or 99% sequence similarity to an amino acid sequence selected from the list consisting of SEQ ID NOs: 9-14, or fragments thereof. In certain embodiments, the pollen, stolon, seed, or achene is from a haploid-inducing strawberry plant of the species Fragaria x ananassa and comprises one or more genetic modifications resulting in decreased expression of 1) a CENH3 protein having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity, or at least 60%, 65%, 70%, 75%, 80%, 85%, 95%, 96%, 97%, 98%, or 99% sequence similarity to the amino acid sequence of SEQ ID NO: 9; 2) a CENH3 protein having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity, or at least 60%, 65%, 70%, 75%, 80%, 85%, 95%, 96%, 97%, 98%, or 99% sequence similarity to the amino acid sequence of SEQ ID NO: 10; 3) a CENH3 protein having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity, or at least 60%, 65%, 70%, 75%, 80%, 85%, 95%, 96%, 97%, 98%, or 99% sequence similarity to the amino acid sequence of SEQ ID NO: 11; and/or 4) a CENH3 protein having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity, or at least 60%, 65%, 70%, 75%, 80%, 85%, 95%, 96%, 97%, 98%, or 99% sequence similarity to the amino acid sequence of SEQ ID NO: 12; 5) a CENH3 protein having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity, or at least 60%, 65%, 70%, 75%, 80%, 85%, 95%, 96%, 97%, 98%, or 99% sequence similarity to the amino acid sequence of SEQ ID NO: 13; and/or 6) a CENH3 protein having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity, or at 64 sf-6744554 197072001240 least 60%, 65%, 70%, 75%, 80%, 85%, 95%, 96%, 97%, 98%, or 99% sequence similarity to the amino acid sequence of SEQ ID NO: 14. [0168] In some embodiments, the pollen, stolon, seed, or achene is from a haploid- inducing strawberry plant of the species Fragaria x ananassa and comprises one or more genetic modifications resulting in decreased expression of an FaCENH3-7a protein, an FaCENH3-7b protein, an FaCENH3-7c protein, an FaCENH3-7d protein, or any combination thereof. In certain embodiments, the pollen, stolon, seed, or achene is from a haploid- inducing strawberry plant of the species Fragaria x ananassa and comprises one or more genetic modifications resulting in decreased expression of one or more CENH3 proteins having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity, or at least 60%, 65%, 70%, 75%, 80%, 85%, 95%, 96%, 97%, 98%, or 99% sequence similarity to an amino acid sequence selected from the list consisting of SEQ ID NOs: 53-56, or fragments thereof. In certain embodiments, the pollen, stolon, seed, or achene is from a haploid-inducing strawberry plant of the species Fragaria x ananassa and comprises one or more genetic modifications resulting in decreased expression of 1) a CENH3 protein having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity, or at least 60%, 65%, 70%, 75%, 80%, 85%, 95%, 96%, 97%, 98%, or 99% sequence similarity to the amino acid sequence of SEQ ID NO: 53; 2) a CENH3 protein having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity, or at least 60%, 65%, 70%, 75%, 80%, 85%, 95%, 96%, 97%, 98%, or 99% sequence similarity to the amino acid sequence of SEQ ID NO: 54; 3) a CENH3 protein having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity, or at least 60%, 65%, 70%, 75%, 80%, 85%, 95%, 96%, 97%, 98%, or 99% sequence similarity to the amino acid sequence of SEQ ID NO: 55; and/or 4) a CENH3 protein having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity, or at least 60%, 65%, 70%, 75%, 80%, 85%, 95%, 96%, 97%, 98%, or 99% sequence similarity to the amino acid sequence of SEQ ID NO: 56. [0169] In some embodiments, the pollen, stolon, seed, or achene is from a haploid- inducing strawberry plant of the species Fragaria x ananassa and comprises one or more 65 sf-6744554 197072001240 genetic modifications of an FaCENH3-1a gene, an FaCENH3-1b gene, an FaCENH3-2a gene, an FaCENH3-2b gene, an FaCENH3-3 gene, an FaCENH3-4 gene, or any combination thereof. In some embodiments, the pollen, stolon, seed, or achene is from a haploid-inducing strawberry plant of the species Fragaria x ananassa and comprises one or more genetic modifications of an FaCENH3-1a gene, an FaCENH3-1b gene, an FaCENH3-2a gene, an FaCENH3-2b gene, an FaCENH3-3 gene, and an FaCENH3-4 gene. In certain embodiments, the pollen, stolon, seed, or achene is from a haploid-inducing strawberry plant of the species Fragaria x ananassa and comprises one or more genetic modifications in a CENH3 gene comprising a nucleotide sequence having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity or complementarity to a nucleotide sequence selected from the group consisting of SEQ ID NOs: 2-7, or a fragment thereof. In certain embodiments, the pollen, stolon, seed, or achene is from a haploid-inducing strawberry plant of the species Fragaria x ananassa and comprises: 1) one or more genetic modifications in a CENH3 gene comprising a nucleotide sequence having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity or complementarity to the nucleotide sequence of SEQ ID NO: 2, or a fragment thereof; 2) one or more genetic modifications in a CENH3 gene comprising a nucleotide sequence having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity or complementarity to the nucleotide sequence of SEQ ID NO: 3, or a fragment thereof; 3) one or more genetic modifications in a CENH3 gene comprising a nucleotide sequence having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity or complementarity to the nucleotide sequence of SEQ ID NO: 4, or a fragment thereof; 4) one or more genetic modifications in a CENH3 gene comprising a nucleotide sequence having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity or complementarity to the nucleotide sequence of SEQ ID NO: 5, or a fragment thereof; 5) one or more genetic modifications in a CENH3 gene comprising a nucleotide sequence having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity or complementarity to the nucleotide sequence of SEQ ID NO: 6, or a fragment thereof; and/or 6) one or more genetic modifications in a CENH3 gene comprising a nucleotide sequence having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 66 sf-6744554 197072001240 98%, 99%, or 100% sequence identity or complementarity to the nucleotide sequence of SEQ ID NO: 7, or a fragment thereof. In some variations, the one or more genetic modifications comprise a modification of an enhancer of an FaCENH3 gene (e.g., an FaCENH3-1a gene, an FaCENH3-1b gene, an FaCENH3-2a gene, an FaCENH3-2b gene, an FaCENH3-3 gene, and/or an FaCENH3-4 gene), a modification of a promoter of an Facenh3 gene (e.g., an FaCENH3-1a gene, an FaCENH3-1b gene, an FaCENH3-2a gene, an FaCENH3-2b gene, an FaCENH3-3 gene, and/or an FaCENH3-4 gene), a modification of a coding region of an Facenh3 gene (e.g., an FaCENH3-1a gene, an FaCENH3-1b gene, an FaCENH3-2a gene, an FaCENH3-2b gene, an FaCENH3-3 gene, and/or an FaCENH3-4 gene), modification of an intron of an Facenh3 gene (e.g., an FaCENH3-1a gene, an FaCENH3-1b gene, an FaCENH3-2a gene, an FaCENH3-2b gene, an FaCENH3-3 gene, and/or an FaCENH3-4 gene), or any combination thereof relative to a pollen, stolon, seed, or achene of a control Fragaria x ananassa plant (e.g., a wild-type or unmodified Fragaria x ananassa plant lacking one or more or all of the genetic modifications). [0170] In some embodiments, the pollen, stolon, seed, or achene is from a haploid- inducing strawberry plant of the species Fragaria x ananassa and comprises one or more genetic modifications of an FaCENH3-7a gene, an FaCENH3-7b gene, an FaCENH3-7c gene, an FaCENH3-7d gene, or any combination thereof. In certain embodiments, the pollen, stolon, seed, or achene is from a haploid-inducing strawberry plant of the species Fragaria x ananassa and comprises one or more genetic modifications in a CENH3 gene comprising a nucleotide sequence having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity or complementarity to a nucleotide sequence selected from the group consisting of SEQ ID NOs: 58-61, or a fragment thereof. In certain embodiments, the pollen, stolon, seed, or achene is from a haploid-inducing strawberry plant of the species Fragaria x ananassa and comprises: 1) one or more genetic modifications in a CENH3 gene comprising a nucleotide sequence having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity or complementarity to the nucleotide sequence of SEQ ID NO: 58, or a fragment thereof; 2) one or more genetic modifications in a CENH3 gene comprising a nucleotide sequence having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity or complementarity to the nucleotide sequence of SEQ ID NO: 59, or a fragment thereof; 3) one or more genetic 67 sf-6744554 197072001240 modifications in a CENH3 gene comprising a nucleotide sequence having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity or complementarity to the nucleotide sequence of SEQ ID NO: 60, or a fragment thereof; and/or 4) one or more genetic modifications in a CENH3 gene comprising a nucleotide sequence having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity or complementarity to the nucleotide sequence of SEQ ID NO: 61, or a fragment thereof. In some variations, the one or more genetic modifications comprise a modification of an enhancer of an FaCENH3 gene (e.g., an FaCENH3-7a gene, an FaCENH3-7b gene, an FaCENH3-7c gene, and/or an FaCENH3-7d gene), a modification of a promoter of an FaCENH3 gene (e.g., an FaCENH3-7a gene, an FaCENH3-7b gene, an FaCENH3-7c gene, and/or an FaCENH3-7d gene), a modification of a coding region of an FaCENH3 gene (e.g., an FaCENH3-7a gene, an FaCENH3-7b gene, an FaCENH3-7c gene, and/or an FaCENH3-7d gene), modification of an intron of an FaCENH3 gene (e.g., an FaCENH3-7a gene, an FaCENH3-7b gene, an FaCENH3-7c gene, and/or an FaCENH3-7d gene), or any combination thereof relative to a pollen, stolon, seed, or achene of a control Fragaria x ananassa plant (e.g., a wild-type or unmodified Fragaria x ananassa plant lacking one or more or all of the genetic modifications). In some embodiments, the one or more genetic modifications comprise one or more deletions, insertions, or nucleotide substitutions in a coding region of an FaCENH3 gene (e.g., an FaCENH3-7a gene, an FaCENH3-7b gene, an FaCENH3-7c gene, and/or an FaCENH3-7d gene). [0171] In some embodiments, the pollen, stolon, seed, or achene is from a haploid- inducing strawberry plant of the species Fragaria x ananassa and has decreased expression of an FaCENH3-1a protein, an FaCENH3-1b protein, an FaCENH3-2a protein, an FaCENH3-2b protein, an FaCENH3-3 protein, an FaCENH3-4 protein, or any combination (e.g., two, three, four, five or all six) thereof relative to a pollen, stolon, seed, or achene of a control Fragaria x ananassa plant (e.g., a Fragaria ananassa plant lacking one or more or all of the genetic modifications). In certain embodiments, the pollen, stolon, seed, or achene is from a haploid-inducing strawberry plant of the species Fragaria x ananassa and lacks detectable expression of an FaCENH3-1a protein, an FaCENH3-1b protein, an FaCENH3-2a protein, an FaCENH3-2b protein, an FaCENH3-3 protein, an FaCENH3-4 protein, or any combination (e.g., two, three, four, five or all six) thereof. 68 sf-6744554 197072001240 [0172] The pollen, stolon, seed, or achene of the haploid-inducing strawberry plant may comprise one or more genetic modifications resulting in decreased expression of any combination of CENH3 genes described herein or known in the art. In some embodiments, the pollen, stolon, seed, or achene of the haploid-inducing strawberry plant may comprise one or more genetic modifications resulting in non-expression of any combination of CENH3 genes described here or known in the art. In further embodiments, the pollen, stolon, seed, or achene of the haploid-inducing strawberry plant may comprise one or more genetic modifications resulting in decreased expression, non-expression, or a combination thereof of any combination of CENH3 genes described here or known in the art. In certain embodiments, the pollen, stolon, seed, or achene of the haploid-inducing strawberry plant lacks detectable expression of CENH3 proteins. [0173] In some embodiments, the pollen, stolon, seed, or achene of the haploid-inducing strawberry plant has decreased expression of CENH3 proteins relative to a control strawberry plant. Control strawberry plants include, for example, a plant of the same species as the haploid- inducing strawberry plant that lacks one or more or all of the genetic modifications comprised by the haploid-inducing strawberry plant, a wild-type plant of the same species as the haploid- inducing strawberry plant, an unmodified plant of the same species as the haploid-inducing strawberry plant, or a null segregant strawberry plant, which underwent the same genetic manipulation, but is lacking one or more or all of the genetic modifications comprised by the haploid-inducing strawberry plant. In certain embodiments, the pollen, stolon, seed, or achene of the haploid-inducing strawberry plant has decreased expression of CENH3 proteins relative to a pollen, stolon, seed, or achene of a strawberry plant of the same species lacking the one or more genetic modifications. Methods and Compositions for Producing Haploid-inducing Strawberry Plants [0174] In one aspect, described herein are methods of producing haploid-inducing strawberry plants. In some embodiments, the method comprises introducing one or more genetic modifications resulting in decreased expression of one or more CENH3 genes described herein into a strawberry plant. In certain embodiments, the method further comprises introducing one or more naturally occurring inactive alleles of one or more CENH3 genes into a strawberry plant by crossing with a plant having the one or more naturally occurring inactive alleles of one or more CENH3 genes. 69 sf-6744554 197072001240 Methods of Introducing Genetic Modifications [0175] In some embodiments, the methods of producing haploid-inducing strawberry plants described herein comprise introducing one or more genetic modifications into a strawberry plant. In certain embodiments, the step of introducing one or more genetic modifications into a strawberry plant results in a decreased expression of one or more CENH3 genes. In certain embodiments, the decreased expression is achieved by gene disruption (e.g., disruption of one or more CENH3 genes), gene knockout (e.g., knockout of one or more CENH3 genes), gene knockdown (e.g., knockdown of one or more CENH3 genes), gene silencing (e.g., silencing of one or more CENH3 genes), RNA interference (e.g., RNA interference of one or more CENH3 genes), induction of methylation (e.g., induction of methylation of one or more CENH3 genes), or any combination thereof. In some embodiments, the method comprises introducing one or more of the genetic modifications by gene editing using a site-directed nuclease. [0176] In certain embodiments, the genetic modifications are introduced by gene editing. Any of several gene editing methods known in the art may be used to introduce the genetic modifications of the one or more CENH3 genes. In some variations, gene editing is performed with one or more natural or engineered site-directed nucleases including, but not limited to, RNA-guided nucleases, meganucleases, zinc finger nucleases (ZFNs), and transcription activator-like effector-based nucleases (TALENs). In further variations, gene editing is performed with RNA-guided nucleases including, but not limited to, Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) associated (Cas) nucleases. In yet additional variations, gene editing may be performed using a deactivated Cas nuclease fused to an engineered reverse transcriptase in a process known as prime editing. Methods of gene editing are numerous, well-known and routine in the art, and are described in US17/045747, US16/977020, and US16/961396, which are herein incorporated in their entirety. [0177] An engineered nuclease may be a guided nuclease, which may function as a ribonucleoprotein (RNP) complex with a guide RNA. According to some embodiments, a guided nuclease may be selected from the group consisting of Cas1, Cas1B, Cas2, Cas3, Cas4, Cas5, Cash, Cas7, Cas8, Cas9 (also known as Csn1 and Csx12), Cas10, Cas12a, Csy1, Csy2, Csy3, Cse1, Cse2, Csc1, Csc2, Csa5, Csn2, Csm2, Csm3, Csm4, Csm5, Csm6, Cmr1, Cmr3, Cmr4, Cmr5, Cmr6, Csb1, Csb2, Csb3, Csx17, Csx14, Csx10, Csx16, CsaX, Csx3, 70 sf-6744554 197072001240 Csx1, Csx15, Csf1, Csf2, Csf3, Csf4, Cpf1, CasX, CasY, CasZ, MAD7, and homologs or modified versions thereof, Argonaute (non-limiting examples of Argonaute proteins include Thermus thermophilus Argonaute (TtAgo), Pyrococcus furiosus Argonaute (PfAgo), Natronobacterium gregoryi Argonaute (NgAgo), and homologs or modified versions thereof). The DNA construct or molecule encoding a guided nuclease may be delivered with or without a guide nucleic acid. [0178] For guided nucleases, a guide nucleic acid molecule may be further provided to direct the guided nuclease to a target site in the genome of the plant via base-pairing or hybridization to cause a DSB or nick at or near the target site. The guide nucleic acid may be transformed or introduced into a plant cell or tissue as a guide nucleic acid molecule, or as a recombinant DNA molecule, construct or vector comprising a transcribable DNA sequence encoding the guide nucleic acid operably linked to a promoter or plant-expressible promoter. The promoter may be a constitutive promoter, a tissue-specific or tissue-preferred promoter, a developmental stage promoter, or an inducible promoter. [0179] In some embodiments, the guide nucleic acid comprises a first segment comprising a nucleotide sequence that is complementary to a sequence in a target nucleic acid (e.g., a target nucleic acid of a CENH3 gene described herein) and a second segment that interacts with a guided nuclease protein. In some embodiments, the first segment of a guide comprising a nucleotide sequence that is complementary to a sequence in a target nucleic acid corresponds to a CRISPR RNA (crRNA or crRNA repeat). In some embodiments, the second segment of a guide comprising a nucleic acid sequence that interacts with a guided nuclease protein corresponds to a trans-acting CRISPR RNA (tracrRNA). In some embodiments, the guide nucleic acid comprises two separate nucleic acid molecules (a polynucleotide that is complementary to a sequence in a target nucleic acid and a polynucleotide that interacts with a guided nuclease protein) that hybridize with one another. In other embodiments, the guide nucleic acid is a single polynucleotide (e.g., a gRNA). In some embodiments, the guide nucleic acid may comprise DNA, RNA or a combination of DNA and RNA. [0180] A protospacer-adjacent motif (PAM) may be present in the genome immediately adjacent and upstream to the 5' end of the genomic target site sequence complementary to the targeting sequence of the guide RNA, immediately downstream (3') to the sense (+) strand of the genomic target site (relative to the targeting sequence of the guide RNA) as known in the art. See, e.g., Wu, X. et al. 2014. "Target specificity of the CRISPR-Cas9 system," Quant 71 sf-6744554 197072001240 Biol. 2(2): 59-70. The genomic PAM sequence on the sense (+) strand adjacent to the target site (relative to the targeting sequence of the guide RNA) may comprise 5'-NGG-3'. However, the corresponding sequence of the guide nucleic acid (immediately downstream (3') to the targeting sequence of the guide RNA) may generally not be complementary to the genomic PAM sequence. [0181] The guide nucleic acid may typically be a non-coding RNA molecule that does not encode a protein. The targeting sequence of the guide nucleic acid may be at least 10 nucleotides in length, such as 12-40 nucleotides, 12-30 nucleotides, 12-20 nucleotides, 12-35 nucleotides, 12-30 nucleotides, 15-30 nucleotides, 17-30 nucleotides, or 17-25 nucleotides in length, or about 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25 or more nucleotides in length. The targeting sequence may be at least 95%, at least 96%, at least 97%, at least 99% or 100% identical or complementary to at least 10, at least 11, at least 12, at least 13, at least 14, at least 15, at least 16, at least 17, at least 18, at least 19, at least 20, at least 21, at least 22, at least 23, at least 24, at least 25, or more consecutive nucleotides of a DNA sequence at the genomic target site. [0182] In addition to the targeting sequence, a guide nucleic acid may further comprise one or more other structural or scaffold sequence(s), which may bind or interact with an RNA-guided endonuclease. Such scaffold or structural sequences may further interact with other RNA molecules (e.g., tracrRNA). Methods and techniques for designing targeting constructs and guide nucleic acids for genome editing and site-directed integration at a target site within the genome of a plant using a guided nuclease are known in the art. [0183] In prime editing, a modified guided nuclease, such as a Cas nickase, is fused to an engineered reverse transcriptase (RT) and complexed with a non-coding RNA typically referred to as a pegRNA. A pegRNA comprises three components, from 5’ to 3’: 1) guide sequence complementary to the protospacer on non-PAM strand of the target site, 2) a template sequence comprising the desired nucleotide edit, and 3) a primer binding site complementary to the protospacer on the PAM strand of the target site. Guide sequence- mediated binding of the nuclease-RT fusion to the target site results in a nick in the PAM strand of the target site. The primer binding site of the pegRNA then binds to the nicked (PAM) strand of the target, which positions the template sequence 3’ of the PAM strand to serve as a template for reverse transcription. Ligation of the newly-synthesized strand and the nicked PAM strand results in successful introduction of the desired edit into the gene. Prime 72 sf-6744554 197072001240 editing allows for precise editing of genomic sequences and is further described, for example, in Anzalone et al. ("Search-and-replace genome editing without double-strand breaks or donor DNA." Nature 576.7785 (2019): 149-157.). [0184] An engineered nuclease may be a site-specific nuclease. Several site-specific nucleases, such as recombinases, zinc finger nucleases (ZFNs), meganucleases, and TALENs, are not nucleic acid-guided and instead rely on their protein structure to determine their target site for causing the DSB or nick, or they are fused, tethered or attached to a DNA- binding protein domain or motif. The protein structure of the site-specific nuclease (or the fused/attached/tethered DNA binding domain) may target the site-specific nuclease to the target site. According to many of these embodiments, non-nucleic acid-guided site-specific nucleases, such as recombinases, zinc finger nucleases (ZFNs), meganucleases, and TALENs, may be designed, engineered and constructed according to known methods to target and bind to a target site at or near the genomic locus of an endogenous gene of a plant to create a DSB or nick at such genomic locus to knockout or knockdown expression of the gene via repair of the DSB or nick, which may lead to the creation of a mutation or insertion of a sequence at the site of the DSB or nick, through cellular repair mechanisms, which may be guided by a donor template molecule. [0185] In some embodiments, a site-specific nuclease is a recombinase. A recombinase may be a serine recombinase attached to a DNA recognition motif, a tyrosine recombinase attached to a DNA recognition motif, or other recombinase enzyme known in the art. A recombinase or transposase may be a DNA transposase or recombinase attached or fused to a DNA binding domain. Non-limiting examples of recombinases include a tyrosine recombinase attached, etc., to a DNA recognition motif provided herein is selected from the group consisting of a Cre recombinase, a Gin recombinase, a Flp recombinase, and a Tnp1 recombinase. In an aspect, a Cre recombinase or a Gin recombinase provided herein is tethered to a zinc-finger DNA-binding domain, or a TALE DNA-binding domain, or a Cas9 nuclease. In another aspect, a serine recombinase attached to a DNA recognition motif provided herein is selected from the group consisting of a PhiC31 integrase, an R4 integrase, and a TP-901 integrase. In another aspect, a DNA transposase attached to a DNA binding domain provided herein is selected from the group consisting of a TALE-piggyBac and TALE-Mutator. 73 sf-6744554 197072001240 [0186] A site-specific nuclease may be a zinc finger nuclease (ZFN). ZFNs are synthetic proteins consisting of an engineered zinc finger DNA-binding domain fused to a cleavage domain (or a cleavage half-domain), which may be derived from a restriction endonuclease (e.g., FokI). The DNA binding domain may be canonical (C2H2) or non-canonical (e.g., C3H or C4). The DNA-binding domain can comprise one or more zinc fingers (e.g., 2, 3, 4, 5, 6, 7, 8, 9 or more zinc fingers) depending on the target site. Multiple zinc fingers in a DNA- binding domain may be separated by linker sequence(s). ZFNs can be designed to cleave almost any stretch of double-stranded DNA by modification of the zinc finger DNA-binding domain. ZFNs form dimers from monomers composed of a non-specific DNA cleavage domain (e.g., derived from the FokI nuclease) fused to a DNA-binding domain comprising a zinc finger array engineered to bind a target site DNA sequence. The DNA-binding domain of a ZFN may typically be composed of 3-4 (or more) zinc-fingers. The amino acids at positions -1, +2, +3, and +6 relative to the start of the zinc finger alpha-helix, which contribute to site-specific binding to the target site, can be changed and customized to fit specific target sequences. The other amino acids may form a consensus backbone to generate ZFNs with different sequence specificities. [0187] Methods and rules for designing ZFNs for targeting and binding to specific target sequences are known in the art. See, e.g., US Patent App. Nos. 2005/0064474, 2009/0117617, and 2012/0142062. The FokI nuclease domain may require dimerization to cleave DNA and therefore two ZFNs with their C-terminal regions are needed to bind opposite DNA strands of the cleavage site (separated by 5-7 bp). The ZFN monomer can cut the target site if the two- ZF-binding sites are palindromic. A ZFN, as used herein, is broad and includes a monomeric ZFN that can cleave double stranded DNA without assistance from another ZFN. The term ZFN may also be used to refer to one or both members of a pair of ZFNs that are engineered to work together to cleave DNA at the same site. Without being limited by any theory, because the DNA-binding specificities of zinc finger domains can be re-engineered using one of various methods, customized ZFNs can theoretically be constructed to target nearly any target sequence (e.g., at or near a gene in a plant genome). Publicly available methods for engineering zinc finger domains include Context-dependent Assembly (CoDA), Oligomerized Pool Engineering (OPEN), and Modular Assembly. In an aspect, a method and/or composition provided herein comprises one or more, two or more, three or more, four or more, or five or more ZFNs. In another aspect, a ZFN provided herein is capable of generating a targeted DSB or nick. 74 sf-6744554 197072001240 [0188] A site-specific nuclease may be a TALEN enzyme. TALENs are artificial restriction enzymes generated by fusing the transcription activator-like effector (TALE) DNA binding domain to a nuclease domain (e.g., FokI). When each member of a TALEN pair binds to the DNA sites flanking a target site, the FokI monomers dimerize and cause a double-stranded DNA break at the target site. Besides the wild-type FokI cleavage domain, variants of the FokI cleavage domain with mutations have been designed to improve cleavage specificity and cleavage activity. The FokI domain functions as a dimer, requiring two constructs with unique DNA binding domains for sites in the target genome with proper orientation and spacing. Both the number of amino acid residues between the TALEN DNA binding domain and the FokI cleavage domain and the number of bases between the two individual TALEN binding sites are parameters for achieving high levels of activity. [0189] TALENs are artificial restriction enzymes generated by fusing the transcription activator-like effector (TALE) DNA binding domain to a nuclease domain. In some aspects, the nuclease is selected from a group consisting of PvuII, MutH, TevI, FokI, AlwI, MlyI, SbfI, SdaI, StsI, CleDORF, Clo051, and Pept071. When each member of a TALEN pair binds to the DNA sites flanking a target site, the FokI monomers dimerize and cause a double- stranded DNA break at the target site. The term TALEN, as used herein, is broad and includes a monomeric TALEN that can cleave double stranded DNA without assistance from another TALEN. The term TALEN also refers to one or both members of a pair of TALENs that work together to cleave DNA at the same site. [0190] Transcription activator-like effectors (TALEs) can be engineered to bind practically any DNA sequence, such as at or near the genomic locus of a gene in a plant. TALE has a central DNA-binding domain composed of 13-28 repeat monomers of 33-34 amino acids. The amino acids of each monomer are highly conserved, except for hypervariable amino acid residues at positions 12 and 13. The two variable amino acids are called repeat-variable diresidues (RVDs). The amino acid pairs NI, NG, HD, and NN of RVDs preferentially recognize adenine, thymine, cytosine, and guanine/adenine, respectively, and modulation of RVDs can recognize consecutive DNA bases. This simple relationship between amino acid sequence and DNA recognition has allowed for the engineering of specific DNA binding domains by selecting a combination of repeat segments containing the appropriate RVDs. 75 sf-6744554 197072001240 [0191] Besides the wild-type FokI cleavage domain, variants of the FokI cleavage domain with mutations have been designed to improve cleavage specificity and cleavage activity. The FokI domain functions as a dimer, requiring two constructs with unique DNA binding domains for sites in the target genome with proper orientation and spacing. Both the number of amino acid residues between the TALEN DNA binding domain and the FokI cleavage domain and the number of bases between the two individual TALEN binding sites are parameters for achieving high levels of activity. PvuII, MutH, and TevI cleavage domains are useful alternatives to FokI and FokI variants for use with TALEs. PvuII functions as a highly specific cleavage domain when coupled to a TALE (see Yank et al. 2013. PLoS One. 8: e82539). MutH is capable of introducing strand-specific nicks in DNA (see Gabsalilow et al. 2013. Nucleic Acids Research. 41: e83). TevI introduces double-stranded breaks in DNA at targeted sites (see Beurdeley et al., 2013. Nature Communications. 4: 1762). [0192] The relationship between amino acid sequence and DNA recognition of the TALE binding domain allows for designable proteins. Software programs such as DNAWorks can be used to design TALE constructs. Other methods of designing TALE constructs are known to those of skill in the art. See Doyle et al., Nucleic Acids Research (2012) 40: W117-122.; Cermak et al., Nucleic Acids Research (2011) 39:e82; and tale-nt.cac.cornelledu/about. In another aspect, a TALEN provided herein is capable of generating a targeted DSB. [0193] A site-specific nuclease may be a meganuclease. Meganucleases, which are commonly identified in microbes, such as the LAGLIDADG family of homing endonucleases, are unique enzymes with high activity and long recognition sequences (>14 bp) resulting in site-specific digestion of target DNA. Engineered versions of naturally occurring meganucleases typically have extended DNA recognition sequences (for example, 14 to 40 bp). According to some embodiments, a meganuclease may comprise a scaffold or base enzyme selected from the group consisting of I-CreI, I-CeuI, I-MsoI, I-SceI, I-AniI, and I-DmoI. The engineering of meganucleases can be more challenging than ZFNs and TALENs because the DNA recognition and cleavage functions of meganucleases are intertwined in a single domain. Specialized methods of mutagenesis and high-throughput screening have been used to create novel meganuclease variants that recognize unique sequences and possess improved nuclease activity. Additionally, meganucleases have been combined with transcription activator-like (TAL) effectors to generate megaTALs, which fuse the DNA- binding region of the TAL to allow recognition of specific sequences to meganucleases to 76 sf-6744554 197072001240 increase the activity and specificity of cleavage (see, e.g., Boissel et al. "megaTALs: a rare- cleaving nuclease architecture for therapeutic genome engineering." Nucleic acids research 42.4 (2014): 2591-2601). Thus, a meganuclease may be selected or engineered to bind to a genomic target sequence in a plant, such as at or near the genomic locus of a gene. In another aspect, a meganuclease provided herein is capable of generating a targeted DSB. [0194] In some embodiments, gene editing comprises (a) inducing a DSB in the genome of a cell at a cleavage site (e.g., a cleavage site within a CENH3 gene) at or near a recognition site (e.g., a recognition site within a CENH3 gene) for a natural or engineered nuclease by expressing in the cell the natural or engineered nuclease recognizing said recognition site and inducing said DSB at the cleavage site; (b) introducing into the cell a repair nucleic acid molecule comprising an upstream flanking region having homology to the DNA region upstream of the preselected site (e.g., the recognition site within the CENH3 gene) and/or a downstream flanking DNA region having homology to the DNA region downstream of the preselected site (e.g., the recognition site within the CENH3 gene) for allowing homologous recombination between said flanking region or regions and said DNA region or regions flanking said preselected site; and (c) selecting a cell wherein said repair nucleic acid molecule has been used as a template for making a modification of said genome at said preselected site. In other embodiments, gene editing comprises (a) inducing a DSB in the genome of a cell at a cleavage site (e.g., a cleavage site within a CENH3 gene) at or near a recognition site (e.g., a recognition site within a CENH3 gene) for a natural or engineered nuclease by introducing into the cell the natural or engineered nuclease recognizing said recognition site and inducing said DSB at the cleavage site; (b) introducing into the cell a repair nucleic acid molecule comprising an upstream flanking region having homology to the DNA region upstream of the preselected site (e.g., the recognition site within the CENH3 gene) and/or a downstream flanking DNA region having homology to the DNA region downstream of the preselected site (e.g., the recognition site within the CENH3 gene) for allowing homologous recombination between said flanking region or regions and said DNA region or regions flanking said preselected site; and (c) selecting a cell wherein said repair nucleic acid molecule has been used as a template for making a modification of said genome at said preselected site. [0195] As used herein, a repair nucleic acid molecule is a single-stranded or double- stranded DNA molecule or RNA molecule that is used as a template for modification of the 77 sf-6744554 197072001240 genomic DNA at the preselected site in the vicinity of or at the cleavage site. As used herein, use as a template for modification of the genomic DNA, means that the repair nucleic acid molecule is copied or integrated at the preselected site by homologous recombination between the flanking region(s) and the corresponding homology region(s) in the target genome flanking the preselected site, optionally in combination with non-homologous end- joining (NHEJ) at one of the two ends of the repair nucleic acid molecule (e.g., in case there is only one flanking region). Integration by homologous recombination will allow precise joining of the repair nucleic acid molecule to the target genome up to the nucleotide level, while NHEJ may result in small insertions/deletions at the junction between the repair nucleic acid molecule and genomic DNA. [0196] In some embodiments, the genetic modifications introduced by gene editing result in the decreased expression (including, e.g., decreased activity or non-expression) of one or more CENH3 genes. In gene editing, the introduction of a DSB or nick may be used to introduce targeted genetic modifications in the genome of a plant. According to this approach, genetic modifications, such as deletions, insertions, inversions and/or substitutions may be introduced at a target site (e.g., a target site within a nucleotide of a CENH3 gene) via imperfect repair of the DSB or nick to produce a knock- out or knock-down of a gene. Such genetic modifications may be generated by imperfect repair of the targeted locus even without the use of a donor template molecule, and can result in decreased expression (including, e.g., decreased activity or non-expression) of an endogenous gene product. For example, genetic modifications may be produced by an indel (insertion or deletion of nucleotide bases in a target DNA sequence through NHEJ), or by specific removal of sequence that reduces or completely destroys the function of sequence at or near the targeting site. A knockout of a CENH3 gene may be achieved by inducing a DSB or nick at or near the endogenous locus of the gene that results in non-expression of the gene product (e.g., a CENH3 protein), whereas a knockdown of a gene may be achieved in a similar manner by inducing a DSB or nick at or near the endogenous locus of the gene that is repaired imperfectly at a site that does not affect the coding sequence of the gene in a manner that would eliminate the function of the gene product (e.g., CENH3 protein). For example, the site of the DSB or nick within the endogenous locus may be in the upstream or 5' region of the CENH3 gene (e.g., a promoter and/or enhancer sequence) to affect or reduce its level of expression. Similarly, such targeted knockout or knockdown mutations of a CENH3 gene may be generated with a donor template molecule to direct a particular or desired mutation at 78 sf-6744554 197072001240 or near the target site via repair of the DSB or nick. The donor template molecule may comprise a homologous sequence with or without an insertion sequence and comprising one or more mutations, such as one or more deletions, insertions, inversions and/or substitutions, relative to the targeted genomic sequence at or near the site of the DSB or nick. For example, targeted knockout mutations of a CENH3 gene may be achieved by substituting, inserting, deleting or inverting at least a portion of the CENH3 gene, including, but not limited to, by introducing a frame shift or premature stop codon into a protein coding sequence of the gene. A deletion of a portion of a CENH3 gene may also be introduced by generating DSBs or nicks at two target sites and causing a deletion of the intervening target region flanked by the target sites. [0197] In some embodiments, the genetic modifications are introduced by transgenesis. Transgenes may include, but are not limited to, one or more protein-coding sequences operably linked to a plant-expressible promoter, one or more transcribable DNA sequences encoding an RNA molecule operably linked to a plant-expressible promoter, a gene of interest, a marker gene, or any combination thereof. Methods for the introduction of transgenes in plants are well-known and routine in the art. In some embodiments, transgenesis comprises (a) inducing a DSB in the genome of a cell at a cleavage site at or near a recognition site for a natural or engineered nuclease by expressing in the cell the natural or engineered nuclease recognizing said recognition site and inducing said DSB at the cleavage site; (b) introducing into the cell a repair nucleic acid molecule comprising an upstream flanking region having homology to the DNA region upstream of the preselected site, a downstream flanking DNA region having homology to the DNA region downstream of the preselected site, and a transgene region flanked by the upstream and downstream flanking DNA regions and comprising the transgene to be inserted at the preselected site; and (c) selecting a cell wherein said repair nucleic acid molecule has been used as a template for making a modification of said genome at said preselected site. In other embodiments, transgenesis comprises (a) inducing a DSB in the genome of a cell at a cleavage site at or near a recognition site for a natural or engineered nuclease by introducing into the cell the natural or engineered nuclease recognizing said recognition site and inducing said DSB at the cleavage site; (b) introducing into the cell a repair nucleic acid molecule comprising an upstream flanking region having homology to the DNA region upstream of the preselected site, a downstream flanking DNA region having homology to the DNA region downstream of the preselected site, and a transgene region flanked by the upstream and downstream flanking 79 sf-6744554 197072001240 DNA regions and comprising the transgene to be inserted at the preselected site; and (c) selecting a cell wherein said repair nucleic acid molecule has been used as a template for making a modification of said genome at said preselected site. In some variations, the recognition site is a site within a CENH3 gene. In other variations, the recognition site is within a neutral (e.g., non-coding) site within the genome. [0198] In some embodiments, the genetic modification comprises introducing proteins, nucleic acids, or a combination thereof into a plant cell. The introduction of the proteins, nucleic acids, or combination thereof into the plant cell may be achieved by any of several means known and routinely-used in the art. In some embodiments, the introduction of the proteins, nucleic acids, or combination thereof into the plant cell comprises isolating protoplasts, transfecting the protoplasts, encapsulating the protoplasts, and regenerating plants from the protoplasts. In other embodiments, the introduction of the proteins, nucleic acids, or combination thereof into the plant cell comprises biolistic transformation. In certain embodiments, the introduction of the proteins, nucleic acids, or combination thereof into the plant cell comprises isolating immature plant embryos, bombarding the embryos with particles comprising nucleic acids, and regenerating plants from the immature embryos. Numerous additional transformation methods may be used to introduce the proteins, nucleic acids, or combination thereof into a suitable plant or plant cell. Transformation methods include the use of liposomes, electroporation, chemicals that increase free DNA uptake, injection of the DNA directly into the plant (cell) such as microinjection, particle gun bombardment, transformation using viruses or pollen and microprojection. Methods may be selected from the calcium/polyethylene glycol method for protoplasts (Krens et al. (1982) Nature 296: 72-74; Negrutiu et al. (1987) Plant. Mol. Biol. 8: 363-373); electroporation of protoplasts (Shillito et al. (1985) Bio/Technol. 3: 1099-1102); microinjection into plant material (Crossway et al. (1986) Mol. Gen. Genet. 202: 179-185); DNA or RNA-coated particle bombardment (Klein et al. (1987) Nature 327: 70) infection with (non-integrative) viruses and the like. [0199] Accordingly, in some embodiments, the method of producing the haploid- inducing strawberry plant comprises contacting a plurality of cells of a strawberry plant with one or more nucleic acid molecules, proteins, or a combination thereof into a plurality of cells of a strawberry plant. In some embodiments, the method comprises introducing one or more nucleic acid molecules, proteins, or a combination thereof into a plurality of cells of a 80 sf-6744554 197072001240 strawberry plant. In certain embodiments, the plurality of cells is a plurality of protoplasts. In some embodiments, the method further comprises, subsequent to the contacting step: allowing the plurality of cells (e.g., protoplasts) of the parent strawberry plant to form calli, plants, or a combination thereof; determining the presence of one or more of the genetic modifications in the calli or plants; and identifying one or more calli or plants as having the genetic modifications resulting in decreased expression of one or more CENH3 genes. Methods of detecting genetic modifications are known in the art and include, for example, PCR amplification, DNA sequencing, hybridization assays (e.g., in situ hybridization), chip- based assays, reporter assays, and the like. A method of detecting genetic modifications in strawberry plants is described in Example 2. [0200] In some embodiments, the method comprises contacting a plurality of cells of a parent strawberry plant with one or more expression vectors comprising an expression cassette for a site-directed nuclease (e.g., a Cas nuclease, a meganuclease, a TALEN, a ZFN, or a megaTAL). In certain embodiments, the method comprises contacting a plurality of cells of a parent strawberry plant with one or more expression vectors comprising an expression cassette for a site-directed nuclease contacting a plurality of cells of a parent strawberry plant with one or more expression vectors together comprising an expression cassette for a Cas nuclease and an expression cassette for a guide RNA molecule. [0201] In some embodiments, the method of producing the haploid-inducing strawberry plant comprises contacting a plurality of cells (e.g., protoplasts) of a parent strawberry plant with one or more expression vectors together comprising an expression cassette for a Cas nuclease and an expression cassette for a guide RNA molecule having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity or complementarity to 15, 16, 17, 18, 19, 20, or more consecutive nucleotides of one or more of the CENH3 genes described herein. In other embodiments, the method of producing the haploid- inducing strawberry plant comprises contacting a plurality of cells (e.g., protoplasts) of a parent strawberry plant with one or more Cas nucleases, wherein each Cas nuclease is complexed with a guide RNA molecule having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity or complementarity to 15, 16, 17, 18, 19, 20, or more consecutive nucleotides of one or more of the CENH3 genes described herein. The guide RNA molecule may be a crRNA, a gRNA, or a pegRNA. In certain embodiments, the guide RNA molecule has at least 80%, at 81 sf-6744554 197072001240 least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity or complementarity to 15, 16, 17, 18, 19, 20, or more consecutive nucleotides of a nucleotide sequence encoding one or more CENH3 proteins having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity, or at least 60%, 65%, 70%, 75%, 80%, 85%, 95%, 96%, 97%, 98%, or 99% sequence similarity to an amino acid sequence selected from the group consisting of SEQ ID NOs: 8-14 and 53-56. In certain embodiments, the guide RNA molecule has at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity or complementarity to 15, 16, 17, 18, 19, 20, or more consecutive nucleotides of a nucleotide sequence having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity or complementarity to a nucleotide sequence selected from the group consisting of SEQ ID NOs: 1-7 and 58-61. In certain embodiments, the guide RNA molecule has at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity or complementarity to 15, 16, 17, 18, 19, 20, or more consecutive nucleotides of an enhancer of one or more CENH3 genes described herein, a promoter of one or more CENH3 genes described herein, a coding region of one or more CENH3 genes, or an intron of one or more of the CENH3 genes described herein. In certain embodiments, the guide RNA has 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity or complementarity to a nucleotide sequence selected from the group consisting of SEQ ID NOs: 25-34. In certain embodiments, the guide RNA has 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity or complementarity to SEQ ID NO: 27, SEQ ID NO: 29, or SEQ ID NO: 31. [0202] In some embodiments, the method of producing the haploid-inducing strawberry plant comprises contacting a plurality of cells (e.g., protoplasts) of a parent strawberry plant of the species Fragaria vesca with one or more expression vectors together comprising an expression cassette for a Cas nuclease and an expression cassette for a guide RNA molecule having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity or complementarity to 15, 16, 17, 18, 19, 20, or more consecutive nucleotides of an FvCENH3 gene. In other embodiments, the method of producing the haploid-inducing strawberry plant comprises contacting a plurality of cells (e.g., protoplasts) of a parent strawberry plant of the species Fragaria vesca with one or more 82 sf-6744554 197072001240 Cas nucleases, wherein each Cas nuclease is complexed with a guide RNA molecule having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity or complementarity to 15, 16, 17, 18, 19, 20, or more consecutive nucleotides of an FvCENH3 gene. The guide RNA molecule may be a crRNA, a gRNA, or a pegRNA. In certain embodiments, the guide RNA molecule has at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity or complementarity to 15, 16, 17, 18, 19, 20, or more consecutive nucleotides of a nucleotide sequence encoding a CENH3 protein having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity, or at least 60%, 65%, 70%, 75%, 80%, 85%, 95%, 96%, 97%, 98%, or 99% sequence similarity to the amino acid sequence of SEQ ID NO: 8. In certain embodiments, the guide RNA molecule has at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity or complementarity to 15, 16, 17, 18, 19, 20, or more consecutive nucleotides of a nucleotide sequence having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity or complementarity to the nucleotide sequence of SEQ ID NO: 1. In certain embodiments, the guide RNA molecule has at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity or complementarity to 15, 16, 17, 18, 19, 20, or more consecutive nucleotides of an enhancer of FvCENH3, a promoter of FvCENH3, a coding region of FvCENH3, or an intron of FvCENH3. [0203] In other embodiments, the method of producing the haploid-inducing strawberry plant comprises contacting a plurality of cells (e.g., protoplasts) of a parent strawberry plant of the species Fragaria vesca with one or more Cas nucleases, wherein each Cas nuclease is complexed with a guide RNA molecule having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity or complementarity to SEQ ID NO: 27, SEQ ID NO: 29, SEQ ID NO: 31, or a combination thereof. In certain embodiments, the method of producing the haploid-inducing strawberry plant comprises contacting a plurality of cells protoplasts of a parent strawberry plant of the species Fragaria vesca with a plurality of Cas nucleases, wherein a first portion of the plurality of the Cas nucleases are each complexed with a guide RNA molecule having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity or complementarity to SEQ ID NO: 27, wherein a second portion of 83 sf-6744554 197072001240 the plurality of the Cas nucleases are each complexed with a guide RNA molecule having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity or complementarity to SEQ ID NO: 29; and wherein a third portion of the plurality of the Cas nucleases are each complexed with a guide RNA molecule having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity or complementarity to SEQ ID NO: 31. [0204] In some embodiments, the method of producing the haploid-inducing strawberry plant comprises contacting a plurality of cells (e.g., protoplasts) of a parent strawberry plant of the species Fragaria x ananassa with one or more expression vectors together comprising an expression cassette for a Cas nuclease and an expression cassette for one or more guide RNA molecules having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity or complementarity to 15, 16, 17, 18, 19, 20, or more consecutive nucleotides of an FaCENH3-1a gene, an FaCENH3-1b gene, an FaCENH3-2a gene, an FaCENH3-2b gene, an FaCENH3-3 gene, an FaCENH3-4 gene, or any combination (e.g., two, three, four, five or all six) thereof. In other embodiments, the method of producing the haploid-inducing strawberry plant comprises contacting a plurality of cells (e.g., protoplasts) of a parent strawberry plant of the species Fragaria x ananassa with one or more Cas nucleases, wherein each Cas nuclease is complexed with a guide RNA molecule having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity or complementarity to 15, 16, 17, 18, 19, 20, or more consecutive nucleotides of an FaCENH3-1a gene, an FaCENH3-1b gene, an FaCENH3-2a gene, an FaCENH3-2b gene, an FaCENH3-3 gene, an FaCENH3-4 gene, or any combination (e.g., two, three, four, five or all six) thereof. The guide RNA molecule may be a crRNA, a gRNA, or a pegRNA. In certain embodiments, the guide RNA molecule has at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity or complementarity to 15, 16, 17, 18, 19, 20, or more consecutive nucleotides of a nucleotide sequence encoding a CENH3 protein having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity, or at least 60%, 65%, 70%, 75%, 80%, 85%, 95%, 96%, 97%, 98%, or 99% sequence similarity to an amino acid sequence selected from the group consisting of SEQ ID NOs: 9-14 and 53-56. In certain embodiments, the guide RNA molecule has at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity or complementarity to 15, 16, 17, 84 sf-6744554 197072001240 18, 19, 20, or more consecutive nucleotides of a nucleotide sequence having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity or complementarity to a nucleotide sequence selected from the group consisting of SEQ ID NOs: 2-7 and 58-61. In certain embodiments, the guide RNA molecule has at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity or complementarity to 15, 16, 17, 18, 19, 20, or more consecutive nucleotides of an enhancer, a promoter, a coding region, or an intron of an FaCENH3-1a gene, an FaCENH3-1b gene, an FaCENH3-2a gene, an FaCENH3-2b gene, an FaCENH3-3 gene, an FaCENH3-4 gene, or any combination (e.g., two, three, four, five or all six) thereof. [0205] In other embodiments, the method of producing the haploid-inducing strawberry plant comprises contacting a plurality of cells (e.g., protoplasts) of a parent strawberry plant of the species Fragaria x ananassa with one or more Cas nucleases, wherein each Cas nuclease is complexed with a guide RNA molecule having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity or complementarity to SEQ ID NO: 27, SEQ ID NO: 29, or SEQ ID NO: 31, or a combination thereof. In certain embodiments, the method of producing the haploid-inducing strawberry plant comprises contacting a plurality of cells protoplasts of a parent strawberry plant of the species Fragaria x ananassa with a plurality of Cas nucleases, wherein a first portion of the plurality of the Cas nucleases are each complexed with a guide RNA molecule having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity or complementarity to SEQ ID NO: 27, and wherein a second portion of the plurality of the Cas nucleases are each complexed with a guide RNA molecule having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity or complementarity to SEQ ID NO: 29, and wherein a third portion of the plurality of the Cas nucleases are each complexed with a guide RNA molecule having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity or complementarity to SEQ ID NO: 31. [0206] In some embodiments, the decreased expression of each of the one or more of the CENH3 genes is independently achieved by introducing an insertion, a deletion, one or more nucleotide changes, or an inversion into the CENH3 gene that that results in decreased expression of the CENH3 gene. In some variations, the insertion, the deletion, the one or 85 sf-6744554 197072001240 more nucleotide changes, or the inversion is introduced into the first 70%, the first 60%, the first 50%, the first 40%, the first 30%, the first 20%, or the first 10% of the nucleotides of the CENH3 gene (e.g., a sequence selected from the group consisting of SEQ ID NOs: 1-7). In certain variations, the insertion, the deletion, the one or more nucleotide changes, or the inversion is introduced into the first 100, the first 200, the first 300, the first 400, the first 500, the first 600, the first 700, the first 800, the first 900, the first 1000, the first 1250, the first 1500, the first 1750, the first 2000, the first 2500, or the first 3000 nucleotides of the CENH3 gene (e.g., a sequence selected from the group consisting of SEQ ID NOs: 1-7). In some variations, the insertion, the deletion, the one or more nucleotide changes, or the inversion decreases, but does not eliminate, expression of the CENH3 gene. In some variations, the insertion, the deletion, the one or more nucleotide changes, or the inversion eliminates expression (e.g., eliminates activity) of the CENH3 gene. In some variations, the activity of the CENH3 gene is eliminated by a premature stop codon introduced into the first 70%, the first 60%, the first 50%, the first 40%, the first 30%, the first 20%, or the first 10% of a sequence selected from the group consisting of SEQ ID NOs: 1-7 or of the nucleotides of the coding sequence of the CENH3 gene following the start codon in the 3’ direction. In certain variations, the activity of the CENH3 gene is eliminated by a premature stop codon introduced into the first 100, the first 200, the first 300, the first 400, the first 500, the first 600, the first 700, the first 800, the first 900, the first 1000, the first 1250, the first 1500, the first 1750, the first 2000, the first 2500, or the first 3000 nucleotides of a sequence selected from the group consisting of SEQ ID NOs: 1-7 or of the coding sequence of the CENH3 gene following the start codon in the 3’ direction. Compositions for Producing Haploid-inducing Strawberry Plants [0207] In some aspects, provided herein are expression vectors, isolated DNA molecules, bacterial cells, and kits for making a haploid-inducing strawberry plant described herein. Also provided are genetically modified plants, plant parts, plant cells, and seeds comprising said expression vectors and isolated DNA molecules. [0208] In one aspect, provided herein are expression vectors for making a haploid- inducing strawberry plant described herein. In some embodiments, the expression vector comprises one or more expression cassettes. The expression cassettes may comprise, for example, a nucleic acid encoding site-directed nuclease operably linked to a promoter, or a nucleic acid encoding a non- coding RNA (e.g., a guide RNA, an siRNA, or the like) 86 sf-6744554 197072001240 operably linked to a promoter. In some embodiments, the expression vector comprises at least a first expression cassette comprising a nucleic acid encoding a guide RNA operably linked to a promoter. In certain embodiments, the expression vector comprises one or more additional expression cassettes, for example, a second expression cassette comprising a nucleic acid encoding a Cas nuclease operably linked to a promoter, and/or one or more additional expression cassettes each comprising a nucleic acid encoding a guide RNA operably linked to a promoter. In certain embodiments, the expression vector comprises a DNA sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity or complementarity to 15, 16, 17, 18, 19, 20, or more consecutive nucleotides of a nucleotide sequence encoding one or more CENH3 proteins having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity, or at least 60%, 65%, 70%, 75%, 80%, 85%, 95%, 96%, 97%, 98%, or 99% sequence similarity to an amino acid sequence selected from the group consisting SEQ ID NOs: 8-14 and 53-56. In certain embodiments, the expression vector comprises a DNA sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity or complementarity to 15, 16, 17, 18, 19, 20, or more consecutive nucleotides of a nucleotide sequence having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity or complementarity to a nucleotide sequence selected from the group consisting of SEQ ID NOs: 1-7 and 58-61. In some embodiments, the expression vector comprises a DNA sequence encoding a site- directed nuclease. In certain embodiments, the site-directed nuclease is a Cas nuclease, a TALEN, ZFN, or a mega-TAL. [0209] In some embodiments, provided herein is an expression vector for making a haploid- inducing strawberry plant, wherein the expression vector comprises at least a first expression cassette comprising a nucleic acid encoding a non-coding RNA molecule operably linked to a promoter. In certain embodiments, the non-coding RNA is a crRNA, a gRNA, a pegRNA, a siRNA, a miRNA, or a dsRNA. In some embodiments, the non-coding RNA molecule (e.g., guide RNA) has at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity or complementarity to 15, 16, 17, 18, 19, 20, or more consecutive nucleotides of a nucleotide sequence encoding one or more CENH3 proteins having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity, 87 sf-6744554 197072001240 or at least 60%, 65%, 70%, 75%, 80%, 85%, 95%, 96%, 97%, 98%, or 99% sequence similarity to an amino acid sequence selected from the group consisting of SEQ ID NOs: 8- 14 and 53-56. In certain embodiments, the non- coding RNA molecule (e.g., guide RNA) has at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity or complementarity to 15, 16, 17, 18, 19, 20, or more consecutive nucleotides of a nucleotide sequence having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity or complementarity to a nucleotide sequence selected from the group consisting of SEQ ID NOs: 1-7 and 58-61. In certain embodiments, the non-coding RNA molecule (e.g., guide RNA) has at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity or complementarity to 15, 16, 17, 18, 19, 20, or more consecutive nucleotides of an enhancer of one or more CENH3 genes described herein, a promoter of one or more CENH3 genes described herein, a coding region of one or more CENH3 genes, or an intron of one or more of the CENH3 genes described herein. In certain embodiments, the non-coding RNA molecule is a guide RNA having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity or complementarity to SEQ ID NO: 17, SEQ ID NO: 19, SEQ ID NO: 21, or any combination thereof. In certain embodiments, the expression vector comprises a first expression cassette comprising a nucleic acid encoding a guide RNA having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity or complementarity to SEQ ID NO: 17, SEQ ID NO: 19, SEQ ID NO: 21, or any combination thereof. In certain embodiments, the expression vector comprises a first expression cassette comprising a nucleic acid encoding a guide RNA having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity or complementarity to SEQ ID NO: 17, a second expression cassette comprising a nucleic acid encoding a guide RNA having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity or complementarity to SEQ ID NO: 19, and a third expression cassette comprising a nucleic acid encoding a guide RNA having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity or complementarity to SEQ ID NO: 21. In some embodiments, the expression vector further comprises a DNA sequence encoding a Cas nuclease operably linked to a promoter. 88 sf-6744554 197072001240 [0210] In some embodiments, provided herein is an expression vector comprising a DNA sequence encoding a site-directed nuclease operably linked to a promoter. In certain embodiments, the site-directed nuclease is a Cas nuclease, a TALEN, ZFN, or a mega-TAL. In some embodiments, the expression vector comprises a DNA sequence encoding a TALEN, ZFN, or a mega-TAL operably linked to a promoter, wherein the TALEN, ZFN, or a mega- TAL comprises an amino acid sequence that confers binding to a polynucleotide sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity or complementarity to 15, 16, 17, 18, 19, 20, or more consecutive nucleotides of a nucleotide sequence encoding one or more CENH3 proteins having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity, or at least 60%, 65%, 70%, 75%, 80%, 85%, 95%, 96%, 97%, 98%, or 99% sequence similarity to an amino acid sequence selected from the group consisting of SEQ ID NOs: 8-14 and 53-56. In some embodiments, the expression vector comprises a DNA sequence encoding a TALEN, ZFN, or a mega-TAL operably linked to a promoter, wherein the TALEN, ZFN, or a mega-TAL comprises an amino acid sequence that confers binding to a polynucleotide sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity or complementarity to 15, 16, 17, 18, 19, 20, or more consecutive nucleotides of a nucleotide sequence having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity or complementarity to a nucleotide sequence selected from the group consisting of SEQ ID NOs: 1-7 and 58-61. [0211] In some embodiments, provided herein is an isolated DNA molecule for making a haploid-inducing strawberry plant, wherein the isolated DNA molecule has at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity or complementarity to at least 15, 16, 17, 18, 19, 20, 50, 100, or more consecutive nucleotides of a nucleotide sequence encoding one or more CENH3 proteins having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity, or at least 60%, 65%, 70%, 75%, 80%, 85%, 95%, 96%, 97%, 98%, or 99% sequence similarity to an amino acid sequence selected from the group consisting of SEQ ID NOs: 8-14 and 53-56. In some embodiments, provided herein is an isolated DNA molecule for making a haploid-inducing strawberry plant, wherein the isolated DNA molecule has at least 80%, at least 85%, at least 89 sf-6744554 197072001240 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity or complementarity to at least 15, 16, 17, 18, 19, 20, 50, 100, or more consecutive nucleotides of a nucleotide sequence having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity or complementarity to a nucleotide sequence selected from the group consisting of SEQ ID NOs: 1-7 and 58-61. In some embodiments, the isolated DNA molecule is a synthetic oligo nucleotide. [0212] In some embodiments, provided herein is an isolated nucleic acid molecule for making a haploid-inducing strawberry plant, wherein the isolated nucleic acid molecule has at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity or complementarity to at least 15, 16, 17, 18, 19, 20, 50, 100, or more consecutive nucleotides of a nucleotide sequence encoding one or more CENH3 proteins having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity, or at least 60%, 65%, 70%, 75%, 80%, 85%, 95%, 96%, 97%, 98%, or 99% sequence similarity to an amino acid sequence selected from the group consisting of SEQ ID NOs: 8-14 and 53-56. In some embodiments, provided herein is an isolated nucleic acid molecule for making a haploid- inducing strawberry plant, wherein the isolated RNA molecule has at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity or complementarity to at least 15, 16, 17, 18, 19, 20, 50, 100, or more consecutive nucleotides of a nucleotide sequence having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity or complementarity to a nucleotide sequence selected from the group consisting of SEQ ID NOs: 25-34. In certain embodiments, the isolated nucleic acid molecule has at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity or complementarity to a nucleotide sequence selected from the group consisting of SEQ ID NOs: 15-24. In certain embodiments, the isolated nucleic acid molecule has at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity or complementarity to SEQ ID NO: 17, SEQ ID NO: 19, or SEQ ID NO: 21, or a combination thereof. In some embodiments, the isolated nucleic acid molecule comprises DNA, RNA, or both. In some embodiments, the isolated nucleic acid molecule is an isolated RNA molecule. In certain embodiments, the isolated RNA molecule is a guide RNA. In certain embodiments, the guide 90 sf-6744554 197072001240 RNA has at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity or complementarity to SEQ ID NO: 27, SEQ ID NO: 29, or SEQ ID NO: 31, or a combination thereof. In certain embodiments, the isolated nucleic acid molecule further comprises a nucleic acid sequence for association with a CRISPR-associated (Cas) enzyme. [0213] In some aspects, provided herein are genetically modified plants, plant parts, plant cells, or seed comprising any of the expression vectors or isolated DNA molecules described herein. In some embodiments, the genetically modified plants, plant parts, plant cells, or seeds are genetically modified strawberry plants, strawberry plant parts (e.g., achenes, stolons, or any strawberry plant part described herein), strawberry plant cells, or strawberry seeds. [0214] In other aspects, provided herein is a bacterial cell comprising any of the expression vectors or isolated DNA molecules described herein. In some embodiments, the bacterial cell is an Agrobacterium cell. In some aspects, provided herein are genetically modified plants, plant parts, plant cells, or seed comprising said bacterial cell. [0215] In yet other aspects, provided herein is a kit for producing a haploid-inducing strawberry plant comprising one or more of the expression vectors, isolated DNA molecules, isolated RNA molecules, bacterial cells, or any combination thereof described herein. In some embodiments, the kit further comprises instructions for using the kit components to produce a haploid-inducing strawberry plant. [0216] In yet other aspects, provided herein is a kit for producing a true homozygous octoploid strawberry plant comprising one or more cells or plant parts of any of the haploid- inducing strawberry plants described herein. In some embodiments, the kit further comprises instructions for using the kit components to produce a true homozygous octoploid strawberry plant. In some embodiments, the kit further comprises a chromosome doubling agent described herein. In certain embodiments, the kit further comprises colchicine, oryzalin, oxide, trifluralin, or any combination thereof. Methods of Producing True Homozygous Octoploid Strawberry Lines [0217] In some aspects, described herein are methods of producing true homozygous octoploid strawberry plant, comprising (a) generating a tetrahaploid strawberry cell or plant 91 sf-6744554 197072001240 through haploid induction, and (b) subjecting the tetrahaploid strawberry plant to genome doubling to produce a doubled tetrahaploid strawberry cell or plant. A doubled tetrahaploid cell or plant may also be described as a homozygous octoploid cell or plant having the same haplotype for each of the chromosomes within each of the subgenomes of the plant. In some embodiments, the method comprises (a) contacting a tetrahaploid cell from a donor octoploid strawberry plant with pollen from a haploid-inducing strawberry plant described herein, and (b) producing a doubled tetrahaploid cell or plant from the tetrahaploid cell. In some embodiments, the method comprises (a) contacting a tetrahaploid cell from a donor octoploid strawberry plant with pollen from a haploid-inducing strawberry plant described herein, and (b) producing an octoploid cell or plant from the doubled tetrahaploid cell or plant. In some embodiments, the method comprises (a1) contacting a reduced egg cell from a donor octoploid strawberry plant with a sperm cell from a haploid-inducing strawberry plant described herein, (a2) allowing a tetrahaploid embryo to form and grow into a tetrahaploid strawberry plant, and (b) subjecting the tetrahaploid strawberry plant to genome doubling to produce a doubled tetrahaploid strawberry plant. In some embodiments, the method comprises (a1) contacting a tetraploid reduced egg cell (a tetrahaploid cell) from a donor octoploid strawberry plant with a sperm cell from a haploid- inducing strawberry plant described herein, (a2) allowing a tetrahaploid embryo to form and grow into a tetrahaploid strawberry plant, and (b) subjecting the tetrahaploid strawberry plant to genome doubling to produce an octoploid strawberry plant from the doubled tetrahaploid. Haploid Induction [0218] In some embodiments, the methods of producing true homozygous octoploid strawberry plant, or plant part thereof, comprise generating a tetrahaploid strawberry plant through haploid induction, or the process of crossing gametes from a haploid-inducing strawberry plant with a donor (non-haploid-inducing) strawberry plant to generate embryos and seeds with half the ploidy of the donor strawberry plant. During a normal cross with two non-haploid-inducing plant plants, two haploid sperm cells migrate down the pollen tube to the ovule. One of the sperm cells fertilizes the central cell of the ovule to form the endosperm, while the other sperm cell fertilizes the haploid reduced egg cell to form an embryo having the ploidy of the parent plant. For example, if both parent plants are octoploid, the haploid (4x) sperm and egg form an octoploid (8x) embryo in a normal fertilization process. During a haploid induction cross beteen a non-haploid inducing plant 92 sf-6744554 197072001240 and a haploid-inducing plants with reduced expression of CENH3, embryogenesis is initiated in the egg cell with only the haploid genome of the male or female gamete, resulting in a haploid embryo. When pollen from a haploid-inducing plant is used to pollenate a non- haploid-inducing plant, the sperm cells from the haploid-inducing plant migrate down the pollen tube to the ovule, where one of the sperm cells fertilizes the central cell to form the endosperm. However, when the other sperm cell contacts the reduced egg cell, embryogenesis is initiated, but the DNA from the sperm cell is not delivered to or retained in the egg cell, resulting in formation of a haploid embryo having a nuclear genome derived from only the maternal plant, and not the paternal plant. For example, in haploid induction using pollen from the haploid-inducing plant, if both parent plants are octoploid, the haploid (4x) egg cell does not incorporate the DNA from the haploid sperm cell (4x), resulting in a haploid (4x) embryo, which may also be referred to herein as a tetrahaploid. By contrast, when pollen from a non-haploid inducing plant is used to pollenate a haploid-inducing plant having decreased expression of CENH3, the sperm cells from the non-haploid-inducing plant migrate down the pollen tube to the ovule, where one of the sperm cells fertilizes the central cell to form the endosperm and the other sperm cell fertilizes the egg. Fertilization of the haploid-inducing egg cell with the non-haploid-inducing sperm cell results in retention of the paternal nuclear DNA present in the sperm cell, but elimination of the maternal nuclear DNA present in the egg cell. This results in resulting in formation of a haploid embryo having a nuclear genome derived from only the paternal plant, and a cytoplasmic genome (e.g., mitochondrial and/or chloroplast genome) derived from the maternal plant. In this scenario, if both the maternal haploid-inducing parent and the paternal parent are octoploid, haploid induction results in a tetrahaploid (4x) embryo. [0219] In some embodiments, the step of generating a tetrahaploid strawberry plant through haploid induction comprises contacting a tetrahaploid cell from a donor octoploid strawberry plant with pollen, or a part thereof, from a haploid-inducing strawberry plant described herein. In certain embodiments, the tetrahaploid cell is a reduced egg cell. In certain embodiments, the tetrahaploid cell is a reduced egg cell and the part of the pollen from the haploid-inducing strawberry plant is a sperm cell. In some embodiments, the step of generating a tetrahaploid strawberry plant through haploid induction comprises contacting pollen of the haploid-inducing strawberry plant with the stigma of a pistil of the donor octoploid strawberry plant comprising the reduced egg cell, allowing formation of a pollen tube and migration of one or more (e.g., two) sperm cells of the pollen to the reduced egg 93 sf-6744554 197072001240 cell, thereby contacting the reduced egg cell with the sperm cells. In some embodiments, the step of generating a tetrahaploid strawberry plant through haploid induction comprises allowing the donor octoploid strawberry plant to form a plurality of seeds. In certain embodiments, the plurality of seeds comprises one or more tetrahaploid seeds. The method may further comprise a step of selecting one or more tetrahaploid seeds of the plurality of seeds by 1) determining the ploidy of the embryo and/or endosperm of one or more seeds of the plurality of seeds, and 2) selecting one or more seeds having a tetrahaploid embryo. In certain embodiments, the step of selecting one or more tetrahaploid seeds of the plurality of seeds comprises 1) determining the ploidy of the embryo and/or endosperm of one or more seeds of the plurality of seeds, and 2) selecting one or more seeds having a tetraploid embryo and an endosperm having a ploidy of greater than 8x (e.g., a ploidy of 9x, 10x, 11x, or 12x). [0220] In some embodiments, the step of generating a tetrahaploid strawberry plant through haploid induction comprises contacting a tetrahaploid cell from a donor octoploid strawberry plant with an egg cell from a haploid-inducing strawberry plant described herein. In certain embodiments, the tetrahaploid cell is a reduced sperm cell. In some embodiments, the step of generating a tetrahaploid strawberry plant through haploid induction comprises contacting the stigma of a pistil of the haploid-inducing strawberry plant with the pollen of the donor octoploid strawberry plant comprising the reduced egg cell, allowing formation of a pollen tube and migration of one or more (e.g., two) reduced sperm cells of the pollen to the egg cell, thereby contacting the egg cell with the reduced sperm cells. In some embodiments, the step of generating a tetrahaploid strawberry plant through haploid induction comprises allowing the haploid-inducing strawberry plant to form a plurality of seeds. In certain embodiments, the plurality of seeds comprises one or more tetrahaploid seeds. The method may further comprise a step of selecting one or more tetrahaploid seeds of the plurality of seeds by 1) determining the ploidy of the embryo and/or endosperm of one or more seeds of the plurality of seeds, and 2) selecting one or more seeds having a tetrahaploid embryo. In certain embodiments, the step of selecting one or more tetrahaploid seeds of the plurality of seeds comprises 1) determining the ploidy of the embryo and/or endosperm of one or more seeds of the plurality of seeds, and 2) selecting one or more seeds having a tetraploid embryo and an endosperm having a ploidy of greater than 8x (e.g., a ploidy of 9x, 10x, 11x, or 12x). In some embodiments, the haploid-inducing strawberry plant and the donor octoploid strawberry plants are from different species of Fragaria. In some embodiments, the haploid- inducing strawberry plant is diploid. In certain embodiments, the haploid-inducing strawberry 94 sf-6744554 197072001240 plant is a plant of the species Fragaria vesca. In some embodiments, the donor octoploid strawberry plant is a plant of the species Fragaria x ananassa. In some embodiments, the embryo of the true homozygous octoploid strawberry seed has a nuclear genome derived from only the donor octoploid strawberry plant and a cytoplasmic genome (e.g., mitochondrial and/or chloroplast genome) derived from the haploid-inducing strawberry plant. [0221] The haploid-inducing strawberry plant and the donor strawberry plant may have the same ploidy or different ploidies. For example, a haploid-inducing strawberry plant may be diploid (2x) and produce haploid (1x) pollen or egg cells that can be crossed with an octoploid (8x) strawberry donor plant, resulting in tetraploid (4x) embryos, also referred to as tetrahaploid embryos. Accordingly, in some embodiments, the method of producing true homozygous octoploid strawberry plant, or plant part thereof, comprises (a) contacting a tetrahaploid cell from a donor octoploid strawberry plant with pollen from a diploid, triploid, tetraploid, pentaploid, hexaploid, septaploid, or octoploid haploid-inducing strawberry plant described herein, and (b) producing an octoploid doubled tetrahaploid cell or plant from the tetrahaploid cell. In certain embodiments, the method comprises (a1) contacting a reduced egg cell from a donor octoploid strawberry plant with a sperm cell from a diploid, triploid, tetraploid, pentaploid, hexaploid, septaploid, or octoploid haploid-inducing strawberry plant described herein, (a2) allowing a tetrahaploid embryo to form and grow into a tetrahaploid strawberry plant, and (b) subjecting the tetrahaploid strawberry plant to genome doubling to produce a doubled tetrahaploid strawberry plant. In certain embodiments, the method comprises (a) contacting a tetrahaploid cell from a donor octoploid strawberry plant with pollen from a diploid, triploid, tetraploid, pentaploid, hexaploid, septaploid, or octoploid haploid-inducing strawberry plant described herein, and (b) producing an octoploid doubled tetrahaploid cell or plant from the tetrahaploid cell. In certain embodiments, the method comprises (a1) contacting a tetrahaploid reduced egg cell from a donor octoploid strawberry plant with a sperm cell from a diploid, triploid, tetraploid, pentaploid, hexaploid, septaploid, or octoploid haploid-inducing strawberry plant described herein, (a2) allowing a tetrahaploid embryo to form and grow into a tetrahaploid strawberry plant, and (b) subjecting the tetrahaploid strawberry plant to genome doubling to produce an octoploid doubled tetrahaploid strawberry plant. 95 sf-6744554 197072001240 [0222] In other embodiments, the method of producing true homozygous octoploid strawberry plant, or plant part thereof, comprises (a) contacting a tetrahaploid cell from a donor octoploid strawberry plant with an egg cell from a diploid, triploid, tetraploid, pentaploid, hexaploid, septaploid, or octoploid haploid-inducing strawberry plant described herein, and (b) producing an octoploid doubled tetrahaploid cell or plant from the tetrahaploid cell. In certain embodiments, the method comprises (a1) contacting a reduced sperm cell from a donor octoploid strawberry plant with an egg cell from a diploid, triploid, tetraploid, pentaploid, hexaploid, septaploid, or octoploid haploid-inducing strawberry plant described herein, (a2) allowing a tetrahaploid embryo to form and grow into a tetrahaploid strawberry plant, and (b) subjecting the tetrahaploid strawberry plant to genome doubling to produce a doubled tetrahaploid strawberry plant. In certain embodiments, the method comprises (a) contacting a tetrahaploid cell from a donor octoploid strawberry plant with an egg cell from a diploid, triploid, tetraploid, pentaploid, hexaploid, septaploid, or octoploid haploid-inducing strawberry plant described herein, and (b) producing an octoploid doubled tetrahaploid cell or plant from the tetrahaploid cell. In certain embodiments, the method comprises (a1) contacting a tetrahaploid reduced sperm cell from a donor octoploid strawberry plant with an egg cell from a diploid, triploid, tetraploid, pentaploid, hexaploid, septaploid, or octoploid haploid-inducing strawberry plant described herein, (a2) allowing a tetrahaploid embryo to form and grow into a tetrahaploid strawberry plant, and (b) subjecting the tetrahaploid strawberry plant to genome doubling to produce an octoploid doubled tetrahaploid strawberry plant. In some embodiments, the haploid-inducing strawberry plant and the donor octoploid strawberry plants are from different species of Fragaria. In some embodiments, the haploid- inducing strawberry plant is diploid. In certain embodiments, the haploid-inducing strawberry plant is a plant of the species Fragaria vesca. In some embodiments, the donor octoploid strawberry plant is a plant of the species Fragaria x ananassa. In some embodiments, the embryo of the true homozygous octoploid strawberry seed has a nuclear genome derived from only the donor octoploid strawberry plant and a cytoplasmic genome (e.g., mitochondrial and/or chloroplast genome) derived from the haploid-inducing strawberry plant. [0223] In some embodiments, the method of producing a true homozygous octoploid strawberry plant, or plant part thereof, comprises (a) contacting a haploid cell from a donor octoploid strawberry plant with pollen from a diploid haploid-inducing strawberry plant described herein, and (b) producing an octoploid cell from a doubled tetrahaploid or plant 96 sf-6744554 197072001240 from the haploid cell. In certain embodiments, the method comprises (a1) contacting a reduced egg cell from a donor octoploid strawberry plant with a sperm cell from a diploid haploid-inducing strawberry plant described herein, (a2) allowing a tetrahaploid embryo to form and grow into a tetrahaploid strawberry plant, and (b) subjecting the tetrahaploid strawberry plant to genome doubling to produce a doubled tetrahaploid strawberry plant. In certain embodiments, the method comprises (a) contacting a tetrahaploid cell from a donor octoploid strawberry plant with pollen from a diploid haploid-inducing strawberry plant described herein, and (b) producing an octoploid doubled tetrahaploid cell or plant from the tetrahaploid cell. In certain embodiments, the method comprises (a1) contacting a tetrahaploid reduced egg cell from a donor octoploid strawberry plant with a sperm cell from a diploid haploid-inducing strawberry plant described herein, (a2) allowing a tetrahaploid embryo to form and grow into a tetrahaploid strawberry plant, and (b) subjecting the tetrahaploid strawberry plant to genome doubling to produce an octoploid doubled tetrahaploid strawberry plant. In some embodiments, the diploid haploid-inducing strawberry plant is a plant of the species Fragaria vesca, Fragaria iinumae, Fragaria nipponica, Fragaria viridis, Fragaria × bifera, Fragaria bucharica, Fragaria chinensis, Fragaria daltoniana, Fragaria emeiensis, Fragaria hayatae, Fragaria iinumae, Fragaria mandshurica, Fragaria viridis, Fragaria nilgerrensis, Fragaria nipponica, Fragaria nubicola, or Fragaria pentaphylla. In certain embodiments, the diploid haploid-inducing strawberry plant is a plant of the species Fragaria vesca. [0224] In some embodiments, the method of producing a true homozygous octoploid strawberry plant, or plant part thereof, comprises (a) contacting a haploid cell from a donor octoploid strawberry plant with an egg cell from a diploid haploid-inducing strawberry plant described herein, and (b) producing an octoploid cell from a doubled tetrahaploid or plant from the haploid cell. In certain embodiments, the method comprises (a1) contacting a reduced sperm cell from a donor octoploid strawberry plant with an egg cell from a diploid haploid-inducing strawberry plant described herein, (a2) allowing a tetrahaploid embryo to form and grow into a tetrahaploid strawberry plant, and (b) subjecting the tetrahaploid strawberry plant to genome doubling to produce a doubled tetrahaploid strawberry plant. In certain embodiments, the method comprises (a) contacting a tetrahaploid cell from a donor octoploid strawberry plant with an egg cell from a diploid haploid-inducing strawberry plant described herein, and (b) producing an octoploid doubled tetrahaploid cell or plant from the tetrahaploid cell. In certain embodiments, the method comprises (a1) contacting a 97 sf-6744554 197072001240 tetrahaploid reduced sperm cell from a donor octoploid strawberry plant with an egg cell from a diploid haploid-inducing strawberry plant described herein, (a2) allowing a tetrahaploid embryo to form and grow into a tetrahaploid strawberry plant, and (b) subjecting the tetrahaploid strawberry plant to genome doubling to produce an octoploid doubled tetrahaploid strawberry plant. In some embodiments, the diploid haploid-inducing strawberry plant is a plant of the species Fragaria vesca, Fragaria iinumae, Fragaria nipponica, Fragaria viridis, Fragaria × bifera, Fragaria bucharica, Fragaria chinensis, Fragaria daltoniana, Fragaria emeiensis, Fragaria hayatae, Fragaria iinumae, Fragaria mandshurica, Fragaria viridis, Fragaria nilgerrensis, Fragaria nipponica, Fragaria nubicola, or Fragaria pentaphylla. In certain embodiments, the diploid haploid-inducing strawberry plant is a plant of the species Fragaria vesca. In some embodiments, the donor octoploid strawberry plant is a plant of the species Fragaria x ananassa. In some embodiments, the embryo of the true homozygous octoploid strawberry seed has a nuclear genome derived from only the donor octoploid strawberry plant (e.g., Fragaria x ananassa) and a cytoplasmic genome (e.g., mitochondrial and/or chloroplast genome) derived from the haploid-inducing strawberry plant (e.g., Fragaria vesca). [0225] In other embodiments, the method of producing true homozygous octoploid strawberry plant, or plant part thereof, comprises (a) contacting a tetrahaploid cell from a donor octoploid strawberry plant with pollen from an octoploid haploid-inducing strawberry plant described herein, and (b) producing an octoploid doubled tetrahaploid cell or plant from the tetrahaploid cell. In certain embodiments, the method comprises (a1) contacting a reduced egg cell from a donor octoploid strawberry plant with a sperm cell from an octoploid haploid- inducing strawberry plant described herein, (a2) allowing a tetrahaploid embryo to form and grow into a tetrahaploid strawberry plant, and (b) subjecting the tetrahaploid strawberry plant to genome doubling to produce a doubled tetrahaploid strawberry plant. In certain embodiments, the method comprises (a) contacting a tetrahaploid cell from a donor octoploid strawberry plant with pollen from an octoploid haploid-inducing strawberry plant described herein, and (b) producing an octoploid doubled tetrahaploid cell or plant from the tetrahaploid cell. In certain embodiments, the method comprises (a1) contacting a tetrahaploid reduced egg cell from a donor octoploid strawberry plant with a sperm cell from an octoploid haploid- inducing strawberry plant described herein, (a2) allowing a tetrahaploid embryo to form and grow into a tetrahaploid strawberry plant, and (b) subjecting the tetrahaploid strawberry plant to genome doubling to produce an octoploid doubled tetrahaploid strawberry plant. In some 98 sf-6744554 197072001240 embodiments, the octoploid haploid-inducing strawberry plant is a plant of the species Fragaria x ananassa or Fragaria chiloensis. In certain embodiments, the octoploid haploid- inducing strawberry plant is a plant of the species Fragaria x ananassa. [0226] In other embodiments, the method of producing true homozygous octoploid strawberry plant, or plant part thereof, comprises (a) contacting a tetrahaploid cell from a donor octoploid strawberry plant with an egg cell from an octoploid haploid-inducing strawberry plant described herein, and (b) producing an octoploid doubled tetrahaploid cell or plant from the tetrahaploid cell. In certain embodiments, the method comprises (a1) contacting a reduced sperm cell from a donor octoploid strawberry plant with an egg cell from an octoploid haploid-inducing strawberry plant described herein, (a2) allowing a tetrahaploid embryo to form and grow into a tetrahaploid strawberry plant, and (b) subjecting the tetrahaploid strawberry plant to genome doubling to produce a doubled tetrahaploid strawberry plant. In certain embodiments, the method comprises (a) contacting a tetrahaploid cell from a donor octoploid strawberry plant with an egg cell from an octoploid haploid- inducing strawberry plant described herein, and (b) producing an octoploid doubled tetrahaploid cell or plant from the tetrahaploid cell. In certain embodiments, the method comprises (a1) contacting a tetrahaploid reduced sperm cell from a donor octoploid strawberry plant with an egg cell from an octoploid haploid-inducing strawberry plant described herein, (a2) allowing a tetrahaploid embryo to form and grow into a tetrahaploid strawberry plant, and (b) subjecting the tetrahaploid strawberry plant to genome doubling to produce an octoploid doubled tetrahaploid strawberry plant. In some embodiments, the octoploid haploid-inducing strawberry plant is a plant of the species Fragaria x ananassa or Fragaria chiloensis. In certain embodiments, the octoploid haploid-inducing strawberry plant is a plant of the species Fragaria x ananassa. In some embodiments, the haploid-inducing strawberry plant and the donor octoploid strawberry plants are from different species of Fragaria. In some embodiments, the donor octoploid strawberry plant is a plant of the species Fragaria x ananassa. In some embodiments, the embryo of the true homozygous octoploid strawberry seed has a nuclear genome derived from only the donor octoploid strawberry plant (e.g., Fragaria x ananassa) and a cytoplasmic genome (e.g., mitochondrial and/or chloroplast genome) derived from the haploid-inducing strawberry plant. [0227] Haploid induction may comprise intraspecific haploid induction (e.g., using pollen of a haploid-inducing Fragaria x ananassa plant to fertilize a donor Fragaria x ananassa 99 sf-6744554 197072001240 plant) or interspecific haploid induction (e.g., using pollen of a haploid-inducing Fragaria vesca plant to fertilize a donor Fragaria x ananassa plant). Accordingly, in some embodiments, the method of producing true homozygous octoploid strawberry plant, or plant part thereof, comprises (a) contacting a tetrahaploid cell from a donor octoploid strawberry plant with pollen from a haploid-inducing strawberry plant described herein, wherein the donor octoploid strawberry plant and the haploid-inducing strawberry plant are plants of different species; and (b) producing an octoploid doubled tetrahaploid cell or plant from the tetrahaploid cell. In certain embodiments, the method comprises (a1) contacting a reduced egg cell from a donor octoploid strawberry plant with a sperm cell from a haploid-inducing strawberry plant described herein, wherein the donor octoploid strawberry plant and the haploid-inducing strawberry plant are plants of different species; (a2) allowing a tetrahaploid embryo to form and grow into a tetrahaploid strawberry plant; and (b) subjecting the tetrahaploid strawberry plant to genome doubling to produce a doubled tetrahaploid strawberry plant. In certain embodiments, the method comprises (a) contacting a tetrahaploid cell from a donor octoploid strawberry plant with pollen from a haploid-inducing strawberry plant described herein, wherein the donor octoploid strawberry plant and the haploid- inducing strawberry plant are plants of different species; and (b) producing an octoploid doubled tetrahaploid cell or plant from the tetrahaploid cell. In certain embodiments, the method comprises (a1) contacting a tetrahaploid reduced egg cell from a donor octoploid strawberry plant with a sperm cell from a haploid-inducing strawberry plant described herein, wherein the donor octoploid strawberry plant and the haploid-inducing strawberry plant are plants of different species; (a2) allowing a tetrahaploid embryo to form and grow into a tetrahaploid strawberry plant; and (b) subjecting the tetrahaploid strawberry plant to genome doubling to produce an octoploid doubled haploid strawberry plant. In certain embodiments, the donor octoploid strawberry plant is a plant of the species Fragaria x ananassa, and the haploid-inducing strawberry plant is a plant of the species Fragaria vesca. [0228] In other embodiments, the method of producing true homozygous octoploid strawberry plant, or plant part thereof, comprises (a) contacting a tetrahaploid cell from a donor octoploid strawberry plant with an egg cell from a haploid-inducing strawberry plant described herein, wherein the donor octoploid strawberry plant and the haploid-inducing strawberry plant are plants of different species; and (b) producing an octoploid doubled tetrahaploid cell or plant from the tetrahaploid cell. In certain embodiments, the method comprises (a1) contacting a reduced sperm cell from a donor octoploid strawberry plant with 100 sf-6744554 197072001240 an egg cell from a haploid-inducing strawberry plant described herein, wherein the donor octoploid strawberry plant and the haploid-inducing strawberry plant are plants of different species; (a2) allowing a tetrahaploid embryo to form and grow into a tetrahaploid strawberry plant; and (b) subjecting the tetrahaploid strawberry plant to genome doubling to produce a doubled tetrahaploid strawberry plant. In certain embodiments, the method comprises (a) contacting a tetrahaploid cell from a donor octoploid strawberry plant with an egg cell from a haploid-inducing strawberry plant described herein, wherein the donor octoploid strawberry plant and the haploid-inducing strawberry plant are plants of different species; and (b) producing an octoploid doubled tetrahaploid cell or plant from the tetrahaploid cell. In certain embodiments, the method comprises (a1) contacting a tetrahaploid reduced sperm cell from a donor octoploid strawberry plant with an egg cell from a haploid-inducing strawberry plant described herein, wherein the donor octoploid strawberry plant and the haploid-inducing strawberry plant are plants of different species; (a2) allowing a tetrahaploid embryo to form and grow into a tetrahaploid strawberry plant; and (b) subjecting the tetrahaploid strawberry plant to genome doubling to produce an octoploid doubled haploid strawberry plant. In certain embodiments, the donor octoploid strawberry plant is a plant of the species Fragaria x ananassa, and the haploid-inducing strawberry plant is a plant of the species Fragaria vesca. In some embodiments, the embryo of the true homozygous octoploid strawberry seed has a nuclear genome derived from Fragaria x ananassa and a cytoplasmic genome (e.g., mitochondrial and/or chloroplast genome) derived from Fragaria vesca. [0229] In other embodiments, the method of producing true homozygous octoploid strawberry plant, or plant part thereof, comprises (a) contacting a tetrahaploid cell from a donor octoploid strawberry plant with pollen from a haploid-inducing strawberry plant described herein, wherein the donor octoploid strawberry plant and the haploid-inducing strawberry plant are plants of the same species; and (b) producing an octoploid doubled tetrahaploid cell or plant from the tetrahaploid cell. In certain embodiments, the method comprises (a1) contacting a reduced egg cell from a donor octoploid strawberry plant with a sperm cell from a haploid-inducing strawberry plant described herein, wherein the donor octoploid strawberry plant and the haploid-inducing strawberry plant are plants of the same species; (a2) allowing a tetrahaploid embryo to form and grow into a tetrahaploid strawberry plant; and (b) subjecting the tetrahaploid strawberry plant to genome doubling to produce a doubled tetrahaploid strawberry plant. In certain embodiments, the method comprises (a) contacting a tetrahaploid cell from a donor octoploid strawberry plant with pollen from a 101 sf-6744554 197072001240 haploid-inducing strawberry plant described herein, wherein the donor octoploid strawberry plant and the haploid-inducing strawberry plant are plants of the same species; and (b) producing an octoploid doubled tetrahaploid cell or plant from the tetrahaploid cell. In certain embodiments, the method comprises (a1) contacting a tetrahaploid reduced egg cell from a donor octoploid strawberry plant with a sperm cell from a haploid-inducing strawberry plant described herein, wherein the donor octoploid strawberry plant and the haploid-inducing strawberry plant are plants of the same species; (a2) allowing a tetrahaploid embryo to form and grow into a tetrahaploid strawberry plant; and (b) subjecting the tetrahaploid strawberry plant to genome doubling to produce an octoploid doubled tetrahaploid strawberry plant. In certain embodiments, both the donor octoploid strawberry plant and the haploid-inducing strawberry plant are plants of the species Fragaria x ananassa. [0230] In other embodiments, the method of producing true homozygous octoploid strawberry plant, or plant part thereof, comprises (a) contacting a tetrahaploid cell from a donor octoploid strawberry plant with an egg cell from a haploid-inducing strawberry plant described herein, wherein the donor octoploid strawberry plant and the haploid-inducing strawberry plant are plants of the same species; and (b) producing an octoploid doubled tetrahaploid cell or plant from the tetrahaploid cell. In certain embodiments, the method comprises (a1) contacting a reduced sperm cell from a donor octoploid strawberry plant with an egg cell from a haploid-inducing strawberry plant described herein, wherein the donor octoploid strawberry plant and the haploid-inducing strawberry plant are plants of the same species; (a2) allowing a tetrahaploid embryo to form and grow into a tetrahaploid strawberry plant; and (b) subjecting the tetrahaploid strawberry plant to genome doubling to produce a doubled tetrahaploid strawberry plant. In certain embodiments, the method comprises (a) contacting a tetrahaploid cell from a donor octoploid strawberry plant with an egg cell from a haploid-inducing strawberry plant described herein, wherein the donor octoploid strawberry plant and the haploid-inducing strawberry plant are plants of the same species; and (b) producing an octoploid doubled tetrahaploid cell or plant from the tetrahaploid cell. In certain embodiments, the method comprises (a1) contacting a tetrahaploid reduced sperm cell from a donor octoploid strawberry plant with an egg cell from a haploid-inducing strawberry plant described herein, wherein the donor octoploid strawberry plant and the haploid-inducing strawberry plant are plants of the same species; (a2) allowing a tetrahaploid embryo to form and grow into a tetrahaploid strawberry plant; and (b) subjecting the tetrahaploid strawberry plant to genome doubling to produce an octoploid doubled tetrahaploid strawberry plant. In 102 sf-6744554 197072001240 certain embodiments, both the donor octoploid strawberry plant and the haploid-inducing strawberry plant are plants of the species Fragaria x ananassa. [0231] In some embodiments, the step of generating a tetrahaploid strawberry plant through haploid induction comprises allowing the donor octoploid strawberry plant to form a plurality of seeds and subsequently allowing one or more seeds of the plurality to germinate and form one or more plants. In certain embodiments, the step of generating a tetrahaploid strawberry plant through haploid induction comprises allowing the donor octoploid strawberry plant to form a plurality of seeds and subsequently allowing two or more seeds of the plurality to germinate and form a plurality of plants. In other embodiments, the step of generating a tetrahaploid strawberry plant through haploid induction comprises collecting one or more embryos from the plurality of seeds, contacting the one or more embryos with a nutrient medium suitable for inducing plant growth, and allowing the one or more embryos to form one or more plants. In certain embodiments, the step of generating a tetrahaploid strawberry plant through haploid induction comprises collecting a plurality of embryos from the plurality of seeds, contacting the plurality of embryos with a nutrient medium suitable for inducing plant growth, and allowing the plurality of embryos to form a plurality of plants. The method may further comprise a step of selecting one or more tetrahaploid plants by 1) determining the ploidy of one or more cells of one or more plants of the plurality of plants, and 2) selecting one or more plants having one or more tetrahaploid cells from the plurality of plants. In some embodiments, the step of selecting one or more tetrahaploid seeds of the plurality of seeds comprises 1) determining the ploidy of one or more cells of one or more plants of the plurality of plants, and 2) selecting one or more plants having one or more tetraploid cells from the plurality of plants. In certain embodiments, determining the ploidy of one or more cells of the one or more plants comprises obtaining a tissue sample of each of the one or more plants and determining the ploidy of one or more cells of each tissue sample. [0232] In some embodiments, the step of generating a tetrahaploid strawberry plant through haploid induction comprises allowing the haploid-inducing strawberry plant to form a plurality of seeds and subsequently allowing one or more seeds of the plurality to germinate and form one or more plants. In certain embodiments, the step of generating a tetrahaploid strawberry plant through haploid induction comprises allowing the haploid-inducing strawberry plant to form a plurality of seeds and subsequently allowing two or more seeds of the plurality to germinate and form a plurality of plants. In other embodiments, the step of 103 sf-6744554 197072001240 generating a tetrahaploid strawberry plant through haploid induction comprises collecting one or more embryos from the plurality of seeds, contacting the one or more embryos with a nutrient medium suitable for inducing plant growth, and allowing the one or more embryos to form one or more plants. In certain embodiments, the step of generating a tetrahaploid strawberry plant through haploid induction comprises collecting a plurality of embryos from the plurality of seeds, contacting the plurality of embryos with a nutrient medium suitable for inducing plant growth, and allowing the plurality of embryos to form a plurality of plants. The method may further comprise a step of selecting one or more tetrahaploid plants by 1) determining the ploidy of one or more cells of one or more plants of the plurality of plants, and 2) selecting one or more plants having one or more tetrahaploid cells from the plurality of plants. In some embodiments, the step of selecting one or more tetrahaploid seeds of the plurality of seeds comprises 1) determining the ploidy of one or more cells of one or more plants of the plurality of plants, and 2) selecting one or more plants having one or more tetraploid cells from the plurality of plants. In certain embodiments, determining the ploidy of one or more cells of the one or more plants comprises obtaining a tissue sample of each of the one or more plants and determining the ploidy of one or more cells of each tissue sample. [0233] Methods of determining the ploidy of a cell are known in the art and are described, for example, in Example 3 of the present disclosure and Galbraith et al (Galbraith et al. 1983. “Rapid flow cytometric analysis of the cell cycle in intact plant tissues.” Science 220, no. 4601: 1049-1051) and reviewed in, for example, Ochatt et al (Ochatt et al. 2011. "Ploidy level determination within the context of in vitro breeding." Plant Cell, Tissue and Organ Culture (PCTOC) 104.3 (2011): 329-341). Haploid Doubling [0234] In some embodiments, the methods of producing true homozygous octoploid strawberry plants, or plant parts thereof, comprise subjecting a tetrahaploid strawberry cell or plant to genome doubling to produce a doubled tetrahaploid strawberry cell or plant. Genome doubling, also known as chromosome doubling, refers to the process of disrupting mitosis in a parent cell or plant in order to generate daughter cells or plants having two clonal copies of the genome of the parent cell or plant. In the case of haploid doubling, mitosis is disrupted in a haploid parent cell or plant in order to generate doubled-haploid daughter cells or plants having two clonal copies of the haploid gene of the parent cell or plant. This results in a true homozygous daughter cell or plant which is monoallelic at all loci across its two haploid sets 104 sf-6744554 197072001240 of chromosomes. For example, if haploid induction in an octoploid (8x) strawberry donor plant to generate a tetrahaploid (4x) cell or plant, genome doubling of the tetrahaploid (4x) cell or plant would result in generating two clonal copies of the 4x haploid chromosomes, resulting in an octoploid (8x) cell or plant having two 4x clonal sets of chromosomes that are monoallelic across all loci. [0235] In some embodiments, the methods of producing true homozygous octoploid strawberry plants, or plant parts thereof, comprise subjecting one or more tetrahaploid cells, tetrahaploid seeds, or tetrahaploid plants to a haploid doubling treatment. The tetrahaploid cells, tetrahaploid seeds, or tetrahaploid plants may have been selected from a plurality of cells, seeds, or plants grown from an octoploid strawberry donor plant that has been contacted with pollen or a part thereof (e.g., sperm cells) from a haploid-inducing strawberry plant, as described herein. In some embodiments, subjecting one or more tetrahaploid cells, tetrahaploid seeds, or tetrahaploid plants to a haploid doubling treatment comprises contacting the one or more tetrahaploid cells, tetrahaploid seeds, or tetrahaploid plants with an agent that causes genome doubling or chromosome doubling, such as an antimitotic agent. In certain embodiments, the antimitotic agent is an anti-microtubule agent. In some embodiments, the antimitotic agent comprises colchicine, oryzalin, oxide, trifluralin, or any combination thereof. In some embodiments, the antimitotic agent comprises colchicine. In some embodiments, the antimitotic agent is an antimitotic herbicide. Antimitotic herbicides include, for example, oryzalin, trifluralin, and amiprofos-methyl (APM). In some embodiments, the haploid doubling treatment comprises contacting the tetrahaploid cell, tetrahaploid seed, or tetrahaploid plant with an anti-microtubule agent to form a doubled tetrahaploid cell, a doubled tetrahaploid seed, or a doubled tetrahaploid plant. [0236] In some embodiments, the doubled tetrahaploid cell or the doubled tetrahaploid seed is allowed to grow into a doubled tetrahaploid plant. In certain embodiments, the doubled tetrahaploid plant is allowed to form seed, thus producing the true homozygous octoploid strawberry seed. [0237] Methods of chromosome doubling in plants are known in the art and reviewed, for example, in Hooghvorst et al. (2021. "Chromosome doubling methods in doubled haploid and haploid inducer-mediated genome-editing systems in major crops." Plant Cell Reports 40.2: 255- 270.). Methods of chromosome doubling include, but are not limited to, treatment with mitotic inhibitors such as colchicine, oryzalin, trifluralin, or nitrous oxide. 105 sf-6744554 197072001240 [0238] In some embodiments, haploid doubling occurs spontaneously. Accordingly, in some embodiments, provided herein are methods of producing true homozygous octoploid strawberry plant, comprising (a) generating a tetrahaploid strawberry cell or plant through haploid induction, and (b) allowing the tetrahaploid strawberry plant to undergo spontaneous genome doubling to produce a doubled tetrahaploid strawberry cell or plant. The method may further comprise a step of selecting one or more doubled tetrahaploid cells, seeds, or plants. From a plurality of tetrahaploid cells, seeds, or plants. In some embodiments, the method comprises selecting one or more doubled tetrahaploid seeds from a plurality of seeds by 1) determining the ploidy of the embryo of one or more seeds of the plurality of seeds, and 2) selecting one or more seeds having an octoploid embryo. In certain embodiments, the step of selecting one or more tetrahaploid seeds of the plurality of seeds comprises 1) determining the ploidy of the embryo and endosperm of one or more seeds of the plurality of seeds, and 2) selecting one or more seeds having an octoploid embryo and an endosperm having a ploidy of greater than 8x (e.g., a ploidy of 9x, 10x, 11x, or 12x). True Homozygous Octoploid Strawberry Seed and Plants [0239] In some aspects, provided herein are true homozygous octoploid strawberry plants, plant parts thereof, plant lines, and seeds produced by the methods described herein. In some embodiments, the true homozygous octoploid strawberry plant is a plant of the species Fragaria x ananassa. In some embodiments, the true homozygous octoploid strawberry plant, or part thereof, comprises two clonal sets of chromosomes. In certain embodiments, the true homozygous octoploid strawberry plant, or part thereof, comprises two clonal sets of chromosomes, each clonal set comprising a haploid set of chromosomes from each of four subgenomes. [0240] In some embodiments, the true homozygous octoploid strawberry plant, plant part, plant line, or seed is monoallelic across a majority of loci in the genome. In some embodiments, the true homozygous octoploid strawberry plant, plant part, plant line, or seed is monoallelic at more than 50%, more than 60%, more than 70%, more than 80%, more than 85%, more than 90%, more than 91%, more than 92%, more than 93%, more than 94%, more than 95%, more than 96%, more than 97%, more than 98%, more than 99%, more than 99.9%, more than 99.99%, or 100% of the loci in the genome. 106 sf-6744554 197072001240 [0241] Methods of measuring homozygosity and heterozygosity within genomes and subgenomes of strawberry plants are known in the art. In one exemplary method, in order to estimate the within-subgenome rate of heterozygosity, whole genome sequencing may be conducted on parent octoploid plants, and doubled tetrahaploids derived from those parent octoploid plants, using paired end next generation sequencing (e.g., Illumina Novaseq with paired end 150nt reads in a 500nt insert library). Read pairs are mapped to a repeat masked version of a phased assembly of the parent octoploid plant (e.g., of the F. x ananassa Camarosa genome (2n=8x=56)) using Burrows-Wheller Alignment (BWA). Read pairs are retained for analysis if they uniquely map to a single chromosome within a single subgenome. Variants are called at specific loci and the rate of within-subgenome heterozygosity is calculated. [0242] In the parental octoploid lines the average rate of within-subgenome heterozygosity is typically calculated to be approximately 1.5%, while the rate of within- subgenome heterozygosity in doubled tetrahaploids is typically less than 0.05% using the same methods, showing that the doubled tetrahaploids are approximately 30 times more homozygous than the octoploid parents after a single generation. [0243] In some embodiments, the true homozygous octoploid strawberry plant, plant part, plant line, or seed has a rate of within-subgenome heterozygosity of less than 1.5%, less than 1.0%, less than 0.5%, less than 0.1%, less than 0.05%, less than 0.01%, or less than 0.005%. [0244] In some aspects, provided herein is a homozygous octoploid strawberry plant, plant part, plant line, or seed comprising at least one cell having a nuclear genome from a first species of Fragaria and a cytoplasmic genome from a second species of Fragaria. The first and second species of Fragaria may each independently be any species of Fragaria known in the art or described herein. In some embodiments, the nuclear genome and the cytoplasmic genome are both from octoploid species of Fragaria. In some embodiments, the nuclear genome is from an octoploid species of Fragaria and the cytoplasmic genome is from a diploid species of Fragaria. In certain embodiments, the nuclear genome is from a plant of the species Fragaria x anannasa. In certain embodiments, the cytoplasmic genome is a from a plant of the species Fragaria vesca. In certain embodiments, the nuclear genome is from a plant of the species Fragaria x anannasa and the cytoplasmic genome is a from a plant of the species Fragaria vesca. 107 sf-6744554 197072001240 [0245] In some aspects, provided herein is a runnerless true homozygous octoploid strawberry plant. In some embodiments, the runnerless true homozygous octoploid strawberry plant comprises one or more genetic modifications resulting in decreased expression of a gibberellin 20-oxidase (ga20ox) gene. In some embodiments, the runnerless true homozygous octoploid strawberry plant does not produce runners or produces fewer runners as compared to a control plant that lacks the one or more genetic modifications resulting in decreased expression of the ga20ox gene. [0246] In some embodiments, provided herein is a population of true homozygous octoploid strawberry seed. In certain embodiments, at least 50% of the true homozygous octoploid strawberry seed are genetically uniform. In some variations, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.9%, or at least 99.99% of the true homozygous octoploid strawberry seed are genetically uniform. Genetic uniformity may be determined by any means known in the art including, for example, DNA-hybridization based methods (e.g., SNP-chip, DNA sequencing methods (e.g., whole genome sequencing or genotyping by sequencing (GBS)), and the like. Methods of Breeding and Producing Uniform Octoploid F1 Hybrid Strawberry Seed [0247] In one aspect, described herein are methods of breeding and producing a uniform octoploid F1 hybrid strawberry seed comprising crossing two true homozygous octoploid strawberry plants. In some embodiments, the methods comprise producing one or more true homozygous octoploid strawberry plants using the methods described herein. In certain embodiments, the methods comprise performing a plurality of crosses between pairs of true homozygous octoploid strawberry lines selected from a plurality of true homozygous octoploid strawberry lines, evaluating the traits of the F1 generation of plants, and selecting one or more of the pairs of true homozygous octoploid strawberry lines that, when crossed, resulted in desirable traits in the F1 generation. In certain embodiments, the method further comprises crossing the one or more selected pairs of true homozygous octoploid strawberry lines to produce the uniform octoploid F1 hybrid strawberry seed. [0248] In some embodiments, the methods of breeding and producing a uniform octoploid F1 hybrid strawberry seed comprise crossing two true homozygous octoploid strawberry plants. In some variations, crossing two true homozygous octoploid strawberry 108 sf-6744554 197072001240 plants comprises contacting pollen from a first true homozygous octoploid strawberry plant with the stigma of a pistil of a second true homozygous octoploid strawberry plant. In some embodiments, the uniform octoploid F1 hybrid strawberry seed is produced by crossing the two true homozygous octoploid strawberry plants and allowing seeds to form. [0249] In some embodiments, the methods of breeding and producing uniform octoploid F1 hybrid strawberry seed comprise maintaining one or more true homozygous octoploid strawberry lines, one or more F1 hybrid strawberry lines, or a combination thereof. In some variations, the inbred and/or hybrid octoploid strawberry lines are maintained via vegetative propagation, selfing, apomixis, or any combination thereof. Additional methods of maintaining strawberry lines are well-known in the art. In some embodiments, the methods of breeding and producing uniform octoploid F1 hybrid strawberry seed comprise maintaining an inventory of true homozygous octoploid strawberry lines from which haplotypes may be selected for rapid deterministic combination of the haplotypes. [0250] In further aspects, provided herein is a uniform F1 hybrid strawberry seed produced by the methods described herein. In some embodiments, provided herein is a population of uniform octoploid F1 hybrid strawberry seed. In certain embodiments, at least 50% of the uniform octoploid F1 hybrid strawberry seed are genetically uniform. In some variations, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.9%, or at least 99.99% of the uniform octoploid F1 hybrid strawberry seed are genetically uniform. Obtaining Plant Lines [0251] In some embodiments, the methods of breeding and producing uniform octoploid F1 hybrid strawberry seed comprise obtaining a set of octoploid strawberry lines. In preferred embodiments, the set of octoploid strawberry lines is genetically diverse and comprises a large and diverse pool of haplotypes. In some embodiments, the methods of breeding and producing uniform octoploid F1 hybrid strawberry seed comprises obtaining a set of lines of the same or related species of octoploid strawberry plant (e.g., Fragaria x ananassa and/or Fragaria chiloensis). The set of strawberry lines may be obtained from any source and by any methods known in the art. In some embodiments, the set of octoploid strawberry lines is 109 sf-6744554 197072001240 obtained from sources including, but not limited to, natural diversity, existing breeding programs, or any combination thereof. [0252] In some embodiments, the set of octoploid strawberry lines is a genetically diverse founder population of plants. These plants may be collected from existent germplasm sources such as wild progenitor species and landraces. In some instances, these plants may have high genetic load and may not have undergone the narrowing of genetic variability attributable to the elite selection practices imposed on modern cultivated materials. As such, many possess traits promoting their fitness within non-agrarian environments in the case of wild progenitors, or have been adapted to vastly differing cultivation practices and agricultural environments in the case of landraces. In either instance, these founding octoploid strawberry lines may contain suites of both desirable and undesirable agronomic characteristics that may be recombined, selected, and complemented to develop a commercializable product. Heterotic Groups [0253] In some embodiments, the methods of breeding and producing uniform octoploid F1 hybrid strawberry seed comprise organizing the set of octoploid strawberry lines obtained into a plurality of heterotic groups, wherein each heterotic group comprises a haplotype, and wherein the haplotypes are grouped based on observed or predicted heterotic performance when combined in the octoploid F1 hybrid strawberry plant. In some variations, the methods of breeding and producing uniform octoploid F1 hybrid strawberry seed comprise organizing the set of octoploid strawberry lines into three or more heterotic groups, four or more heterotic groups, five or more heterotic groups, six or more heterotic groups, seven or more heterotic groups, or eight or more heterotic groups. The obtained set of octoploid strawberry lines may be organized by assigning their membership into complementary heterotic groups based upon heterotic patterns observed from preliminary estimation or prediction of their combining ability for traits and environments of interest. In this context, the combining ability is an estimation of the value of an octoploid strawberry plant as a parent as inferred by progeny testing in an established factorial or hierarchical mating design. [0254] In defining heterotic groups, the main goal is identifying subpopulations of the set of octoploid strawberry lines based on employment of a clustering procedure that maximizes some measure of interpopulation combining ability. Exhaustive evaluation of all possible parental combinations for the final octoploid F1 hybrid strawberry, as in the case of a 110 sf-6744554 197072001240 factorial mating scheme, is infeasible for all but a trivial number of potential parents (i.e. even a partial diallel ignoring reciprocal crosses scales at (n+3)!/(4!(n-1)!) crosses per n parents). Yet, hierarchical mating schemes necessitate an understanding and judgment of which set of “testers” or analogous constructs should serve as a relevant and efficient basis or frame of reference for inferring combining ability. The sample of testers selected invariably biases perceptions of existing heterotic patterns. Furthermore, the relative importance of traits and environments of interest used to infer these combining abilities are dynamic and depend upon market trends. As such, predictive modeling is essential and the process of assigning and refining heterotic groups and testers to represent them is one of iterative improvement and refinement throughout repeated cycles of the breeding process. Nonetheless, once preliminary heterotic group membership is assigned, interpopulation improvement of the true homozygous octoploid strawberry lines and development of hybrid octoploid strawberry plants may proceed. Evaluating Characteristics of Octoploid F1 Hybrid Strawberry Plants [0255] In some embodiments, the methods of breeding and producing uniform octoploid F1 hybrid strawberry seed comprise evaluating one or more characteristics of an octoploid F1 hybrid plant grown from the uniform octoploid F1 hybrid strawberry seed. Methods for evaluating strawberry plant characteristics are numerous and well-known in the art. The one or more characteristics evaluated may include, but are not limited to, plant size, plant vigor, fruit yield, fruit color, fruit flavor, fruit sweetness, fruit aroma, abiotic stress resistance, disease resistance, pest resistance, and the like. The strawberry plants to be evaluated may then be grown under different geographical, climatic, and soil conditions, and further selections can be made during, and at the end of, the growing season. Promising advanced breeding strawberry lines are thoroughly tested and compared to appropriate standards in environments representative of the commercial target area(s) for three years at least. The best strawberry lines are candidates for new commercial strawberry cultivars. These processes, which lead to the final step of marketing and distribution, usually take from five to ten years from the time the first cross or selection is made. [0256] In some embodiments, the method of breeding a uniform octoploid F1 hybrid strawberry seed comprises repeating the steps of the method using the one or more characteristics of one or more octoploid F1 hybrid strawberry plants grown from the uniform octoploid F1 hybrid strawberry seed evaluated to guide the breeding of candidate octoploid 111 sf-6744554 197072001240 strawberry lines, the selecting of candidate octoploid strawberry lines, or both. In some variations, the method comprises repeating the steps of the method two, three, four, five, six, seven, eight, nine, or ten times or more. In additional variations, the repeating of the steps of the method iteratively informs a genome prediction model for improved prediction of heterotic performance. In certain variations, the improved prediction of heterotic performance allows for rapid combination of haplotypes with strong heterotic performance and acceleration of breeding programs. Genetically Modified Plants, Plant Parts, Plant Cells, and Processed Plant Products [0257] In yet another aspect, provided herein are genetically modified plants, plant parts, and plant cells grown from haploid-inducing strawberry plants, true homozygous octoploid strawberry seed, and uniform octoploid F1 strawberry seed described herein. Also provided herein are processed plant products derived from the genetically modified plants, plant parts, or plant cells provided herein. In some embodiments, the genetically modified plant parts, genetically modified plant cells, and processed plant products provided herein are non- regenerable. [0258] In some embodiments, genetically modified strawberry plants and genetically modified strawberry plant parts are provided herein. The genetically modified strawberry plants and strawberry plant parts may be grown from true homozygous octoploid strawberry seed or uniform octoploid F1 strawberry seed described herein. Alternatively, the genetically modified strawberry plants and strawberry plant parts may be regenerated from a genetically modified cell of a haploid-inducing strawberry plant, a true homozygous octoploid strawberry plant, or a uniform octoploid F1 strawberry plant described herein. In certain embodiments, the genetically modified strawberry plants and strawberry plant parts may be regenerated from a genetically modified cell of a haploid-inducing strawberry plant, wherein the genetically modified cell comprises a genetic modification resulting in decreased expression of one or more CENH3 genes described herein. Genetically modified strawberry plants can be obtained from a genetically modified strawberry seed. Genetically modified strawberry plant parts can be obtained by cutting, snapping, grinding or otherwise disassociating the part from the strawberry plant. The strawberry plant part may be any strawberry plant part known in the art, including, but not limited to, a flower, a pistil, a leaf, a stem, a petiole, a cutting, a tissue, a seed coat, an ovule, pollen, a sperm cell, a root, a fruit (e.g., an aggregate fruit), a cotyledon, a hypocotyl, a protoplast, an embryo, an anther, a seed, an achene, a stolon (also 112 sf-6744554 197072001240 known as a runner) or any portion thereof. In certain embodiments, a genetically modified strawberry plant part provided herein is a non-regenerable portion of a genetically modified strawberry plant part. As used in this context, a “non-regenerable” portion of a genetically modified strawberry plant part refers to a portion that cannot be induced to form a whole strawberry plant or that cannot be induced to form a whole strawberry plant (e.g., through in vitro culture) that is capable of sexual and/or asexual reproduction. A non-regenerable portion of a genetically modified strawberry plant part may be a portion of a flower, a pistil, a leaf, a stem, a petiole, a cutting, a tissue, a seed coat, an ovule, pollen, a sperm cell, a root, a fruit (e.g., an aggregate fruit), a cotyledon, a hypocotyl, a protoplast, an embryo, an anther, an achene, a stolon (also known as a runner) or any portion thereof. [0259] In some embodiments, a non-regenerable or non-propagating strawberry plant cell is provided herein. As used in this context, a “non-regenerable strawberry plant cell” is a cell from a strawberry plant which cannot be regenerated into a whole strawberry plant that is capable of sexual and/or asexual reproduction through in vitro culture. The non-regenerable strawberry plant cell may be in a strawberry plant or strawberry plant part described herein. The non-regenerable strawberry plant cell may be a cell in a stolon, an achene, the hull or pericarp of said achene, a seed, or in the seedcoat of said seed. Mature strawberry plant organs, including a mature leaf, a mature stem, a mature root, or a mature stolon contain at least one non-regenerable cell. In certain embodiments, the non-regenerable strawberry plant cell is a somatic cell. [0260] Also provided herein is a cell culture or tissue culture of non-regenerable or regenerable cells or tissue of a genetically modified strawberry plant or genetically modified strawberry plant part described herein, wherein the non-regenerable or regenerable cells have one or more genetic modifications resulting in decreased expression CENH3 described herein. Preferably, the regenerable cells are derived from embryos, protoplasts, meristematic cells, callus, pollen, leaves, anthers, pistils, ovules, stems, petioles, roots, root tips, stolons (also known as runners), fruits (e.g., aggregate fruits), seeds, achenes, flowers, cotyledons, and/or hypocotyls of a genetically modified plant or a genetically modified plant part grown from a haploid-inducing strawberry plant embryo or seed, a true homozygous octoploid strawberry seed, or a uniform octoploid F1 strawberry seed described herein. [0261] In some embodiments, provided herein is a processed strawberry plant product derived from a genetically modified strawberry plant, plant part, or plant cell described 113 sf-6744554 197072001240 herein. In certain embodiments, the processed plant product contains sufficient nucleic acid (e.g., DNA or RNA) and/or protein material from the genetically modified strawberry plant, plant part, or plant cell to detect nucleic acid and/or protein sequences corresponding to the haplotypes of one or two true homozygous octoploid strawberry plants described herein, the haplotypes of a uniform octoploid F1 strawberry seed described herein, or both. In some embodiments, the processed strawberry plant product is non-regenerable, i.e., cannot be induced to form a whole strawberry plant or that cannot be induced to form a whole strawberry plant that is capable of sexual and/or asexual reproduction. [0262] A processed strawberry plant product may be a seed, an achene, a fruit (e.g., an aggregate fruit), a root, a vegetable, or any plant part described herein, and may be blended as a commodity or other product which moves through commerce and is derived from a genetically modified strawberry plant or a genetically modified plant part. In some embodiments, the commodity or other product can be tracked through commerce by detecting nucleic acid and/or protein sequences of the genetically modified plant or plant part from which they were obtained. In certain embodiments, the processed strawberry plant product comprises a detectable amount of nucleotide and/or protein sequences corresponding to the haplotypes of one or two true homozygous octoploid strawberry plants described herein, the haplotypes of a uniform octoploid F1 strawberry seed described herein, or both. In certain embodiments, the commodity or other product is produced in or maintained in the genetically modified strawberry plant or plant part from which the commodity or other product has been obtained. Such commodities or other products of commerce include, but are not limited to, plant parts, fruit, biomass, oil, meal, food starch, syrup, sugar, animal feed, flour, flakes, bran, lint, hull, processed seed, seed, seedless fruit, puree, juice, juice concentrate, pulp, pomace, preserve, or sauce. The processed plant product may be a food product that is processed by any means known in the art, e.g., canned, steamed, boiled, fried, dried, blanched, juiced, pureed, and/or frozen etc. [0263] In some embodiments, provided herein is a genetically modified strawberry plant, strawberry plant part, or strawberry plant cell comprising one or more genetic modifications resulting in decreased expression of one or more CENH3 proteins described herein. In some embodiments, genetically modified strawberry plant, strawberry plant part, or strawberry plant cell comprises one or more genetic modifications of one or more CENH3 genes. In some embodiments, genetically modified strawberry plant, strawberry plant part, or 114 sf-6744554 197072001240 strawberry plant cell comprises one or more genetic modifications resulting in decreased expression of one or more CENH3 proteins having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity, or at least 60%, 65%, 70%, 75%, 80%, 85%, 95%, 96%, 97%, 98%, or 99% sequence similarity to an amino acid sequence selected from the list consisting of SEQ ID NOs: 8-14 and 53-56, or fragments thereof. In some embodiments, the genetically modified strawberry plant, strawberry plant part, or strawberry plant cell comprises one or more genetic modifications of one or more CENH3 genes comprising a nucleotide sequence having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity or complementarity to a nucleotide sequence selected from the group consisting of SEQ ID NOs: 1-7 and 58-61, or a fragment thereof. In certain embodiments, the one or more genetic modifications comprise a modification of an enhancer of one or more of the CENH3 genes, a modification of a promoter of one or more of the CENH3 genes, a modification of a coding region of one or more of the CENH3 genes, a modification of an intron of one or more of the CENH3 genes, a modification of methylation status of one or more of the CENH3 genes, expression of a repressor protein that targets the DNA or an mRNA of one or more of the CENH3 genes, expression of an RNA interference construct that targets an mRNA of one or more of the CENH3 genes, or any combination thereof. In some embodiments, genetically modified strawberry plant, strawberry plant part, or strawberry plant cell has decreased expression of one or more CENH3 genes described herein relative to a strawberry plant (e.g., a control strawberry plant, e.g., a strawberry plant of the same species) lacking the one or more genetic modifications. In certain embodiments, genetically modified strawberry plant, strawberry plant part, or strawberry plant cell lacks detectable expression of one or more CENH3 genes described herein relative to a strawberry plant (e.g., a control strawberry plant, e.g., a strawberry plant of the same species) lacking the one or more genetic modifications. In certain embodiments, genetically modified strawberry plant, strawberry plant part, or strawberry plant cell lacks detectable expression of any CENH3 genes. In certain embodiments, genetically modified strawberry plant, strawberry plant part, or strawberry plant cell further comprises one or more naturally occurring inactive alleles of one or more CENH3 genes. [0264] In some embodiments, the genetically modified strawberry plant, strawberry plant part, or strawberry plant cell is a plant or from a plant of the species Fragaria vesca and 115 sf-6744554 197072001240 comprises one or more genetic modifications resulting in decreased expression of one or more CENH3 genes. In certain embodiments, the genetically modified strawberry plant, strawberry plant part, or strawberry plant cell is a plant or from a plant of the species Fragaria vesca and comprises one or more genetic modifications resulting in decreased expression of an FvCENH3 gene. In certain embodiments, the genetically modified strawberry plant, strawberry plant part, or strawberry plant cell is a plant or from a plant of the species Fragaria vesca and comprises one or more genetic modifications resulting in decreased expression of an FvCENH3 protein. In certain embodiments, the genetically modified strawberry plant, strawberry plant part, or strawberry plant cell is a plant or from a plant of the species Fragaria vesca and comprises one or more genetic modifications resulting in decreased expression of a CENH3 protein having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity, or at least 60%, 65%, 70%, 75%, 80%, 85%, 95%, 96%, 97%, 98%, or 99% sequence similarity to the FvCENH3 protein of SEQ ID NO. 8, or a fragment thereof. [0265] In some embodiments, the genetically modified strawberry plant, strawberry plant part, or strawberry plant cell is a plant or from a plant of the species Fragaria vesca and comprises one or more genetic modifications of an FvCENH3 gene or gene product. In certain embodiments, the genetically modified strawberry plant, strawberry plant part, or strawberry plant cell is a plant or from a plant of the species Fragaria vesca and comprises one or more genetic modifications in a FvCENH3 gene comprising a nucleotide sequence having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity or complementarity to the FvCENH3 gene nucleotide sequence of SEQ ID NO. 1, or a fragment thereof. In certain embodiments, the genetically modified strawberry plant, strawberry plant part, or strawberry plant cell is a plant or from a plant of the species Fragaria vesca and comprises genetic modifications in both alleles of an FvCENH3 gene. In some variations, the one or more genetic modifications comprise a modification of an enhancer of FvCENH3, a modification of a promoter of FvCENH3, a modification of a coding region of FvCENH3, modification of an intron of FvCENH3, or any combination thereof relative to a control Fragaria vesca plant (e.g., a Fragaria vesca plant lacking one or more of the genetic modifications). In some embodiments, the genetically modified strawberry plant, strawberry plant part, or strawberry plant cell is a plant or from a plant of the species Fragaria vesca and has decreased 116 sf-6744554 197072001240 expression of FvCENH3 protein relative to a control Fragaria vesca plant (e.g., a Fragaria vesca plant lacking one or more of the genetic modifications). In certain embodiments, the genetically modified strawberry plant, strawberry plant part, or strawberry plant cell is a plant or from a plant of the species Fragaria vesca and lacks detectable expression of FvCENH3 protein. [0266] In some embodiments, the genetically modified strawberry plant, strawberry plant part, or strawberry plant cell is a plant or from a plant of the species Fragaria x ananassa and comprises one or more genetic modifications resulting in decreased expression of one or more FaCENH3 genes. In certain embodiments, the genetically modified strawberry plant, strawberry plant part, or strawberry plant cell is a plant or from a plant of the species Fragaria x ananassa and comprises one or more genetic modifications resulting in decreased expression of an FaCENH3-1a gene, an FaCENH3-1b gene, an FaCENH3-2a gene, an FaCENH3-2b gene, an FaCENH3-3 gene, an FaCENH3-4 gene, or any combination thereof. In certain embodiments, the genetically modified strawberry plant, strawberry plant part, or strawberry plant cell is a plant or from a plant of the species Fragaria x ananassa and comprises one or more genetic modifications resulting in decreased expression of an FaCENH3-1a gene, an FaCENH3-1b gene, an FaCENH3-2a gene, an FaCENH3-2b gene, an FaCENH3-3 gene, and an FaCENH3-4 gene. [0267] In some embodiments, the genetically modified strawberry plant, strawberry plant part, or strawberry plant cell is a plant or from a plant of the species Fragaria x ananassa and comprises one or more genetic modifications resulting in decreased expression of an FaCENH3-1a protein, an FaCENH3-1b protein, an FaCENH3-2a protein, an FaCENH3-2b protein, an FaCENH3-3 protein, an FaCENH3-4 protein, or any combination thereof. In certain embodiments, the genetically modified strawberry plant, strawberry plant part, or strawberry plant cell is a plant or from a plant of the species Fragaria x ananassa and comprises one or more genetic modifications resulting in decreased expression of an FaCENH3-1a protein, an FaCENH3-1b protein, an FaCENH3-2a protein, an FaCENH3-2b protein, an FaCENH3-3 protein, and an FaCENH3-4 protein. In certain embodiments, the genetically modified strawberry plant, strawberry plant part, or strawberry plant cell is a plant or from a plant of the species Fragaria x ananassa and comprises one or more genetic modifications resulting in decreased expression of one or more CENH3 proteins having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 117 sf-6744554 197072001240 95%, 96%, 97%, 98%, 99%, or 100% sequence identity, or at least 60%, 65%, 70%, 75%, 80%, 85%, 95%, 96%, 97%, 98%, or 99% sequence similarity to an amino acid sequence selected from the list consisting of SEQ ID NOs: 9-14, or fragments thereof. In certain embodiments, the genetically modified strawberry plant, strawberry plant part, or strawberry plant cell is a plant or from a plant of the species Fragaria x ananassa and comprises one or more genetic modifications resulting in decreased expression of 1) a CENH3 protein having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity, or at least 60%, 65%, 70%, 75%, 80%, 85%, 95%, 96%, 97%, 98%, or 99% sequence similarity to the amino acid sequence of SEQ ID NO: 9; 2) a CENH3 protein having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity, or at least 60%, 65%, 70%, 75%, 80%, 85%, 95%, 96%, 97%, 98%, or 99% sequence similarity to the amino acid sequence of SEQ ID NO: 10; 3) a CENH3 protein having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity, or at least 60%, 65%, 70%, 75%, 80%, 85%, 95%, 96%, 97%, 98%, or 99% sequence similarity to the amino acid sequence of SEQ ID NO: 11; 4) a CENH3 protein having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity, or at least 60%, 65%, 70%, 75%, 80%, 85%, 95%, 96%, 97%, 98%, or 99% sequence similarity to the amino acid sequence of SEQ ID NO: 12; 5) a CENH3 protein having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity, or at least 60%, 65%, 70%, 75%, 80%, 85%, 95%, 96%, 97%, 98%, or 99% sequence similarity to the amino acid sequence of SEQ ID NO: 13; and/or 6) a CENH3 protein having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity, or at least 60%, 65%, 70%, 75%, 80%, 85%, 95%, 96%, 97%, 98%, or 99% sequence similarity to the amino acid sequence of SEQ ID NO: 14. [0268] In some embodiments, the genetically modified strawberry plant, strawberry plant part, or strawberry plant cell is a plant or from a plant of the species Fragaria x ananassa and comprises one or more genetic modifications resulting in decreased expression of an FaCENH3-7a protein, an FaCENH3-7b protein, an FaCENH3-7c protein, an FaCENH3-7d protein, or any combination thereof. In certain embodiments, the genetically modified 118 sf-6744554 197072001240 strawberry plant, strawberry plant part, or strawberry plant cell is a plant or from a plant of the species Fragaria x ananassa and comprises one or more genetic modifications resulting in decreased expression of one or more CENH3 proteins having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity, or at least 60%, 65%, 70%, 75%, 80%, 85%, 95%, 96%, 97%, 98%, or 99% sequence similarity to an amino acid sequence selected from the list consisting of SEQ ID NOs: 53-56, or fragments thereof. In certain embodiments, the genetically modified strawberry plant, strawberry plant part, or strawberry plant cell is a plant or from a plant of the species Fragaria x ananassa and comprises one or more genetic modifications resulting in decreased expression of 1) a CENH3 protein having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity, or at least 60%, 65%, 70%, 75%, 80%, 85%, 95%, 96%, 97%, 98%, or 99% sequence similarity to the amino acid sequence of SEQ ID NO: 53; 2) a CENH3 protein having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity, or at least 60%, 65%, 70%, 75%, 80%, 85%, 95%, 96%, 97%, 98%, or 99% sequence similarity to the amino acid sequence of SEQ ID NO: 54; 3) a CENH3 protein having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity, or at least 60%, 65%, 70%, 75%, 80%, 85%, 95%, 96%, 97%, 98%, or 99% sequence similarity to the amino acid sequence of SEQ ID NO: 55; and/or 4) a CENH3 protein having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity, or at least 60%, 65%, 70%, 75%, 80%, 85%, 95%, 96%, 97%, 98%, or 99% sequence similarity to the amino acid sequence of SEQ ID NO: 56. [0269] In some embodiments, the genetically modified strawberry plant, strawberry plant part, or strawberry plant cell is a plant or from a plant of the species Fragaria x ananassa and comprises one or more genetic modifications of an FaCENH3-1a gene or gene product, an FaCENH3-1b gene or gene product, an FaCENH3-2a gene or gene product, an FaCENH3-2b gene or gene product, an FaCENH3-3 gene or gene product, an FaCENH3-4 gene or gene product, or any combination thereof. In some embodiments, the genetically modified strawberry plant, strawberry plant part, or strawberry plant cell is a plant or from a plant of the species Fragaria x ananassa and comprises one or more genetic modifications of an FaCENH3-1a gene or gene product, an FaCENH3-1b gene or gene product, an FaCENH3-2a 119 sf-6744554 197072001240 gene or gene product, an FaCENH3-2b gene or gene product, an FaCENH3-3 gene or gene product, and an FaCENH3-4 gene or gene product. In certain embodiments, the genetically modified strawberry plant, strawberry plant part, or strawberry plant cell is a plant or from a plant of the species Fragaria x ananassa and comprises one or more genetic modifications in a CENH3 gene comprising a nucleotide sequence having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity or complementarity to a nucleotide sequence selected from the group consisting of SEQ ID NOs: 2-7, or a fragment thereof. In certain embodiments, the genetically modified strawberry plant, strawberry plant part, or strawberry plant cell is a plant or from a plant of the species Fragaria x ananassa and comprises: 1) one or more genetic modifications in a CENH3 gene comprising a nucleotide sequence having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity or complementarity to the nucleotide sequence of SEQ ID NO: 2, or a fragment thereof; 2) one or more genetic modifications in a CENH3 gene comprising a nucleotide sequence having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity or complementarity to the nucleotide sequence of SEQ ID NO: 3, or a fragment thereof; 3) one or more genetic modifications in a CENH3 gene comprising a nucleotide sequence having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity or complementarity to the nucleotide sequence of SEQ ID NO: 4, or a fragment thereof; 4) one or more genetic modifications in a CENH3 gene comprising a nucleotide sequence having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity or complementarity to the nucleotide sequence of SEQ ID NO: 5, or a fragment thereof; 5) one or more genetic modifications in a CENH3 gene comprising a nucleotide sequence having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity or complementarity to the nucleotide sequence of SEQ ID NO: 6, or a fragment thereof; 6) one or more genetic modifications in a CENH3 gene comprising a nucleotide sequence having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity or complementarity to the nucleotide sequence of SEQ ID NO: 7, or a fragment thereof. In some variations, the one or more genetic modifications comprise a modification of an enhancer of an FaCENH3 gene (e.g., an FaCENH3-1a gene, an FacCENH3-1b gene, an 120 sf-6744554 197072001240 FaCENH3-2a gene, an FaCENH3-2b gene, an FaCENH3-3 gene, and/or an FaCENH3-4 gene), a modification of a promoter of an FaCENH3 gene (e.g., an FaCENH3-1a gene, an FacCENH3-1b gene, an FaCENH3-2a gene, an FaCENH3-2b gene, an FaCENH3-3 gene, and/or an FaCENH3-4 gene), a modification of a coding region of an FaCENH3 gene (e.g., an FaCENH3-1a gene, an FacCENH3-1b gene, an FaCENH3-2a gene, an FaCENH3-2b gene, an FaCENH3-3 gene, and/or an FaCENH3-4 gene), modification of an intron of an FaCENH3 gene (e.g., an FaCENH3-1a gene, an FacCENH3-1b gene, an FaCENH3-2a gene, an FaCENH3-2b gene, an FaCENH3-3 gene, and/or an FaCENH3-4 gene), or any combination thereof relative to a control Fragaria x ananassa plant (e.g., a wild-type or unmodified Fragaria x ananassa plant lacking one or more or all of the genetic modifications). [0270] In some embodiments, the genetically modified strawberry plant, strawberry plant part, or strawberry plant cell is a plant or from a plant of the species Fragaria x ananassa and comprises one or more genetic modifications of an FaCENH3-7a gene or gene product, an FaCENH3-7b gene or gene product, an FaCENH3-7c gene or gene product, an FaCENH3-7d gene or gene product, or any combination thereof. In certain embodiments, the genetically modified strawberry plant, strawberry plant part, or strawberry plant cell is a plant or from a plant of the species Fragaria x ananassa and comprises one or more genetic modifications in a CENH3 gene comprising a nucleotide sequence having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity or complementarity to a nucleotide sequence selected from the group consisting of SEQ ID NOs: 58-61, or a fragment thereof. In certain embodiments, the genetically modified strawberry plant, strawberry plant part, or strawberry plant cell is a plant or from a plant of the species Fragaria x ananassa and comprises: 1) one or more genetic modifications in a CENH3 gene comprising a nucleotide sequence having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity or complementarity to the nucleotide sequence of SEQ ID NO: 58, or a fragment thereof; 2) one or more genetic modifications in a CENH3 gene comprising a nucleotide sequence having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity or complementarity to the nucleotide sequence of SEQ ID NO: 59, or a fragment thereof; 3) one or more genetic modifications in a CENH3 gene comprising a nucleotide sequence having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 121 sf-6744554 197072001240 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity or complementarity to the nucleotide sequence of SEQ ID NO: 60, or a fragment thereof; 4) one or more genetic modifications in a CENH3 gene comprising a nucleotide sequence having at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity or complementarity to the nucleotide sequence of SEQ ID NO: 61, or a fragment thereof. In some variations, In some variations, the one or more genetic modifications comprise a modification of an enhancer of an FaCENH3 gene (e.g., an FaCENH3-7a gene, an FaCENH3-7b gene, an FaCENH3-7c gene, and/or an FaCENH3-7d gene), a modification of a promoter of an FaCENH3 gene (e.g., an FaCENH3-7a gene, an FaCENH3-7b gene, an FaCENH3-7c gene, and/or an FaCENH3-7d gene), a modification of a coding region of an FaCENH3 gene (e.g., an FaCENH3-7a gene, an FaCENH3-7b gene, an FaCENH3-7c gene, and/or an FaCENH3-7d gene), modification of an intron of an FaCENH3 gene (e.g., an FaCENH3-7a gene, an FaCENH3-7b gene, an FaCENH3-7c gene, and/or an FaCENH3-7d gene), or any combination thereof relative to a pollen, stolon, seed, or achene of a control Fragaria x ananassa plant (e.g., a wild-type or unmodified Fragaria x ananassa plant lacking one or more or all of the genetic modifications). In some embodiments, the one or more genetic modifications comprise one or more deletions, insertions, or nucleotide substitutions in a coding region of an FaCENH3 gene (e.g., an FaCENH3-7a gene, an FaCENH3-7b gene, an FaCENH3-7c gene, and/or an FaCENH3-7d gene). [0271] In some embodiments, the genetically modified strawberry plant, strawberry plant part, or strawberry plant cell is a plant or from a plant of the species Fragaria x ananassa and has decreased expression of an FaCENH3-1a protein, an FaCENH3-1b protein, an FaCENH3-2a protein, an FaCENH3-2b protein, an FaCENH3-3 protein, an FaCENH3-4 protein, or any combination (e.g., one, two, three, four, five or all six) thereof relative to a control Fragaria x ananassa plant (e.g., a Fragaria ananassa plant lacking one or more or all of the genetic modifications). In certain embodiments the genetically modified strawberry plant, strawberry plant part, or strawberry plant cell is a plant or from a plant of the species Fragaria x ananassa and lacks detectable expression of FaCENH3-1a protein, FaCENH3-1b protein, FaCENH3-2a protein, FaCENH3-2b protein, FaCENH3-3 protein, FaCENH3-4 protein, or any combination (e.g., two, three, four, five or all six) thereof. 122 sf-6744554 197072001240 [0272] In some embodiments, provided herein is a processed strawberry plant product derived from any of the foregoing embodiments of genetically modified strawberry plants, plant parts, or plant cells, wherein the processed strawberry plant product comprises a detectable amount of nucleotide and/or protein sequences corresponding to the haplotypes of one or two true homozygous octoploid strawberry plants described herein, the haplotypes of a uniform octoploid F1 strawberry seed described herein, or both of the genetically modified plant, plant part, or plant cell. In some embodiments, the product is selected from the group consisting of plant biomass, oil, meal, food starch, syrup, animal feed, flour, flakes, bran, lint, hulls, processed seed, puree, juice, juice concentrate, pulp, pomace, preserve, or sauce. In certain embodiments, the processed plant product is non-regenerable. Table 1. Nucleotide and amino acid sequences of CENH3 genes and gene products from Fragaria vesca and Fragaria x ananassa, and associated target protospacer site sequences and guide RNA sequences. 123 sf-6744554 197072001240 124 sf-6744554 197072001240 125 sf-6744554 197072001240 126 sf-6744554 197072001240 127 sf-6744554 197072001240 128 sf-6744554 197072001240 129 sf-6744554 197072001240 130 sf-6744554 197072001240 131 sf-6744554 197072001240 132 sf-6744554 197072001240 133 sf-6744554 197072001240 134 sf-6744554 197072001240 135 sf-6744554 197072001240 136 sf-6744554 197072001240 137 sf-6744554 197072001240 138 sf-6744554 197072001240 139 sf-6744554 197072001240 140 sf-6744554 197072001240 141 sf-6744554 197072001240 142 sf-6744554 197072001240 143 sf-6744554 197072001240 144 sf-6744554 197072001240 145 sf-6744554 197072001240 146 sf-6744554 197072001240 147 sf-6744554 197072001240 148 sf-6744554 197072001240 149 sf-6744554 197072001240 150 sf-6744554 197072001240 151 sf-6744554 197072001240 152 sf-6744554 197072001240 153 sf-6744554 197072001240 154 sf-6744554 197072001240 155 sf-6744554 197072001240 156 sf-6744554 197072001240 157 sf-6744554 197072001240 158 sf-6744554 197072001240 159 sf-6744554 197072001240 160 sf-6744554 197072001240 161 sf-6744554 197072001240 162 sf-6744554 197072001240 163 sf-6744554 197072001240 164 sf-6744554 197072001240 165 sf-6744554 197072001240 166 sf-6744554 197072001240 167 sf-6744554 197072001240 ENUMERATED EMBODIMENTS [0273] The following enumerated embodiments are representative of some aspects of the invention. 1. A strawberry plant, or a plant part thereof, comprising one or more genetic modifications resulting in decreased expression of one or more CENH3 genes. 2. The strawberry plant, or plant part thereof, of embodiment 1, wherein one or more of the CENH3 genes comprise a polynucleotide sequence encoding an amino acid sequence having at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity to an amino acid sequence selected from the group consisting of SEQ ID NOs: 8-14 and 53-56. 3. The strawberry plant, or plant part thereof, of embodiment 1 or 2, wherein one or more of the CENH3 genes comprises a polynucleotide sequence having at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity to a polynucleotide sequence selected from the group consisting of SEQ ID NOs: 1-7 and 58-61. 4. The strawberry plant, or plant part thereof, of any one of embodiments 1-3, wherein the one or more genetic modifications comprise a modification of an enhancer of one or more of the CENH3 genes, a modification of a promoter of one or more of the CENH3 genes, a modification of a coding region of one or more of the CENH3 genes, a modification of an intron of one or more of the CENH3 genes, a modification of methylation status of one or more of the CENH3 genes, expression of a repressor protein that targets the DNA or an mRNA of one or more of the CENH3 genes, expression of an RNA interference construct that targets an mRNA of one or more of the CENH3 genes, or any combination thereof. 5. The strawberry plant, or plant part thereof, of any one of embodiments 1-3, wherein the one or more genetic modifications comprise a modification of an enhancer of one or more of the CENH3 genes, a modification of a promoter of one or more of the CENH3 genes, a modification of a coding region of one or more of the CENH3 genes, modification of an intron of one or more of the CENH3 genes, or any combination thereof relative to an unmodified strawberry plant of the same species or an unmodified control strawberry plant. 6. The strawberry plant, or plant part thereof, of any one of embodiments 1-5, wherein the strawberry plant or plant part has decreased expression of CENH3 proteins relative to a strawberry plant of the same species lacking the one or more genetic modifications. 168 sf-6744554 197072001240 7. The strawberry plant, or plant part thereof, of any one of embodiments 1-6, wherein the strawberry plant is diploid. 8. The strawberry plant, or plant part thereof, of embodiment 7, wherein the strawberry plant is a plant of the species Fragaria vesca, Fragaria iinumae, Fragaria nipponica, Fragaria viridis, Fragaria × bifera, Fragaria bucharica, Fragaria chinensis, Fragaria daltoniana, Fragaria emeiensis, Fragaria hayatae, Fragaria iinumae, Fragaria mandshurica, Fragaria viridis, Fragaria nilgerrensis, Fragaria nipponica, Fragaria nubicola, or Fragaria pentaphylla. 9. The strawberry plant, or plant part thereof, of embodiment 8, wherein the strawberry plant is a plant of the species Fragaria vesca. 10. The strawberry plant, or plant part thereof, of embodiment 9, wherein the strawberry plant is a Fragaria vesca plant of the subspecies Fragaria vesca ssp. vesca, Fragaria vesca ssp. americana, or Fragaria vesca ssp. bracteate. 11. The strawberry plant, or plant part thereof, of any one of embodiments 7-10, wherein the one or more CENH3 genes comprise a polynucleotide sequence encoding an amino acid sequence having at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity the amino acid sequence of SEQ ID NO: 8. 12. The strawberry plant, or plant part thereof, of any one of embodiments 7-11, wherein the one or more CENH3 genes comprise a polynucleotide sequence having at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity to the polynucleotide sequence of SEQ ID NO: 1. 13. The strawberry plant, or plant part thereof, of any one of embodiments 7-12, wherein the strawberry plant is a plant of the species Fragaria vesca and wherein the one or more CENH3 genes comprise FvCENH3. 14. The strawberry plant, or plant part thereof, of embodiment 13, wherein the one or more genetic modifications comprise a modification of an enhancer of FvCENH3, a modification of a promoter of FvCENH3, a modification of a coding region of FvCENH3, modification of an intron of FvCENH3, or any combination thereof relative to a wild-type Fragaria vesca plant. 15. The strawberry plant, or plant part thereof, of any one of embodiments 1-6, wherein the strawberry plant is octoploid. 16. The strawberry plant, or plant part thereof, of embodiment 15, wherein the strawberry plant is a plant of the species Fragaria x ananassa, Fragaria chiloensis, Fragaria virginiana, or Fragaria iturupensis. 169 sf-6744554 197072001240 17. The strawberry plant, or plant part thereof, of embodiment 16, wherein the strawberry plant is Fragaria chiloensis plant of the subspecies Fragaria chiloensis subsp. chiloensis, Fragaria chiloensis subsp. lucida, Fragaria chiloensis subsp. pacifica, or Fragaria chiloensis subsp. sandwicensis. 18. The strawberry plant, or plant part thereof, of embodiment 17, wherein the strawberry plant is a plant of the species Fragaria x ananassa. 19. The strawberry plant, or plant part thereof, of embodiment 18, wherein the strawberry plant is a plant of the variety ‘Chandler’ 20. The strawberry plant, or plant part thereof, of any one of embodiments 15-19, wherein one or more of the CENH3 genes comprises a polynucleotide sequence encoding an amino acid sequence having at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity to an amino acid sequence selected from the group consisting of SEQ ID NOs: 9-14 and 53-56. 21. The strawberry plant, or plant part thereof, of any one of embodiments 15-20, wherein one or more of the CENH3 genes comprises a polynucleotide sequence having at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity to a polynucleotide sequence selected from the group consisting of SEQ ID NOs: 2-7 and 58-61. 22. The strawberry plant, or plant part thereof, of any one of embodiments 15-21, wherein the one or more CENH3 genes comprise: a. a polynucleotide sequence encoding an amino acid sequence having at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity to the amino acid sequence of SEQ ID NO: 53; b. a polynucleotide sequence encoding an amino acid sequence having at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity to the amino acid sequence of SEQ ID NO: 54; c. a polynucleotide sequence encoding an amino acid sequence having at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity to the amino acid sequence of SEQ ID NO: 55; and/or d. a polynucleotide sequence encoding an amino acid sequence having at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity to the amino acid sequence of SEQ ID NO: 56. 23. The strawberry plant, or plant part thereof, of any one of embodiments 15-22, wherein the one or more CENH3 genes comprise: 170 sf-6744554 197072001240 a. a polynucleotide sequence having at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity to the polynucleotide sequence of SEQ ID NO: 58; b. a polynucleotide sequence having at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity to the polynucleotide sequence of SEQ ID NO: 59; c. a polynucleotide sequence having at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity to the polynucleotide sequence of SEQ ID NO: 60; and/or d. a polynucleotide sequence having at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity to the polynucleotide sequence of SEQ ID NO: 61. 24. The strawberry plant, or plant part thereof, of embodiment 15-23, wherein the strawberry plant is a plant of the species Fragaria x ananassa and the one or more CENH3 genes comprise one, two, three, four, five, or all six of FaCENH3-7a, FaCENH3-7b, FaCENH3-7c, and FaCENH3-7d. 25. The strawberry plant, or plant part thereof, of embodiment 24, wherein the one or more genetic modifications comprise: a. a modification of an enhancer of FaCENH3-7a, a modification of a promoter of FaCENH3-7a, a modification of a coding region of FaCENH3-7a, a modification of an intron of FaCENH3-7a, or any combination thereof relative to an unmodified Fragaria x ananassa plant; b. a modification of an enhancer of FaCENH3-7b, a modification of a promoter of FaCENH3-7b, a modification of a coding region of FaCENH3-7b, a modification of an intron of FaCENH3-7b, or any combination thereof relative to an unmodified Fragaria x ananassa plant; c. a modification of an enhancer of FaCENH3-7c, a modification of a promoter of FaCENH3-7c, a modification of a coding region of FaCENH3-7c, a modification of an intron of FaCENH3-7c, or any combination thereof relative to an unmodified Fragaria x ananassa plant; and/or d. a modification of an enhancer of FaCENH3-7d, a modification of a promoter of FaCENH3-7d, a modification of a coding region of FaCENH3-7d, a modification of an intron of FaCENH3-7d, or any combination thereof relative to an unmodified Fragaria x ananassa plant. 171 sf-6744554 197072001240 26. The strawberry plant, or plant part thereof, of any one of embodiments 1-25, wherein the expression or activity of one or more CENH3 genes is decreased by no more than about 50%, about 60%, about 70%, about 80%, about 90%, about 95%, or about 99% as compared to a control plant, plant part, or plant cell lacking the one or more genetic modifications. 27. The strawberry plant, or plant part thereof, of any one of embodiments 1-26, wherein the expression or activity of one or more CENH3 genes is decreased by at least about 5%, 10%, 20%, 30%, 40%, or 50% as compared to a control plant, plant part, or plant cell lacking the one or more genetic modifications. 28. The strawberry plant of any one of embodiments 1-27, wherein the strawberry plant has decreased pollen germination, flower diameter, anther width, filament length, or any combination thereof as compared to a control strawberry plant lacking the one or more genetic modifications. 29. The strawberry plant of any one of embodiments 1-28, wherein the strawberry plant has delayed microspore formation as compared to a control strawberry plant lacking the one or more genetic modifications. 30. The strawberry plant of any one of embodiments 1-29, wherein the strawberry plant has a larger bud size at which microspores form as compared to a control strawberry plant lacking the one or more genetic modifications. 31. The strawberry plant of any one of embodiments 28-30, wherein the control strawberry plant is a strawberry plant of the same ploidy as the strawberry plant, the same species as the strawberry plant, or both. 32. The strawberry plant, or plant part thereof, of any one of embodiments 1-31, wherein the plant part is selected from the group consisting of pollen, an anther, a seed, an achene, a leaf, a flower, a fruit, and a stolon. 33. The strawberry plant, or plant part thereof, of any one of embodiments 1-32, wherein the plant part is pollen. 34. The strawberry plant, or plant part thereof, of any one of embodiments 1-33, wherein the plant part is a seed. 35. A haploid-inducing strawberry plant, or a plant part thereof, comprising one or more genetic modifications resulting in decreased expression of one or more CENH3 genes. 36. The haploid-inducing strawberry plant, or plant part thereof, of embodiment 35, wherein one or more of the CENH3 genes comprise a polynucleotide sequence encoding an amino acid sequence having at least 90%, at least 95%, at least 96%, at least 97%, at least 172 sf-6744554 197072001240 95%, at least 99%, or 100% identity to an amino acid sequence selected from the group consisting of SEQ ID NOs: 8-14 and 53-56. 37. The haploid-inducing strawberry plant, or plant part thereof, of embodiment 35 or 36, wherein one or more of the CENH3 genes comprises a polynucleotide sequence having at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity to a polynucleotide sequence selected from the group consisting of SEQ ID NOs: 1- 7 and 58-61. 38. The haploid-inducing strawberry plant, or plant part thereof, of any one of embodiments 35-37, wherein the one or more genetic modifications comprise a modification of an enhancer of one or more of the CENH3 genes, a modification of a promoter of one or more of the CENH3 genes, a modification of a coding region of one or more of the CENH3 genes, a modification of an intron of one or more of the CENH3 genes, a modification of methylation status of one or more of the CENH3 genes, expression of a repressor protein that targets the DNA or an mRNA of one or more of the CENH3 genes, expression of an RNA interference construct that targets an mRNA of one or more of the CENH3 genes, or any combination thereof. 39. The haploid-inducing strawberry plant, or plant part thereof, of any one of embodiments 35-37, wherein the one or more genetic modifications comprise a modification of an enhancer of one or more of the CENH3 genes, a modification of a promoter of one or more of the CENH3 genes, a modification of a coding region of one or more of the CENH3 genes, modification of an intron of one or more of the CENH3 genes, or any combination thereof relative to an unmodified strawberry plant of the same species or an unmodified control strawberry plant. 40. The haploid-inducing strawberry plant, or plant part thereof, of any one of embodiments 35-39, wherein the haploid-inducing strawberry plant or plant part has decreased expression of CENH3 proteins relative to a strawberry plant of the same species lacking the one or more genetic modifications. 41. The haploid-inducing strawberry plant, or plant part thereof, of any one of embodiments 35-40, wherein the haploid-inducing strawberry plant or plant part lacks detectable expression of CENH3 proteins. 42. The haploid-inducing strawberry plant, or plant part thereof, of any one of embodiments 35-41, wherein the haploid-inducing strawberry plant is diploid, triploid, tetraploid, pentaploid, hexaploid, septaploid, octoploid, or nonaploid, or has a ploidy of 10x, 11x, 12x, 13x, 14x, 15x, 16x, 17x, 18x, 19x, or 20x. 173 sf-6744554 197072001240 43. The haploid-inducing strawberry plant, or plant part thereof, of any one of embodiments 35-42, wherein the haploid-inducing strawberry plant is diploid. 44. The haploid-inducing strawberry plant, or plant part thereof, of embodiment 43, wherein the haploid-inducing strawberry plant is a plant of the species Fragaria vesca, Fragaria iinumae, Fragaria nipponica, Fragaria viridis, Fragaria × bifera, Fragaria bucharica, Fragaria chinensis, Fragaria daltoniana, Fragaria emeiensis, Fragaria hayatae, Fragaria iinumae, Fragaria mandshurica, Fragaria viridis, Fragaria nilgerrensis, Fragaria nipponica, Fragaria nubicola, or Fragaria pentaphylla. 45. The haploid-inducing strawberry plant, or plant part thereof, of embodiment 43, wherein the haploid-inducing strawberry plant is a plant of the species Fragaria vesca. 46. The haploid-inducing strawberry plant, or plant part thereof, of embodiment 45, wherein the haploid-inducing strawberry plant is a Fragaria vesca plant of the subspecies Fragaria vesca ssp. vesca, Fragaria vesca ssp. americana, or Fragaria vesca ssp. bracteate. 47. The haploid-inducing strawberry plant, or plant part thereof, of any one of embodiments 43-46, wherein the one or more CENH3 genes comprise a polynucleotide sequence encoding an amino acid sequence having at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity the amino acid sequence of SEQ ID NO: 8. 48. The haploid-inducing strawberry plant, or plant part thereof, of any one of embodiments 43-47, wherein the one or more CENH3 genes comprise a polynucleotide sequence having at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity to the polynucleotide sequence of SEQ ID NO: 1. 49. The haploid-inducing strawberry plant, or plant part thereof, of any one of embodiments 45-48, wherein the haploid-inducing strawberry plant is a plant of the species Fragaria vesca and wherein the one or more CENH3 genes comprise FvCENH3. 50. The haploid-inducing strawberry plant, or plant part thereof, of embodiment 49, wherein the one or more genetic modifications comprise a modification of an enhancer of FvCENH3, a modification of a promoter of FvCENH3, a modification of a coding region of FvCENH3, modification of an intron of FvCENH3, or any combination thereof relative to a wild-type Fragaria vesca plant. 51. The haploid-inducing strawberry plant, or plant part thereof, of any one of embodiments 35-42, wherein the haploid-inducing strawberry plant is octoploid. 174 sf-6744554 197072001240 52. The haploid-inducing strawberry plant, or plant part thereof, of embodiment 51, wherein the haploid-inducing strawberry plant is a plant of the species Fragaria x ananassa, Fragaria chiloensis, Fragaria virginiana, or Fragaria iturupensis. 53. The haploid-inducing strawberry plant, or plant part thereof, of embodiment 52, wherein the haploid-inducing strawberry plant is Fragaria chiloensis plant of the subspecies Fragaria chiloensis subsp. chiloensis, Fragaria chiloensis subsp. lucida, Fragaria chiloensis subsp. pacifica, or Fragaria chiloensis subsp. sandwicensis. 54. The haploid-inducing strawberry plant, or plant part thereof, of embodiment 53, wherein the haploid-inducing strawberry plant is a plant of the species Fragaria x ananassa. 55. The haploid-inducing strawberry plant, or plant part thereof, of embodiment 54, wherein the haploid-inducing strawberry plant is a plant of the variety ‘Chandler’ 56. The haploid-inducing strawberry plant, or plant part thereof, of any one of embodiments 51-55, wherein one or more of the CENH3 genes comprises a polynucleotide sequence encoding an amino acid sequence having at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity to an amino acid sequence selected from the group consisting of SEQ ID NOs: 9-14 and 53-56. 57. The haploid-inducing strawberry plant, or plant part thereof, of any one of embodiments 51-56, wherein one or more of the CENH3 genes comprises a polynucleotide sequence having at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity to a polynucleotide sequence selected from the group consisting of SEQ ID NOs: 2-7 and 58-61. 58. The haploid-inducing strawberry plant, or plant part thereof, of any one of embodiments 51-57, wherein the one or more CENH3 genes comprise: a. a polynucleotide sequence encoding an amino acid sequence having at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity to the amino acid sequence of SEQ ID NO: 9; b. a polynucleotide sequence encoding an amino acid sequence having at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity to the amino acid sequence of SEQ ID NO: 10; c. a polynucleotide sequence encoding an amino acid sequence having at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity to the amino acid sequence of SEQ ID NO: 11; 175 sf-6744554 197072001240 d. a polynucleotide sequence encoding an amino acid sequence having at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity to the amino acid sequence of SEQ ID NO: 12; e. a polynucleotide sequence encoding an amino acid sequence having at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity to the amino acid sequence of SEQ ID NO: 13; and/or f. a polynucleotide sequence encoding an amino acid sequence having at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity to the amino acid sequence of SEQ ID NO: 14. 59. The haploid-inducing strawberry plant, or plant part thereof, of any one of embodiments 51-58, wherein the one or more CENH3 genes comprise: a. a polynucleotide sequence having at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity to the polynucleotide sequence of SEQ ID NO: 2; b. a polynucleotide sequence having at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity to the polynucleotide sequence of SEQ ID NO: 3; c. a polynucleotide sequence having at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity to the polynucleotide sequence of SEQ ID NO: 4; d. a polynucleotide sequence having at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity to the polynucleotide sequence of SEQ ID NO: 5; e. a polynucleotide sequence having at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity to the polynucleotide sequence of SEQ ID NO: 6; and/or f. a polynucleotide sequence having at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity to the polynucleotide sequence of SEQ ID NO: 7. 60. The haploid-inducing strawberry plant, or plant part thereof, of any one of embodiments 51-59, wherein the one or more CENH3 genes comprise: 176 sf-6744554 197072001240 a. a polynucleotide sequence encoding an amino acid sequence having at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity to the amino acid sequence of SEQ ID NO: 53; b. a polynucleotide sequence encoding an amino acid sequence having at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity to the amino acid sequence of SEQ ID NO: 54; c. a polynucleotide sequence encoding an amino acid sequence having at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity to the amino acid sequence of SEQ ID NO: 55; and/or d. a polynucleotide sequence encoding an amino acid sequence having at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity to the amino acid sequence of SEQ ID NO: 56. 61. The haploid-inducing strawberry plant, or plant part thereof, of any one of embodiments 51-60, wherein the one or more CENH3 genes comprise: a. a polynucleotide sequence having at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity to the polynucleotide sequence of SEQ ID NO: 58; b. a polynucleotide sequence having at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity to the polynucleotide sequence of SEQ ID NO: 59; c. a polynucleotide sequence having at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity to the polynucleotide sequence of SEQ ID NO: 60; and/or d. a polynucleotide sequence having at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity to the polynucleotide sequence of SEQ ID NO: 61. 62. The haploid-inducing strawberry plant, or plant part thereof, of embodiment 51-61, wherein the haploid-inducing strawberry plant is a plant of the species Fragaria x ananassa and the one or more CENH3 genes comprise one, two, three, four, five, or all six of FaCENH3-1a, FaCENH3-1b, FaCENH3-2a, FaCENH3-2b, FaCENH3-3, and FaCENH3-4. 63. The haploid-inducing strawberry plant, or plant part thereof, of embodiment 62, wherein the one or more CENH3 genes comprise FaCENH3-1a, FaCENH3-1b, FaCENH3- 2a, FaCENH3-2b, FaCENH3-3, and FaCENH3-4. 177 sf-6744554 197072001240 64. The haploid-inducing strawberry plant, or plant part thereof, of embodiment 62 or 63, wherein the one or more genetic modifications comprise: a. a modification of an enhancer of FaCENH3-1a, a modification of a promoter of FaCENH3-1a, a modification of a coding region of FaCENH3-1a, a modification of an intron of FaCENH3-1a, or any combination thereof relative to an unmodified Fragaria x ananassa plant; b. a modification of an enhancer of FaCENH3-1b, a modification of a promoter of FaCENH3-1b, a modification of a coding region of FaCENH3-1b, a modification of an intron of FaCENH3-1b, or any combination thereof relative to an unmodified Fragaria x ananassa plant; c. a modification of an enhancer of FaCENH3-2a, a modification of a promoter of FaCENH3-2a, a modification of a coding region of FaCENH3-2a, a modification of an intron of FaCENH3-2a, or any combination thereof relative to an unmodified Fragaria x ananassa plant; d. a modification of an enhancer of FaCENH3-2b, a modification of a promoter of FaCENH3-2b, a modification of a coding region of FaCENH3-2b, a modification of an intron of FaCENH3-2b, or any combination thereof relative to an unmodified Fragaria x ananassa plant; e. a modification of an enhancer of FaCENH3-3, a modification of a promoter of FaCENH3-3, a modification of a coding region of FaCENH3-3, a modification of an intron of FaCENH3-3, or any combination thereof relative to an unmodified Fragaria x ananassa plant; and/or f. a modification of an enhancer of FaCENH3-4, a modification of a promoter of FaCENH3-4, a modification of a coding region of FaCENH3-4, a modification of an intron of FaCENH3-3, or any combination thereof relative to an unmodified Fragaria x ananassa plant. 65. The haploid-inducing strawberry plant, or plant part thereof, of embodiment 51-61, wherein the haploid-inducing strawberry plant is a plant of the species Fragaria x ananassa and the one or more CENH3 genes comprise one, two, three, four, five, or all six of FaCENH3-7a, FaCENH3-7b, FaCENH3-7c, and FaCENH3-7d. 66. The haploid-inducing strawberry plant, or plant part thereof, of embodiment 65, wherein the one or more genetic modifications comprise: a. a modification of an enhancer of FaCENH3-7a, a modification of a promoter of FaCENH3-7a, a modification of a coding region of FaCENH3-7a, a 178 sf-6744554 197072001240 modification of an intron of FaCENH3-7a, or any combination thereof relative to an unmodified Fragaria x ananassa plant; b. a modification of an enhancer of FaCENH3-7b, a modification of a promoter of FaCENH3-7b, a modification of a coding region of FaCENH3-7b, a modification of an intron of FaCENH3-7b, or any combination thereof relative to an unmodified Fragaria x ananassa plant; c. a modification of an enhancer of FaCENH3-7c, a modification of a promoter of FaCENH3-7c, a modification of a coding region of FaCENH3-7c, a modification of an intron of FaCENH3-7c, or any combination thereof relative to an unmodified Fragaria x ananassa plant; and/or d. a modification of an enhancer of FaCENH3-7d, a modification of a promoter of FaCENH3-7d, a modification of a coding region of FaCENH3-7d, a modification of an intron of FaCENH3-7d, or any combination thereof relative to an unmodified Fragaria x ananassa plant. 67. The haploid-inducing strawberry plant, or plant part thereof, of any one of embodiments 35-66, wherein the expression or activity of one or more CENH3 genes is decreased by no more than about 50%, about 60%, about 70%, about 80%, about 90%, about 95%, or about 99% as compared to a control plant, plant part, or plant cell lacking the one or more genetic modifications. 68. The haploid-inducing strawberry plant, or plant part thereof, of any one of embodiments 35-67, wherein the expression or activity of one or more CENH3 genes is decreased by at least about 5%, 10%, 20%, 30%, 40%, or 50% as compared to a control plant, plant part, or plant cell lacking the one or more genetic modifications. 69. The haploid-inducing strawberry plant of any one of embodiments 35-68, wherein the haploid-inducing strawberry plant has decreased pollen germination, flower diameter, anther width, filament length, or any combination thereof as compared to a control strawberry plant lacking the one or more genetic modifications. 70. The haploid-inducing strawberry plant of any one of embodiments 35-69, wherein the haploid-inducing strawberry plant has delayed microspore formation as compared to a control strawberry plant lacking the one or more genetic modifications. 71. The haploid-inducing strawberry plant of any one of embodiments 35-67, wherein the haploid-inducing strawberry plant has a larger bud size at which microspores form as compared to a control strawberry plant lacking the one or more genetic modifications. 179 sf-6744554 197072001240 72. The haploid-inducing strawberry plant of any one of embodiments 69-71, wherein the control strawberry plant is a strawberry plant of the same ploidy as the haploid-inducing strawberry plant, the same species as the haploid-inducing strawberry plant, or both. 73. The haploid-inducing strawberry plant, or plant part thereof, of any one of embodiments 35-72, wherein the plant part is selected from the group consisting of pollen, an anther, a seed, an achene, a leaf, a flower, a fruit, and a stolon. 74. The haploid-inducing strawberry plant, or plant part thereof, of any one of embodiments 35-73, wherein the plant part is pollen. 75. The haploid-inducing strawberry plant, or plant part thereof, of any one of embodiments 35-74, wherein the plant part is a seed. 76. A method of producing the strawberry plant of any one of embodiments 1-34 or the haploid-inducing strawberry plant of any one of embodiments 35-75. 77. The method embodiment 76, wherein the decreased expression of the one or more CENH3 genes is achieved by gene disruption, gene knockout, gene knockdown, gene silencing, RNA interference, induction of methylation, or any combination thereof. 78. The method of embodiment 76 or 78, comprising introducing one or more of the genetic modifications by mutagenesis, gene editing, transgenesis, or a combination thereof into a strawberry plant, plant part, or plant cell to produce the strawberry plant or the haploid- inducing strawberry plant. 79. The method of any one of embodiments 76-78, further comprising selecting a strawberry plant, plant part, or plant cell having decreased expression of the one or more CENH3 genes to produce the strawberry plant or the haploid-inducing strawberry plant. 80. The method of embodiment 76-79, comprising introducing one or more of the genetic modifications by gene editing using a site-directed nuclease. 81. The method of embodiment 80, wherein the site-directed nuclease is a CRISPR- associated (Cas) nuclease, a zinc finger nuclease (ZFN), a transcription activator-like effector nuclease (TALEN), or a megaTAL. 82. The method of any one of embodiments 76-81, comprising contacting a plurality of cells of a parent strawberry plant with one or more expression vectors together comprising an expression cassette for a Cas nuclease and an expression cassette for an RNA molecule having at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% complementarity to 15, 16, 17, 18, 19, or 20 consecutive nucleotides of one or more of the CENH3 genes. 180 sf-6744554 197072001240 83. The method of any one of embodiments 76-81, comprising contacting a plurality of cells of a parent strawberry plant with one or more Cas nucleases, wherein each Cas nuclease is complexed with a RNA molecule having at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% complementarity to 15, 16, 17, 18, 19, or 20 consecutive nucleotides of one or more of the CENH3 genes. 84. The method of embodiment 82 or 83, wherein the RNA molecule is a crRNA, a gRNA, or a pegRNA. 85. The method of embodiment 84, wherein the RNA molecule is complementary to one or more nucleotide sequences consisting of SEQ ID NOs: 15-24 or comprises a nucleotide sequence selected from the group consisting of SEQ ID NOs: 25-34. 86. The method of embodiment 84, wherein the RNA molecule is complementary to one or more nucleotide sequences consisting of SEQ ID NOs: 17, 19, and 21, or comprises a nucleotide sequence selected from the group consisting of SEQ ID NOs: 27, 29, or 31. 87. The method any one of embodiments 82-86, wherein the plurality of cells is a plurality of protoplasts. 88. The method of any one of embodiments 82087, wherein the method further comprises, subsequent to the contacting step, allowing the plurality of cells of the parent strawberry plant to form calli, plants, or a combination thereof, and identifying one or more calli or plants having the genetic modifications resulting in decreased expression of one or more CENH3 genes. 89. A method of producing true homozygous octoploid strawberry seed, the method comprising (a) contacting a tetrahaploid reduced egg cell from a donor octoploid strawberry plant with a sperm cell of the haploid-inducing strawberry plant of any one of embodiments 35-75, and (b) producing a doubled tetrahaploid cell from the tetrahaploid reduced egg cell. 90. The method of embodiment 89, wherein step (a) comprises contacting pollen of the haploid-inducing strawberry plant with the stigma of a pistil of the donor octoploid strawberry plant comprising the reduced egg cell and allowing formation of a pollen tube and migration of the pollen tube to the reduced egg cell, thereby contacting the reduced egg cell with the sperm cell. 91. The method of embodiment 90, further comprising allowing the donor octoploid strawberry plant to form a plurality of seeds. 92. The method of embodiment 91, further comprising selecting one or more tetrahaploid seeds by 1) determining the ploidy of the embryo and endosperm of one or more seeds of the 181 sf-6744554 197072001240 plurality of seeds, and 2) selecting one or more seeds having tetraploid embryo and an endosperm having a ploidy of greater than 8x. 93. A method of producing true homozygous octoploid strawberry seed, the method comprising (a) contacting an egg cell from the haploid-inducing strawberry plant of any one of embodiments 35-75 with a tetrahaploid reduced sperm cell from a donor octoploid strawberry plant, and (b) producing a doubled tetrahaploid cell from the egg cell, wherein the doubled tetrahaploid cell comprises the nuclear genome of the donor octoploid strawberry plant and the cytoplasmic genome of the haploid-inducing strawberry plant. 94. The method of embodiment 93, wherein the haploid-inducing strawberry plant and the donor octoploid strawberry plants are from different species of Fragaria. 95. The method of embodiment 94, wherein the haploid-inducing strawberry plant is diploid. 96. The method of embodiment 95, wherein the haploid-inducing strawberry plant is a plant of the species Fragaria vesca. 97. The method of embodiment 95, wherein the haploid-inducing strawberry plant is a plant of the species Fragaria x ananassa. 98. The method of any one of embodiments 93-97, wherein the donor octoploid strawberry plant is a plant of the species Fragaria x ananassa. 99. The method of any one of embodiments 93-98, wherein step (a) comprises contacting the stigma of a pistil of the haploid-inducing strawberry plant with the pollen from the donor octoploid strawberry plant and allowing formation of a pollen tube and migration of the pollen tube to the egg cell, thereby contacting the egg cell with the tetrahaploid reduced sperm cell. 100. The method of embodiment 99, further comprising allowing the haploid-inducing strawberry plant to form a plurality of seeds. 101. The method of embodiment 91, 92, or 99, further comprising collecting the plurality of seeds and allowing them to germinate and form a plurality of seedlings. 102. The method of embodiment 91, 92, or 99, further comprising collecting a plurality of embryos from the plurality of seeds, contacting the plurality of embryos with a nutrient medium suitable for inducing seedling growth, and allowing them to form a plurality of seedlings. 103. The method of embodiment 101 or 102, further comprising selecting one or more tetrahaploid seedlings by 1) determining the ploidy of the cells of one or more seedlings of 182 sf-6744554 197072001240 the plurality of seedlings, and 2) selecting one or more tetrahaploid seedlings from the plurality of seedling. 104. The method of embodiment 103, wherein determining the ploidy of the cells of the one or more seedlings comprises obtaining a tissue sample of one or more seedlings and determining the ploidy of the cells of the tissue sample. 105. The method of any one of embodiments 89-104, wherein step (b) comprises subjecting the selected tetrahaploid cell, tetrahaploid seed, or tetrahaploid seedling to a tetrahaploid doubling treatment. 106. The method of embodiment 105, wherein the tetrahaploid doubling treatment comprises contacting the tetrahaploid cell, tetrahaploid seed, or tetrahaploid seedling with an anti-microtubule agent to form a doubled tetrahaploid cell, a doubled tetrahaploid seed, or a doubled tetrahaploid seedling. 107. The method of embodiment 106, wherein the anti-microtubule agent is an anti-mitotic herbicide. 108. The method of embodiment 106, wherein the anti-microtubule agent comprises colchicine, oryzalin, oxide, trifluralin, or any combination thereof. 109. The method of any one of embodiments 106-108, wherein the doubled tetrahaploid cell or the doubled tetrahaploid seed is allowed to grow into a doubled tetrahaploid seedling. 110. The method of any one of embodiments 106-109, wherein the doubled tetrahaploid seedling is allowed to form seed, thus producing the true homozygous octoploid strawberry seed. 111. The method of any one of embodiments 89-110, wherein the donor octoploid strawberry plant is a plant of the species Fragaria x ananassa, Fragaria chiloensis, Fragaria virginiana, or Fragaria iturupensis, or a hybrid of any combination thereof. 112. The method of embodiment 111, wherein the donor octoploid strawberry plant is Fragaria chiloensis plant of the subspecies Fragaria chiloensis subsp. chiloensis, Fragaria chiloensis subsp. lucida, Fragaria chiloensis subsp. pacifica, or Fragaria chiloensis subsp. sandwicensis. 113. The method of embodiment 111, wherein the donor octoploid strawberry plant is a plant of the species Fragaria x ananassa. 114. The method of embodiment 113, wherein the donor octoploid strawberry plant is a Fragaria x ananassa plant of the variety Camarosa or Albion. 115. The method of any one of embodiments 89-114, wherein the method further comprises introducing one or more genetic modifications resulting in decreased expression of 183 sf-6744554 197072001240 a gibberellin 20-oxidase (GA20ox) gene into the true homozygous octoploid strawberry plant or seed. 116. A method of producing a runnerless true homozygous octoploid strawberry plant comprising the steps of the method of any one of embodiments 89-115 and further comprising introducing one or more genetic modifications resulting in decreased expression of a gibberellin 20-oxidase (GA20ox) gene into the true homozygous octoploid strawberry plant, wherein the runnerless true homozygous octoploid strawberry plant does not produce runners. 117. A true homozygous octoploid strawberry seed produced according to the method of any one of embodiments 89-116. 118. The true homozygous octoploid strawberry seed of embodiment 117, wherein the true homozygous octoploid strawberry seed comprises one or more genetic modifications resulting in decreased expression of a gibberellin 20-oxidase (GA20ox) gene. 119. A true homozygous octoploid strawberry plant or a plant part thereof produced according to the method of any one of embodiments 89-116. 120. The true homozygous octoploid strawberry plant of embodiment 119, wherein the true homozygous octoploid strawberry plant comprises one or more genetic modifications resulting in decreased expression of a gibberellin 20-oxidase (GA20ox) gene. 121. The true homozygous octoploid strawberry plant of embodiment 120, wherein the true homozygous octoploid strawberry plant does not produce runners. 122. The true homozygous octoploid strawberry plant, or plant part thereof, of any one of embodiments 119-121, wherein the plant part is selected from the group consisting of pollen, an anther, a seed, an achene, a leaf, a flower, a fruit, and a stolon. 123. A true homozygous octoploid strawberry seed produced according to the method of any one of embodiments 47-68, comprising an embryo having the nuclear genome of the donor octoploid strawberry plant and the cytoplasmic genome of the haploid-inducing strawberry plant, wherein the nuclear genome and the cytoplasmic genome are from plants of different varieties of strawberry. 124. The true homozygous octoploid strawberry seed of embodiment 123, wherein the different varieties of strawberry are different species of strawberry. 125. The true homozygous octoploid strawberry seed of embodiment 123, wherein the different varieties of strawberry are different varieties of Fragaria x ananassa. 126. A true homozygous octoploid strawberry seed produced according to the method of any one of embodiments 89-116, comprising an embryo having the nuclear genome of the 184 sf-6744554 197072001240 donor octoploid strawberry plant and the cytoplasmic genome of the haploid-inducing strawberry plant, wherein the nuclear genome and the cytoplasmic genome are from plants of different species of Fragaria. 127. The true homozygous octoploid strawberry seed of embodiment 126, wherein the cytoplasmic genome is from a plant of the species Fragaria vesca. 128. The true homozygous octoploid strawberry seed of embodiment 126 or 127, wherein the nuclear genome is from a plant of the species Fragaria x ananassa. 129. The true homozygous octoploid strawberry seed of any one of embodiments 126-128, wherein the true homozygous octoploid strawberry seed comprises one or more genetic modifications resulting in decreased expression of a gibberellin 20-oxidase (GA20ox) gene. 130. A true homozygous octoploid strawberry plant or a plant part thereof produced according to the method of any one of embodiments 89-116, comprising at least one somatic cell having the nuclear genome of the donor octoploid strawberry plant and the cytoplasmic genome of the haploid-inducing strawberry plant, wherein the nuclear genome and the cytoplasmic genome are from plants of different varieties of strawberry. 131. The true homozygous octoploid strawberry seed of embodiment 130, wherein the different varieties of strawberry are different species of strawberry. 132. The true homozygous octoploid strawberry seed of embodiment 130, wherein the different varieties of strawberry are different varieties of Fragaria x ananassa. 133. A true homozygous octoploid strawberry plant or a plant part thereof produced according to the method of any one of embodiments 89-116, comprising at least one somatic cell having the nuclear genome of the donor octoploid strawberry plant and the cytoplasmic genome of the haploid-inducing strawberry plant, wherein the nuclear genome and the cytoplasmic genome are from plants of different species of Fragaria. 134. The true homozygous octoploid strawberry plant of embodiment 133, wherein the cytoplasmic genome is from a plant of the species Fragaria vesca. 135. The true homozygous octoploid strawberry plant of embodiment 133 or 134, wherein the nuclear genome is from a plant of the species Fragaria x ananassa. 136. The true homozygous octoploid strawberry plant of any one of embodiments 133-135, wherein the true homozygous octoploid strawberry plant comprises one or more genetic modifications resulting in decreased expression of a gibberellin 20-oxidase (GA20ox) gene. 137. The true homozygous octoploid strawberry plant of embodiment 136, wherein the true homozygous octoploid strawberry plant does not produce runners. 185 sf-6744554 197072001240 138. The true homozygous octoploid strawberry plant, or plant part thereof, of any one of embodiments 130-137, wherein the plant part is selected from the group consisting of pollen, an anther, a seed, an achene, a leaf, a flower, a fruit, and a stolon. 139. A true homozygous octoploid strawberry seed comprising an embryo having the nuclear genome of a first variety of strawberry and the cytoplasmic genome of a variety of strawberry. 140. The true homozygous octoploid strawberry seed of embodiment 139, wherein the first and second varieties of strawberry are different species of strawberry. 141. The true homozygous octoploid strawberry seed of embodiment 139, wherein the first and second varieties of strawberry are two different varieties of Fragaria x ananassa. 142. A true homozygous octoploid strawberry seed comprising an embryo having the nuclear genome of a first species of Fragaria and the cytoplasmic genome of a second plant species of Fragaria. 143. The true homozygous octoploid strawberry seed of embodiment 142, wherein the first species of Fragaria is Fragaria x ananassa. 144. The true homozygous octoploid strawberry seed of embodiment 142 or 143, wherein the second species of Fragaria is a diploid species of Fragaria. 145. The true homozygous octoploid strawberry seed of embodiment 144, wherein the second species of Fragaria is Fragaria vesca. 146. The true homozygous octoploid strawberry seed of any one of embodiments 142-145, wherein the true homozygous octoploid strawberry seed comprises one or more genetic modifications resulting in decreased expression of a gibberellin 20-oxidase (GA20ox) gene. 147. A true homozygous octoploid strawberry plant or plant part thereof comprising an embryo having the nuclear genome of a first variety of strawberry and the cytoplasmic genome of a second variety of strawberry. 148. The true homozygous octoploid strawberry seed of embodiment 147, wherein the first and second varieties of strawberry are different species of strawberry. 149. The true homozygous octoploid strawberry seed of embodiment 147, wherein the first and second varieties of strawberry are two different varieties of Fragaria x ananassa. 150. A true homozygous octoploid strawberry plant or plant part thereof comprising an embryo having the nuclear genome of a first species of Fragaria and the cytoplasmic genome of a second plant species of Fragaria. 151. The true homozygous octoploid strawberry plant of embodiment 150, wherein the first species of Fragaria is Fragaria x ananassa. 186 sf-6744554 197072001240 152. The true homozygous octoploid strawberry plant of embodiment 150 or 151, wherein the second species of Fragaria is a diploid species of Fragaria 153. The true homozygous octoploid strawberry plant of embodiment 152, wherein the second species of Fragaria is Fragaria vesca. 154. The true homozygous octoploid strawberry plant of any one of embodiments 150-153, wherein the true homozygous octoploid strawberry seed comprises one or more genetic modifications resulting in decreased expression of a gibberellin 20-oxidase (GA20ox) gene. 155. The true homozygous octoploid strawberry plant of embodiment 154, wherein the true homozygous octoploid strawberry plant does not produce runners. 156. The true homozygous octoploid strawberry plant, or plant part thereof, of any one of embodiments 147-155, wherein the plant part is selected from the group consisting of pollen, an anther, a seed, an achene, a leaf, a flower, a fruit, and a stolon. 157. A method of producing uniform octoploid F1 hybrid strawberry seed comprising crossing two true homozygous octoploid strawberry plants of any one of embodiments 119- 122, 130-138, and 147-156. 158. A uniform octoploid F1 hybrid strawberry seed produced according to the method of embodiment 157. 159. The uniform octoploid F1 hybrid strawberry seed of embodiment 158, wherein the uniform octoploid F1 hybrid strawberry seed comprises one or more genetic modifications resulting in decreased expression of a gibberellin 20-oxidase (GA20ox) gene. 160. A uniform octoploid F1 hybrid strawberry plant or a plant part thereof produced according to the method of embodiment 157. 161. The uniform octoploid F1 hybrid strawberry plant of embodiment 160, wherein the uniform octoploid F1 hybrid strawberry plant comprises one or more genetic modifications resulting in decreased expression of a gibberellin 20-oxidase (GA20ox) gene. 162. The uniform octoploid F1 hybrid strawberry plant of embodiment 161, wherein the uniform octoploid F1 hybrid strawberry plant does not produce runners. 163. The uniform octoploid F1 hybrid strawberry plant, or plant part thereof, of any one of embodiments 160-162, wherein the plant part is selected from the group consisting of pollen, an anther, a seed, an achene, a leaf, a flower, a fruit, and a stolon. 164. An expression vector or isolated DNA molecule for making the strawberry plant, or plant part thereof, of any one of embodiments 1-34, or the haploid-inducing strawberry plant, or a plant part thereof, of any one of embodiments 35-75. 187 sf-6744554 197072001240 165. The expression vector or isolated DNA molecule of embodiment 164, comprising a DNA sequence having at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity or complementarity to 15, 16, 17, 18, 19, or 20 consecutive nucleotides of a polynucleotide sequence encoding an amino acid sequence having at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity to an amino acid sequence selected from the group consisting of SEQ ID NOs: 8-14 and 53-56. 166. The expression vector or isolated DNA molecule of embodiment 164 or 165, comprising a DNA sequence having at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity or complementarity to 15, 16, 17, 18, 19, or 20 consecutive nucleotides of a polynucleotide having at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity to an polynucleotide sequence selected from the group consisting of SEQ ID NOs: 1-7 and 58-61. 167. The expression vector or isolated DNA molecule of any one of embodiments 164- 166, comprising a DNA sequence encoding a non-coding RNA. 168. The expression vector or isolated DNA molecule of embodiment 167, wherein the non-coding RNA comprises a polynucleotide sequence having at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity or complementarity to 15, 16, 17, 18, 19, or 20 consecutive nucleotides of a polynucleotide sequence encoding an amino acid sequence having at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity to an amino acid sequence selected from the group consisting of SEQ ID NOs: 8-14 and 53-56. 169. The expression vector or isolated DNA molecule of embodiment 167 or 168, wherein the non-coding RNA comprises a polynucleotide sequence having at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity or complementarity to 15, 16, 17, 18, 19, or 20 consecutive nucleotides of a polynucleotide having at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity to an polynucleotide sequence selected from the group consisting of SEQ ID NOs: 1-7 and 58-61. 170. The expression vector or isolated DNA molecule of embodiment 167-169, wherein the RNA molecule is complementary to one or more nucleotide sequences consisting of SEQ ID NOs: 15-24 or comprises a nucleotide sequence selected from the group consisting of SEQ ID NOs: 25-34. 171. The expression vector or isolated DNA molecule of embodiment 167-169, wherein the RNA molecule is complementary to one or more nucleotide sequences consisting of SEQ 188 sf-6744554 197072001240 ID NOs: 17, 19, and 21, or comprises a nucleotide sequence selected from the group consisting of SEQ ID NOs: 27, 29, or 31. 172. The expression vector or isolated DNA molecule of any one of embodiments 167- 171, wherein the non-coding RNA is a crRNA, a gRNA, a pegRNA, a siRNA, a miRNA, or a dsRNA. 173. The expression vector or isolated DNA molecule of any one of embodiments 164- 172, comprising a DNA sequence encoding a site-directed nuclease. 174. The expression vector or isolated DNA molecule of embodiment 173 wherein the site- directed nuclease is a Cas nuclease, a TALEN, ZFN, or a mega-TAL. 175. The expression vector or isolated DNA molecule of embodiment 173 or 174, wherein the site-directed nuclease comprises an amino acid sequence that confers binding to a polynucleotide sequence having at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity or complementarity to 15, 16, 17, 18, 19, or 20 consecutive nucleotides of a polynucleotide sequence encoding an amino acid sequence having at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity to an amino acid sequence selected from the group consisting of SEQ ID NOs: 8-14 and 53-56. 176. The expression vector or isolated DNA molecule of any one of embodiments 173- 175, wherein the site-directed nuclease comprises an amino acid sequence that confers binding to a polynucleotide sequence having at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity or complementarity to 15, 16, 17, 18, 19, or 20 consecutive nucleotides of a polynucleotide having at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity to an polynucleotide sequence selected from the group consisting of SEQ ID NOs: 1-7 and 58-61. 177. A bacterial cell comprising the expression vector or isolated DNA molecule of any one of embodiments 164-176. 178. The bacterial cell of embodiment 177, wherein the bacterial cell is an Agrobacterium cell. 179. A genetically modified plant, plant part, plant cell, or seed comprising the expression vector or isolated DNA molecule of any one of embodiments 164-176. 180. A kit comprising the expression vector or isolated DNA molecule of 164-176 or the bacterial cell of embodiment 177 or 178. 181. A genetically modified non-regenerable plant cell of the strawberry plant of any one of embodiments 1-34 or the haploid-inducing strawberry plant of any one of embodiments 35-75. 189 sf-6744554 197072001240 182. A genetically modified plant genome of the strawberry plant of any one of embodiments 1-34 or the haploid-inducing strawberry plant of any one of embodiments 35- 75. EXAMPLES [0274] The presently disclosed subject matter will be better understood by reference to the following Examples, which are provided as exemplary of the invention, and not by way of limitation. Example 1: General Methods Plants [0275] An octoploid Fragaria x ananassa ‘Chandler' (PED524) and a diploid Fragaria vesca Hawaii 4 (PED505) and Alexandria (PED503) were initiated in vitro by meristem extraction using hormone free ½ MS media (Murashige & Skoog (1962) “A revised medium for rapid growth and bio assays with tobacco tissue cultures” Physiologia Plantarum, 15(3): 473-497). The hormone free ½ MS media was supplemented with 30 g/L sucrose and plants were then grown under 16 hour day / 8 hour night lighting regime with cool white fluorescent lighting. After establishment, plants were micropropagated on the same medium. RNPs [0276] RNPs were prepared by standard methods known in the art. In short, 2 µL of NEB buffer 2.1 (10x stock) was placed into a 1.5 mL microcentrifuge tube with 10-40 µg of crRNA (CRISPR RNA) and an equal mass of Cas nuclease. The final volume was adjusted to 20 µL using nuclease free water. Protoplasts [0277] Approximately 0.3 g of leaves from 3 to 4 week old explants were removed under aseptic conditions and sliced into thin sections 0.5 mm to 1 mm in width. Protoplasts were prepared from the sliced leaf tissue essentially as described in Barceló et al. (Barceló et al. (2019) “Isolation and culture of strawberry protoplasts and field evaluation of regenerated plants” Scientia Horticulturae 256: 108552). Protoplast cells were quantified using a Bürker hemocytometer before transfection. 190 sf-6744554 197072001240 Transfection [0278] Protoplasts were combined with freshly prepared RNPs and transformed using standard PEG transfection methods, essentially as described in Y. Gou et al. (Gou et al. (2020) “Optimization of the protoplast transient expression system for gene functional studies in strawberry (Fragaria vesca)” Plant Cell, Tissue, and Organ Culture 141(1): 41-53). In short, RNPs were added to 100 µL of isolated protoplast solution (approximately 2 x 105 protoplasts per mL) in 2 mL microfuge tubes. An equal volume of PEG solution (40% PEG 4000 (wt/vol), 0.2 M mannitol, and 100 mM CaCl2) was immediately mixed with the protoplasts by shaking gently and thoroughly. The amount of plasmid DNA was determined experimentally between 5-80 µg. Transfection mixtures were incubated at room temperature for 2-40 minutes, as determined experimentally, and then diluted with 440 µL W5 solution (2 mM MES (pH 5.7), 154 mM NaCl, 5 mM glucose, 125 mM CaCl2, and 5 mM KCl) at room temperature, and the transfection process terminated by gentle shaking. Protoplasts were centrifuged at 100 x g for 1 minute at room temperature. Finally, the protoplasts were gently resuspended with 200 µL WI solution (4 mM MES (pH 5.7), 0.5 M mannitol, and 20 mM KCl) and incubated in the dark for 20-25 hours at room temperature. Plant regeneration [0279] Protoplasts were plated and maintained under sterile conditions until callus formation. Callus with confirmed edits was regenerated essentially as described in Barceló et al. (Barceló et al. (2019) “Isolation and culture of strawberry protoplasts and field evaluation of regenerated plants” Scientia Horticulturae 256: 108552). [0280] Alternatively protoplasts were encapsulated using alginate and cultured in callus induction media. Once microcallus were approximately 1-3 mm in size, microcallus were transferred to shoot induction media. Shoots from protoplasts with confirmed edits was regenerated essentially as described in Barceló et al. (Barceló et al. (2019) “Isolation and culture of strawberry protoplasts and field evaluation of regenerated plants” Scientia Horticulturae 256: 108552). Example 2: Identification of target sites and guide RNA design Identification of cenh3 orthologs 191 sf-6744554 197072001240 [0281] Candidate CENH3 orthologs from strawberry were identified by comparing CENH3 protein sequences from Zea mays (ZmCENH3) (SEQ ID NO: 44) and Arabidopsis thaliana (AtCENH3) (SEQ ID NO: 35) using Protein Blast, tblastn, and Clustal Omega workflows. [0282] FvCENH3 (SEQ ID NO: 8) from diploid strawberry Fragaria vesca and six FaCENH3 homeologs (SEQ ID NOs: 9-14) from octoploid strawberry Fragaria x ananassa (Gene IDs based on Fragaria x ananassa Camarosa Genome v1.0.a2: www[dot]rosaceae[dot]org/Analysis/9642085) were identified as putative orthologs of ZmCENH3 (SEQ ID NO: 44) and AtCENH3 (SEQ ID NO: 35). [0283] The configuration of the CENH3 loci in Fragaria x ananassa is illustrated in FIG. 3. The configuration of the CENH3 loci in Fragaria x ananassa is illustrated in FIG. 4. [0284] Additional CENH3 orthologs were identified from Potentilla micrantha CENH3 (SEQ ID NO: 42) and closely related Fragaria species, Fragaria chiloensis CENH3-AV (SEQ ID NO: 36), Fragaria chiloensis CENH3-B1 (SEQ ID NO: 37), Fragaria chiloensis CENH3-B2 (SEQ ID NO: 38), Fragaria chiloensis CENH3-Bi (SEQ ID NO: 39), Fragaria iinumae CENH3 (SEQ ID NO: 40), and Fragaria viridis CENH3 (SEQ ID NO: 41) were identified. The protein sequence alignment and a phylogenetic tree of CENH3 orthologs from Fragaria species, Zea mays, Arabidopsis thaliana are shown in FIG. 5A-5C. In FIG. 6, the phylogenetic tree is a UPGMA consensus tree. The scale bar represents phylogenetic distance of 0.05 nucleotide substitutions per site. The tree is generated using software Geneious Prime 2023.2.1, with Genetic Distance Model: Jukes-Cantor. Guide RNA design [0285] Protospacer sequences targeting FvCENH3 (SEQ ID NO: 1) or FaCENH3 homeologs (SEQ ID NO: 2-7) in Table 2 were selected. Guide RNAs were synthesized based on protospacer sequence and scaffold sequence and were screened in vivo. Table 2: Protospacer sequences 192 sf-6744554 197072001240 Editing in protoplasts [0286] RNPs with the synthesized guide RNAs and a Cas nuclease were prepared as in Example 1. Protoplasts were generated, transfected with the RNPs, and regenerated as described in Example 1. Sequence-based confirmation of editing [0287] Primers were designed to amplify each candidate edited cenh3 region based on the reference genome, and one or more long range direct PCRs were performed using crude lysate from transfected protoplasts. Alternatively, DNA was extracted from callus, leaf or other plant material. PCR products were pooled by transfection sample and a seqWell library preparation performed to generate an Illumina library. Samples were loaded onto an Illumina iSeq and sequenced with a Paired End 150 nt sequencing kit. Sequences were analyzed by aligning fastq files to reference sequences, and mutations adjacent to target PAM sites for each targeted cenh3 gene were tabulated relative to a control. Editing efficiency was calculated based on the frequency of observed mutations as illustrated in FIG. 7, and used to calculate how many plants should be screened to identify the cenh3 gene knockouts required to confer a haploid induction phenotype. Guides targets protospacers PRS874, PRS876 and PRS878 were selected based on the observed editing efficiencies in protoplasts (FIG. 7). Example 3: Haploid inducer system for strawberry Creation of haploid inducer lines [0288] An octoploid Fragaria x ananassa (PED524) and a diploid Fragaria vesca (PED505) were edited with guide RNAs targeting PRS874 (SEQ ID NO. 27), PRS876 193 sf-6744554 197072001240 (SEQ ID NO: 29), and PRS878 (SEQ ID NO. 31) which were selected based on their unexpectedly high editing efficiency. These guide RNAs are specific to FvCENH3 and all six homeologs of FaCENH3 in Fragaria x ananassa Camarosa (FIGS. 8A, 9A, and 10A), as well as all four homeologs of FaCENH3 in Fragaria x ananassa Royal Royce (FIGS. 8B, 9B, and 10B). [0289] Table 3A provides exemplary Fragaria x ananassa plants designated PED524- Fa-I-01 through PED524-Fa-I-33 which bear the indicated combinations of edited, nonfunctional cenh3 alleles, and functional CENH3 alleles, respectively. Edited, non- functional cenh3 alleles are denoted by a 0 and functional CENH3 alleles are denoted by a 1. Each allele is given using the same terminology as FIG. 3 (-1a, -1b, -2a, -2b, -3 and 4), wherein “m” designates the maternal copy of that respective allele and “p” designates the paternal copy of that respective allele. Additional Fragaria x ananassa plants are recovered, such that enough plants are recovered to cover all possible combinations of functional and edited, non-functional alleles (Table 3A provides an exemplary list and does not provide all possible combinations, all of which are recovered and within the scope of this example). Table 3A: Fragaria x ananassa haploid inducer lines. 194 sf-6744554 197072001240 [0290] Table 3B provides the Fragaria vesca plants designated PED505-Fv-Inducer-01 through PED505-Fv-Inducer-03 which bear the indicated combinations of edited, nonfunctional cenh3 alleles, and functional CENH3 alleles, respectively. Edited, non- functional cenh3 alleles are denoted by a 0, a weak allele (resulting in reduced, but not eliminated, expression of CENH3) is denoted by a 1, and a functional CENH3 alleles are 195 sf-6744554 197072001240 denoted by a 2. The letter “m” designates the maternal copy of the respective allele and “p” designates the paternal copy of that respective allele. Table 3B: Fragaria vesca haploid inducer lines [0291] After in vitro propagation, the populations of plants indicated the groups in Table 4 (below) are grown to maturity. Pollen is harvested from mature flowers of the plants from Groups A-i, C, E-i and G and used to pollinate the emasculated flowers of the plants from Groups B-i, D and F-i. All combinations for haploid induction of the maternal and paternal genome are tested including those described in Table 5 (below). For the unedited groups C and D, other strawberry varieties may be used, both cultivated and wild. Table 4: Exemplary groups of strawberry plants for haploid induction 196 sf-6744554 197072001240 Table 5: Exemplary crosses for haploid induction [0292] Upon maturation of the berries all seeds (achenes) of the unmodified plants are collected, surface disinfected and placed into hormone free ½ MS basal salts for in vitro germination. Germinated seeds are grown in vitro until the second true leaf forms, at which point a leaf is harvested for crude lysate DNA extraction. Potential tetraploids are first screened by one of two molecular marker assays depending on the ploidy level of the inducer. When a diploid Fragaria vesca inducer is used, a presence-absence marker between Fragaria vesca and Fragaria x ananassa is used for the screening. The marker is uniquely present in Fragaria vesca but not in any octoploid Fragaria x ananassa. F1 plants with the presence of this marker will be hybrids and F1 plants with the absence of this marker will be candidate 197 sf-6744554 197072001240 tetraploids. When an octoploid Fragaria x ananassa inducer is used, a drop-off assay detecting CENH3 mutation at the editing site will be used for screening. F1 plants with edited CENH3 alleles at the editing site will be hybrids and F1 plants with all WT CENH3 at the editing site will be candidate induced tetraploids or spontaneously doubled tetrahaploids. Confirmation of tetraploids is through flow cytometry performed essentially as described in Galbraith et al. (1983. “Rapid flow cytometric analysis of the cell cycle in intact plant tissues” Science 220(4601): 1049-1051). In short, intact nuclei are extracted into Galbraith buffer, then filtered and stained with propidium iodide. DNA content of nuclei is determined by applying the samples to a BD Accuri™ C6 Flow Cytometer. Gating is performed and genomic DNA content is calculated by comparing the peak area for the sample to the known position of the internal control. Tetrahaploids are identified based on relative DNA content of nuclei between samples and internal controls, i.e. samples with approximately half the DNA content of unmodified octoploid Fragaria x ananassa controls are classified as tetrahaploid. [0293] The tetrahaploid induction rate varies from among the ten populations of plants recovered, as determined by the number of haploid seedlings observed (including spontaneously doubled tetrahaploid) divided by the total number of germinated seedlings that develop. Haploid doubling [0294] Tetrahaploids are propagated in vitro on ½ MS medium with 0.5 mg/L 6- benzylaminopurine (BAP) and are subsequently doubled to produce fully homozygous octoploid plants with treatment of 0.1 to 0.3% colchicine solution in vitro for 4 to 24 hours. Successful doubling of tetrahaploids is initially identified by plant morphology (larger and thicker leaves) and is then confirmed by flow cytometry using the same methods as above. Resulting fully homozygous octoploid plants have approximately twice the DNA content of the unmodified tetrahaploid plants and are similar in DNA content to unmodified octoploid parents. Production of uniform F1 hybrid octoploid strawberry [0295] Fully homozygous octoploid plants are crossed with one another to produce uniform true seed populations of F1 hybrid octoploid Fragaria x ananassa. Crosses are performed to identify the best breeding parents for commercial production. Seeds are 198 sf-6744554 197072001240 collected and grown to maturity, where they exhibit phenotypic uniformity and increased heterotic vigor. Production of “runner-less” uniform F1 hybrid octoploid strawberry [0296] Plants are also edited to disrupt the expression of GA20ox, to reduce or eliminate the production of stolons (also referred to as “runners” in the industry). Editing occurs at any stage in the process including in the parent lines or in the tetrahaploid lines. [0297] In some examples, an octoploid Fragaria x ananassa (PED524) is edited to disrupt expression of FaGa20ox4-1 (SEQ. ID 46) and FaGa20ox4-3 (SEQ. ID 47), while simultaneously being edited for one or more FvCenh3 gene as described above. In other examples, a tetrahaploid (which has already undergone cenh3-induced haploid induction) is edited to disrupt expression of FaGa20ox4-1 (SEQ. ID 46) and FaGa20ox4-3 (SEQ. ID 47). Example 4: Evaluation of CENH3-Edited Strawberry Lines Methods Sequence-based confirmation of editing [0298] Primers were designed to amplify each candidate edited cenh3 region based on the Fragaria x ananassa Camarosa or Royal Royce reference genome, and one or more long range direct PCRs were performed using crude lysate front transfected protoplasts. Alternatively, DNA was extracted from callus, leaf, or other plant material. PCR products were pooled either by transfection sample or plant material, and a seqWell library preparation was performed to generate Illumina libraries. Samples were sequenced on either an Illumina iSeq or MiSeq i100 using either a paired end 150 nt or paired end 300nt sequencing kit. [0299] To quantify editing rates in transfection samples, reads were aligned to a reference genome, and the frequencies of mutations within a window adjacent to the target PAM sites were calculated relative to a control. Editing efficiency was calculated based on the frequency of observed mutations as illustrated in FIG. 7, and used to calculate how many plants should be screened to identify the cenh3 knockouts required to confer a haploid induction phenotype. Guide targeting PRS874, PRS876, and PRS878 were selected based on the observed editing efficiencies in protoplasts (FIG. 7). To characterize and phase edited haplotypes in regenerated shoots, reads were aligned to a reference genome, and local haplotypes 199 sf-6744554 197072001240 surrounding each cut site were determined based on nucleotide differences between each homoeolog of cenh3. Lines were prioritized based on the number of edits, their size, and predicted functional impact. Identification of cenh3 mutant lines [0300] Fragaria x ananassa and Fragaria vesca lines were edited at cenh3 loci as described in Examples 1-3. The total number of callus and callus with shoots was recorded. Tissue from callus with shoots was sampled for sequencing. Edited shoots were identified, excised from the callus, elongated, multiplied, and rooted for maintenance in tissue culture for cell and growth evaluation or transplanted in the greenhouse for gene expression, flower morphology and ploidy evaluation. Cell density, size and division evaluation [0301] Five plantlets of each cenh3 mutant were maintained in tissue culture on rooting media at temperature between 20-25℃, a 16-hour photoperiod with a photon flux density of 40 µmol m-2 s-1, and a relative humidity of 60-75%. Leaves from 3-week old plantlets of 15 cenh3 mutant F. ×ananassa (variety “Chandler”; PED524) were harvested and protoplasts were isolated, following methods described in Example 1. In addition to quantifying protoplast cells using a Bürker hemocytometer, protoplast size was measured. High- resolution microscopic images of protoplasts were annotated and used to train a custom object detection model capable of identifying viable protoplasts and generating bounding boxes around each cell. Protoplasts without bounding boxes are non-viable. The bounding box dimensions were then used to determine cell size (FIG. 11A). Isolated protoplasts were encapsulated and maintained under sterile conditions, following methods described in Example 1.Six encapsulated lens were made for each cenh3 mutant line. The number of microcallus 0.2–0.4 mm in size per encapsulated lens was recorded 21 days after encapsulation. (FIG. 11B). Experiments evaluating cell density, size, and division were repeated twice. Flower morphology, fruit set and ploidy evaluation [0302] Six plantlets of each cenh3 mutant were transplanted in the greenhouse. At least three plants of each cenh3 mutant were maintained in the greenhouse for evaluation of gene flower morphology and used as females for crosses with wild type F. ×ananassa (variety “Albion”) to evaluate ploidy of progeny. 200 sf-6744554 197072001240 [0303] One F. x ananassa ‘Chandler’ wild-type and 26 cenh3 mutants were transplanted in the greenhouse. Greenhouse conditions were optimized for floral induction. The minimum and maximum day and night temperatures were 15–24℃. The minimum and maximum day and night humidity were 54–90% (average 77%) and 32–90% (average 68%). The photoperiod was 12 hours. [0304] Emerging, folded leaflets were destructively harvested from healthy plants at the same developmental stage and time of day. Leaf tissue was immediately flash-frozen in liquid nitrogen and stored at -80℃ until RNA extraction. Total RNA was extracted using an RNA isolation kit following the manufacturer’s protocol. RNA quality and quantity were assessed and high-quality RNA samples were used to construct sequencing libraries for transcriptome analysis. [0305] Shortly after the cenh3 mutants flowered, between 3–5 months after transplanting, reproductive morphology, pollen germination, and flower architecture was evaluated. Three fully expanded primary or secondary flowers with bright-yellow anthers were destructively sampled from three plants. The flower was categorized as either perfect (male and female reproductive structures), staminate (male only) or pistillate (female only). Number of flowers per truss, , petal color, filament length, anther width and flower diameter were evaluated. Calipers were used to measure filament length, anther width, and flower diameter in mm. Pollen germination was evaluated by removing three bright-yellow anthers from each of the three flowers. Anthers were placed in a 24-well plate and incubated at room temperature for an hour. After, the plate was gently tapped to release the pollen. Then, 400 µL of pollen germination media (20 mg/L boric acid, 100 g/L sucrose, pH 5.7) was added to each well and the plate was incubated at room temperature in the dark for four hours. Pollen germination was evaluated by counting the number of pollen grains with pollen tubes greater than the length of pollen grains. Either 50 or 100 pollen grains were observed, depending on the abundance of pollen grains. [0306] Flowers that were not destructively harvested were emasculated 3–4 days prior to opening. Then, 3–4 days after emasculation, receptive, glossy stigmas were pollinated using dried pollen collected from bright-yellow anthers of a wild type F. ×ananassa (variety “Albion”). Approximately 4–6 weeks after pollination, ripe fruits were harvested and seeds were removed. The number of seeds were recorded. Seeds were sanitized and germinated in- vitro by cutting the seed in half using a size 10 scalpel blade, placing the cut seed on 201 sf-6744554 197072001240 germination media (2.2 g/L MS basal salts with vitamins, 10 g/L sucrose, 3 g/L agar, pH 5.7), and incubating the seed at temperature between 20–25℃, a 16-hour photoperiod with a photon flux density of 40 µmol m-2 s-1, and a relative humidity of 60–75%. Microspore development microscopy [0307] Strawberry buds at a variety of sizes were picked off the plant and measured. Six cenh3 mutant lines were analyzed (3 - 4 buds selected per plant) and 2 WT lines (3 buds/plant). Buds were then fixed in Farmers Fixative 1:3 acetic acid:95% EtOH) overnight at 4C. Buds were washed in 70% EtOH and stored in fresh 70% EtOH at -20C. Each bud was then dissected to expose the anthers. Two anthers were extracted from each bud and slides were prepped for microscopy by squashing the tissue between a cover slip and microscope slide with a drop of acetocarmine solution. Slides were heated at 95C for 5 min and then visualized on a light microscope. Presence or absence of microspores was determined visually. Haploid induction rate [0308] Ploidy was determined by sampling cotyledon tissue, approximately 0.2 cm2, from germinated seedlings 7-21 days after germination by flow cytometry. Tissue from F. vesca ‘Hawaii 4’ (2n), interspecific hybrid (5n), F. x ananassa ‘Chandler’ (8n) were collected as controls. Tissue was added to a 96-well plate with 30 μl of nuclei extraction buffer (Cystain kit). The tissue was finely cut using tattoo gun for 40-60 seconds. The instrument was rinsed thoroughly between samples. After, 170 ul of nuclei extraction buffer was added to each sample. Then, four samples were pooled together by dispensing 50 ul of each sample through the Celltrics filters (total volume 200 ul). The remaining 150 ul of each sample was stored was stored in the refrigerator. The filters were removed and 500 μl of extraction buffer (60 μl RNase A, 20 ml staining buffer, and 120 μl propidium iodide) was added to the sample. The pooled samples incubated for 30-60 minutes at room temperature. The BD CSampler Plus software was used to analyze the results. The settings were as follows: run with limits, medium for fluidics, and threshold value for 2,000. The “A” detector channel was used to determine the mean phycoerythrin fluorescent protein-chromophore (PE) labeled molecules bound to the nuclei. The mean PE-A values of the peak(s) were recorded. If a pooled sample had two or more peaks, each individual was re-sampled individually by dispensing the remaining 150 ul of the sample stored in the refrigerator through the Celltric filters, removing the filters, adding 500 ml of the extraction buffer and incubating for 30-60 202 sf-6744554 197072001240 minutes at room temperature The number of aneuploids and tetrahaploids was recorded. The frequency of aneuploids was calculated as the number of aneuploid individuals divided by the total number of germinated seedlings. The haploid induction rate was calculated as the number of tetrahaploids divided by the total number of germinated seedlings. Results Identification of cenh3 mutant lines [0309] The regeneration rate, the number of calli with shoots of the total number of calli of F. × ananassa (variety “Chandler”) was 38.0% (Table 6). A total of 26 edited shoots of Chandler were identified from callus with shoots derived from protoplasts, respectively. The editing rate was, the number of calli with shoots that were edited of the total number of calli with shoots, 10.2%. The F. ×ananassa cenh3 mutant lines had between 1–6 alleles of the total 8 alleles edited. The editing profiles of the F. ×ananassa cenh3 mutant lines is shown in Table 7, and edited FaCENH3 allele sequences for a subset of lines are shown in FIGS. 12A- 12F (FaCENH3-7a), FIGS. 13A-13C (FaCENH3-7b), FIGS. 14A-14F (FaCENH3-7c), and FIGS. 15A-15C (FaCENH3-7d). Table 6. The number of calli, number of calli with shoots, regeneration rate (percentage of callus with shoots by the total callus) edited shoots, and editing rate (percentage of edited callus with shoots by the total callus with shoots) of F. ×ananassa Chandler protoplasts transfected with gRNA targeting either cenh3. Table 7. Summary of phased edits in mutant cenh3 lines. Bracketed values correspond to deletion sizes (nt) in the alleles of either Subgenome A, Subgenome B, Subgenome C, or Subgenome D, as indicated at the top of the respective column. ‘0’ corresponds to an unedited wild-type allele with no deletion. The two bracketed values in each cell correspond to the two homologous chromosomes of each subgenome. For example, row 2 under Subgenome A “['0', '9']” indicates that the line has an unedited cenh3 allele on one 203 sf-6744554 197072001240 homologous chromosome of Subgenome A and a 9-nt deletion on the other homologous chromosome of Subgenome A. Subgenome A Subgenome B Subgenome C Subgenome D 204 sf-6744554 197072001240 Cell density, size and division evaluation [0310] Fifteen cenh3 mutant lines were evaluated for cell density (cells/g of tissue), size, and growth. Differences were observed between cenh3 mutant lines and wild type. There were two cenh3 mutant lines, cenh3-13, cenh3-06, cenh3-13 withlower cell density compared to wild type. Two cenh3 mutant lines, cenh3-001 and cenh3-13, had significantly larger cells compared to wild type. These lines differed in cell growth (microcallus/ encapsulated lens. Four cenh3 mutant lines, cenh3-001, cenh3-005, cenh3-009, and cenh3-12 had fewer microcallus compared to wild type while on cenh3 mutant line, cenh3-010, had more microcallus compared to wild type. (Table 8). Table 8. Cell density (protoplasts/g of leaf tissue), cell area, and cell growth (microcallus/ encapsulated lens) of wild type and cenh3 mutant lines. Statistical comparisons between group means were performed using Tukey’s Honestly Significant Difference (HSD) post-hoc test following ANOVA. Differences were considered statistically significant at p < 0.05.. 205 sf-6744554 197072001240 Microspore development microscopy [0311] The average bud size at which microspores were present for WT samples was 4.6mm and the average bud size at which cenh3 mutants developed microspheres was 5.5mm (FIG. 16A). This suggests that more growth was necessary before reaching this stage of sexual maturity in the cenh3 mutant lines. Photographs of an exemplary squashed anther with microspheres is shown at various magnifications in FIGS. 16B-16D. In one cenh3 mutant plant (E-PED524-4145, cenh3-006), tetrads were observed in approximately two thirds of the buds. Tetrads are a normal stage prior to formation of microspores. Exemplary images of microspores observed in cenh3-006 are shown in FIGS. 16E and 16F. Flower morphology, fruit set and ploidy evaluation [0312] Observable differences between pollen germination, flower architecture, and fruit set are expected in cenh3 mutant lines – more so than differences in cell density, cell size, and cell division, as errors in meiosis are far less tolerated than in mitosis. Vegetative tissues are more resilient due to meristem plasticity, redundant growth pathways, and compensatory cell expansion. Reproductive tissues are less tolerant to defects in centrosome function and chromosome segregation as developmental timing is fixed and terminal. Defects can result in misshapen anthers, pollen viability and low seed set (Demidov et al. (2019). Deregulated phosphorylation of CENH3 at Ser65 affects the development of floral meristems in Arabidopsis thaliana. Frontiers in Plant Science, 10, 928.). Therefore, these characteristics 206 sf-6744554 197072001240 were evaluated to determine if the cenh3 mutants have the expected phenotypic traits consistent with other cenh3 mutants. [0313] Six of the fifteen F. x ananassa ‘Chandler’ cenh3 mutant lines evaluated for cell density, size, and growth were also evaluated for plant vigor, pollen germination and flower architecture. All the cenh3 mutant lines had good vigor (FIGS. 17A, 18A, 19A, 20A, 21A, 22A, and 23A) and an average of five perfect flowers per truss with white petals (FIGS. 17B, 18B, 19B, 20B, 21B, 22B, and 23B). There were no obvious differences in vigor from the wild type. The flower diameter was smaller for the cenh3 mutant lines compared to wild type. Two cenh3 mutant lines, cenh3-003 and cenh3-006 had a significantly smaller flower diameter compared to WT (p < 0.05). cenh3-006 also had smaller anther width and filament length (FIG. 22B). The anther width and filament length was 0.6 and 0.4 mm compared to the wild type, which was 1.1 and 2.1 mm, respectively. The rate of pollen germination, or the fraction of pollen grains which form a pollen tube after contacting a stigma, ranged from 3.8 - 64.2%. The cenh3 mutant line, cenh3-006, with lowest pollen germination also had the smallest flower diameter, anther width, and filament length. This line also had lower cell density and cell growth. cenh3-003 also had low pollen germination, small flower diameter and filament length (Table 9). Photos of exemplary plants, flowers, and anthers of cenh3 mutant lines are shown in FIGS. 17A-23B. [0314] The cenh3 mutant lines were then pollinated with pollen from F. x ananassa (‘Albion’), and pollination success rate was measured as the rate of fruit set. Of the flowering cenh3 mutant lines pollination success rate was greater than 80%, with the exception of cenh3-003 and cenh3-004 which was 58.9 and 0%, respectively. The seed per berry was lower for all the cenh3 mutant lines compared to wild type. Seed viability ranged from 40 - 80% (Table 10). Table 9. Mean pollen germination (%), flower diameter (mm), anther width (mm), and filament length (mm) ± standard error of wild type and cenh3 mutant lines. Statistical comparisons between group means were performed using Tukey’s Honestly Significant Difference (HSD) post-hoc test following ANOVA. Differences were considered statistically significant at p < 0.05.. 207 sf-6744554 197072001240 Table 10. Number of crosses made, number of successful crosses, average number of seed per berry, and percentage of viable seed of wild type and cenh3 mutant lines. Haploid induction rate [0315] The PE-A value for F. vesca ‘Hawaii 4’ (2n), interspecific hybrid (5n), and F. x ananassa ‘Chandler’ (8n) is approximately 44,000, 94,000, 140,000, respectively Two peaks were observed for the pooled sample of an interspecific hybrid (5n) and F. x ananassa ‘Chandler’ (8n) at a ratio of 1:3. The PE-values were approximately 94,000 and 140,000. There were pooled samples from a F. x ananassa ‘Chandler’ cenh3-007 x F. x ananassa ‘Albion’ cross where a peak was observed between 44,000 – 94,000, between 94,000 – 140,000 or greater than 140,000 PA-E value. When the samples were analyzed individually seedlings were confirmed to have a PA-E value between 44,000 – 94,000 were characterized as a tetrahaploid. Individual seedlings confirmed to have a PA-E value between 94,000- 140,000 or greater than 140,000 were characterized as aneuploid. The haploid induction and aneuploid rate for F. x ananassa ‘Chandler’ cenh3-006 crossed with F. x ananassa ‘Albion’ 208 sf-6744554 197072001240 is shown in Table 11. The Cenh3-006 parent plant that resulted in a tetrahaploid seedling is the same plant that was observed to have tetrads in about two thirds of the buds, as described above and exemplified in FIGS. 16E-16F. Table 11. Number of seedlings evaluated for ploidy from crosses with F. x ananassa ‘Chandler’ wild type or cenh3 mutant lines as the female and F. x ananassa ‘Albion’ as the male. 209 sf-6744554

Claims

197072001240 CLAIMS What is claimed is: 1. A strawberry plant, or a plant part thereof, comprising one or more genetic modifications resulting in decreased expression of one or more CENH3 genes. 2. The strawberry plant, or plant part thereof, of claim 1, wherein one or more of the CENH3 genes comprise a polynucleotide sequence encoding an amino acid sequence having at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity to an amino acid sequence selected from the group consisting of SEQ ID NOs: 8-14 and 53-56. 3. The strawberry plant, or plant part thereof, of claim 1 or 2, wherein one or more of the CENH3 genes comprises a polynucleotide sequence having at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity to a polynucleotide sequence selected from the group consisting of SEQ ID NOs: 1-7 and 58-61. 4. The strawberry plant, or plant part thereof, of any one of claims 1-3, wherein the one or more genetic modifications comprise a modification of an enhancer of one or more of the CENH3 genes, a modification of a promoter of one or more of the CENH3 genes, a modification of a coding region of one or more of the CENH3 genes, modification of an intron of one or more of the CENH3 genes, or any combination thereof relative to an unmodified strawberry plant of the same species or an unmodified control strawberry plant, optionally wherein the one or more genetic modifications comprise a deletion, insertion, or one or more nucleotide changes in a coding region of one or more of the CENH3 genes. 5. The strawberry plant, or plant part thereof, of any one of claims 1-4, wherein the strawberry plant is diploid. 6. The strawberry plant, or plant part thereof, of claim 5, wherein the strawberry plant is a plant of the species Fragaria vesca. 7. The strawberry plant, or plant part thereof, of claim 5 or 6, wherein the one or more CENH3 genes comprise a polynucleotide sequence encoding an amino acid sequence having at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity the amino acid sequence of SEQ ID NO: 8. 210 sf-6744554 197072001240 8. The strawberry plant, or plant part thereof, of any one of claims 5-7, wherein the one or more CENH3 genes comprise a polynucleotide sequence having at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity to the polynucleotide sequence of SEQ ID NO: 1. 9. The strawberry plant, or plant part thereof, of any one of claims 1-4, wherein the strawberry plant is octoploid. 10. The strawberry plant, or plant part thereof, of claim 9, wherein the strawberry plant is a plant of the species Fragaria x ananassa. 11. The strawberry plant, or plant part thereof, of claim 9 or 10, wherein one or more of the CENH3 genes comprises a polynucleotide sequence encoding an amino acid sequence having at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity to an amino acid sequence selected from the group consisting of SEQ ID NOs: 9-14 and 53-56. 12. The strawberry plant, or plant part thereof, of any one of claims 9-11, wherein one or more of the CENH3 genes comprises a polynucleotide sequence having at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity to a polynucleotide sequence selected from the group consisting of SEQ ID NOs: 2-7 and 58-61. 13. The strawberry plant, or plant part thereof, of any one of claims 9-12, wherein the one or more CENH3 genes comprise: a. a polynucleotide sequence encoding an amino acid sequence having at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity to the amino acid sequence of SEQ ID NO: 53; b. a polynucleotide sequence encoding an amino acid sequence having at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity to the amino acid sequence of SEQ ID NO: 54; c. a polynucleotide sequence encoding an amino acid sequence having at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity to the amino acid sequence of SEQ ID NO: 55; and/or 211 sf-6744554 197072001240 d. a polynucleotide sequence encoding an amino acid sequence having at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity to the amino acid sequence of SEQ ID NO: 56. 14. The strawberry plant, or plant part thereof, of any one of claims 9-13, wherein the one or more CENH3 genes comprise: a. a polynucleotide sequence having at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity to the polynucleotide sequence of SEQ ID NO: 58; b. a polynucleotide sequence having at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity to the polynucleotide sequence of SEQ ID NO: 59; c. a polynucleotide sequence having at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity to the polynucleotide sequence of SEQ ID NO: 60; and/or d. a polynucleotide sequence having at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity to the polynucleotide sequence of SEQ ID NO: 61. 15. The strawberry plant, or plant part thereof, of any one of claims 1-14, wherein the expression or activity of one or more CENH3 genes is decreased by no more than about 50%, about 60%, about 70%, about 80%, about 90%, about 95%, or about 99% as compared to a control plant, plant part, or plant cell lacking the one or more genetic modifications. 16. The strawberry plant, or plant part thereof, of any one of claims 1-15, wherein the expression or activity of one or more CENH3 genes is decreased by at least about 1%, 5%, 10%, 20%, 30%, 40%, or 50% as compared to a control plant, plant part, or plant cell lacking the one or more genetic modifications. 17. The strawberry plant of any one of claims 1-16, wherein the strawberry plant has: 212 sf-6744554 197072001240 (a) decreased pollen germination, flower diameter, anther width, filament length, fruit per berry, cell growth rate, or any combination thereof as compared to a control strawberry plant lacking the one or more genetic modifications; (b) delayed microspore formation as compared to a control strawberry plant lacking the one or more genetic modifications; and/or (c) the strawberry plant has a larger bud size at which microspores form as compared to a control strawberry plant lacking the one or more genetic modifications as compared to a control strawberry plant lacking the one or more genetic modifications. 18. The strawberry plant, or plant part thereof, of any one of claims 1-17, wherein the plant part is selected from the group consisting of pollen, an anther, a seed, an achene, a leaf, a flower, a fruit, and a stolon. 19. The strawberry plant, or plant part thereof, of any one of claims 1-18, wherein the strawberry plant is a haploid-inducing strawberry plant. 20. A method of producing the strawberry plant of any one of claims 1-19. 21. The method claim 20, wherein the decreased expression of the one or more CENH3 genes is achieved by gene disruption, gene knockout, gene knockdown, gene silencing, RNA interference, induction of methylation, or any combination thereof. 22. The method of claim 20 or 21, comprising introducing one or more of the genetic modifications by mutagenesis, gene editing, transgenesis, or a combination thereof into a strawberry plant, plant part, or plant cell to produce the strawberry plant. 23. The method of any one of claims 20-22, further comprising selecting a strawberry plant, plant part, or plant cell having decreased expression of the one or more CENH3 genes to produce the strawberry plant. 24. The method of claim 20-23, comprising introducing one or more of the genetic modifications by gene editing using a site-directed nuclease. 213 sf-6744554 197072001240 25. The method of any one of claims 20-24, comprising contacting a plurality of cells of a parent strawberry plant with one or more expression vectors together comprising an expression cassette for a Cas nuclease and an expression cassette for an RNA molecule having at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% complementarity to 15, 16, 17, 18, 19, or 20 consecutive nucleotides of one or more of the CENH3 genes, optionally wherein the RNA molecule is a crRNA, a gRNA, or a pegRNA. 26. The method of any one of claims 20-25, comprising contacting a plurality of cells of a parent strawberry plant with one or more Cas nucleases, wherein each Cas nuclease is complexed with a RNA molecule having at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% complementarity to 15, 16, 17, 18, 19, or 20 consecutive nucleotides of one or more of the CENH3 genes, optionally wherein the RNA molecule is a crRNA, a gRNA, or a pegRNA. 27. A method of producing true homozygous octoploid strawberry seed, the method comprising (a) contacting a tetrahaploid reduced egg cell from a donor octoploid strawberry plant with a sperm cell of the haploid-inducing strawberry plant of claim 19, and (b) producing a doubled tetrahaploid cell from the tetrahaploid reduced egg cell. 28. The method of claim 27, wherein step (a) comprises contacting pollen of the haploid- inducing strawberry plant with the stigma of a pistil of the donor octoploid strawberry plant comprising the reduced egg cell and allowing formation of a pollen tube and migration of the pollen tube to the reduced egg cell, thereby contacting the reduced egg cell with the sperm cell. 29. The method of claim 28, further comprising allowing the donor octoploid strawberry plant to form a plurality of seeds. 30. A method of producing true homozygous octoploid strawberry seed, the method comprising (a) contacting an egg cell from the haploid-inducing strawberry plant of claim 19 with a tetrahaploid reduced sperm cell from a donor octoploid strawberry plant, and (b) producing a doubled tetrahaploid cell from the egg cell, wherein the doubled tetrahaploid cell comprises the nuclear genome of the donor octoploid strawberry plant and the cytoplasmic genome of the haploid-inducing strawberry plant. 214 sf-6744554 197072001240 31. The method of claim 30, wherein step (a) comprises contacting the stigma of a pistil of the haploid-inducing strawberry plant with the pollen from the donor octoploid strawberry plant and allowing formation of a pollen tube and migration of the pollen tube to the egg cell, thereby contacting the egg cell with the tetrahaploid reduced sperm cell. 32. The method of claim 31, further comprising allowing the haploid-inducing strawberry plant to form a plurality of seeds. 33. The method of claim 29 or 32, further comprising: (a) collecting the plurality of seeds and allowing them to germinate and form a plurality of seedlings; or (b) collecting a plurality of embryos from the plurality of seeds, contacting the plurality of embryos with a nutrient medium suitable for inducing seedling growth, and allowing them to form a plurality of seedlings. 34. The method of claim 33, further comprising selecting one or more tetrahaploid seedlings by 1) determining the ploidy of the cells of one or more seedlings of the plurality of seedlings, and 2) selecting one or more tetrahaploid seedlings from the plurality of seedling. 35. The method of any one of claims 27-34, wherein step (b) comprises subjecting the selected tetrahaploid cell, tetrahaploid seed, or tetrahaploid seedling to a tetrahaploid doubling treatment. 36. The method of claim 35, wherein the tetrahaploid doubling treatment comprises contacting the tetrahaploid cell, tetrahaploid seed, or tetrahaploid seedling with an anti- microtubule agent to form a doubled tetrahaploid cell, a doubled tetrahaploid seed, or a doubled tetrahaploid seedling, optionally wherein the anti-microtubule agent comprises colchicine, oryzalin, oxide, trifluralin, or any combination thereof. 37. The method of any one of claims 27-36, wherein the donor octoploid strawberry plant is a plant of the species Fragaria x ananassa. 38. A method of producing a runnerless true homozygous octoploid strawberry plant comprising the steps of the method of any one of claims 27-37 and further comprising introducing one or more genetic modifications resulting in decreased expression of a 215 sf-6744554 197072001240 gibberellin 20-oxidase (GA20ox) gene into the true homozygous octoploid strawberry plant, wherein the runnerless true homozygous octoploid strawberry plant does not produce runners. 39. A true homozygous octoploid strawberry seed produced according to the method of any one of claims 27-38. 40. A true homozygous octoploid strawberry plant or a plant part thereof produced according to the method of any one of claims 27-38. 41. A true homozygous octoploid strawberry seed produced according to the method of any one of claims 27-38, comprising an embryo having the nuclear genome of the donor octoploid strawberry plant and the cytoplasmic genome of the haploid-inducing strawberry plant, wherein the nuclear genome and the cytoplasmic genome are from plants of different varieties of strawberry. 42. A true homozygous octoploid strawberry plant or a plant part thereof produced according to the method of any one of claims 27-38, comprising at least one somatic cell having the nuclear genome of the donor octoploid strawberry plant and the cytoplasmic genome of the haploid-inducing strawberry plant, wherein the nuclear genome and the cytoplasmic genome are from plants of different varieties of strawberry. 43. The true homozygous octoploid strawberry seed of claim 41 or the true homozygous octoploid strawberry plant or plant part of claim 42, wherein the different varieties of strawberry are different species of Fragaria, optionally wherein the first species of Fragaria is Fragaria x ananassa and the second species of Fragaria is Fragaria vesca. 44. The true homozygous octoploid strawberry seed of claim 41 or the true homozygous octoploid strawberry plant or plant part of claim 42, wherein the different varieties of strawberry are different varieties of Fragaria x ananassa. 45. A true homozygous octoploid strawberry seed comprising an embryo having the nuclear genome of a first variety of strawberry and the cytoplasmic genome of a variety of strawberry. 46. A true homozygous octoploid strawberry plant or plant part thereof comprising an embryo having the nuclear genome of a first variety of strawberry and the cytoplasmic genome of a second variety of strawberry. 216 sf-6744554 197072001240 47. The true homozygous octoploid strawberry seed of claim 56 or the true homozygous octoploid strawberry plant or plant part of claim 46, wherein the first and second varieties of strawberry are different species of Fragaria, optionally wherein the first species of Fragaria is Fragaria x ananassa and the second species of Fragaria is Fragaria vesca. 48. The true homozygous octoploid strawberry seed of claim 56 or the true homozygous octoploid strawberry plant or plant part of claim 46, wherein the first and second varieties of strawberry are two different varieties of Fragaria x ananassa. 49. The true homozygous octoploid strawberry plant, or plant part thereof, of any one of claims 40, 42-44, and 46-48, wherein the plant part is selected from the group consisting of pollen, an anther, a seed, an achene, a leaf, a flower, a fruit, and a stolon. 50. A method of producing uniform octoploid F1 hybrid strawberry seed comprising crossing two true homozygous octoploid strawberry plants of any one of claims 40, 42-44, and 46-48. 51. A uniform octoploid F1 hybrid strawberry seed produced according to the method of claim 50. 52. A uniform octoploid F1 hybrid strawberry plant or a plant part thereof produced according to the method of claim 50. 53. The uniform octoploid F1 hybrid strawberry plant, or plant part thereof, of claim 52, wherein the plant part is selected from the group consisting of pollen, an anther, a seed, an achene, a leaf, a flower, a fruit, and a stolon. 54. An expression vector or isolated DNA molecule for making the strawberry plant, or plant part thereof, of any one of claims 1-19. 55. The expression vector or isolated DNA molecule of claim 54, comprising a DNA sequence having at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity or complementarity to 15, 16, 17, 18, 19, or 20 consecutive nucleotides of a polynucleotide sequence encoding an amino acid sequence having at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity to an amino acid sequence selected from the group consisting of SEQ ID NOs: 8-14 and 53-56. 217 sf-6744554 197072001240 56. The expression vector or isolated DNA molecule of claim 54 or 44, comprising a DNA sequence having at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity or complementarity to 15, 16, 17, 18, 19, or 20 consecutive nucleotides of a polynucleotide having at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity to an polynucleotide sequence selected from the group consisting of SEQ ID NOs: 1-7 and 58-61. 57. The expression vector or isolated DNA molecule of any one of claims 54-56, comprising a DNA sequence encoding a non-coding RNA, optionally wherein the non-coding RNA is a crRNA, a gRNA, a pegRNA, a siRNA, a miRNA, or a dsRNA. 58. The expression vector or isolated DNA molecule of claim 57, wherein the non-coding RNA comprises a polynucleotide sequence having at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity or complementarity to 15, 16, 17, 18, 19, or 20 consecutive nucleotides of a polynucleotide sequence encoding an amino acid sequence having at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity to an amino acid sequence selected from the group consisting of SEQ ID NOs: 8-14 and 53-56. 59. The expression vector or isolated DNA molecule of claim 57 or 58, wherein the non- coding RNA comprises a polynucleotide sequence having at least 90%, at least 95%, at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity or complementarity to 15, 16, 17, 18, 19, or 20 consecutive nucleotides of a polynucleotide having at least 96%, at least 97%, at least 95%, at least 99%, or 100% identity to an polynucleotide sequence selected from the group consisting of SEQ ID NOs: 1-7 and 58-61. 60. A bacterial cell comprising the expression vector or isolated DNA molecule of any one of claims 54-59, optionally wherein the bacterial cell is an Agrobacterium cell. 61. A genetically modified plant, plant part, plant cell, or seed comprising the expression vector or isolated DNA molecule of any one of claims 54-59. 62. A kit comprising the expression vector or isolated DNA molecule of 54-59 or the bacterial cell of claim 60. 63. A genetically modified non-regenerable plant cell of the strawberry plant of any one of claims 1-19. 218 sf-6744554 197072001240 64. A genetically modified plant genome of the strawberry plant of any one of claims 1- 219 sf-6744554
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