CN114262713A

CN114262713A - Application of E41 gene in regulation and control of plant embryonic development

Info

Publication number: CN114262713A
Application number: CN202210127737.4A
Authority: CN
Inventors: 潘荣辉; 张钰婵; 李未然; 阿赫塔尔德拉拉
Original assignee: ZJU Hangzhou Global Scientific and Technological Innovation Center
Current assignee: ZJU Hangzhou Global Scientific and Technological Innovation Center
Priority date: 2022-02-11
Filing date: 2022-02-11
Publication date: 2022-04-01
Anticipated expiration: 2042-02-11
Also published as: CN114262713B; WO2023151322A1

Abstract

The invention discloses application of an E41 gene and an E41 gene encoding protein in regulation and control of plant embryonic development, and belongs to the technical field of plant genetic engineering, wherein the nucleotide sequence of the E41 gene is shown as any one of SEQ ID No. 1-5, and the amino acid sequence of the E41 gene encoding protein is shown as any one of SEQ ID No. 6-9. The invention discloses the influence of plant peroxisome neoprotein coded by E41 gene on embryonic development for the first time, and the lethal character of homozygous mutant is caused by mutation of E41 gene; the invention provides new gene resources and theoretical guidance for deepening peroxisome function research and application of characters such as plant embryonic development and the like.

Description

Application of E41 gene in regulation and control of plant embryonic development

Technical Field

The invention relates to the technical field of plant genetic engineering, in particular to application of a peroxisome gene E41 in regulation and control of plant embryonic development.

Background

Peroxisomes are small, ubiquitous organelles in eukaryotic cells, consisting of a single membrane coating. Although peroxisome is small in size and simple in structure, it has high diversity in morphology, protein content, metabolism and the like, can respond to various signals, and plays an important role in growth and development of animals and plants.

Plant peroxisomes contain at least 200 proteins, which are involved in a wide range of physiological functions, including phytohormone biosynthesis, lipid catabolism, reactive oxygen metabolism, etc. (Pan and Hu, 2018). Mutant analysis shows that peroxisomes play a key role in plant propagation, seed development, seed germination and seedling morphogenesis, however, the potential mechanism of peroxisomes still needs to be fully elucidated.

Beta-oxidation is one of the major metabolic functions of plant peroxisomes and is critical for seed development. Arabidopsis mutants severely deficient in beta-oxidation, such as the aim1 mfp2 double mutant of two multifunctional proteins and the acx3 acx4 double mutant of acyl-CoA oxidase, exhibit a trait of stunted embryonic development (Rylott et al, 2006; Rylott et al, 2003; Khan et al, 2012). In addition, arabidopsis and barley RNA interference (RNAi) lines of the transporter CTS of fatty acids into peroxisomes produce small seeds (mendiiodo et al, 2014). In the peroxisome biogenesis mutant, the metabolic function of peroxisome cannot be normally exerted, and the seed development is influenced. Arabidopsis peroxidase protein mutants pex19a, pex19b, pex2, pex10, pex12, pex26/apem9/dayu, pex5-10pex7-2 and pex5-1 pex7-2 mutants are all embryonic lethal, with the embryos shrinking when the heart-shaped embryonic stage is arrested and/or matured (McDonnell et al, 2016; Hu et al, 2002; Sparks et al, 2003; Fan et al, 2005; Li et al, 2014; Bartel, 2010). The T-DNA insertion mutants AtPEX16 and sse1 seeds died after drying and shrinking (Lin et al, 1999).

In many other cases, it is unclear how peroxisome gene mutations lead to certain developmental defect molecular mechanisms, particularly those involved in embryonic development. The search and analysis of the functions of the novel peroxisome proteins contribute to the deepening of the understanding of peroxisomes, the improvement of the understanding of eukaryote biology and metabolism, and the improvement of crop quality, yield and stress resistance.

Disclosure of Invention

The invention provides application of the E41 gene in regulation and control of plant embryonic development, can be used for improving the quality of crops, and provides new gene resources and theoretical guidance for deepening the research on the functions of peroxisomes and the research and application of characters such as plant embryonic development and the like.

The specific technical scheme is as follows:

the invention provides an application of an E41 gene in regulation and control of plant embryonic development, wherein a nucleotide sequence of the E41 gene is shown as any one of SEQ ID No. 1-5.

The invention also provides an application of the E41 protein in regulating and controlling plant embryonic development, wherein the E41 protein is obtained by encoding the nucleotide sequence shown by any one of SEQ ID NO. 1-5.

Wherein the nucleotide sequence shown in SEQ ID NO.1 is a genome full-length sequence of the E41 gene, and the sequence length is 3393 bp; the nucleotide sequences shown in SEQ ID No. 2-5 are nucleotide sequences of different spliceosome coding regions of the E41 gene, and the sequence lengths are 2981bp, 2997bp, 2983bp and 2802bp in sequence; the length of the encoded amino acid sequence is 803aa, 766aa, 604aa and 570aa in sequence.

The invention finds a gene capable of influencing the embryonic development of plants through research, is named as E41, and determines that the protein coded by the gene is positioned in peroxisome through subcellular localization analysis. Furthermore, T-DNA insertion in Arabidopsis thalianaPrepared byE41 gene mutant plants, heterozygous mutants can only be obtained, and wild type and heterozygous in progeny plants correspond to 1: 2 separation ratio. Compared with the wild type, the hybrid mutant has the defects that partial seed development is influenced and the phenotype of shriveling and dry shrinkage is generated. The E41 gene is shown to be a coding gene of plant peroxisome new protein, which can affect the development of plant seeds.

The protein can promote the embryonic development of plants, and the reduction or loss of the activity of the protein can influence the embryonic development of seeds, so that viable seeds cannot be obtained.

Further, the plant is a dicot. Still further, the plant is arabidopsis thaliana.

The plant embryo abnormal development caused by E41 gene deletion shows that the tender seed is white transparent, after maturation, it is shriveled and has no vitality, and the progeny can not obtain the plant of homozygous mutant. The research of the invention shows that the E41 gene is introduced into the plant with abnormal embryonic development to obtain the transgenic plant with normal embryonic development.

The E41 gene provided by the invention has potential application value in crop breeding.

In scientific research, a model plant with embryo dysplasia needs to be constructed, and based on the characteristic that E41 gene function loss causes the embryo dysplasia of the plant, the invention provides the application of the gene in constructing an embryo dysplasia plant model. Specifically, the expression of the plant E41 gene is reduced or not expressed by using a genetic engineering technology.

Further, the application comprises the following steps:

(1) the deletion or the expression quantity reduction of the E41 gene is caused by utilizing gene mutation, gene knockout, gene interference or gene silencing technology, so that a plant model with embryonic dysplasia is constructed.

(2) Introducing the E41 gene into a receptor plant, and culturing to obtain a transgenic plant with acquired function; the recipient plant is a plant with abnormal embryonic development caused by deletion of the E41 gene.

Still further, the plant is arabidopsis thaliana. The constructed arabidopsis thaliana with abnormal embryo development is slow in embryo development or incapable of forming embryo, and gradually shrinks and shrinks along with the maturation of the silique.

Compared with the prior art, the invention has the following beneficial effects:

(1) the invention discloses the influence of plant peroxisome neoprotein coded by E41 gene on embryonic development for the first time, and the lethal character of homozygous mutant caused by mutation of E41 gene.

(2) The invention provides new gene resources and theoretical guidance for deepening peroxisome function research and application of characters such as plant embryonic development and the like.

Drawings

FIG. 1 is a map of the localization of tobacco transient expression subcellular;

wherein CFP-PTS1 is a peroxisome marker, E41-YFP, YFP-E41, mVenus-E41.2, mVenus-E41.3 and mVenus-E41.4 are respectively E41 main spliceosome proteins fused at the N end and the C end of YFP fluorescent protein and other three spliceosome proteins of E41 fused at the C end of mVenus fluorescent protein; bar 10 μ M.

FIG. 2 is a schematic diagram showing the position of insertion of mutant T-DNA;

wherein white boxes represent UTR regions, black boxes represent exon regions, and black lines represent intron regions.

FIG. 3 shows the seed traits of mutants e41-1 and e 41-2;

wherein, A is the shape of the seeds in the young siliques, and the arrows indicate abnormal white transparent seeds and slightly shrunken seeds; B. c, showing the shape of the seeds in the maturation process, wherein the arrow indicates the abnormal development and the early yellowing and shrinking seeds; d, the graph is the mature seed form, and arrows indicate shrivelled abnormal seeds; bar is 1 mm.

FIG. 4 is a schematic diagram of the structure of a complementary vector;

wherein, A is E41 genome sequence driven by endogenous promoter; the B picture is the sequence of the E41 coding region driven by the endogenous promoter.

FIG. 5 shows the T-DNA insertion detection and genotype identification of positive seedlings;

wherein, A picture is the insertion detection result of T-DNA of a complementary strain of an E41 coding region sequence vector and a complementary strain of an E41 genome sequence vector which are started by an endogenous promoter; and B is the genotype identification result.

Detailed Description

In order to make the technical solution of the present invention better understood by those skilled in the art, the following technical solution of the present invention is clearly and completely described with reference to the specific embodiments. It is to be understood that the following detailed description is exemplary, and is intended to cover only some embodiments, but not all embodiments, of the invention.

All other embodiments, which can be obtained by a person skilled in the art without any inventive step based on the embodiments of the present invention, shall fall within the scope of protection of the present invention.

Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this application belongs. The experimental materials used in the examples of the present invention are all conventional experimental materials in the art, and are commercially available. Experimental procedures without specification of details were carried out according to the conventional experimental procedures or according to the instructions recommended by the supplier.

Example 1

1. Construction of E41 gene subcellular localization vector

The pCAMBIA1300-YFP vector with the YFP fluorescent protein label is obtained by modifying the pCAMBIA1300 vector. A primer is designed according to a CDS sequence of a main spliceosome (shown as SEQ ID NO.2) of the E41 gene, and the CDS sequence is amplified by taking arabidopsis cDNA as a template. The amplified fragment is ligated to the N-or C-terminus of YFP. The primers used were E41-Y-F and E41-Y-R, Y-E41-F and Y-E41-R, respectively.

The primer sequence is as follows:

E41-Y-F：5’-cacgggggactctagaATGGCGAGAAAAGCTAACAATAGTTTTTTC-3’；

E41-Y-R：5’-cttgctcaccattgttggatccCCGAGTCGAGCAGCCAACTGGTTC-3’；

Y-E41-F：5’-gtacaagagatctgcgtcgactATGGCGAGAAAAGCTAACAATAGTTTTTTC-3’；

Y-E41-R：5’-cgatcggggaaattcgagctcTCAGAGTCGAGCAGCCAACTGGTTC-3’；

in addition, pEarleyGate100 vector with mVenus fluorescent protein tag is obtained by modifying pEarleyGate100 vector. Designing a primer according to a CDS sequence of a non-main spliceosome (SEQ ID NO.3, SEQ ID NO.4 and SEQ ID NO.5) of the E41 gene, and amplifying the CDS sequence by taking arabidopsis cDNA as a template; connecting the amplified fragment to the C end of YFP; the primers are V-E41.2-F and Y-E41.2-R, V-E41.2-F and Y-E41.3-R, V-E41.4-F and Y-E41.4-R, respectively.

The primer sequence is as follows:

V-E41.2-F：5’-GACGAGCTGTACAAGtctagaATGGCGAGAAAAGCTAACAATAG-3’；

Y-E41.2-R：5’-GGTCTTAATTAACTCTCTAGATCATAAGATATGGCAACACC-3’；

V-E41.3-R：5’-GGTCTTAATTAACTCTCTAGATCAAGGATACAAAAACATGGTTGATGCAACC-3’；

V-E41.4-F：5’-GACGAGCTGTACAAGtctagaATGATTCTTCATTTGATTGAGTGGG-3’；

V-E41.4-R：5’-GGTCTTAATTAACTCTCTAGATCAGAGTCGAGCAGCCAACTGGTTC-3’。

the PCR reaction conditions are as follows:

95 deg.C for 3 min; 95 deg.C, 15sec, 58 deg.C, 15min, 72 deg.C, 2min, 24sec, 35 cycles; 72 ℃ for 5 min.

The two fragments were recovered by agarose gel electrophoresis.

For a carrier fused at the C end of the gene by YFP, XbaI and BamHI are used for double enzyme digestion of pCAMBIA1300-YFP carrier, and a target fragment and a linear carrier are connected by a homologous recombination method to obtain a 35S-E41-YFP recombinant carrier; and for the carrier of YFP fusion at the N end of the gene, carrying out double digestion on the pCAMBIA1300-YFP carrier by SalI and SacI, and connecting a target fragment and a linearized carrier by using a homologous recombination method to obtain a 35S-YFP-E41 recombinant carrier. For the vector fused at the N end of the gene mVenus, XbaI is used for cutting pEarleyGate100-mVenus vector, and a target fragment and a linearized vector are connected by a homologous recombination method to obtain three recombinant vectors of mVenus-E41.2, mVenus-E41.3 and mVenus-E41.4 respectively.

Mu.l of the recombinant product was taken and transformed into E.coli DH 5. alpha. by freeze-thaw method, and the transformed product was spread on LB medium resistant to kanamycin (50. mu.g/ml). Cultured overnight at 37 ℃ and the single clone was picked up to extract the plasmid. And the constructed vector is stored for later use after sequencing verification.

2. Subcellular localization experiments

Agrobacterium was transformed with the sequenced vector obtained in example 1 by methods according to the kit instructions.

Injecting the fusion expression vector into the Nicotiana benthamiana leaf by an agrobacterium infiltration method, simultaneously injecting a 35S-CFP-SKL vector as peroxisome control, and observing a fluorescence signal by using a laser confocal microscope after 48 hours.

The results of the experiment are shown in FIG. 1. YFP protein fusion is positioned at the N-terminal (the peroxisome positioning signal peptide PTS1 of E41 protein is not blocked) or the C-terminal (YFP blocks the peroxisome positioning signal peptide) of the main spliceosome protein and is positioned to peroxisome; the mVenus protein is fused at the N-terminal of the E41 non-major spliceosome protein and is positioned to peroxisomes. All the above results indicate that E41 is a peroxisomal protein.

Example 2

1. Identification of T-DNA insertion mutants

Two T-DNA insertion-causing E41 mutants were purchased from Arabidopsis Biological Resource Center (ABRC, https:// ABRC. osu. edu /), designated E41-1 and E41-2, respectively, and the T-DNA insertion positions are shown in FIG. 2.

The mutant genotype identification is carried out by a three-primer method. Primers were designed by the T-DNA primer design website (http:// signal. salk. edu/tdnaprimers.2. html). The primers designed for the website are respectively marked as e41-1-LP and e41-1-RP, e41-2-LP and e 41-2-RP. The LB primer is LBb1.3 provided by the website.

The primer sequence is as follows:

e41-1-LP：5’-TACCAGTCACGTGAATCGTTG-3’；

e41-1-RP：5’-ACTCTTCTTCCAAAAGCCTGC-3’；

e41-2-LP：5’-GCAGACTTGTTTACGCAAAGG-3’；

e41-2-RP：5’-GCAGACTTGTTTACGCAAAGG-3’；

LBb1.3：5’-ATTTTGCCGATTTCGGAAC-3’；

the PCR reaction conditions are as follows:

95 deg.C for 3 min; 95 deg.C, 15sec, 58 deg.C, 15min, 72 deg.C, 15sec, 32 cycles; 72 ℃ for 5 min.

The mutant progeny cannot obtain homozygous mutants.

And (3) carrying out genotype identification on the heterozygous mutant progeny, and analyzing that the genotype separation ratio of the progeny wild type and the heterozygous mutant accords with 1: 2 (table 1), suggesting that deletion of the E41 protein affects the development of homozygous mutant progeny.

TABLE 1 Caliper test of suitability of segregating populations of heterozygous mutant progeny

Note: chi shape² _0.05(1)＝3.84

2. Character analysis of T-DNA insertion mutant

As shown in fig. 3, the siliques during development of the wild type and the mutant were opened and dysplastic seeds were observed in the mutant. The abnormal seeds are white and transparent when young and tender, and gradually shriveled along with the mature seeds of the siliques, and the abnormal seeds with small weight exist in the seeds during final harvest.

Example 3

1.E41 complementary vector construction and genetic transformation experiments

In order to verify that the character deletion of the T-DNA mutant line which cannot obtain homozygous offspring is caused by the fact that E41 gene mutant offspring seeds cannot develop smoothly, but is not caused by unknown T-DNA insertion, a vector for endogenously expressing E41 is constructed for complementation. The complementary vector is schematically shown in FIG. 4.

The vector for endogenously expressing E41 consists of an endogenous promoter (SEQ ID NO.10) for promoting an E41 genome sequence (SEQ ID NO.1) and an endogenous promoter (SEQ ID NO.6) for promoting an E41 coding region sequence (SEQ ID NO. 2). The promoter of E41 is amplified by primers E41-pro-F and E41-pro-R, the recovered target fragment is connected with pCAMBIA1300-YFP vector which is double digested by HindIII and XbaI, after escherichia coli DH5 alpha competence is transformed, single colony is selected to extract plasmid. The constructed vector is stored for later use after sequencing verification and is marked as pCAMBIA 1300-pro-YFP.

The genome DNA of arabidopsis thaliana is taken as a template, the genome sequence of E41 is amplified by using primers E41-g-F and E41-g-R, the recovered target fragment is connected with a pCAMBIA1300-pro-YFP vector which is subjected to double enzyme digestion by XbaI and SalI, and after the escherichia coli DH5 alpha is transformed, a single colony is picked to extract a plasmid. The constructed vector is stored for later use after sequencing verification and is marked as pCAMBIA 1300-pro-genomic.

The reverse transcription cDNA of arabidopsis thaliana is taken as a template, a coding region sequence (SEQ ID NO.2) of E41 is amplified by primers E41-c-F and E41-c-R, a recovered target fragment is connected with a pCAMBIA1300-pro-YFP vector which is subjected to double enzyme digestion by XbaI and SalI, and after escherichia coli DH5 alpha competence is transformed, a single colony is picked to extract a plasmid. The constructed vector is stored for later use after sequencing verification and is marked as pCAMBIA 1300-pro-cDNA.

The vector with correct sequencing is transformed into agrobacterium by referring to the method of the kit instruction.

Agrobacterium is transferred into e41-1 mutant by inflorescence infection method to obtain T0 generation transgenic seed.

The PCR reaction conditions are as follows: 95 deg.C for 3 min; 95 deg.C, 15sec, 58 deg.C, 15min, 72 deg.C, 15sec, 32 cycles; 72 ℃ for 5 min.

The primer sequence is as follows:

E41-pro-F：5’-acgacggccagtgccaagcttaacttcaaatttaatgaggcctcctc-3’；

E41-pro-R：5’-accattgttggatcctctagaggatccctttaatacaactaattcag-3’；

E41-g-F：5’-gtattaaagggatcctctagaATGGCGAGAAAAGCTAACAATAG-3’；

E41-g-R：5’-ggccgctacactcgagtcgaccctgattcgtttcttaccctttcaa-3’；

E41-c-F：5’-gtattaaagggatcctctagaATGGCGAGAAAAGCTAACAATAG-3’；

E41-c-R：5’-ggccgctacactcgagtcgaccctgattcgtttcttaccctttcaa-3’。

2. screening and identification of complementary transgenic positive seedlings

Transgenic seeds of T0 generation were selected by sowing in 1/2MS medium containing hygromycin (50. mu.g/ml) resistance, cultured under normal light for about 1 week, and positive seedlings were observed and selected. The resistant seedlings are transplanted into nutrient soil, and the positive seedlings are subjected to T-DNA insertion detection by using primers T1-F and T1-R. The positive seedlings complemented by the endogenous promoter vector were genotyped with primers Ne41-1-LP, Ne41-1-RP, LBb1.3.

The primer sequence is as follows:

T1-F：5’-AACAGAAAAGTGGCGCAAGC-3’；

T1-R：5’-atcgcaagaccggcaacagga-3’；

Ne41-1-LP：5’-cccctgtgtttcctatgctgcttg-3’；

Ne41-1-RP：5’-TTGGAGATTACGCCTGTCATC-3’；

the PCR reaction conditions are as follows:

The results are shown in FIG. 5, where homozygous T-DNA insertion mutants with successful complementation were selected, indicating that mutant silique dysplasia is caused by the deletion of E41.

The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention, and various modifications and changes may be made by those skilled in the art. Any modification, equivalent replacement, or improvement made within the spirit and principle of the present invention should be included in the protection scope of the present invention.

Sequence listing

<110> Hangzhou international scientific center of Zhejiang university

Application of <120> E41 gene in regulation and control of plant embryonic development

<160> 30

<170> SIPOSequenceListing 1.0

<210> 1

<211> 3393

<212> DNA

<213> Arabidopsis thaliana (Arabidopsis thaliana)

<400> 1

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gaactcgctg cgtagccaaa gcctcttcaa ttttgctctt ttgttcaatt ccgtcgctta 180

attatctggt tttgcgtatt ttgaagagtc agtttgacga atttatgctg aaatttcgtt 240

ctttctctgt ttatctttca gcgacgattt ttctgggttt ttcagttttg gatcaatcca 300

taatctgtcg tgattagggt tttatacctt gatgaggatt tactgaatta gttgtattaa 360

agatggcgag aaaagctaac aatagttttt tccttgaaga atggttaagg actgtgagtg 420

gaagtagtgt ctctggtgat ctagtgaagc aaaattctgc tccatctgct aggtcgatta 480

ttcaagcatg gtctgagatt cgtgaatctc ttcaaaacca aaactttgat tcacgttacc 540

ttcaagcttt gagagctttg gttagctctg agtccactat ccatgttgca gatccccaag 600

caaagctact tatttctata ctagcttttc aagatgtatc tcttccgtct gagtcttaca 660

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ttgttggtgt agcggttcag gccattcgtg gcgttgttga tgacaggcgt aatctccaac 780

cagccttagt agcgcagagt gttctggttt ctggtgcgtt tgcgtgtgtt ccttctctgt 840

caggagacgt gaaagtctta tgcttggaat tgttatgcag gcttttggaa gaagagtact 900

ctttagtggg atctcaagaa gaacttgttc ctgtagtgct tgcaggaatt gggtatgctt 960

tatcttcttc gttggatgtt cattacgtca gactattgga tttattgttt ggcatttggt 1020

taaaagacga gggtcctcgt ggcactgtca cttacggtct gatgattctt catttgattg 1080

agtgggttgt gtcaggttat atgcggtcta attctatcaa caagatgtct ctttttgcga 1140

acgaggtact agagacttct aaggaaaagt atgctgtttt cgctgtcttc atggcagcag 1200

ctggggtagt gagagcttcc acggcaggat tcagtagtgg tgcacagagt ttggagattt 1260

ctaaactaag aaattcagct gaaaagcgaa tagaatttgt agctcaaatt ttagtttcta 1320

atggtaatgt tgttacactt ccaaccacac agagagaagg tcccctgttg aagtgctttg 1380

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tcgagaaaat tgcagaatca tctttcctta tggttgtcgt ctttgcgcta gctgttacga 1800

aacaatggct aaaaccaata gtgtctaaag aaagaaaaat ggtgacatca gttaagatat 1860

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agtccattcc tgcctatgat agcttgacga atccgaaagg tttgagaaat cacttgtttc 2040

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ggaggtaact aagcaaaaaa agaatatgaa tgttacgtag ctctttttct gtttatttct 2580

tatctctaat gtatatggtt ggttgaaata actgcaggtt taccctgaga tcacaccttt 2640

tgaaggttta gcttccggcg ttgcaacctt ggtccagcat ctacctgcgg gaagtcctgc 2700

tatattttat tctgttcata gccttgttga gaaggcttct acatttagca ccgaatcatt 2760

gcaaggcaga aagtctgatc ctggtaatca gattcttgag ttgcttttgc gacttgtctc 2820

tttggttgat atacaagtaa gtttcttttt tggtcatctc catctacttg agctactaga 2880

aaatggttac tgtgctcata cgttaatgtt ctctccgtct ttgtatacat ttttgttttg 2940

gtttaggtgt tgccatatct tatgaagtca ttggcacagc tagttataaa gttaccaaaa 3000

gaaagacaga acgttgtgct tggtgaattg tatggtcaag tggctgagtc agacgatgtg 3060

attcgcaagc cttctttggt ctcttggcta cagtcactga actacttatg ttctaataac 3120

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ctctagggat ctcctcatag tttacaaaca tgtgaaatag aataaagttt gtatgacttg 3240

tatcatggaa ttatgttgaa aagaattata aattaacatc atctccattt tttgatcagt 3300

agccaaaaat agtcaatcct gttaaatgtg ttgcggttta aatttatttg gttacgaaga 3360

agaaccggat tgaaagggta agaaacgaat cag 3393

<210> 2

<211> 2981

<212> DNA

<213> Arabidopsis thaliana (Arabidopsis thaliana)

<400> 2

gtttatcctt aaccacaaaa caaacgtcac aactccgtca ccgcaaacta ggcagattat 60

ctggttttcc ggcggtttct cgttatctac gagagagtga gagaggaaat acctcggggt 120

gaactcgctg cgtagccaaa gcctcttcaa ttttgctctt ttgttcaatt ccgtcgctta 180

attatctggt tttgcgtatt ttgaagagtc agtttgacga atttatgctg aaatttcgtt 240

ctttctctgt ttatctttca gcgacgattt ttctgggttt ttcagttttg gatcaatcca 300

taatctgtcg tgattagggt tttatacctt gatgaggatt tactgaatta gttgtattaa 360

agatggcgag aaaagctaac aatagttttt tccttgaaga atggttaagg actgtgagtg 420

gaagtagtgt ctctggtgat ctagtgaagc aaaattctgc tccatctgct aggtcgatta 480

ttcaagcatg gtctgagatt cgtgaatctc ttcaaaacca aaactttgat tcacgttacc 540

ttcaagcttt gagagctttg gttagctctg agtccactat ccatgttgca gatccccaag 600

caaagctact tatttctata ctagcttttc aagatgtatc tcttccgtct gagtcttaca 660

cacttgttct cagactactc tatgtgtgga ttaggaaagc atttcgaccg tctcaagcac 720

ttgttggtgt agcggttcag gccattcgtg gcgttgttga tgacaggcgt aatctccaac 780

cagccttagt agcgcagagt gttctggttt ctggtgcgtt tgcgtgtgtt ccttctctgt 840

caggagacgt gaaagtctta tgcttggaat tgttatgcag gcttttggaa gaagagtact 900

ctttagtggg atctcaagaa gaacttgttc ctgtagtgct tgcaggaatt gggtatgctt 960

tatcttcttc gttggatgtt cattacgtca gactattgga tttattgttt ggcatttggt 1020

taaaagacga gggtcctcgt ggcactgtca cttacggtct gatgattctt catttgattg 1080

agtgggttgt gtcaggttat atgcggtcta attctatcaa caagatgtct ctttttgcga 1140

acgaggtact agagacttct aaggaaaagt atgctgtttt cgctgtcttc atggcagcag 1200

ctggggtagt gagagcttcc acggcaggat tcagtagtgg tgcacagagt ttggagattt 1260

ctaaactaag aaattcagct gaaaagcgaa tagaatttgt agctcaaatt ttagtttcta 1320

atggtaatgt tgttacactt ccaaccacac agagagaagg tcccctgttg aagtgctttg 1380

ctattgcatt ggctcgatgt gggtctgttt cttcctctgc tcctctgctt ttgtgtctca 1440

cttctgcatt gttaactcag gtatttcctt taggccagat atatgaatca ttttgcaatg 1500

cttttggcaa agagcctatt ggaccaagac tcatttgggt cagggagcat ctttctgatg 1560

ttcttttcaa ggaatcaggg gccatatctg gggctttctg caatcaatat tcatcagcaa 1620

gtgaagagaa caagtatatt gtggagaata tgatttggga tttctgtcaa aatctctact 1680

tacagcaccg tcaaattgct atgttgcttt gtggtataga agatacattg cttggagaca 1740

tcgagaaaat tgcagaatca tctttcctta tggttgtcgt ctttgcgcta gctgttacga 1800

aacaatggct aaaaccaata gtgtctaaag aaagaaaaat ggtgacatca gttaagatat 1860

tagtttcgtt ctcctgtgta gagtatttca ggcatattcg tttacctgaa tacatggaga 1920

caattagaga ggtgatttca tgtgtccagg agaatgatgc tccttgtgtt tcatttgtag 1980

agtccattcc tgcctatgat agcttgacga atccgaaaga cttgtttacg caaaggataa 2040

aatatgaatg gtcaagagat gatgtgcaga catctcgaat attgttttat cttcgagtaa 2100

tcccaacttg catcggacgg ttatctgctt ctgccttcag gggagtggtt gcatcaacca 2160

tgtttttata tattggacat cctaacagaa aagtggcgca agcatctcat acgttgttgg 2220

cagcatttct ctcttcagca aaagaatcag aggaggacga acgaactcaa ttcaaagaac 2280

aacttgtttt ctattacatg caacgatctt tggaggttta ccctgagatc acaccttttg 2340

aaggtttagc ttccggcgtt gcaaccttgg tccagcatct acctgcggga agtcctgcta 2400

tattttattc tgttcatagc cttgttgaga aggcttctac atttagcacc gaatcattgc 2460

aaggcagaaa gtctgatcct ggtaatcaga ttcttgagtt gcttttgcga cttgtctctt 2520

tggttgatat acaagtgttg ccatatctta tgaagtcatt ggcacagcta gttataaagt 2580

taccaaaaga aagacagaac gttgtgcttg gtgaattgta tggtcaagtg gctgagtcag 2640

acgatgtgat tcgcaagcct tctttggtct cttggctaca gtcactgaac tacttatgtt 2700

ctaataaccg tactgaagtt ttagcttctg gatcaacgat agacacttcg aaccagttgg 2760

ctgctcgact ctagggatct cctcatagtt tacaaacatg tgaaatagaa taaagtttgt 2820

atgacttgta tcatggaatt atgttgaaaa gaattataaa ttaacatcat ctccattttt 2880

tgatcagtag ccaaaaatag tcaatcctgt taaatgtgtt gcggtttaaa tttatttggt 2940

tacgaagaag aaccggattg aaagggtaag aaacgaatca g 2981

<210> 3

<211> 2997

<212> DNA

<213> Arabidopsis thaliana (Arabidopsis thaliana)

<400> 3

ttatccttaa ccacaaaaca aacgtcacaa ctccgtcacc gcaaactagg cagattatct 60

ggttttccgg cggtttctcg ttatctacga gagagtgaga gaggaaatac ctcggggtga 120

actcgctgcg tagccaaagc ctcttcaatt ttgctctttt gttcaattcc gtcgcttaat 180

tatctggttt tgcgtatttt gaagagtcag tttgacgaat ttatgctgaa atttcgttct 240

ttctctgttt atctttcagc gacgattttt ctgggttttt cagttttgga tcaatccata 300

atctgtcgtg attagggttt tataccttga tgaggattta ctgaattagt tgtattaaag 360

atggcgagaa aagctaacaa tagttttttc cttgaagaat ggttaaggac tgtgagtgga 420

agtagtgtct ctggtgatct agtgaagcaa aattctgctc catctgctag gtcgattatt 480

caagcatggt ctgagattcg tgaatctctt caaaaccaaa actttgattc acgttacctt 540

caagctttga gagctttggt tagctctgag tccactatcc atgttgcaga tccccaagca 600

aagctactta tttctatact agcttttcaa gatgtatctc ttccgtctga gtcttacaca 660

cttgttctca gactactcta tgtgtggatt aggaaagcat ttcgaccgtc tcaagcactt 720

gttggtgtag cggttcaggc cattcgtggc gttgttgatg acaggcgtaa tctccaacca 780

gccttagtag cgcagagtgt tctggtttct ggtgcgtttg cgtgtgttcc ttctctgtca 840

ggagacgtga aagtcttatg cttggaattg ttatgcaggc ttttggaaga agagtactct 900

ttagtgggat ctcaagaaga acttgttcct gtagtgcttg caggaattgg gtatgcttta 960

tcttcttcgt tggatgttca ttacgtcaga ctattggatt tattgtttgg catttggtta 1020

aaagacgagg gtcctcgtgg cactgtcact tacggtctga tgattcttca tttgattgag 1080

tgggttgtgt caggttatat gcggtctaat tctatcaaca agatgtctct ttttgcgaac 1140

gaggtactag agacttctaa ggaaaagtat gctgttttcg ctgtcttcat ggcagcagct 1200

ggggtagtga gagcttccac ggcaggattc agtagtggtg cacagagttt ggagatttct 1260

aaactaagaa attcagctga aaagcgaata gaatttgtag ctcaaatttt agtttctaat 1320

ggtaatgttg ttacacttcc aaccacacag agagaaggtc ccctgttgaa gtgctttgct 1380

attgcattgg ctcgatgtgg gtctgtttct tcctctgctc ctctgctttt gtgtctcact 1440

tctgcattgt taactcaggt atttccttta ggccagatat atgaatcatt ttgcaatgct 1500

tttggcaaag agcctattgg accaagactc atttgggtca gggagcatct ttctgatgtt 1560

cttttcaagg aatcaggggc catatctggg gctttctgca atcaatattc atcagcaagt 1620

gaagagaaca agtatattgt ggagaatatg atttgggatt tctgtcaaaa tctctactta 1680

cagcaccgtc aaattgctat gttgctttgt ggtatagaag atacattgct tggagacatc 1740

gagaaaattg cagaatcatc tttccttatg gttgtcgtct ttgcgctagc tgttacgaaa 1800

caatggctaa aaccaatagt gtctaaagaa agaaaaatgg tgacatcagt taagatatta 1860

gtttcgttct cctgtgtaga gtatttcagg catattcgtt tacctgaata catggagaca 1920

attagagagg tgatttcatg tgtccaggag aatgatgctc cttgtgtttc atttgtagag 1980

tccattcctg cctatgatag cttgacgaat ccgaaagact tgtttacgca aaggataaaa 2040

tatgaatggt caagagatga tgtgcagaca tctcgaatat tgttttatct tcgagtaatc 2100

ccaacttgca tcggacggtt atctgcttct gccttcaggg gagtggttgc atcaaccatg 2160

tttttatata ttggacatcc taacagaaaa gtggcgcaag catctcatac gttgttggca 2220

gcatttctct cttcagcaaa agaatcagag gaggacgaac gaactcaatt caaagaacaa 2280

cttgttttct attacatgca acgatctttg gaggtttacc ctgagatcac accttttgaa 2340

ggtttagctt ccggcgttgc aaccttggtc cagcatctac ctgcgggaag tcctgctata 2400

ttttattctg ttcatagcct tgttgagaag gcttctacat ttagcaccga atcattgcaa 2460

ggcagaaagt ctgatcctgg taatcagatt cttgagttgc ttttgcgact tgtctctttg 2520

gttgatatac aagtaagttt cttttttggt catctccatc tacttgagct actagaaaat 2580

ggttactgtg ctcatacgtt aatgttctct ccgtctttgt atacattttt gttttggttt 2640

aggtgttgcc atatcttatg aagtcattgg cacagctagt tataaagtta ccaaaagaaa 2700

gacagaacgt tgtgcttggt gaattgtatg gtcaagtggc tgagtcagac gatgtgattc 2760

gcaagccttc tttggtctct tggctacagt cactgaacta cttatgttct aataaccgta 2820

ctgaagtttt agcttctgga tcaacgatag acacttcgaa ccagttggct gctcgactct 2880

agggatctcc tcatagttta caaacatgtg aaatagaata aagtttgtat gacttgtatc 2940

atggaattat gttgaaaaga attataaatt aacatcatct ccattttttg atcagta 2997

<210> 4

<211> 2983

<212> DNA

<213> Arabidopsis thaliana (Arabidopsis thaliana)

<400> 4

ttatccttaa ccacaaaaca aacgtcacaa ctccgtcacc gcaaactagg cagattatct 60

ggttttccgg cggtttctcg ttatctacga gagagtgaga gaggaaatac ctcggggtga 120

actcgctgcg tagccaaagc ctcttcaatt ttgctctttt gttcaattcc gtcgcttaat 180

tatctggttt tgcgtatttt gaagagtcag tttgacgaat ttatgctgaa atttcgttct 240

ttctctgttt atctttcagc gacgattttt ctgggttttt cagttttgga tcaatccata 300

atctgtcgtg attagggttt tataccttga tgaggattta ctgaattagt tgtattaaag 360

atggcgagaa aagctaacaa tagttttttc cttgaagaat ggttaaggac tgtgagtgga 420

agtagtgtct ctggtgatct agtgaagcaa aattctgctc catctgctag gtcgattatt 480

caagcatggt ctgagattcg tgaatctctt caaaaccaaa actttgattc acgttacctt 540

caagctttga gagctttggt tagctctgag tccactatcc atgttgcaga tccccaagca 600

aagctactta tttctatact agcttttcaa gatgtatctc ttccgtctga gtcttacaca 660

cttgttctca gactactcta tgtgtggatt aggaaagcat ttcgaccgtc tcaagcactt 720

gttggtgtag cggttcaggc cattcgtggc gttgttgatg acaggcgtaa tctccaacca 780

gccttagtag cgcagagtgt tctggtttct ggtgcgtttg cgtgtgttcc ttctctgtca 840

ggagacgtga aagtcttatg cttggaattg ttatgcaggc ttttggaaga agagtactct 900

ttagtgggat ctcaagaaga acttgttcct gtagtgcttg caggaattgg gtatgcttta 960

tcttcttcgt tggatgttca ttacgtcaga ctattggatt tattgtttgg catttggtta 1020

aaagacgagg gtcctcgtgg cactgtcact tacggtctga tgattcttca tttgattgag 1080

tgggttgtgt caggttatat gcggtctaat tctatcaaca agatgtctct ttttgcgaac 1140

gaggtactag agacttctaa ggaaaagtat gctgttttcg ctgtcttcat ggcagcagct 1200

ggggtagtga gagcttccac ggcaggattc agtagtggtg cacagagttt ggagatttct 1260

aaactaagaa attcagctga aaagcgaata gaatttgtag ctcaaatttt agtttctaat 1320

ggtaatgttg ttacacttcc aaccacacag agagaaggtc ccctgttgaa gtgctttgct 1380

attgcattgg ctcgatgtgg gtctgtttct tcctctgctc ctctgctttt gtgtctcact 1440

tctgcattgt taactcaggt atttccttta ggccagatat atgaatcatt ttgcaatgct 1500

tttggcaaag agcctattgg accaagactc atttgggtca gggagcatct ttctgatgtt 1560

cttttcaagg aatcaggggc catatctggg gctttctgca atcaatattc atcagcaagt 1620

gaagagaaca agtatattgt ggagaatatg atttgggatt tctgtcaaaa tctctactta 1680

cagcaccgtc aaattgctat gttgctttgt ggtatagaag atacattgct tggagacatc 1740

gagaaaattg cagaatcatc tttccttatg gttgtcgtct ttgcgctagc tgttacgaaa 1800

caatggctaa aaccaatagt gtctaaagaa agaaaaatgg tgacatcagt taagatatta 1860

gtttcgttct cctgtgtaga gtatttcagg catattcgtt tacctgaata catggagaca 1920

attagagagg tgatttcatg tgtccaggag aatgatgctc cttgtgtttc atttgtagag 1980

tccattcctg cctatgatag cttgacgaat ccgaaagact tgtttacgca aaggataaaa 2040

tatgaatggt caagagatga tgtgcagaca tctcgaatat tgttttatct tcgagtaatc 2100

ccaacttgca tcggacggtt atctgcttct gccttcaggg gagtggttgc atcaaccatg 2160

tttttgtatc cttgagaaaa gatcgagaag tcttttactc tgttttactt aactacgagt 2220

actaaactat ctttcctatt ccttaacaat tctgtgacta gatatattgg acatcctaac 2280

agaaaagtgg cgcaagcatc tcatacgttg ttggcagcat ttctctcttc agcaaaagaa 2340

tcagaggagg acgaacgaac tcaattcaaa gaacaacttg ttttctatta catgcaacga 2400

tctttggagg tttaccctga gatcacacct tttgaaggtt tagcttccgg cgttgcaacc 2460

ttggtccagc atctacctgc gggaagtcct gctatatttt attctgttca tagccttgtt 2520

gagaaggctt ctacatttag caccgaatca ttgcaaggca gaaagtctga tcctggtaat 2580

cagattcttg agttgctttt gcgacttgtc tctttggttg atatacaagt gttgccatat 2640

cttatgaagt cattggcaca gctagttata aagttaccaa aagaaagaca gaacgttgtg 2700

cttggtgaat tgtatggtca agtggctgag tcagacgatg tgattcgcaa gccttctttg 2760

gtctcttggc tacagtcact gaactactta tgttctaata accgtactga agttttagct 2820

tctggatcaa cgatagacac ttcgaaccag ttggctgctc gactctaggg atctcctcat 2880

agtttacaaa catgtgaaat agaataaagt ttgtatgact tgtatcatgg aattatgttg 2940

aaaagaatta taaattaaca tcatctccat tttttgatca gta 2983

<210> 5

<211> 2802

<212> DNA

<213> Arabidopsis thaliana (Arabidopsis thaliana)

<400> 5

caaacgtcac aactccgtca ccgcaaacta ggcagattat ctggttttcc ggcggtttct 60

cgttatctac gagagagtga gagaggaaat acctcggggt gaactcgctg cgtagccaaa 120

gcctcttcaa ttttgctctt ttgttcaatt ccgtcgctta attatctggt tttgcgtatt 180

ttgaagagtc agtttgacga atttatgctg aaatttcgtt ctttctctgt ttatctttca 240

gcgacgattt ttctgggttt ttcagttttg gatcaatcca taatctgtcg tgattagggt 300

tttatacctt gatgaggatt tactgaatta gttgtattaa agatggcgag aaaagctaac 360

aatagttttt tccttgaaga atggttaagg actgtgagtg gaagtagtgt ctctggtgat 420

ctagtgaagc aaaattctgc tccatctgct aggtcgatta ttcaagcatg gtctgagatt 480

cgtgaatctc ttcaaaacca aaactttgat tcacgttacc ttcaagcttt gagagctttg 540

gttagctctg agtccactat ccatgttgca gatccccaag caaagctact tatttctata 600

ctagcttttc aagatgtatc tcttccgtct gagtcttaca cacttgttct cagactactc 660

tatgtgtgga ttaggaaagc atttcgaccg tctcaagcac ttgttggtgt agcggttcag 720

gccattcgtg gcgttgttga tgacaggcgt aatctccaac cagccttagt agcgcagagt 780

gttctggttt ctggcttttg gaagaagagt actctttagt gggatctcaa gaagaacttg 840

ttcctgtagt gcttgcagga attgggtatg ctttatcttc ttcgttggat gttcattacg 900

tcagactatt ggatttattg tttggcattt ggttaaaaga cgagggtcct cgtggcactg 960

tcacttacgg tctgatgatt cttcatttga ttgagtgggt tgtgtcaggt tatatgcggt 1020

ctaattctat caacaagatg tctctttttg cgaacgaggt actagagact tctaaggaaa 1080

agtatgctgt tttcgctgtc ttcatggcag cagctggggt agtgagagct tccacggcag 1140

gattcagtag tggtgcacag agtttggaga tttctaaact aagaaattca gctgaaaagc 1200

gaatagaatt tgtagctcaa attttagttt ctaatggtaa tgttgttaca cttccaacca 1260

cacagagaga aggtcccctg ttgaagtgct ttgctattgc attggctcga tgtgggtctg 1320

tttcttcctc tgctcctctg cttttgtgtc tcacttctgc attgttaact caggtatttc 1380

ctttaggcca gatatatgaa tcattttgca atgcttttgg caaagagcct attggaccaa 1440

gactcatttg ggtcagggag catctttctg atgttctttt caaggaatca ggggccatat 1500

ctggggcttt ctgcaatcaa tattcatcag caagtgaaga gaacaagtat attgtggaga 1560

atatgatttg ggatttctgt caaaatctct acttacagca ccgtcaaatt gctatgttgc 1620

tttgtggtat agaagataca ttgcttggag acatcgagaa aattgcagaa tcatctttcc 1680

ttatggttgt cgtctttgcg ctagctgtta cgaaacaatg gctaaaacca atagtgtcta 1740

aagaaagaaa aatggtgaca tcagttaaga tattagtttc gttctcctgt gtagagtatt 1800

tcaggcatat tcgtttacct gaatacatgg agacaattag agaggtgatt tcatgtgtcc 1860

aggagaatga tgctccttgt gtttcatttg tagagtccat tcctgcctat gatagcttga 1920

cgaatccgaa agacttgttt acgcaaagga taaaatatga atggtcaaga gatgatgtgc 1980

agacatctcg aatattgttt tatcttcgag taatcccaac ttgcatcgga cggttatctg 2040

cttctgcctt caggggagtg gttgcatcaa ccatgttttt atatattgga catcctaaca 2100

gaaaagtggc gcaagcatct catacgttgt tggcagcatt tctctcttca gcaaaagaat 2160

cagaggagga cgaacgaact caattcaaag aacaacttgt tttctattac atgcaacgat 2220

ctttggaggt ttaccctgag atcacacctt ttgaaggttt agcttccggc gttgcaacct 2280

tggtccagca tctacctgcg ggaagtcctg ctatatttta ttctgttcat agccttgttg 2340

agaaggcttc tacatttagc accgaatcat tgcaaggcag aaagtctgat cctggtaatc 2400

agattcttga gttgcttttg cgacttgtct ctttggttga tatacaagtg ttgccatatc 2460

ttatgaagtc attggcacag ctagttataa agttaccaaa agaaagacag aacgttgtgc 2520

ttggtgaatt gtatggtcaa gtggctgagt cagacgatgt gattcgcaag ccttctttgg 2580

tctcttggct acagtcactg aactacttat gttctaataa ccgtactgaa gttttagctt 2640

ctggatcaac gatagacact tcgaaccagt tggctgctcg actctaggga tctcctcata 2700

gtttacaaac atgtgaaata gaataaagtt tgtatgactt gtatcatgga attatgttga 2760

aaagaattat aaattaacat catctccatt ttttgatcag ta 2802

<210> 6

<211> 1500

<212> DNA

<213> Arabidopsis thaliana (Arabidopsis thaliana)

<400> 6

aacttcaaat ttaatgaggc ctcctcttcc ttgaacgcaa aggaagtttc ttcgttggaa 60

aacgctgtct ccaattcaca tctctctctt tctctttccc tctgtacaca catacgtata 120

cgtatatata ttaacactat atttcattta tgtcttcgat ggtcctatga tctttaatct 180

ccatggcgac gactagagtc tcacatgtct tagggtttct tctatggatc tctcttctca 240

tcttcgtctc catagggttg tttggcaatt tcagctccaa acccataaac ccttttcctt 300

ctccggttat taccttgccg gccctttact accggccggg aagaagagct ttggcggtga 360

aaacgtttga cttcacaccg ttcttgaaag atcttcgccg gagtaatcat cggaaagctt 420

tgccggcggg aggatccgag attgacccga gatatggcgt tgaaaaacga ctcgtcccat 480

ctggtccaaa ccccttgcac cactgatttg tcttcttaat ttttgcttta agttttcttt 540

attttttcaa agatatagta aaacacgtat attttactaa tactagaaga ttaattcata 600

agatattagc taatattctt tcttggtatg tatatgtctt gaattattta ttcttctttg 660

tcatgtgatt tatttttttt cacttctggt gtaatgatgt aatttactca tctttttatt 720

ctttcttcat ttgatactaa tcgttgtaat aataaagcat catgtaagca gaaataaaag 780

aaaagatcat gtacacattt gtttaaatgc caaaaaagaa gttagcacca cagtaatcta 840

tactagtttt cgattaagaa gtgtttgtta agggaaagag aaattaaatt agcttcgatg 900

ttaccagtca cgtgaatcgt tgacttaatt agtttccttt tccaggtttt gtgcaaaact 960

ttttctgaat tttgtgggaa gtagacaacg aggaaatgag ctcatgtgat ttcgaaacct 1020

gcaattacag tttgtttacg tattatgttg acctcattga agttgaaaaa gacatttaac 1080

aattctgtag ctggtttact aaccgtgcgg ttttatctcc ggttttctgg tttatttggt 1140

ttatccttaa ccacaaaaca aacgtcacaa ctccgtcacc gcaaactagg cagattatct 1200

ggttttccgg cggtttctcg ttatctacga gagagtgaga gaggaaatac ctcggggtga 1260

actcgctgcg tagccaaagc ctcttcaatt ttgctctttt gttcaattcc gtcgcttaat 1320

tatctggttt tgcgtatttt gaagagtcag tttgacgaat ttatgctgaa atttcgttct 1380

ttctctgttt atctttcagc gacgattttt ctgggttttt cagttttgga tcaatccata 1440

atctgtcgtg attagggttt tataccttga tgaggattta ctgaattagt tgtattaaag 1500

<210> 7

<211> 46

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 7

cacgggggac tctagaatgg cgagaaaagc taacaatagt tttttc 46

<210> 8

<211> 46

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 8

cttgctcacc attgttggat ccccgagtcg agcagccaac tggttc 46

<210> 9

<211> 52

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 9

gtacaagaga tctgcgtcga ctatggcgag aaaagctaac aatagttttt tc 52

<210> 10

<211> 46

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 10

cgatcgggga aattcgagct ctcagagtcg agcagccaac tggttc 46

<210> 11

<211> 44

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 11

gacgagctgt acaagtctag aatggcgaga aaagctaaca atag 44

<210> 12

<211> 41

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 12

ggtcttaatt aactctctag atcataagat atggcaacac c 41

<210> 13

<211> 52

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 13

ggtcttaatt aactctctag atcaaggata caaaaacatg gttgatgcaa cc 52

<210> 14

<211> 46

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 14

gacgagctgt acaagtctag aatgattctt catttgattg agtggg 46

<210> 15

<211> 46

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 15

ggtcttaatt aactctctag atcagagtcg agcagccaac tggttc 46

<210> 16

<211> 21

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 16

taccagtcac gtgaatcgtt g 21

<210> 17

<211> 21

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 17

actcttcttc caaaagcctg c 21

<210> 18

<211> 21

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 18

gcagacttgt ttacgcaaag g 21

<210> 19

<211> 21

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 19

gcagacttgt ttacgcaaag g 21

<210> 20

<211> 19

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 20

attttgccga tttcggaac 19

<210> 21

<211> 47

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 21

acgacggcca gtgccaagct taacttcaaa tttaatgagg cctcctc 47

<210> 22

<211> 47

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 22

accattgttg gatcctctag aggatccctt taatacaact aattcag 47

<210> 23

<211> 44

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 23

gtattaaagg gatcctctag aatggcgaga aaagctaaca atag 44

<210> 24

<211> 46

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 24

ggccgctaca ctcgagtcga ccctgattcg tttcttaccc tttcaa 46

<210> 25

<211> 44

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 25

gtattaaagg gatcctctag aatggcgaga aaagctaaca atag 44

<210> 26

<211> 46

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 26

ggccgctaca ctcgagtcga ccctgattcg tttcttaccc tttcaa 46

<210> 27

<211> 20

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 27

aacagaaaag tggcgcaagc 20

<210> 28

<211> 21

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 28

atcgcaagac cggcaacagg a 21

<210> 29

<211> 24

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 29

cccctgtgtt tcctatgctg cttg 24

<210> 30

<211> 21

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 30

ttggagatta cgcctgtcat c 21

Claims

The application of the E41 gene in regulating and controlling the plant embryonic development is characterized in that the nucleotide sequence of the E41 gene is shown as any one of SEQ ID NO. 1-5.
The application of the E41 protein in regulation and control of plant embryonic development is characterized in that the E41 protein is obtained by encoding a nucleotide sequence shown by any one of SEQ ID NO. 1-5.
3. The use of any one of claims 1 or 2, wherein the plant is a dicot.
4. Use according to claim 3, wherein the plant is Arabidopsis thaliana.
5. The use of any of claims 1 or 2, wherein the route of application comprises:

(1) utilizing gene mutation, gene knockout, gene interference or gene silencing technology to cause deletion or reduction of expression quantity of the E41 gene, thereby constructing a plant model with abnormal embryonic development;

(2) introducing the E41 gene into a receptor plant, and culturing to obtain a transgenic plant with acquired function; the recipient plant is a plant with abnormal embryonic development caused by deletion of the E41 gene.
6. The use of claim 5, wherein the plant is Arabidopsis thaliana.