CN113527455A - Grain development related protein TaGSR1 and application thereof in wheat plant type breeding - Google Patents

Abstract

The invention discloses an application of a grain development related protein TaGSR1 or an encoding gene thereof in wheat plant type breeding; wherein the amino acid sequence of the grain development related protein TaGSR1 is shown as SEQ ID NO.1, and the nucleotide sequence of the coding gene thereof is shown as SEQ ID NO. 2; the application in the wheat plant type breeding refers to the application of regulating and controlling the wheat plant height or the application of regulating and controlling the included angle of wheat leaves. The invention defines the relationship between the TaGSR1 gene of wheat and the plant height and leaf angle of wheat, and verifies that the high expression of TaGSR1 can reduce the plant height of wheat and the leaf angle. The invention realizes the rapid breeding of the ideal plant type of the wheat by utilizing the genetic engineering technology, provides a feasible method for improving the lodging resistance of the wheat and reasonably close planting, and has important breeding application value and wide market application prospect.

Description

Grain development related protein TaGSR1 and application thereof in wheat plant type breeding

Technical Field

The invention relates to a seed grain development related protein and application thereof, in particular to application of a seed grain development related protein TaGSR1 or an encoding gene thereof in wheat plant type breeding; belongs to the technical field of plant genetic engineering and molecular breeding.

Background

Wheat is the second largest food crop in the world, providing 20% carbohydrate and 23% protein to humans. The human food safety problem faces the problems of reduction of cultivated land area, global climate warming, continuous population growth, aggravation of environmental pollution and the like, and how to improve the wheat yield under the condition of limited resources has important significance on food safety. The green revolution of wheat reduces the plant height of wheat, increases the lodging resistance of wheat, greatly increases the yield of wheat, and proves the importance of the ideal plant type of crops to the yield.

Brassinolide (BR) as the sixth plant hormone plays an important role in the processes of cell elongation and division, leaf senescence, plant morphogenesis, plant stress resistance and the like. Due to the wide role of BR in the growth and development of plants, its application in agricultural production is also gaining increasing attention. BRI1 (Brassicasteroid-inducing 1) is used as a BR receptor located on a cell membrane, after being combined with BR, the BR receptor is mutually phosphorylated with a co-receptor BAK1(BRI1-associated receptor) to initiate downstream signal transduction and further phosphorylate BSK/CDG1 kinase, the phosphorylated BSK/CDG1 further activates BSU1(BRI 1-repressisor 1) downstream through phosphorylation, the BSU1 is used as a phosphatase, and negative regulatory factors in the BR signal channel of BIN2 are inactivated through dephosphorylation BIN2 (Brassicasteroid-inducing 2) to further regulate the expression of downstream genes.

The applicant researches and discovers that TaGSR1(Grain Size Regulator 1of Triticum aestivum L.) is a transcription factor downstream of BR signals, has a function of regulating and controlling the Size of wheat grains, and has an important regulating and controlling effect on plant types such as wheat leaf included angles and plant heights. The regulation and control function provides very important information for researching growth and development of BR in wheat and breeding of yield. Through retrieval, the coding sequence of the wheat TaGSR1 gene, the grain development related protein TaGSR1 coded by the coding sequence and the application of the protein TaGSR1 in the regulation and control of wheat plant type breeding are not reported.

Disclosure of Invention

Aiming at the defects of the prior art, the invention aims to provide the application of a seed grain development related protein TaGSR1 or a coding gene thereof in wheat plant type breeding.

The grain development related protein is characterized in that: the protein is named as protein TaGSR1, is derived from common wheat, and is the protein described in the following 1) or 2):

1) a protein consisting of an amino acid sequence shown by SEQ ID NO.1 in a sequence table;

2) the protein which is obtained by substituting and/or deleting and/or increasing one or more amino acid residues of the amino acid sequence shown by SEQ ID NO.1 in the sequence table, is related to grain development and is derived from the SEQ ID NO. 1.

Further preferred embodiments are: the amino acid sequence of the protein TaGSR1 is composed of 203 amino acids, and the amino acid sequence is shown in SEQ ID NO. 1.

The invention discloses a coding gene of the grain development related protein TaGSR1, which is characterized in that: the nucleic acid sequence of the coding gene is the nucleotide sequence of a) or b) or c) as follows:

a) a deoxyribonucleotide sequence shown by SEQ ID NO.2 in the sequence table;

b) a deoxyribonucleotide sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% homology to the deoxyribonucleotide sequence represented by SEQ ID No.2 and encoding the protein;

c) a ribonucleotide sequence that is complementary to the deoxyribonucleotide sequence defined in a) or b) and encodes said protein under stringent conditions.

Further preferred embodiments are: the coding gene consists of 612 deoxyribonucleotides, and the nucleotide sequence of the coding gene is shown as SEQ ID NO. 2.

The invention also discloses a binary vector with the function of ectopically highly expressing the TaGSR1 gene in wheat, which is characterized in that: the binary vector is named as pLGY02-TaGSR1, and the nucleotide sequence of the binary vector is shown in SEQ ID NO. 4; the binary vector contains an expression cassette E1 of a TaGSR1 gene playing a main role in the genetic transformation process of wheat, wherein the nucleotide sequence of the expression cassette E1 is shown as SEQ ID NO.3, and the expression cassette E1 comprises the following components in sequence from upstream to downstream: ubiquitin promoter Ubi from corn, coding sequence of TaGSR1 gene, fluorescence enhanced green fluorescent protein coding gene GFP from multipipecacer, and terminator T1.

Wherein: the nucleotide sequence of the ubiquitin promoter Ubi is shown as SEQ ID NO.5, the coding sequence of the TaGSR1 gene is shown as SEQ ID NO.2, and the nucleotide sequence of the fluorescence enhanced green fluorescent protein coding gene GFP from the aequorea is shown as SEQ ID NO. 6; the nucleotide sequence of the terminator T1 is shown as SEQ ID NO. 7.

The invention further discloses application of the grain development related protein TaGSR1 or the coding gene thereof in wheat plant type breeding; wherein the amino acid sequence of the grain development related protein TaGSR1 is shown as SEQ ID NO.1, and the nucleotide sequence of the coding gene thereof is shown as SEQ ID NO. 2; the application in the wheat plant type breeding refers to the application of regulating and controlling the wheat plant height or the application of regulating and controlling the included angle of wheat leaves.

Wherein, the application method comprises the following steps: constructing a binary expression vector pLGY02-TaGSR 1of ectopic expression TaGSR1 gene, and transforming the binary expression vector into wheat to realize the ectopic high expression of TaGSR1 gene; or the binary expression vector is transformed into common wheat through agrobacterium to obtain transgenic wheat with TaGSR1 gene over-expression, so that the reduction of the plant height of the wheat and the reduction of the included angle of leaves are realized; wherein the nucleotide sequence of the TaGSR1 gene is shown in SEQ ID NO. 2.

The invention provides a grain development related protein TaGSR1 or a coding gene thereof and application thereof in optimizing wheat ideal plant type breeding; wherein, the grain development related protein TaGSR1 or the coding gene thereof is disclosed for the first time. The invention further determines that TaGSR1 is a transcription factor of BR signal downstream, has the function of regulating and controlling the size of wheat grains, and has important regulation and control effects on plant types such as wheat leaf included angle, plant height and the like. Meanwhile, experiments are utilized to confirm the relationship between the TaGSR1 gene of the wheat and the plant height and the leaf included angle of the wheat, and the high expression of the TaGSR1 is verified to reduce the plant height reduction of the wheat and the leaf included angle. The invention has the following outstanding effects: by constructing a binary expression vector pLGY02-TaGSR1 capable of ectopically expressing the TaGSR1 gene and genetically transforming the binary expression vector into common wheat to obtain a descendant with high expression of the TaGSR1 gene, the overexpression of the TaGSR1 gene is realized. Experiments prove that the obtained TaGSR1 gene overexpression transgenic wheat realizes the reduction of the plant height and the upright leaves of the wheat, and the plant type is more suitable for close planting. The invention realizes the rapid breeding of the ideal plant type of the wheat by utilizing the genetic engineering technology, provides a feasible method for improving the lodging resistance of the wheat and reasonably close planting, and has important breeding application value and wide market application prospect.

Drawings

FIG. 1 is the electrophoresis diagram of the TaGSR1 gene amplification of Shixin 828.

Wherein: lane 1 is Transgen 2K plus marker, and lane 2 is TaGSR1 band.

FIG. 2 sequencing result of the cloning vector pDOOR 221 into which the TaGSR1 gene had been ligated.

FIG. 3TaGSR1 gene was ligated into binary expression vector pLGY02 vector by LR reaction.

Wherein: 1 is Transgen 2K plus marker, 2-6 is a positive clone of TaGSR1 successfully ligated into pLGY02 vector.

FIG. 4 PCR identification of transgenic progeny overexpressing TaGSR 1.

Wherein: 1 is a Transgen 2K plus marker, and 2-9 are transgenic strains 1^#～8^#The target band detection result of (1).

FIG. 5 protein expression quantity identification of transgenic offspring overexpressing TaGSR 1.

FIG. 6OE-TaGSR1 overexpression transgenic line and its gene editing knockout line TaGSR1-ko seed size comparison.

Wherein: a & B, compared with wild type JW1 wheat, the length and width of seeds of OE-TaGSR1 overexpression lines are obviously reduced; c & D, compared with wild JW1 wheat, the homozygous gene knockout strain tagsr1-ko has obviously increased kernel length and no obvious change in kernel width. Bar is 1 cm.

FIG. 7 shows the comparison of the height of OE-TaGSR1 overexpression transgenic line and TaGSR1-ko gene editing knockout line.

Wherein: the plant height of the TaGSR1 overexpression transgenic line is obviously reduced, and the plant height of the TaGSR1-ko knockout line is not obviously changed. Bar is 5 cm.

FIG. 8OE-TaGSR1 overexpression transgenic line and its gene editing knockout line TaGSR1-ko leaf angle trait comparison.

Wherein: the included angle of the leaves of the TaGSR1 overexpression transgenic line is obviously reduced, and the included angle of the leaves of the TaGSR1-ko knockout line is obviously increased. Bar is 1 cm.

Detailed Description

The present invention will be described in detail with reference to the following detailed drawings and examples. The following examples are only preferred embodiments of the present invention, and it should be noted that the following descriptions are only for explaining the present invention and not for limiting the present invention in any form, and any simple modifications, equivalent changes and modifications made to the embodiments according to the technical spirit of the present invention are within the scope of the technical solution of the present invention.

The experimental procedures in the following examples are conventional unless otherwise specified.

The test materials used in the following examples were purchased from conventional biochemicals, unless otherwise specified.

The high fidelity enzyme required for PCR amplification is KOD-FXNEO (Toyobo); the vector used for the BP reaction was pDonor221 (Invitrogen); 2 × PCR master mix was purchased from Beijing Convergence scientific and biological Co., Ltd; t4 ligase was purchased from NEB; BPClonase and LR Clonase for homologous recombination, gel recovery kit, plasmid extraction kit and western blotting antibody required for enzyme digestion fragment recovery were purchased from Saimer Feishell science and technology. The inorganic salts required for the preparation of the culture medium were purchased from the national pharmaceutical group, vitamins and antibiotics and from Sigma. The plasmid pLGY02 was obtained from the crop research institute of agronomy academy of Shandong province. Coli Transgen5 α, purchased from tokyo holotype gold, inc; the primers used were synthesized by Qingdao Optimaki Biotechnology Limited, and the sequences of the relevant primers are shown in Table 1:

table 1: primers used in the present invention

The wheat variety JW1 used in the invention is a new germplasm with good tissue culture capability, which is self-bred by crops of agricultural academy of Shandong province, and can be obtained by the public from the crop research institute of agricultural academy of Shandong province; shixin 828 is a wheat variety which is bred in 2001 by 8 generations of pedigree breeding method at New wheat New breed New technology institute in Shijiazhuang City, examined and rated by the variety examination committee of Hebei province in 2005 and has excellent properties of cold resistance, drought resistance, lodging resistance and the like, and can be obtained by Jiafeng variety Co Ltd.

EXAMPLE 1 construction of expression vectors

Cloning of the coding sequence of TaGSR1

In order to obtain a coding sequence of TaGSR1 protein, the sequence of a TaGSR1 gene is amplified in the wheat variety Shixin 828 by the experiment, primers are TaGSR1-F and TaGSR1-R, and a PCR reaction system is as follows: KOD-FX NEO buffer: 25 μ L of dNTP (2 mM): 10 μ L, TaGSR1-F (10 μ M): 1.5 μ L, TaGSR1-F (10 μ M): 1.5 μ L, Shixin 828gDNA (50 ng/. mu.L): 1 μ L, KOD-FXNEO: 1 μ L, ddH₂The total amount of O is 50 μ L. The PCR reaction program is: pre-denaturation at 98 ℃ for 2min, and denaturation at 98 ℃ for 12 sec; annealing at 58 ℃ for 20sec, extension at 68 ℃ for 25sec, and reacting for 35 cycles; renaturation at 68 ℃ for 5 min.

The PCR results are shown in FIG. 1, lane 1 is Transgen 2K plus marker, and lane 2 is TaGSR1 band.

The PCR product is sent to Qingdao Zhixi biotechnology Limited company for sequencing, and the similarity of the TaGSR1 gene in Shixin 828 wheat and the known sequence of wheat in Ensembl Plants database (http:// Plants. ensemble. org/index. html) is 93.33%. The gene coding sequence has a total length of 612 bases and does not contain introns. The nucleotide sequence is shown in SEQ ID NO. 2.

BP reaction of TaGSR1 with pDonor221

A homojunction attB1 and attB2 were added to the TaGSR1 gene.

Taking TaGSR1 amplified in the wheat variety Shixin 828 as a template, using primers of TaGSR1-P1 and TaGSR1-P2, and adopting a PCR reaction system as follows: KOD-FX NEO buffer: 25 μ L of dNTP (2 mM): 10 μ L, TaGSR1-P1(10 μ M): 1.5 μ L, TaGSR1-P2(10 μ M): 1.5. mu.L, TaGSR1 PCR gel product DNA (10 ng/. mu.L): 1 μ L, KOD-FXNEO: 1 μ L, ddH₂The total amount of O is 50 μ L. The PCR reaction program is: pre-denaturation at 98 ℃ for 2min, and denaturation at 98 ℃ for 12 sec; annealing at 58 ℃ for 20sec, extension at 68 ℃ for 25sec, and reacting for 35 cycles; renaturation at 68 ℃ for 5 min.

And recovering the PCR product through running gel, and carrying out BP reaction with pDOnor221 plasmid by using a reaction system: gel recovery product (about 50 ng/. mu.L): 1 μ L, pDOnor221(50 ng/. mu.L): 1 μ L, BP clonase: 0.5. mu.L, ddH₂Make up to 10. mu.L of O. The reaction system reacts for 1h at 25 ℃, the reaction product is transformed into Escherichia coli Transgen5 alpha, the Escherichia coli Transgen5 alpha is cultured on an LB (containing kanamycin) plate until a clone grows out, a single clone is selected for carrying out PCR identification and sequencing on bacterial liquid, and primers are M13F and M13R.

The positive recombinant strain is sent to Qingdao Hingxi biotechnology Limited company for sequencing, the sequencing result is shown in figure 2, and the result shows that the TaGSR1 gene is connected into an entry vector pDonor221, and the recombinant vector is named as pDonor221-TaGSR 1.

3. Transformation of

The PCR product was recovered by running gel, and subjected to LR reaction with pLGY02 plasmid in the following reaction system: pDOnor221-TaGSR1 plasmid (10 ng/. mu.L): 1 μ L, pLGY02(50 ng/. mu.L): 1 μ L, LR clonase: 0.5. mu.L, ddH₂Make up to 10. mu.L of O. The reaction system reacts for 1h at 25 ℃, the reaction product is transformed into Escherichia coli Transgen5 alpha, the Escherichia coli Transgen5 alpha is cultured on an LB (containing kanamycin) plate until a clone grows out, a single clone is selected for carrying out PCR identification and sequencing on bacterial liquid, and TaGSR1-F and CCFP-R are used as primers.

The electrophoresis results are shown in FIG. 3, and the clone having the objective band is a positive clone carrying the binary vector pLGY02-TaGSR 1. Wherein the nucleotide sequence of the binary expression vector pLGY02-TaGSR1 is shown in SEQ ID NO. 4.

Example 2 transgenic progeny acquisition and characterization

Obtaining transgenic progeny of TaGSR1

The binary expression vector pLGY02-TaGSR1 constructed in the experimental example 1 is transformed into agrobacterium EHA105 competent cells, specifically: adding a recombinant binary expression vector pLGY02-TaGSR1 into agrobacterium EHA105 competence, performing ice bath for 5min, performing liquid nitrogen quick freezing for 5min, performing heat shock reaction at 37 ℃ for 5min, performing ice bath for 5min, adding nonresistant LB, putting into a shaker at 28 ℃ for recovery for 2h, then coating on an LB (containing rifampicin, streptomycin and kanamycin) plate by using an applicator, and performing inverted culture at 28 ℃ until clone grows out. The single clone was selected and inoculated into LB medium containing the corresponding resistance, shaken at 28 ℃ and cultured at 160rpm for 24 hours.

Taking JW1 wheat seeds about 15 days after pollination, and stripping young embryos. The Agrobacterium suspension was prepared by placing 1ml of the suspension in a 1.5ml centrifuge tube, adding 1.4ul acetosyringone (0.1M), and mixing. Adding prepared bacterial liquid to infect for 5 minutes, placing on a co-culture medium, and culturing in dark at 23 ℃ for 3 days. After co-cultivation, the cells were cultured in the dark at 25 ℃ for 5 days in a resting medium. The callus was transferred to the screening medium 1, sealed with a sealing film, and cultured in the dark in an incubator at 25.5 ℃ for 2 weeks. The callus was cut and transferred to screening medium 2, the petri dish was sealed again with a sealing film, and dark culture was continued in an incubator at 25.5 ℃ for 2 weeks. After 2 weeks of callus cutting and selection, resistant calli exhibiting green shoots were transferred to regeneration medium. The dishes were sealed and placed in an incubator at 25 ℃ for 2 weeks in light/dark (16h/8 h). After 2 weeks of regeneration, healthy growing plantlets were transferred to new resistant regeneration pods. When the seedlings grow to a certain size, sampling detection can be carried out.

The various culture media involved in the genetic transformation of wheat and the preparation thereof are disclosed in the following documents:

Kan Wang(ed.),Agrobacterium Protocals:Volume 1,Methods in Molecular Biology,vol.1223DOI10.007/978-1-4939-1695-5_15,Spring Science+Businedd Media New York 2015。

identification of transgenic progeny of TaGSR1

DNA of young leaves of T0 generation wheat plants is extracted by a CTAB method, and PCR identification is carried out on transgenic wheat by taking gDNA as a template and LGY-DF and LGY-DR as primers. The reaction system is as follows: 2 × PCR master mix: 10 μ L, LGY-DF (10 μ M): 0.5 μ L, LGY-DR (10 μ M): 0.5. mu.L, gDNA (50 ng/. mu.L): 1 μ L, ddH₂Make up to 20. mu.L of O. The PCR reaction conditions are as follows: pre-denaturation at 94 ℃ for 2 min; denaturation at 94 ℃ for 30s, annealing at 60 ℃ for 30s, and extension at 72 ℃ for 30s for 33 cycles; finally, extension is carried out for 5min at 72 ℃. The electrophoresis result is shown in figure 4, and the total number of transgenic positive plants of 6T 0 generations is.

Selecting a positive transgenic strain OE-TaGSR1-1^#、OE-TaGSR1-5^#And OE-TaGSR1-6^#And (3) carrying out western blotting protein expression quantity identification. The main experimental operations were: SDS method extracts total leaf protein, using 12% SDS-PAGE glue to make electrophoretic separation, using GFP antibody fused with TaGSR1 protein to make western blotting detection to express transgenic wheat, the result is shown in figure 5, showing positive transgenic line OE-TaGSR1-1^#With OE-TaGSR1-6^#Has high expression amount.

Example 3 phenotypic identification of wheat progeny overexpressing the TaGSR1 Gene

Wild type receptor variety JW1, over-expression transgenic wheat OE-TaGSR1 and TaGSR1 and homozygous knockout strains TaGSR1-ko of homologous copies thereof are planted in a climatic chamber of Qingdao school district of Shandong university under the culture conditions: illuminating for 16h and darkness for 8 h; the temperature in the daytime is 22 ℃, and the temperature at night is 16 ℃; humidity is 40% -50%; CO 2₂The concentration is 500ppm-700 ppm.

Grain length, width and thousand kernel weight were analyzed for JW, OE-TaGSR1 and TaGSR 1-ko.

Compared with the wild JW1, the grain length and width of the over-expression strain OE-TaGSR1 are obviously reduced, while the grain length of the mutant strain TaGSR1-ko is obviously increased, and the grain width is not obviously changed (as shown in Table 2 and figure 6). Measurement of thousand kernel weight of grains showed that the thousand kernel weight of the OE-TaGSR1 overexpression transgenic line was significantly reduced, and the thousand kernel weight of TaGSR1-ko was significantly increased (see table 2). The result shows that the TaGSR1 gene has negative regulation and control effect on wheat grain development.

TABLE 2 statistics of the yield traits of homozygous three-copy knockout mutant tagsr1-ko

Note: different lower case superscripts indicate significant differences at the 95% confidence level.

And analyzing the plant height and the leaf angle of a transgenic strain OE-TaGSR1 and a gene knockout strain TaGSR1-ko of the over-expression TaGSR1 gene.

The analysis result of the plant height is shown in FIG. 7, compared with JW1, the plant height of OE-TaGSR1 is obviously reduced, and the plant height of the gene knockout strain TaGSR1-ko has no obvious difference compared with JW1, which indicates that the TaGSR1 gene has obvious inhibition effect on the wheat plant height. Meanwhile, compared with the wild plant JW1, the included angle of leaves of OE-TaGSR1 is obviously reduced, the leaves are upright, and the included angle of leaves of the gene knockout strain TaGSR1-ko is obviously larger than that of JW1, as shown in FIG. 8. The reduction of the included angle of the leaves leads to the erect leaves, so that the plant type of the OE-TaGSR1 transgenic plant is more compact, and the plant type of the gene knockout strain TaGSR1-ko is looser than that of JW1, as shown in FIG. 7.

In the experiment, TaGSR1 element is transferred into wheat by constructing a TaGSR1 overexpression vector and infecting wheat immature embryo callus through agrobacterium, and the Ubi is verified that although TaGSR1 has a negative effect on the grain size, the overexpression of the TaGSR1 obviously reduces the plant height of the wheat and reduces the included angle of leaves, so that the generated wheat plant type with short stems and compact plant type has important significance for lodging-resistant and high-density planting in crop production, and an excellent candidate gene site and a feasible method are provided for plant type breeding of wheat.

Sequence listing

<110> Shandong university

<120> seed grain development related protein TaGSR1 and application thereof in wheat plant type breeding

<141> 2021-08-06

<160> 7

<210> 1

<211> 203

<212> PRT

<213> wheat (Triticum aestivum L.)

<221> amino acid sequence of protein TaGSR1

<222>（1）…（203）

<400> 1

MET Asp Ala Lys Gly Ala Thr Glu Ser Ser Asn Pro Asn Arg Arg Pro Ala Ser Val Pro

5 10 15 20

Ser Ala Ala Val Ala Ser Ala Ser Ala Pro Thr Lys Arg MET Leu Ala Phe His Phe Leu

25 30 35 40

Arg Ala Leu Ser Arg Ile His Gly Ala Ala Gly Pro Ala Arg Arg Arg Thr Arg Thr Ile

45 50 55 60

Arg Arg Ala Ala Tyr Ser Ser MET Ala Arg Ala Thr Gly Pro Arg Arg Ala Trp Ser Arg

65 70 75 80

Ala Leu Leu Leu Gln Ala Gln Ala Arg Ala Arg Arg Ser Arg Ala Glu Thr Ser Arg Arg

85 90 95 100

Ala Ala Val Leu Val Arg Arg Arg Val Val Ala Gly Pro Ala Val Ala Ala Ala Pro Ala

105 110 115 120

Pro Ala Arg Ala Ala Ser Val Gly Gly Gln Thr Ser Ser Ala Ala Ala Ile Arg Ala Ala

125 130 135 140

Leu Val Pro Pro Pro Pro Pro Ala Arg Gln Ala Gly Glu Pro Ala Arg Ser Asp Ala Leu

145 150 155 160

Arg Arg Leu Val Pro Gly Gly Ala Gly MET Glu Tyr Cys Ser Leu Leu Glu Glu Thr Ala

165 170 175 180

Asp Tyr Val Arg Cys Leu Arg Ala Gln Val Gln Leu MET Gln Gly Leu Ala Asp Leu Phe

185 190 195 200

Ser Cys Gln

203

<210> 2

<211> 612

<212> DNA

<213> wheat (Triticum aestivum L.)

<221> nucleotide sequence encoding protein TaGSR1

<222>（1）…（612）

<400> 2

atggacgcca agggcgcgac ggagagctca aaccctaacc gccgtccagc cagcgtaccg 60

tcggccgccg tggcgtcggc gtcggcgccg acgaagcgca tgctggcgtt ccacttcctg 120

cgcgcgctgt cccggatcca cggcgcggcc ggcccggcga ggcgccgcac gcgcaccatc 180

cgccgcgcgg cctactcctc catggcgcgc gccaccggcc cgcgccgcgc ctggagccgc 240

gcgctgctgc tccaggccca ggcgcgcgcg cggagatcca gggcggagac gtcgaggcgg 300

gccgccgtgc tcgtacggcg gcgcgtcgtc gccggaccgg cggtagcagc agcaccagca 360

ccggcacgcg ccgcttctgt cggcggacag acgtcgtcgg cggctgctat tcgcgcggcg 420

ctggtccctc cgccccctcc ggcgcggcag gcgggggagc cggccaggag cgacgcgctg 480

cggcggctcg tccccggagg cgccgggatg gagtactgca gcctgctgga ggagaccgcc 540

gactacgtcc gctgcctccg cgcgcaggtg cagctcatgc agggcctcgc cgacctcttc 600

tcctgccaat ga 612

<210> 3

<211> 3705

<212> DNA

<213> Artificial sequence

<221> nucleotide sequence of expression cassette E1

<222>（1）…（3705）

<400> 3

ctgcagtgca gcgtgacccg gtcgtgcccc tctctagaga taatgagcat tgcatgtcta 60

agttataaaa aattaccaca tatttttttt gtcacacttg tttgaagtgc agtttatcta 120

tctttataca tatatttaaa ctttactcta cgaataatat aatctatagt actacaataa 180

tatcagtgtt ttagagaatc atataaatga acagttagac atggtctaaa ggacaattga 240

gtattttgac aacaggactc tacagtttta tctttttagt gtgcatgtgt tctccttttt 300

ttttgcaaat agcttcacct atataatact tcatccattt tattagtaca tccatttagg 360

gtttagggtt aatggttttt atagactaat ttttttagta catctatttt attctatttt 420

agcctctaaa ttaagaaaac taaaactcta ttttagtttt tttatttaat aatttagata 480

taaaatagaa taaaataaag tgactaaaaa ttaaacaaat accctttaag aaattaaaaa 540

aactaaggaa acatttttct tgtttcgagt agataatgcc agcctgttaa acgccgtcga 600

cgagtctaac ggacaccaac cagcgaacca gcagcgtcgc gtcgggccaa gcgaagcaga 660

cggcacggca tctctgtcgc tgcctctgga cccctctcga gagttccgct ccaccgttgg 720

acttgctccg ctgtcggcat ccagaaattg cgtggcggag cggcagacgt gagccggcac 780

ggcaggcggc ctcctcctcc tctcacggca ccggcagcta cgggggattc ctttcccacc 840

gctccttcgc tttcccttcc tcgcccgccg taataaatag acaccccctc cacaccctct 900

ttccccaacc tcgtgttgtt cggagcgcac acacacacaa ccagatctcc cccaaatcca 960

cccgtcggca cctccgcttc aaggtacgcc gctcgtcctc cccccccccc cctctctacc 1020

ttctctagat cggcgttccg gtccatggtt agggcccggt agttctactt ctgttcatgt 1080

ttgtgttaga tccgtgtttg tgttagatcc gtgctgctag cgttcgtaca cggatgcgac 1140

ctgtacgtca gacacgttct gattgctaac ttgccagtgt ttctctttgg ggaatcctgg 1200

gatggctcta gccgttccgc agacgggatc gatttcatga ttttttttgt ttcgttgcat 1260

agggtttggt ttgccctttt cctttatttc aatatatgcc gtgcacttgt ttgtcgggtc 1320

atcttttcat gctttttttt gtcttggttg tgatgatgtg gtctggttgg gcggtcgttc 1380

tagatcggag tagaattctg tttcaaacta cctggtggat ttattaattt tggatctgta 1440

tgtgtgtgcc atacatattc atagttacga attgaagatg atggatggaa atatcgatct 1500

aggataggta tacatgttga tgcgggtttt actgatgcat atacagagat gctttttgtt 1560

cgcttggttg tgatgatgtg gtgtggttgg gcggtcgttc attcgttcta gatcggagta 1620

gaatactgtt tcaaactacc tggtgtattt attaattttg gaactgtatg tgtgtgtcat 1680

acatcttcat agttacgagt ttaagatgga tggaaatatc gatctaggat aggtatacat 1740

gttgatgtgg gttttactga tgcatataca tgatggcata tgcagcatct attcatatgc 1800

tctaaccttg agtacctatc tattataata aacaagtatg ttttataatt attttgatct 1860

tgatatactt ggatgatggc atatgcagca gctatatgtg gattttttta gccctgcctt 1920

catacgctat ttatttgctt ggtactgttt cttttgtcga tgctcaccct gttgtttggt 1980

gttacttctg caggtcgact ctagaggatc cacaagtttg tacaaaaaag caggctctat 2040

ggacgccaag ggcgcgacgg agagctcaaa ccctaaccgc cgtccagcca gcgtaccgtc 2100

ggccgccgtg gcgtcggcgt cggcgccgac gaagcgcatg ctggcgttcc acttcctgcg 2160

cgcgctgtcc cggatccacg gcgcggccgg cccggcgagg cgccgcacgc gcaccatccg 2220

ccgcgcggcc tactcctcca tggcgcgcgc caccggcccg cgccgcgcct ggagccgcgc 2280

gctgctgctc caggcccagg cgcgcgcgcg gagatccagg gcggagacgt cgaggcgggc 2340

cgccgtgctc gtacggcggc gcgtcgtcgc cggaccggcg gtagcagcag caccagcacc 2400

ggcacgcgcc gcttctgtcg gcggacagac gtcgtcggcg gctgctattc gcgcggcgct 2460

ggtccctccg ccccctccgg cgcggcaggc gggggagccg gccaggagcg acgcgctgcg 2520

gcggctcgtc cccggaggcg ccgggatgga gtactgcagc ctgctggagg agaccgccga 2580

ctacgtccgc tgcctccgcg cgcaggtgca gctcatgcag ggcctcgccg acctcttctc 2640

ctgccaaagt attggggata agggtgggcg cgccgaccca gctttcttgt acaaagtggt 2700

gcctatggtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct 2760

ggacggcgac gtaaacggcc acaagttcag cgtgtccggc gagggcgagg gcgatgccac 2820

ctacggcaag ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc 2880

caccctcgtg accaccttcg gctacggcct gcagtgcttc gcccgctacc ccgaccacat 2940

gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat 3000

cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac 3060

cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg 3120

gcacaagctg gagtacaact acaacagcca caacgtctat atcatggccg acaagcagaa 3180

gaacggcatc aaggtgaact tcaagatccg ccacaacatc gaggacggca gcgtgcagct 3240

cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa 3300

ccactacctg agctaccagt ccgccctgag caaagacccc aacgagaagc gcgatcacat 3360

ggtcctgctg gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa 3420

gtagactagt tctagagagc tcgaatttcc ccgatcgttc aaacatttgg caataaagtt 3480

tcttaagatt gaatcctgtt gccggtcttg cgatgattat catataattt ctgttgaatt 3540

acgttaagca tgtaataatt aacatgtaat gcatgacgtt atttatgaga tgggttttta 3600

tgattagagt cccgcaatta tacatttaat acgcgataga aaacaaaata tagcgcgcaa 3660

actaggataa attatcgcgc gcggtgtcat ctatgttact agatc 3705

<210> 4

<211> 12699

<212> DNA

<213> Artificial sequence

<221> nucleotide sequence of recombinant vector pLGY02-TaGSR1

<222>（1）…（12699）

<400> 4

cgtcagacac gttctgattg ctaacttgcc agtgtttctc tttggggaat cctgggatgg 60

ctctagccgt tccgcagacg ggatcgattt catgattttt tttgtttcgt tgcatagggt 120

ttggtttgcc cttttccttt atttcaatat atgccgtgca cttgtttgtc gggtcatctt 180

ttcatgcttt tttttgtctt ggttgtgatg atgtggtctg gttgggcggt cgttctagat 240

cggagtagaa ttctgtttca aactacctgg tggatttatt aattttggat ctgtatgtgt 300

gtgccataca tattcatagt tacgaattga agatgatgga tggaaatatc gatctaggat 360

aggtatacat gttgatgcgg gttttactga tgcatataca gagatgcttt ttgttcgctt 420

ggttgtgatg atgtggtgtg gttgggcggt cgttcattcg ttctagatcg gagtagaata 480

ctgtttcaaa ctacctggtg tatttattaa ttttggaact gtatgtgtgt gtcatacatc 540

ttcatagtta cgagtttaag atggatggaa atatcgatct aggataggta tacatgttga 600

tgtgggtttt actgatgcat atacatgatg gcatatgcag catctattca tatgctctaa 660

ccttgagtac ctatctatta taataaacaa gtatgtttta taattatttt gatcttgata 720

tacttggatg atggcatatg cagcagctat atgtggattt ttttagccct gccttcatac 780

gctatttatt tgcttggtac tgtttctttt gtcgatgctc accctgttgt ttggtgttac 840

ttctgcaggt cgactctaga ggatccacaa gtttgtacaa aaaagcaggc tctatggacg 900

ccaagggcgc gacggagagc tcaaacccta accgccgtcc agccagcgta ccgtcggccg 960

ccgtggcgtc ggcgtcggcg ccgacgaagc gcatgctggc gttccacttc ctgcgcgcgc 1020

tgtcccggat ccacggcgcg gccggcccgg cgaggcgccg cacgcgcacc atccgccgcg 1080

cggcctactc ctccatggcg cgcgccaccg gcccgcgccg cgcctggagc cgcgcgctgc 1140

tgctccaggc ccaggcgcgc gcgcggagat ccagggcgga gacgtcgagg cgggccgccg 1200

tgctcgtacg gcggcgcgtc gtcgccggac cggcggtagc agcagcacca gcaccggcac 1260

gcgccgcttc tgtcggcgga cagacgtcgt cggcggctgc tattcgcgcg gcgctggtcc 1320

ctccgccccc tccggcgcgg caggcggggg agccggccag gagcgacgcg ctgcggcggc 1380

tcgtccccgg aggcgccggg atggagtact gcagcctgct ggaggagacc gccgactacg 1440

tccgctgcct ccgcgcgcag gtgcagctca tgcagggcct cgccgacctc ttctcctgcc 1500

aaagtattgg ggataagggt gggcgcgccg acccagcttt cttgtacaaa gtggtgccta 1560

tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc gagctggacg 1620

gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg 1680

gcaagctgac cctgaagttc atctgcacca ccggcaagct gcccgtgccc tggcccaccc 1740

tcgtgaccac cttcggctac ggcctgcagt gcttcgcccg ctaccccgac cacatgaagc 1800

agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc accatcttct 1860

tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg 1920

tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc ctggggcaca 1980

agctggagta caactacaac agccacaacg tctatatcat ggccgacaag cagaagaacg 2040

gcatcaaggt gaacttcaag atccgccaca acatcgagga cggcagcgtg cagctcgccg 2100

accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact 2160

acctgagcta ccagtccgcc ctgagcaaag accccaacga gaagcgcgat cacatggtcc 2220

tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg tacaagtaga 2280

ctagttctag agagctcgaa tttccccgat cgttcaaaca tttggcaata aagtttctta 2340

agattgaatc ctgttgccgg tcttgcgatg attatcatat aatttctgtt gaattacgtt 2400

aagcatgtaa taattaacat gtaatgcatg acgttattta tgagatgggt ttttatgatt 2460

agagtcccgc aattatacat ttaatacgcg atagaaaaca aaatatagcg cgcaaactag 2520

gataaattat cgcgcgcggt gtcatctatg ttactagatc gggaattcgt aatcatgtca 2580

tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga 2640

agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg 2700

cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc 2760

caacgcgcgg ggagaggcgg tttgcgtatt ggctagagca gcttgccaaa catggtggag 2820

cacgacactc tcgtctactc caagaatatc aaagatacag tctcagaaga ccaaagggct 2880

attgagactt ttcaacaaag ggtaatatcg ggaaacctcc tcggattcca ttgcccagct 2940

atctgtcact tcatcaaaag gacagtagaa aaggaaggtg gcacctacaa atgccatcat 3000

tgcgataaag gaaaggctat cgttcaagat gcctctgccg acagtggtcc caaagatgga 3060

cccccaccca cgaggagcat cgtggaaaaa gaagacgttc caaccacgtc ttcaaagcaa 3120

gtggattgat gtgataacat ggtggagcac gacactctcg tctactccaa gaatatcaaa 3180

gatacagtct cagaagacca aagggctatt gagacttttc aacaaagggt aatatcggga 3240

aacctcctcg gattccattg cccagctatc tgtcacttca tcaaaaggac agtagaaaag 3300

gaaggtggca cctacaaatg ccatcattgc gataaaggaa aggctatcgt tcaagatgcc 3360

tctgccgaca gtggtcccaa agatggaccc ccacccacga ggagcatcgt ggaaaaagaa 3420

gacgttccaa ccacgtcttc aaagcaagtg gattgatgtg atatctccac tgacgtaagg 3480

gatgacgcac aatcccacta tccttcgcaa gaccttcctc tatataagga agttcatttc 3540

atttggagag gacacgctga aatcaccagt ctctctctac aaatctatct cctcgagctt 3600

tcgcagatcc cggggggcaa tgagatatga aaaagcctga actcaccgcg acgtctgtcg 3660

agaagtttct gatcgaaaag ttcgacagcg tctccgacct gatgcagctc tcggagggcg 3720

aagaatctcg tgctttcagc ttcgatgtag gagggcgtgg atatgtcctg cgggtaaata 3780

gctgcgccga tggtttctac aaagatcgtt atgtttatcg gcactttgca tcggccgcgc 3840

tcccgattcc ggaagtgctt gacattgggg agtttagcga gagcctgacc tattgcatct 3900

cccgccgtgc acagggtgtc acgttgcaag acctgcctga aaccgaactg cccgctgttc 3960

tacaaccggt cgcggaggct atggatgcga tcgctgcggc cgatcttagc cagacgagcg 4020

ggttcggccc attcggaccg caaggaatcg gtcaatacac tacatggcgt gatttcatat 4080

gcgcgattgc tgatccccat gtgtatcact ggcaaactgt gatggacgac accgtcagtg 4140

cgtccgtcgc gcaggctctc gatgagctga tgctttgggc cgaggactgc cccgaagtcc 4200

ggcacctcgt gcacgcggat ttcggctcca acaatgtcct gacggacaat ggccgcataa 4260

cagcggtcat tgactggagc gaggcgatgt tcggggattc ccaatacgag gtcgccaaca 4320

tcttcttctg gaggccgtgg ttggcttgta tggagcagca gacgcgctac ttcgagcgga 4380

ggcatccgga gcttgcagga tcgccacgac tccgggcgta tatgctccgc attggtcttg 4440

accaactcta tcagagcttg gttgacggca atttcgatga tgcagcttgg gcgcagggtc 4500

gatgcgacgc aatcgtccga tccggagccg ggactgtcgg gcgtacacaa atcgcccgca 4560

gaagcgcggc cgtctggacc gatggctgtg tagaagtact cgccgatagt ggaaaccgac 4620

gccccagcac tcgtccgagg gcaaagaaat agagtagatg ccgaccggat ctgtcgatcg 4680

acaagctcga gtttctccat aataatgtgt gagtagttcc cagataaggg aattagggtt 4740

cctatagggt ttcgctcatg tgttgagcat ataagaaacc cttagtatgt atttgtattt 4800

gtaaaatact tctatcaata aaatttctaa ttcctaaaac caaaatccag tactaaaatc 4860

cagatccccc gaattaattc ggcgttaatt cagtacatta aaaacgtccg caatgtgtta 4920

ttaagttgtc taagcgtcaa tttgtttaca ccacaatata tcctgccacc agccagccaa 4980

cagctccccg accggcagct cggcacaaaa tcaccactcg atacaggcag cccatcagtc 5040

cgggacggcg tcagcgggag agccgttgta aggcggcaga ctttgctcat gttaccgatg 5100

ctattcggaa gaacggcaac taagctgccg ggtttgaaac acggatgatc tcgcggaggg 5160

tagcatgttg attgtaacga tgacagagcg ttgctgcctg tgatcaccgc ggtttcaaaa 5220

tcggctccgt cgatactatg ttatacgcca actttgaaaa caactttgaa aaagctgttt 5280

tctggtattt aaggttttag aatgcaagga acagtgaatt ggagttcgtc ttgttataat 5340

tagcttcttg gggtatcttt aaatactgta gaaaagagga aggaaataat aaatggctaa 5400

aatgagaata tcaccggaat tgaaaaaact gatcgaaaaa taccgctgcg taaaagatac 5460

ggaaggaatg tctcctgcta aggtatataa gctggtggga gaaaatgaaa acctatattt 5520

aaaaatgacg gacagccggt ataaagggac cacctatgat gtggaacggg aaaaggacat 5580

gatgctatgg ctggaaggaa agctgcctgt tccaaaggtc ctgcactttg aacggcatga 5640

tggctggagc aatctgctca tgagtgaggc cgatggcgtc ctttgctcgg aagagtatga 5700

agatgaacaa agccctgaaa agattatcga gctgtatgcg gagtgcatca ggctctttca 5760

ctccatcgac atatcggatt gtccctatac gaatagctta gacagccgct tagccgaatt 5820

ggattactta ctgaataacg atctggccga tgtggattgc gaaaactggg aagaagacac 5880

tccatttaaa gatccgcgcg agctgtatga ttttttaaag acggaaaagc ccgaagagga 5940

acttgtcttt tcccacggcg acctgggaga cagcaacatc tttgtgaaag atggcaaagt 6000

aagtggcttt attgatcttg ggagaagcgg cagggcggac aagtggtatg acattgcctt 6060

ctgcgtccgg tcgatcaggg aggatatcgg ggaagaacag tatgtcgagc tattttttga 6120

cttactgggg atcaagcctg attgggagaa aataaaatat tatattttac tggatgaatt 6180

gttttagtac ctagaatgca tgaccaaaat cccttaacgt gagttttcgt tccactgagc 6240

gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat 6300

ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga 6360

gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt 6420

ccttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata 6480

cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac 6540

cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg 6600

ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg 6660

tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag 6720

cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct 6780

ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc 6840

aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt 6900

ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg 6960

tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga 7020

gtcagtgagc gaggaagcgg aagagcgcct gatgcggtat tttctcctta cgcatctgtg 7080

cggtatttca caccgcatat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt 7140

aagccagtat acactccgct atcgctacgt gactgggtca tggctgcgcc ccgacacccg 7200

ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa 7260

gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc 7320

gcgaggcagg gtgccttgat gtgggcgccg gcggtcgagt ggcgacggcg cggcttgtcc 7380

gcgccctggt agattgcctg gccgtaggcc agccattttt gagcggccag cggccgcgat 7440

aggccgacgc gaagcggcgg ggcgtaggga gcgcagcgac cgaagggtag gcgctttttg 7500

cagctcttcg gctgtgcgct ggccagacag ttatgcacag gccaggcggg ttttaagagt 7560

tttaatagtt ttccgtctgt cgaagcgtga ccgacgagct ggcgaggtga tccgctacga 7620

gcttccagac gggcacgtag aggtttccgc agggccggcc ggcatggcca gtgtgtggga 7680

ttacgacctg gtactgatgg cggtttccca tctaaccgaa tccatgaacc gataccggga 7740

agggaaggga gacaagcccg gccgcgtgtt ccgtccacac gttgcggacg tactcaagtt 7800

ctgccggcga gccgatggcg gaaagcagaa agacgacctg gtagaaacct gcattcggtt 7860

aaacaccacg cacgttgcca tgcagcgtac gaagaaggcc aagaacggcc gcctggtgac 7920

ggtatccgag ggtgaagcct tgattagccg ctacaagatc gtaaagagcg aaaccgggcg 7980

gccggagtac atcgagatcg agctagctga ttggatgtac cgcgagatca cagaaggcaa 8040

gaacccggac gtgctgacgg ttcaccccga ttactttttg atcgatcccg gcatcggccg 8100

ttttctctac cgcctggcac gccgcgccgc aggcaaggca gaagccagat ggttgttcaa 8160

gacgatctac gaacgcagtg gcagcgccgg agagttcaag aagttctgtt tcaccgtgcg 8220

caagctgatc gggtcaaatg acctgccgga gtacgatttg aaggaggagg cggggcaggc 8280

tggcccgatc ctagtcatgc gctaccgcaa cctgatcgag ggcgaagcat ccgccggttc 8340

ctaatgtacg gagcagatgc tagggcaaat tgccctagca ggggaaaaag gtcgaaaagg 8400

tctctttcct gtggatagca cgtacattgg gaacccaaag ccgtacattg ggaaccggaa 8460

cccgtacatt gggaacccaa agccgtacat tgggaaccgg tcacacatgt aagtgactga 8520

tataaaagag aaaaaaggcg atttttccgc ctaaaactct ttaaaactgg ccacgtccat 8580

gatgctgcga ctatcgcggg tgcccacgtc atagagcatc ggaacgaaaa aatctggttg 8640

ctcgtcgccc ttgggcggct tcctaatcga cggcgcaccg gctgccggcg gttgccggga 8700

ttctttgcgg attcgatcag cggccgcttg ccacgattca ccggggcgtg cttctgcctc 8760

gatgcgttgc cgctgggcgg cctgcgcggc cttcaacttc tccaccaggt catcacccag 8820

cgccgcgccg atttgtaccg ggccggatgg tttgcgaccg tcacgccgat tcctcgggct 8880

tgggggttcc agtgccattg cagggccggc agacaaccca gccgcttacg cctggccaac 8940

cgcccgttcc tccacacatg gggcattcca cggcgtcggt gcctggttgt tcttgatttt 9000

ccatgccgcc tcctttagcc gctaaaattc atctactcat ttattcattt gctcatttac 9060

tctggtagct gcgcgatgta ttcagatagc agctcggtaa tggtcttgcc ttggcgtacc 9120

gcgtacatct tcagcttggt gtgatcctcc gccggcaact gaaagttgac ccgcttcatg 9180

gctggcgtgt ctgccaggct ggccaacgtt gcagccttgc tgctgcgtgc gctcggacgg 9240

ccggcactta gcgtgtttgt gcttttgctc attttctctt tacctcatta actcaaatga 9300

gttttgattt aatttcagcg gccagcgcct ggacctcgcg ggcagcgtcg ccctcgggtt 9360

ctgattcaag aacggttgtg ccggcggcgg cagtgcctgg gtagctcacg cgctgcgtga 9420

tacgggactc aagaatgggc agctcgtacc cggccagcgc ctcggcaacc tcaccgccga 9480

tgcgcgtgcc tttgatcgcc cgcgacacga caaaggccgc ttgtagcctt ccatccgtga 9540

cctcaatgcg ctgcttaacc agctccacca ggtcggcggt ggcccatatg tcgtaagggc 9600

ttggctgcac cggaatcagc acgaagtcgg ctgccttgat cgcggacaca gccaagtccg 9660

ccgcctgggg cgctccgtcg atcactacga agtcgcgccg gccgatggcc ttcacgtcgc 9720

ggtcaatcgt cgggcggtcg atgccgacaa cggttagcgg ttgatcttcc cgcacggccg 9780

cccaatcgcg ggcactgccc tggggatcgg aatcgactaa cagaacatcg gccccggcga 9840

gttgcagggc gcgggctaga tgggttgcga tggtcgtctt gcctgacccg cctttctggt 9900

taagtacagc gataaccttc atgcgttccc cttgcgtatt tgtttattta ctcatcgcat 9960

catatacgca gcgaccgcat gacgcaagct gttttactca aatacacatc acctttttag 10020

acggcggcgc tcggtttctt cagcggccaa gctggccggc caggccgcca gcttggcatc 10080

agacaaaccg gccaggattt catgcagccg cacggttgag acgtgcgcgg gcggctcgaa 10140

cacgtacccg gccgcgatca tctccgcctc gatctcttcg gtaatgaaaa acggttcgtc 10200

ctggccgtcc tggtgcggtt tcatgcttgt tcctcttggc gttcattctc ggcggccgcc 10260

agggcgtcgg cctcggtcaa tgcgtcctca cggaaggcac cgcgccgcct ggcctcggtg 10320

ggcgtcactt cctcgctgcg ctcaagtgcg cggtacaggg tcgagcgatg cacgccaagc 10380

agtgcagccg cctctttcac ggtgcggcct tcctggtcga tcagctcgcg ggcgtgcgcg 10440

atctgtgccg gggtgagggt agggcggggg ccaaacttca cgcctcgggc cttggcggcc 10500

tcgcgcccgc tccgggtgcg gtcgatgatt agggaacgct cgaactcggc aatgccggcg 10560

aacacggtca acaccatgcg gccggccggc gtggtggtgt cggcccacgg ctctgccagg 10620

ctacgcaggc ccgcgccggc ctcctggatg cgctcggcaa tgtccagtag gtcgcgggtg 10680

ctgcgggcca ggcggtctag cctggtcact gtcacaacgt cgccagggcg taggtggtca 10740

agcatcctgg ccagctccgg gcggtcgcgc ctggtgccgg tgatcttctc ggaaaacagc 10800

ttggtgcagc cggccgcgtg cagttcggcc cgttggttgg tcaagtcctg gtcgtcggtg 10860

ctgacgcggg catagcccag caggccagcg gcggcgctct tgttcatggc gtaatgtctc 10920

cggttctagt cgcaagtatt ctactttatg cgactaaaac acgcgacaag aaaacgccag 10980

gaaaagggca gggcggcagc ctgtcgcgta acttaggact tgtgcgacat gtcgttttca 11040

gaagacggct gcactgaacg tcagaagccg actgcactat agcagcggag gggttggatc 11100

aaagtacttt gatcccgagg ggaaccctgt ggttggcatg cacatacaaa tggacgaacg 11160

gataaacctt ttcacgccct tttaaatatc cgttattcta ataaacgctc ttttctctta 11220

ggtttacccg ccaatatatc ctgtcaaaca ctgatagttt aattcccgat ctagtaacat 11280

agatgacacc gcgcgcgata atttatccta gtttgcgcgc tatattttgt tttctatcgc 11340

gtattaaatg tataattgcg ggactctaat cataaaaacc catctcataa ataacgtcat 11400

gcattacatg ttaattatta catgcttaac gtaattcaac agaaattata tgataatcat 11460

cgcaagaccg gcaacaggat tcaatcttaa gaaactttat tgccaaatgt ttgaacgatc 11520

ggggaaattc gagctggtca ccaagcttgc atgcctgcag tgcagcgtga cccggtcgtg 11580

cccctctcta gagataatga gcattgcatg tctaagttat aaaaaattac cacatatttt 11640

ttttgtcaca cttgtttgaa gtgcagttta tctatcttta tacatatatt taaactttac 11700

tctacgaata atataatcta tagtactaca ataatatcag tgttttagag aatcatataa 11760

atgaacagtt agacatggtc taaaggacaa ttgagtattt tgacaacagg actctacagt 11820

tttatctttt tagtgtgcat gtgttctcct ttttttttgc aaatagcttc acctatataa 11880

tacttcatcc attttattag tacatccatt tagggtttag ggttaatggt ttttatagac 11940

taattttttt agtacatcta ttttattcta ttttagcctc taaattaaga aaactaaaac 12000

tctattttag tttttttatt taataattta gatataaaat agaataaaat aaagtgacta 12060

aaaattaaac aaataccctt taagaaatta aaaaaactaa ggaaacattt ttcttgtttc 12120

gagtagataa tgccagcctg ttaaacgccg tcgacgagtc taacggacac caaccagcga 12180

accagcagcg tcgcgtcggg ccaagcgaag cagacggcac ggcatctctg tcgctgcctc 12240

tggacccctc tcgagagttc cgctccaccg ttggacttgc tccgctgtcg gcatccagaa 12300

attgcgtggc ggagcggcag acgtgagccg gcacggcagg cggcctcctc ctcctctcac 12360

ggcaccggca gctacggggg attcctttcc caccgctcct tcgctttccc ttcctcgccc 12420

gccgtaataa atagacaccc cctccacacc ctctttcccc aacctcgtgt tgttcggagc 12480

gcacacacac acaaccagat ctcccccaaa tccacccgtc ggcacctccg cttcaaggta 12540

cgccgctcgt cctccccccc cccccctctc taccttctct agatcggcgt tccggtccat 12600

ggttagggcc cggtagttct acttctgttc atgtttgtgt tagatccgtg tttgtgttag 12660

atccgtgctg ctagcgttcg tacacggatg cgacctgta 12699

<210> 5

<211> 1992

<212> DNA

<213> corn (Zea mays L.)

<221> nucleotide sequence of ubiquitin promoter Ubi of Zea mays

<222>（1）…（1992）

<400> 5

ctgcagtgca gcgtgacccg gtcgtgcccc tctctagaga taatgagcat tgcatgtcta 60

agttataaaa aattaccaca tatttttttt gtcacacttg tttgaagtgc agtttatcta 120

tctttataca tatatttaaa ctttactcta cgaataatat aatctatagt actacaataa 180

tatcagtgtt ttagagaatc atataaatga acagttagac atggtctaaa ggacaattga 240

gtattttgac aacaggactc tacagtttta tctttttagt gtgcatgtgt tctccttttt 300

ttttgcaaat agcttcacct atataatact tcatccattt tattagtaca tccatttagg 360

gtttagggtt aatggttttt atagactaat ttttttagta catctatttt attctatttt 420

agcctctaaa ttaagaaaac taaaactcta ttttagtttt tttatttaat aatttagata 480

taaaatagaa taaaataaag tgactaaaaa ttaaacaaat accctttaag aaattaaaaa 540

aactaaggaa acatttttct tgtttcgagt agataatgcc agcctgttaa acgccgtcga 600

cgagtctaac ggacaccaac cagcgaacca gcagcgtcgc gtcgggccaa gcgaagcaga 660

cggcacggca tctctgtcgc tgcctctgga cccctctcga gagttccgct ccaccgttgg 720

acttgctccg ctgtcggcat ccagaaattg cgtggcggag cggcagacgt gagccggcac 780

ggcaggcggc ctcctcctcc tctcacggca cggcagctac gggggattcc tttcccaccg 840

ctccttcgct ttcccttcct cgcccgccgt aataaataga caccccctcc acaccctctt 900

tccccaacct cgtgttgttc ggagcgcaca cacacacaac cagatctccc ccaaatccac 960

ccgtcggcac ctccgcttca aggtacgccg ctcgtcctcc cccccccccc ctctctacct 1020

tctctagatc ggcgttccgg tccatggtta gggcccggta gttctacttc tgttcatgtt 1080

tgtgttagat ccgtgtttgt gttagatccg tgctgctagc gttcgtacac ggatgcgacc 1140

tgtacgtcag acacgttctg attgctaact tgccagtgtt tctctttggg gaatcctggg 1200

atggctctag ccgttccgca gacgggatcg atttcatgat tttttttgtt tcgttgcata 1260

gggtttggtt tgcccttttc ctttatttca atatatgccg tgcacttgtt tgtcgggtca 1320

tcttttcatg cttttttttg tcttggttgt gatgatgtgg tctggttggg cggtcgttct 1380

agatcggagt agaattctgt ttcaaactac ctggtggatt tattaatttt ggatctgtat 1440

gtgtgtgcca tacatattca tagttacgaa ttgaagatga tggatggaaa tatcgatcta 1500

ggataggtat acatgttgat gcgggtttta ctgatgcata tacagagatg ctttttgttc 1560

gcttggttgt gatgatgtgg tgtggttggg cggtcgttca ttcgttctag atcggagtag 1620

aatactgttt caaactacct ggtgtattta ttaattttgg aactgtatgt gtgtgtcata 1680

catcttcata gttacgagtt taagatggat ggaaatatcg atctaggata ggtatacatg 1740

ttgatgtggg ttttactgat gcatatacat gatggcatat gcagcatcta ttcatatgct 1800

ctaaccttga gtacctatct attataataa acaagtatgt tttataatta ttttgatctt 1860

gatatacttg gatgatggca tatgcagcag ctatatgtgg atttttttag ccctgccttc 1920

atacgctatt tatttgcttg gtactgtttc ttttgtcgat gctcaccctg ttgtttggtg 1980

ttacttctgc ag 1992

<210> 6

<211> 720

<212> DNA

<213> Artificial sequence

<221> nucleotide sequence of Green fluorescent protein GFP

<222>（1）…（720）

<400> 6

atggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac 60

ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac 120

ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc 180

ctcgtgacca ccttcggcta cggcctgcag tgcttcgccc gctaccccga ccacatgaag 240

cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc 300

ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg 360

gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac 420

aagctggagt acaactacaa cagccacaac gtctatatca tggccgacaa gcagaagaac 480

ggcatcaagg tgaacttcaa gatccgccac aacatcgagg acggcagcgt gcagctcgcc 540

gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac 600

tacctgagct accagtccgc cctgagcaaa gaccccaacg agaagcgcga tcacatggtc 660

ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaagtag 720

<210> 7

<211> 253

<212> DNA

<213> Artificial sequence

<221> nucleotide sequence of terminator T1

<222>（1）…（253）

<400> 7

gatcgttcaa acatttggca ataaagtttc ttaagattga atcctgttgc cggtcttgcg 60

atgattatca tataatttct gttgaattac gttaagcatg taataattaa catgtaatgc 120

atgacgttat ttatgagatg ggtttttatg attagagtcc cgcaattata catttaatac 180

gcgatagaaa acaaaatata gcgcgcaaac taggataaat tatcgcgcgc ggtgtcatct 240

atgttactag atc 253

Claims (8)

1. A seed development-associated protein, characterized by: the protein is named as protein TaGSR1, is derived from common wheat, and is the protein described in the following 1) or 2):

1) a protein consisting of an amino acid sequence shown by SEQ ID NO.1 in a sequence table;

2) the protein which is obtained by substituting and/or deleting and/or increasing one or more amino acid residues of the amino acid sequence shown by SEQ ID NO.1 in the sequence table, is related to grain development and is derived from the SEQ ID NO. 1.

2. The grain development associated protein of claim 1, wherein: the amino acid sequence of the protein TaGSR1 is composed of 203 amino acids, and the amino acid sequence is shown in SEQ ID NO. 1.

3. A gene encoding the grain development associated protein TaGSR 1of claim 1, wherein the gene comprises the following genes: the nucleic acid sequence of the coding gene is the nucleotide sequence of a) or b) or c) as follows:

a) a deoxyribonucleotide sequence shown by SEQ ID NO.2 in the sequence table;

b) a deoxyribonucleotide sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% homology to the deoxyribonucleotide sequence represented by SEQ ID No.2 and encoding the protein;

c) a ribonucleotide sequence that is complementary to the deoxyribonucleotide sequence defined in a) or b) and encodes said protein under stringent conditions.

4. The coding gene according to claim 3, characterized in that: the coding gene consists of 612 deoxyribonucleotides, and the nucleotide sequence of the coding gene is shown as SEQ ID NO. 2.

5. A binary vector with the function of ectopically and highly expressing TaGSR1 gene in wheat is characterized in that: the binary vector is named as pLGY02-TaGSR1, and the nucleotide sequence of the binary vector is shown in SEQ ID NO. 4; the binary vector contains an expression cassette E1 of a TaGSR1 gene playing a main role in the genetic transformation process of wheat, wherein the nucleotide sequence of the expression cassette E1 is shown as SEQ ID NO.3, and the expression cassette E1 comprises the following components in sequence from upstream to downstream: ubiquitin promoter Ubi from corn, coding sequence of TaGSR1 gene, fluorescence enhanced green fluorescent protein coding gene GFP from multipipecacer, and terminator T1.

6. The binary vector having the function of ectopically highly expressing TaGSR1 gene in wheat according to claim 5, wherein: the nucleotide sequence of the ubiquitin promoter Ubi is shown as SEQ ID NO.5, the coding sequence of the TaGSR1 gene is shown as SEQ ID NO.2, and the nucleotide sequence of the fluorescence enhanced green fluorescent protein coding gene GFP from the aequorea is shown as SEQ ID NO. 6; the nucleotide sequence of the terminator T1 is shown as SEQ ID NO. 7.

7. The application of grain development related protein TaGSR1 or the coding gene thereof in wheat plant type breeding; wherein the amino acid sequence of the grain development related protein TaGSR1 is shown as SEQ ID NO.1, and the nucleotide sequence of the coding gene thereof is shown as SEQ ID NO. 2; the application in the wheat plant type breeding refers to the application of regulating and controlling the wheat plant height or the application of regulating and controlling the included angle of wheat leaves.

8. The application according to claim 7, wherein the application method is: constructing a binary expression vector pLGY02-TaGSR 1of ectopic expression TaGSR1 gene, and transforming the binary expression vector into wheat to realize the ectopic high expression of TaGSR1 gene; or the binary expression vector is transformed into common wheat through agrobacterium to obtain transgenic wheat with TaGSR1 gene over-expression, so that the reduction of the plant height of the wheat and the reduction of the included angle of leaves are realized; wherein the nucleotide sequence of the TaGSR1 gene is shown in SEQ ID NO. 2.

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