CN116042576B

CN116042576B - Application of soybean protein GmSFR2 related to seed weight regulation and its related biomaterials

Info

Publication number: CN116042576B
Application number: CN202211206132.0A
Authority: CN
Inventors: 张劲松; 姜志浩; 张万科; 韦伟; 陶建军; 阴翠翠; 陈受宜
Original assignee: Institute of Genetics and Developmental Biology of CAS
Current assignee: Institute of Genetics and Developmental Biology of CAS
Priority date: 2022-09-30
Filing date: 2022-09-30
Publication date: 2025-03-04
Anticipated expiration: 2042-09-30
Also published as: CN116042576A

Abstract

The application discloses a soybean protein GmSFR2 related to seed weight regulation and application of a related biological material, and aims to solve the technical problem of how to increase the weight of plant seeds. The application discloses GmSFR protein, substance for up-regulating or enhancing or increasing the expression of a coding gene of the protein or substance for up-regulating or enhancing or increasing the content of the protein, which is applied to any one of A1) increasing the weight of plant seeds, A2) preparing a product for increasing the weight of plant seeds, A3) cultivating high-grain-weight seed plants and A4) preparing a product for cultivating high-grain-weight seed plants. The weight of the over-expressed plant of the over-expressed GmSFR protein coding gene is obviously higher than that of a wild receptor plant. Substances which up-regulate or enhance or increase GmSFR gene expression can be used to increase plant seed weight or for plant high-seed re-breeding.

Description

Soybean protein GmSFR2 related to seed weight regulation and application of related biological material thereof

Technical Field

The invention particularly relates to a soybean protein GmSFR2 related to seed weight regulation and application of a related biological material thereof.

Background

Soybean is an important traditional crop, contains rich nutritional value, is an important economic crop for providing grain and oil and feed, and has a plurality of applications in industrial production, such as biofuel, surfactant, softener and the like

The weight (grain weight) of seeds is an important index in crop production and is one of important agronomic traits affecting crop yield. The plant can achieve the aim of increasing yield by increasing the grain weight.

The soybean yield is composed of plant type, pod bearing rate, pod number, seed hundred grain weight and other factors, wherein grain weight is the highest genetic power factor. The influence of grain weight on yield is not limited to leguminous plants, but is also an important factor for yield potential of other monocotyledonous plants, and therefore becomes an important selection trait to be considered in the process of crop variety breeding. Previous studies have shown that grain weight is affected by cultivation environment and genetics. Under normal cultivation conditions, inheritance, i.e. the relevant genes, plays an important role. Thus, research on molecular mechanisms related to particle weight has become a hotspot.

Glycoside hydrolases (glycoside hydrolase), also known as glycosidases, are a broad class of hydrolase families, belonging to glycosyl enzymes, and are a generic term for enzymes that act on various glycosides or oligosaccharides to hydrolyze glycosidic bonds. The enzymology number is EC 3.2.1. In the glycoside hydrolase family, there are 195 different types of enzymes, depending on the substrate hydrolyzed. Many kinds of glycoside hydrolases are known to exist in organisms, and they play an important role in plant growth, development, stress tolerance and the like. For example, the glycoside hydrolase 32 family (GH 32) is responsible for hydrolysis or synthesis of levan glycosidic bonds, plays a role in cell wall metabolism, cell differentiation and development, plant defense and the like, and GH7 acts on beta-1, 4-bonds of cellulose crystalline regions or amorphous regions, and can be used for efficiently degrading cellulose to convert into fermentable sugars, GH61 in combination with cellulase can degrade cellulose to fermentable sugars and the like. The application locates the grain-weight related QTL through Kefeng 1 (small grain weight)/Nannong 1138-1 (large grain weight) recombinant inbred line, and identifies the gene with the gene number Glyma.17G037100 encoding the glycoside hydrolase superfamily protein in the QTL interval of chromosome 17. Through examination, the gene response is frozen and related to freezing tolerance, so the gene response is named GmSFR2. The related report of the protein and the enzyme and the regulation of seed grain weight is not seen.

Disclosure of Invention

The invention discloses a soybean protein GmSFR2 related to seed weight regulation and application of related biological materials.

The invention aims to solve the technical problem of how to increase the weight of plant seeds, in particular to arabidopsis thaliana.

In order to solve the above problems, the present application provides the following applications:

A protein, a substance that up-regulates or enhances or increases expression of a gene encoding the protein, or a substance that up-regulates or enhances or increases activity or content of the protein, for use in any of the following:

a1 Increasing the weight of plant seeds;

a2 Preparing a product for increasing the weight of plant seeds;

A3 Cultivating a high grain weight seed plant;

A4 Preparing and cultivating a high-grain-weight seed plant product.

The protein is any one of the following:

B1 Amino acid sequence is a protein shown in sequence 2;

B2 Protein which is obtained by substituting and/or deleting and/or adding amino acid residues of the protein B1), has more than 80 percent of identity with the protein B1), and has the function of up-regulating or enhancing or increasing the weight of plant seeds;

B3 Fusion proteins obtained by ligating the N-terminal or/and C-terminal of B1) or B2) with a protein tag.

Among the above proteins, the protein tag (protein-tag) refers to a polypeptide or protein that is fusion expressed together with a target protein by using a DNA in vitro recombination technique, so as to facilitate the expression, detection, tracing and/or purification of the target protein. The protein tag may be a Flag tag, his tag, MBP tag, HA tag, myc tag, GST tag, and/or SUMO tag, etc.

In the above proteins, the identity refers to the identity of amino acid sequences. The identity of amino acid sequences can be determined using homology search sites on the internet, such as BLAST web pages of the NCBI homepage website. For example, in advanced BLAST2.1, by using blastp as a program, expect values are set to 10, all filters are set to OFF, BLOSUM62 is used as Matrix, gap existence cost, per residue gap cost and Lambda ratio are set to 11,1 and 0.85 (default values), respectively, and identity of a pair of amino acid sequences is searched for and calculated, and then the value (%) of identity can be obtained.

In the above protein, the 80% or more identity may be at least 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 95%, 96%, 98%, 99% or 100% identity.

In the above protein, sequence 2 (SEQ ID No. 2) consists of 637 amino acid residues

In the above, the substance regulating the expression of a gene may be a substance which performs at least one of 1) regulation at the level of transcription of the gene, 2) regulation after transcription of the gene (i.e., regulation of splicing or processing of the primary transcript of the gene), 3) regulation of RNA transport of the gene (i.e., regulation of nuclear to cytoplasmic transport of mRNA of the gene), 4) regulation of translation of the gene, 5) regulation of mRNA degradation of the gene, and 6) regulation after translation of the gene (i.e., regulation of activity of protein translated by the gene).

In the above application, the protein is derived from soybean.

The protein may be named GmSFR2 and may be derived from soybean. The soybean may be nannong 1138-2.

Herein, the substance that regulates the activity and/or content of the protein may be a substance that regulates the expression of a gene encoding a protein of any one of the following:

c1 Amino acid sequence is a protein shown in sequence 2;

C2 Protein obtained by substituting and/or deleting and/or adding amino acid residues of the protein of C1) and having more than 80% identity with the protein shown in C1) and having the function of increasing the weight of plant seeds.

The application also provides application of the related biological material.

The use of a related biomaterial in any one of the following;

the biological material is any one of the following:

D1 A nucleic acid molecule encoding the protein of claim 1 or 2;

D2 An expression cassette comprising D1) said nucleic acid molecule;

D3 A recombinant vector comprising D1) said nucleic acid molecule, or a recombinant vector comprising D2) said expression cassette;

d4 A recombinant microorganism comprising D1) said nucleic acid molecule, or a recombinant microorganism comprising D2) said expression cassette, or a recombinant microorganism comprising D3) said recombinant vector;

D5 A transgenic plant cell line comprising D1) said nucleic acid molecule, or a transgenic plant cell line comprising D2) said expression cassette, or a transgenic plant cell line comprising D3) said recombinant vector;

D6 A transgenic plant tissue comprising D1) said nucleic acid molecule, or a transgenic plant tissue comprising D2) said expression cassette, or a transgenic plant tissue comprising D3) said recombinant vector;

d7 A transgenic plant organ comprising D1) said nucleic acid molecule, or a transgenic plant organ comprising D2) said expression cassette, or a transgenic plant organ comprising D3) said recombinant vector;

The application is any one of the following:

a1 Increasing the weight of plant seeds;

a2 Preparing a product for increasing the weight of plant seeds;

A3 Cultivating a high grain weight seed plant;

A4 Preparing and cultivating a high-grain-weight seed plant product. In the above application, the protein is a coding gene of any one of the following:

e1 A nucleic acid sequence is a DNA shown in sequence 1;

E2 DNA obtained by substitution and/or deletion and/or addition of nucleotide residues of the DNA of E1) and having 80% or more identity with the DNA shown in E1) and having an increased weight of plant seeds.

The substance which up-regulates or enhances or increases the expression of the gene encoding the protein may be the above-mentioned D1) nucleic acid molecule encoding the above-mentioned protein, D2) expression cassette containing the above-mentioned nucleic acid molecule D1) or D3) recombinant vector containing the above-mentioned nucleic acid molecule D1) or recombinant vector containing the above-mentioned expression cassette D2).

B1 In the nucleic acid molecules, the person skilled in the art can easily mutate the nucleotide sequence encoding the protein GmSFR of the invention by known methods, for example directed evolution or point mutation. Those artificially modified nucleotides having 80% or more identity to the nucleotide sequence of the protein GmSFR2 isolated by the present invention are derived from the nucleotide sequence of the present invention and are equivalent to the sequence of the present invention as long as they encode the protein GmSFR2 and have the function of the protein GmSFR.

The 80% or more identity may be 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity.

Herein, identity refers to identity of an amino acid sequence or a nucleotide sequence. The identity of amino acid sequences can be determined using homology search sites on the internet, such as BLAST web pages of the NCBI homepage website. For example, in advanced BLAST2.1, by using blastp as a program, expect values are set to 10, all filters are set to OFF, BLOSUM62 is used as Matrix, gap existence cost, per residue gap cost and Lambda ratio are set to 11,1 and 0.85 (default values), respectively, and search is performed to calculate the identity of amino acid sequences, and then the value (%) of identity can be obtained.

In the above biological material, the nucleic acid molecule of B1) may be a gene encoding the protein. B1 The nucleic acid molecule may specifically be a DNA molecule in which the coding sequence of the coding strand is as shown in SEQ ID No.1.

Herein, the vectors are well known to those skilled in the art and include, but are not limited to, plasmids, phages (e.g., lambda phage or M13 filamentous phage, etc.), cosmids (i.e., cosmids), ti plasmids, or viral vectors. Specifically, the vector pCR8/GW/TOPO and/or pGWB411;

In the above biological material, the expression cassette of B2) means a DNA capable of expressing the gene in a host cell, and the DNA may include not only a promoter for promoting transcription of the gene but also a terminator for terminating transcription of the gene. Further, the expression cassette may also include an enhancer sequence. Promoters useful in the present invention include, but are not limited to, constitutive promoters, tissue, organ and development specific promoters, and inducible promoters. Examples of promoters include, but are not limited to, the constitutive promoter 35S of cauliflower mosaic virus, the wound-inducible promoter from tomato, leucine aminopeptidase ("LAP", chao et al (1999) Plant Physiol 120:979-992), the chemically inducible promoter from tobacco, pathogenesis-related 1 (PR 1) (induced by salicylic acid and BTH (benzothiadiazole-7-thiol S-methyl ester)), the tomato protease inhibitor II promoter (PIN 2) or LAP promoter (both inducible with jasmonic acid a ester), the heat shock promoter (U.S. Pat. No. 5,187,267), the tetracycline inducible promoter (U.S. Pat. No. 5,057,422), seed-specific promoters such as the millet seed-specific promoter pF128 (CN 101063139B (Chinese patent 200710099169.7)), the seed storage protein specific promoters (e.g., phaseolin, na, oleosin and the promoter of soybean beta conglycin (Beach et al (1985: J.3047-3047)). They may be used alone or in combination with other plant promoters. All references cited herein are incorporated by reference in their entirety. Suitable transcription terminators include, but are not limited to, agrobacterium nopaline synthase terminator (NOS terminator), cauliflower mosaic virus CaMV 35S terminator, tml terminator, pea rbcS E9 terminator and nopaline and octopine synthase terminators (see, e.g., odell et al (I985) Nature 313:810; rosenberg et al (1987) Gene,56:125; guerineau et al (1991) mol. Gen. Genet. 262:141; proudfoot (1991) Cell,64:671; sanfacon et al Genes Dev.,5:141; mogen et al (1990) PLANT CELL,2:1261; munroe et al (1990) Gene,91:151; ballad et al (1989) Nucleic Acids Res. 17:9691; jo et al (1987) Nucleic Acids Res. 15:27).

In B3) above, a recombinant expression vector containing the gene expression cassette may be constructed using a plant expression vector. The plant expression vector can be a Gateway system vector or a binary agrobacterium vector, etc., such as pGWB411、pGWB412、pGWB405、pBin438、pCAMBIA1302、pCAMBIA2301、pCAMBIA1301、pCAMBIA1300、pBI121、pCAMBIA1391-Xa or pCAMBIA1391-Xb. When TaHsfC aL is used to construct a recombinant expression vector, any one of an enhanced, constitutive, tissue-specific or inducible promoter such as a cauliflower mosaic virus (CAMV) 35S promoter, a ubiquitin gene Ubiqutin promoter (pUbi) and the like may be added before the transcription initiation nucleotide thereof, and they may be used alone or in combination with other plant promoters, and in addition, when the plant expression vector is constructed using the gene of the present invention, enhancers including a translation enhancer or a transcription enhancer may be used, and these enhancer regions may be ATG initiation codons or adjacent region initiation codons and the like, but are necessarily identical to the reading frame of the coding sequence to ensure proper translation of the entire sequence. The sources of the translational control signals and initiation codons are broad, and can be either natural or synthetic. The translation initiation region may be derived from a transcription initiation region or a structural gene.

The microorganism of B4) may be Agrobacterium. The agrobacterium is GV3101.

In the above application, the substance that up-regulates or enhances or increases expression of a gene encoding the protein is the above-mentioned biological material.

In the above application, the plant is any one of the following F1) to F4):

f1 Dicotyledonous or monocotyledonous plants;

f2 Cruciferous and leguminous plants.

In order to solve the problems, the application also provides a method for increasing the weight of plant seeds.

The method for increasing the weight of the plant seeds provided by the application comprises the step of up-regulating or enhancing or increasing the weight of the plant seeds by up-regulating or enhancing or increasing the expression of the coding gene of the protein or the content of the protein.

In order to solve the problems, the application also provides a plant breeding method.

The method for plant breeding provided by the application comprises the step of up-regulating or enhancing or increasing the expression level of the coding gene of the protein or the activity or the content of the protein in a target plant to obtain a high-seed-weight plant, wherein the seed weight of the high-seed-weight plant is higher than that of the target plant.

In the above, the seed weight is the dry weight of the seed.

In the above method, the method of up-regulating or enhancing or increasing the expression level of a gene encoding the above protein in a target plant or the activity or content of the above protein is to introduce the above biological material into the target plant.

In the above, the biological material is as follows:

D1 A nucleic acid molecule encoding the above protein.

D2 An expression cassette comprising D1) said nucleic acid molecule.

D3 A recombinant vector comprising the nucleic acid molecule of D1), or a recombinant vector comprising the expression cassette of D2). The recombinant vector may be as described above, and may specifically be pGWB411,411.

In the above, the introduction may be effected by microbial infection. The microorganism may be agrobacterium. The agrobacterium is GV3101. The method comprises the following steps:

d4 A recombinant microorganism comprising D1) said nucleic acid molecule, or a recombinant microorganism comprising D2) said expression cassette, or a recombinant microorganism comprising D3) said recombinant vector.

The nucleic acid molecules encoding the above proteins may be:

e1 A nucleic acid sequence is a DNA shown in sequence 1;

E2 DNA obtained by substitution and/or deletion and/or addition of nucleotide residues of the DNA described in E1) and having 80% or more identity with the DNA described in E1) and having an increased weight of plant seeds.

In the above method, the plant is any one of the following H1) -H4):

h1 Dicotyledonous or monocotyledonous plants;

H2 Cruciferous and leguminous plants;

The beneficial effects are that:

The coding gene GmSFR is introduced into model plant Arabidopsis thaliana (Columbia) to obtain transgenic GmSFR2 Arabidopsis thaliana plant with over-expressed GmSFR2 gene. The seed weight of the transgenic GmSFR gene Arabidopsis plant was weighed against a wild type recipient plant (Columbia) and found to be increased by the transgenic GmSFR gene Arabidopsis plant compared to the wild type Arabidopsis plant. The GmSFR protein and the coding gene thereof provided by the invention play an important role in regulating the weight of plant seeds, and can be used for regulating the weight of the seeds of plants.

Drawings

FIG. 1 is a schematic diagram of cloning vectors and plant expression vectors pGWB-GmSFR, A is a pCR8/GW/TOPO vector diagram, and B is a partial map of pGWB411-GmSFR 2.

FIG. 2 shows molecular characterization of GmSFR over-expressed Arabidopsis lines, with different lines on the abscissa and relative amounts of expression on the ordinate.

FIG. 3 shows the comparison of the thousand kernel weight of GmSFR transgenic lines with that of control seeds, with the different lines on the abscissa and the thousand kernel weight on the ordinate.

Detailed Description

The following detailed description of the invention is provided in connection with the accompanying drawings that are presented to illustrate the invention and not to limit the scope thereof. The examples provided below are intended as guidelines for further modifications by one of ordinary skill in the art and are not to be construed as limiting the invention in any way.

The experimental methods in the following examples are conventional methods unless otherwise specified. Materials, reagents and the like used in the examples described below are commercially available unless otherwise specified.

The following examples were run using SPSS11.5 statistical software and the experimental results were expressed as mean ± standard deviation, using One-way ANOVA test, P <0.05 (x) indicated significant differences, P <0.01 (x) indicated very significant differences, and P <0.001 (x) indicated very significant differences.

Soybean [ Glycine max (l.) Merr ] nannong 1138-2 (soybean improvement center germplasm pool, provided by the national soybean improvement center of nanjing university of agriculture) in the following examples.

The expression vector pGWB411(Tsuyoshi Nakagawa,et al.,Gatway Vectors for Plant Transformation,Plant Biotechnology,2009,26,275-284) in the examples described below was provided by the Tsuyoshi Nakagawa doctor, and was obtained from the national academy of sciences genetic and developmental biology study after approval by the public Tsuyoshi Nakagawa doctor, and was used only for repeated experiments related to the invention, and was not used for other purposes.

Agrobacterium GV3101 (Lee CW et al ,Agrobacterium tumefaciens promotes tumor induction by modulating pathogen defense in Arabidopsis thaliana,Plant Cell,2009,21(9),2948-62),, publicly available from the national academy of sciences genetic and developmental biology research) was used only for repeated experiments related to the invention and was not used for other purposes.

EXAMPLE 1 cDNA clone of seed weight-related protein GmSFR Gene (GmSFR 2) derived from Soybean and construction of plant expression vector

Recombinant inbred lines with Kefeng 1 (small grain) and Nannong 1138-2 (large grain) as parents are taken as positioning groups, and QTL related to grain weight is positioned on chromosome, including QTL region positioned on chromosome 17. Among genes related to QTL region of chromosome 17, a gene having a site of glyma.17g037100 (Locus name: glyma.17g037100) encoding a glycoside hydrolase superfamily protein was selected as a candidate gene for controlling grain weight. Through examination, the gene response is frozen and related to freezing tolerance, so the gene response is named GmSFR2. The related report of the protein and the enzyme and the regulation of seed grain weight is not seen.

Total RNA of Nannong 1138-2 seedlings is extracted, and the RNA is reversely transcribed into cDNA by reverse transcriptase.

Based on the information of the full-length cDNA sequence of GmSFR in the soybean genomic sequence of PlantGDB and the determined genomic sequence of Nannong 1138-2, primers were designed, and the sequences of the primers are as follows:

GmSFR2-up:5’-ATGACGGTGGTCGGACTCTT

GmSFR2-dp:5’-AGTTTCAAGAGGCTGGAGAATCA

PCR amplification was performed using Taq DNA polymerase with Nannong 1138-2cDNA as a template and GmSFR-up and GmSFR-dp as primers, yielding a PCR product of about 1.9 kb. Through sequencing, the PCR product is 1914bp (abbreviated as fragment 1), the sequence is nucleotide shown in the 1 st position to 1911 st position of the sequence 1, the sequence 1 is the CDS sequence of GmSFR gene, the coding sequence of GmSFR is the sequence 1 in the sequence table, the protein coded by the gene is GmSFR2, and the amino acid sequence of GmSFR protein is the sequence 2 in the sequence table.

Gene cloning used the Gateway system provided by the company Invitrogen, the 3' -T overhang of the vector, for direct ligation to the PCR product of Taq enzymatic amplification. Using the principle of TA cloning, the 1914bp PCR product amplified above was ligated to cloning vector pCR8/GW/TOPO (Invitrogen corporation, FIG. 1A) using DNA ligase to give recombinant vector pTOPO-GmSFR2.pTOPO-GmSFR and the target expression vector pGWB411 are provided with recombination sites attR1 and attR2 pTOPO-GmSFR containing GmSFR2 and the expression vector pGWB (GenBank: AB294435.1, 24-AUG-2007) are subjected to LR recombination reaction under the action of recombinase, finally the target gene GmSFR2 is successfully constructed on the expression vector pGWB411, and the obtained recombinant vector is named pGWB411-GmSFR (B in FIG. 1).

PGWB 411A 411-GmSFR is a recombinant expression vector obtained by replacing the fragment between attR1 site and attR2 site of pGWB A vector with attB1-GmSFR2-attB2, and keeping other nucleotides of pGWB A unchanged.

EXAMPLES 2, gmSFR obtaining of overexpression of Arabidopsis thaliana

1. Acquisition of recombinant Agrobacterium

Recombinant vector pGWB411 to GmSFR2 containing GmSFR2 obtained in example 1 was introduced into Agrobacterium GV3101 by electric shock to obtain recombinant Agrobacterium containing pGWB411 to GmSFR2, which was designated as recombinant Agrobacterium GV3101/GmSFR2.

2. Acquisition and identification of transgenic GmSFR A.thaliana

Recombinant Agrobacterium GV3101/GmSFR was cultured to the logarithmic phase, then transformed into Columbia ecological Arabidopsis thaliana (Col-0) (seed from Arabidopsis Biological Resource Center (ABRC)) by the vacuum method, seeds were harvested after cultivation (T ₁ generation), the seeds were sown on MS screening medium containing kanamycin (50 mg/L), T ₁ generation plants to be screened were grown on vermiculite when growing to 4-6 leaves, T ₁ generation individual plants were harvested, each individual plant seed (T ₂ generation) was sown separately, selection was continued with the same MS screening medium to observe the segregation of the T ₂ generation, and the multiple generations were repeated until a genetically stable transgenic homozygous line was obtained, obtaining 9 transgenic GmSFR Arabidopsis thaliana pure lines (T ₄ generation). The gene expression level of GmSFR genes was examined by randomly taking 3 strains of 9 strains, namely OE1, OE15 and OE 26.

The total RNA of seedlings of the 3 strains and Columbia ecological arabidopsis thaliana (Col-0, which is a wild type arabidopsis thaliana control, abbreviated as a control) is extracted respectively, reverse transcription is performed, cDNA obtained by reverse transcription is used as a template, and primers are GmSFR-up:5 '-ATGACGGTGGTCGGACTCTT and GmSFR-dp:5' -AGTTTCAAGAGGCTGGAGAATCA, respectively, and Real Time-PCR identification is performed. The Arabidopsis AtActin2 gene is an internal standard, and the primers used are Primer-TF:5' -ATGCCCAGAAGTCTTGTTCC and Primer-TR:5'-TGCTCATACGGTCAGCGATA-3'. And determining the relative expression quantity of GmSFR coding gene GmSFR2 by taking the expression quantity of the internal standard AtActin2 gene as 1. The experiment was repeated three times and the results averaged. The relative expression levels of GmSFR in OE1, OE15 and OE26 were 0.002, 0.0015 and 0.11, respectively, and the expression level of GmSFR was not detected in the wild type arabidopsis control (fig. 2).

The above results further demonstrate that GmSFR2 was transferred into Arabidopsis and expressed. The OE1, OE15 and OE26 3 strains were GmSFR2 over-expressed strains.

3. Phenotypic analysis of transgenic GmSFR Gene Arabidopsis thaliana

The phenotype of the 3 GmSFR over-expressed strains of control and OE1, OE15 and OE26 under normal conditions was first examined. Under normal conditions, the phenotypes of the control and GmSFR over-expressed lines, such as rosettes, plant height, etc., were not significantly different from the control.

The thousand seed weights of the GmSFR over-expressed strains OE1, OE15 and OE26, i.e., thoroughly dried seeds, were measured against the wild-type Arabidopsis Col. The preparation method comprises harvesting mature Arabidopsis seeds, drying at room temperature for 7 days, weighing, drying for 5 days, and weighing until the weight is unchanged.

20 Seeds of each strain of wild arabidopsis Col, OE1, OE15 and OE26 are taken, 200 seeds of each strain are weighed, and the seeds are weighed by a precise balance to obtain thousand seed weight data of the strain. Biological experiments were repeated three times and the results averaged ± standard deviation.

As a result, as shown in FIG. 3, the thousand seed weights of the wild type Arabidopsis thaliana Col, gmSFR2 overexpressing strains OE1, OE15 and OE26 were 16.7.+ -. 2.3,23.4.+ -. 6.9,23.6.+ -. 3.2 and 20.1.+ -. 2.1 mg, respectively. The results showed that the thousand seed weight of the seeds of the 3 GmSFR2 overexpressing transgenic lines was significantly or very significantly higher than that of the wild-type control (fig. 3). In fig. 3, significant differences are shown compared to wild type arabidopsis thaliana, and very significant differences are shown compared to wild type arabidopsis thaliana.

The experiment shows that the excessive expression of GmSFR < 2 > improves the thousand seed weight of the transgenic plant seeds, and the excessive expression does not influence the normal growth of the plants while improving the seed weight (grain weight) of the transgenic plant. Therefore, the gene can be used as a target gene for improving the seed yield of plants. GmSFR2 is a protein related to seed weight.

The present application is described in detail above. It will be apparent to those skilled in the art that the present application can be practiced in a wide range of equivalent parameters, concentrations, and conditions without departing from the spirit and scope of the application and without undue experimentation. While the application has been described with respect to specific embodiments, it will be appreciated that the application may be further modified. In general, this application is intended to cover any variations, uses, or adaptations of the application following, in general, the principles of the application and including such departures from the present disclosure as come within known or customary practice within the art to which the application pertains. The application of some of the basic features may be done in accordance with the scope of the claims that follow.

Claims

1. The use of a related biomaterial in any one of the following;

the biological material is any one of the following:

d1 A nucleic acid molecule encoding a protein;

D2 An expression cassette comprising D1) said nucleic acid molecule;

The application is any one of the following:

a1 Increasing the weight of plant seeds;

a2 Preparing a product for increasing the weight of plant seeds;

A3 Cultivating a high grain weight seed plant;

a4 Preparing and cultivating a high-grain-weight seed plant product;

the protein has an amino acid sequence shown in a sequence 2, and the plants are cruciferous plants and leguminous plants.

2. The use according to claim 1, wherein the protein is a coding gene whose nucleic acid sequence is the DNA shown in sequence 1.

3. A method for increasing the seed weight of a plant, comprising introducing the biological material of claim 1 or 2 into the plant of interest to increase the seed weight of the plant of interest, said plant being a cruciferous or leguminous plant.

4. A method of plant breeding comprising introducing the biological material of claim 1 or 2 into the plant of interest to obtain a high seed weight plant having a higher seed weight than the plant of interest, the plant being a crucifer and leguminous plant.