CN107058348B

CN107058348B - Wheat gene for improving plant scab resistance and application thereof

Info

Publication number: CN107058348B
Application number: CN201611148584.2A
Authority: CN
Inventors: 吴磊; 丁彬彬; 张旭; 姜朋; 张瑜; 马鸿翔
Original assignee: Jiangsu Academy of Agricultural Sciences
Current assignee: Jiangsu Academy of Agricultural Sciences
Priority date: 2016-12-13
Filing date: 2016-12-13
Publication date: 2020-12-18
Anticipated expiration: 2036-12-13
Also published as: CN107058348A

Abstract

The invention discloses a wheat gene for improving the resistance of plant scab and application thereof, wherein the amino acid sequence of the wheat resistance TaXAT gene is shown as SEQ ID NO.2, and the gene belongs to glycosyltransferase family 61; the gene is subjected to functional verification by transforming arabidopsis thaliana, and proves that the gene can improve the resistance of gibberellic disease, and the gibberellic disease resistance of plants is obviously enhanced by over-expressing the gene.

Description

Wheat gene for improving plant scab resistance and application thereof

Technical Field

The invention belongs to the technical field of biological gene engineering, and particularly relates to cloning of a wheat xylan-arabinosyltransferase gene TaXAT and application of the cloning to improvement of plant gibberellic disease resistance.

Background

Gibberellic disease (Fusarium Head light) is caused by Fusarium graminearum and is one of important diseases which harm cereal crops such as wheat, barley, oat, corn, rice, rye and the like, and the yield and the quality of grain production in China are seriously influenced. Gibberellic disease occurs in large areas in the middle and lower reaches of Yangtze river and northeast areas of China, and the yield is reduced by 20-40% in a pandemic year. In recent years, with the change of farming modes and climate change, head blight has spread to other main producing areas in China. In addition, mycotoxin carried by diseased wheat grains affects the quality of seeds, harms the health of people and animals, and becomes one of the hidden dangers threatening the safety of food. The change of farming mode and cultivation technique is difficult to solve the infection and spread of disease, and the use of pesticide for preventing and controlling the occurrence and prevalence of gibberellic disease has certain effect, but increases the production cost, and the use of chemical pesticide inevitably causes environmental pollution. The utilization of gibberellic disease resistance genes for variety improvement is an effective way to reduce the harm of gibberellic disease.

Studies have shown that resistance to wheat scab is a complex quantitative trait controlled by a few major genes plus numerous minor genes, and more than 100 scab-associated resistance sites have been located in common wheat or closely related species, distributed on all 20 chromosomes except 7D. Analysis methods such as genome, transcriptome and proteome show that the gibberellic disease candidate genes are distributed in various biological processes of wheat responding to fusarium graminearum, such as secondary metabolic biosynthesis, cell wall defense, reduction of intracellular peroxides and conversion toxins and the like.

The TaFROG gene (Alexandra et al,2015) is located on chromosome 4 of the A genome, is cloned from an anti-sex germplasm 'CM 82036', encodes a protein with unknown function, and can slow down the occurrence of gibberellic disease to a certain extent after interacting with TaSnRK1 alpha.

The TaABCC3 gene (Stephanie et al,2015) has one copy on chromosome 3 of A, B, D genome, the total length of the gene is 4503bp, the coded protein contains an ABC transporter C family structural domain, and virus-induced gene silencing is utilized to confirm that the TaACT gene participates in the degradation of the TaABCC3 gene, so that the tolerance of the plant to the toxin can be enhanced.

The TaACT gene (Kage et al,2016) is located on the long arm of chromosome 2 of the D genome, the cDNA has total length of 1326bp, and the coded protein contains one transferase domain.

PFTThe gene (Rawat et al,2016) is located in the short arm of chromosome 3 of the B genome and cloned from the scab resistant variety Sumai No.3 in China. The PFT gene has a total length of 3427bp and contains 15 'untranslated region, 2 exons, 1 intron and 1 3' untranslated regionThe encoded protein contains 2 lectin domains and 1 ETX/MTX2 domain. The resistance of resistant plants can be reduced by frame shift mutation caused by single nucleotide mutation in the gene region.

Gibberellins infect wheat spikelets during the flowering phase, producing large amounts of cell wall degrading enzymes such as: pectinase, xylanase, cellulase and the like to degrade cell wall polysaccharide, thereby achieving the purpose of infection and expansion on a host. The extent of degradation of the cell wall by degradative enzymes is influenced by the components of the cell wall and their relative amounts and plays an important role in host-pathogen interactions, but this has been less studied in cereal crops.

Disclosure of Invention

In view of the above problems, the present invention provides a method for encoding cell wall xylan-arabinosyltransferaseTaXATThe gene belongs to glycosyltransferase family 61, participates in arabinose modification reaction of xylan to influence components and content of cell walls, improves plant gibberellic disease resistance in the process of cell wall defense of fusarium graminearum, and is realized as follows:

a wheat gene for improving the resistance of plant scab has a nucleotide sequence shown in SEQ ID N0.1.

The protein coded by the gene shown in SEQ ID N0.1 has the amino acid sequence shown in SEQ ID N0.2, namely the sequence coded by the gene from 264 th position to 1844 th position in the sequence of SEQ ID NO. 1.

Plant expression vector pCAMBIA2301-CaMV35S-TaXAT。

The application of the wheat gene for improving the resistance of the plant to the gibberellic disease as shown in SEQ ID N0.1 in enhancing the resistance of the plant to the gibberellic disease is specifically that the wheat gene for improving the resistance of the plant to the gibberellic disease as shown in SEQ ID N0.1 is transferred into a target plant through an expression vector so as to improve the disease resistance of the plant to the gibberellic disease.

The wheat gene provided by the invention for improving the resistance of plant scabTaXATHas important application value in enhancing the resistance of plant scab, and can connect the gene coding region sequence shown as SEQ ID N0.1 to plants by an enzyme digestion connection methodOn the expression vector, Arabidopsis thaliana, wheat, corn or other plant cells are introduced by agrobacterium-mediated transformation to obtain transgenic disease-resistant material for expressing the gene, so that the transgenic disease-resistant material is applied to production. The gene is constructed on a plant transformation vector, the gene or the regulatory sequence thereof can be properly modified, and other promoters can be used for replacing the original promoter of the gene, so that the disease resistance spectrum is widened or the resistance to pathogenic bacteria is enhanced.

The invention has the beneficial effects that: the cloned disease-resistant gene is transferred into a susceptible plant, which is beneficial to obtaining a new disease-resistant plant. The cloned disease-resistant gene can be transferred and utilized among different species, thereby overcoming the difficulty of distant hybridization in the traditional disease-resistant breeding. In addition, a plurality of disease-resistant genes can be accumulated in the plant by using a hybridization and transformation method without generating linkage drag problem in the genome accompanying with the traditional breeding technology, and the breeding time can be shortened, and the variety breeding efficiency can be improved.

Drawings

FIG. 1 is an amplification by RACETaXATDetection of 5 'and 3' fragments of the Gene,

lanes

1 and 2 respectivelyTaXAT5 'and 3' fragments of the gene.

FIG. 2 shows the plant expression vector pCAMBIA2301-CaMV35S-TaXATConstructing a detection electrophoretogram;

wherein, FIG. 2A is the electrophoresis diagram of the recombinant plasmid PCR detection, Lane 1 is the electrophoresis diagram of the recombinant plasmid PCR detection, Lane 2 is the electrophoresis diagram of the non-recombinant plasmid PCR detection;

FIG. 2B shows the recombinant plasmid pCAMBIA2301-CaMV35S-TaXATLane 1 is the electrophoretogram after digestion of the recombinant plasmid, and lane 2 is the electrophoretogram without digestion of the recombinant plasmid.

FIG. 3 shows transgenic plantsTaXATPCR identification of the gene.

Wherein, lanes 1-3 are PCR amplification products of leaves of transgenic plants, lane 4 is PCR products of leaves of control wild plants, lane 5 is recombinant plasmid pCAMBIA2301-CaMV35S-TaXATLane 6 is the double distilled PCR product.

FIG. 4 is a schematic diagram showing the expression results of three transgenic lines;

FIG. 5 is a diagram showing the identification result of the resistance of the floral sequences of the transgenic plants to pathogenic bacteria;

wherein, FIG. 5A is the identification chart of the resistance of the floral sequences of the transgenic plants to pathogenic bacteria;

FIG. 5B is the identification chart of the resistance of the inflorescence of the control wild type plant to pathogenic bacteria.

FIG. 6 is a diagram showing the identification result of the resistance of the transgenic plant silique to pathogenic bacteria;

wherein, FIG. 6A is the identification chart of the resistance of the transgenic plant silique to pathogenic bacteria;

FIG. 6B is a chart of identification of resistance of control wild type plants to pathogenic bacteria.

Detailed Description

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

The nucleotide and amino acid sequences related to the following embodiments are shown in sequence tables SEQ ID NO.1-SEQ ID NO. 10.

Examples relate to material sources:

sumai No. 3: the wheat is preserved by the wheat research institute of agricultural biotechnology of agricultural academy of sciences of Jiangsu province;

arabidopsis plants: the wheat is preserved by the wheat research institute of agricultural biotechnology of agricultural academy of sciences of Jiangsu province;

gibberella: the wheat is preserved by the wheat research institute of agricultural biotechnology of agricultural academy of sciences of Jiangsu province;

PT19-T vector: from Biotechnology engineering (Dalian) Co Ltd

pCAMBIA2301-CaMV 35S: the wheat is preserved by the wheat research institute of agricultural biotechnology of agricultural academy of sciences of Jiangsu province;

example 1TaXATGene acquisition and analysis

Primers for 5 'and 3' RACE nested amplification were designed using Primer Premier 5.0 based on the EST sequence of the resulting gene of interest: 5 '-Primer (the sequence of which is shown in SEQ ID NO. 3) and 3' -Primer (the sequence of which is shown in SEQ ID NO. 4);

3 'and 5' RACE amplifications were performed using cDNA Amplification kit, respectively, using Perilla 3 ear cDNA inoculated with gibberella for 8 hours:

and (3) PCR reaction system: 2.0. mu.l of SeqAmp Buffer, 1.0. mu.l of SeqAmp DNA Polymerase, 2.5. mu.l of cDNA template, 5. mu.l of 10 XUniversal Primer A Mix, 1. mu.l of 5 'or 3' -Primer (10. mu.M), ddH₂O 15.5 μl。

PCR reaction procedure: (vii) 3 min 94; extends for 30 cycles at 94C 30 sec, 55C 30 sec, and 72C 2 min; (vii) 5min extension at 72 ℃;

the PCR product was subjected to gel electrophoresis on 1% agarose gel, and the electrophoresis results are shown in FIG. 1, wherein in FIG. 1,

lanes

1 and 2 are 5 'and 3' fragments of the TaXAT gene, respectively, and M is DNA Marker.

And cutting the target strip, recovering the cut strip, cloning the cut strip into a PT19-T vector, and sequencing the obtained product by Nanjing Jinslei Biotech Co. The complete gene is obtained by sequence splicing, and the applicant has the name ofTaXATThe nucleotide sequence is shown as SEQ ID NO.1, and the total length of the gene sequence is 2030 bp.

The complete open reading frame is predicted by the software ORF Finder (http:// www.ncbi.nlm.nih.gov/gorf. html) to have the total length of 1581bp (i.e. the sequence from 264 to 1844 in the sequence of SEQ ID NO. 1), and the coded amino acid sequence is shown in SEQ ID NO. 2.

EXAMPLE 2 construction of plant expression vectors

Using the Perilla frutescens ear part cDNA as a template, amplification was performed using the primers TaXATFXba (shown in SEQ ID NO. 5) and TaXATRSac (shown in SEQ ID NO. 6)TaXATComplete open reading frame, and introduction of restriction sitesXbaI andSaci, recovering the target fragment.

And (3) PCR reaction system: 10 XEx buffer 5 μ l, MgCl2(25mM) 4 μ l, dNTP (2.5 mM) 1.6 μ l, TaXATFXba primer (10 uM) 1 μ l, TaXATRSac primer (10 uM) 1 μ l, ExTaq 0.25 μ l, ddH2O 13.7.7 μ l.

PCR reaction procedure: (vii) 3 min 94; (ii) 94C 15 sec, 55C 30 sec, 2 min extension from 72C, 30 cycles; (vii) 5min extension at 72 ℃;

using restriction enzymesXbaI andSaci, respectively carrying out enzyme digestion on pCAMBIA2301-CaMV35S and a target gene fragment, carrying out enzyme digestion at 37 ℃ for 15min, and recycling again.

Enzyme digestion system: 200ng of the target fragment or 1000ng of the vector, 10 Xbuffer 2. mu.l,XbaI 1μl, SacI 1μl,ddH₂o is supplemented to 20 μ l;

fragments were recovered by ligation with ligase SolutionI:

a connection system: solution I5 mul, pCAMBIA2301 digested product 1 mul, target gene digested product 4 mul.

And transforming escherichia coli competent cells after connecting the products, placing the flat plate in a front direction in a 37 ℃ incubator until liquid is completely absorbed by a solid culture medium, then inverting the flat plate in a dark place, and culturing for 12-14 h at 37 ℃.

The recombinant plasmid was identified by electrophoresis, and the electrophoresis result is shown in FIG. 2A, wherein lane 1 is the PCR detection electropherogram of the recombinant plasmid, and lane 2 is the PCR detection electropherogram of the non-recombinant plasmid.

And (3) carrying out bacterial liquid PCR screening on the recombinant transformant by adopting gene specific primers TaXATFXba and TaXATRSac.

And (3) PCR reaction system: 10 × buffer (containing Mg)²⁺) 2.0 μ l, dNTP (2.5 mM) 1.6 μ l, bacterial liquid 2.0 μ l (OD)₆₀₀Between 0.4 and 0.6), 0.1 μ l of primer TaXATFXba (10 uM), 0.1 μ l of primer TaXATRSac (10 uM), rTaq, 0.5 μ l, ddH₂And O is complemented to 20 mu l.

The PCR reaction conditions are as follows: 5min at 94 ℃; (ii) 94 ℃ 30 sec, 55 ℃ 30 sec, 72 ℃ 2 min, 30 cycles; 72C for 5 min.

The amplification products were detected by electrophoresis on a 1.0% agarose gel and the results recorded by a gel imaging system.

Extracting positive recombinant plasmid with plasmid miniextraction kit, and using restriction endonucleaseXbaI andSaci restriction enzyme identification of the recombinant pCAMBIA2301-CaMV35S-TaXATFIG. 2B shows the recombinant plasmid pCAMBIA2301-CaMV35S-TaXATThe enzyme cleavage identification electrophoretogram of (a), whichIn lane 1, the electrophoretogram of the recombinant plasmid after digestion is shown, and lane 2, the electrophoretogram of the recombinant plasmid without digestion is shown.

The above results indicate that the plant expression vector pCAMBIA2301-CaMV35S-TaXATThe construction is successful, and the method can be applied to plant genetic transformation.

Example 3 Arabidopsis genetic transformation and Gene function verification

The experiment was divided into transgenic and control wild groups:

transforming the genome: genetic transformation of Arabidopsis thaliana was carried out by the Floral dip method (see Clough SJ, Bent AF (1998) Floral dip: a simplified method for Agrobacterium-mediated transformation of Arabidopsis thaliana. Plant J16: 735-TaXATTransferring into Arabidopsis thaliana plant, screening with MS culture medium containing kanamycin (final concentration 50mg/L), transplanting Arabidopsis thaliana plant with true leaf and green color onto soil matrix, and culturing in incubator for 6-8 weeks. The culture conditions were: relative humidity of 80%, constant temperature of 20-24 deg.C, and illumination intensity of 80-200umol/M²And S, culturing in 8h of darkness and 16h of light.

Control wild group: the wild seeds germinate on antibiotic-free plates, grow true leaves, are transplanted on a soil matrix, and are placed in an incubator for normal culture for 6-8 weeks under the culture conditions consistent with those of the transgenic group.

1. Plant leaf validation

Collecting leaves, extracting genome DNA, performing PCR verification by using gene specific primers TaXATFXba (SEQ ID NO. 5) and TaXATRSac (SEQ ID NO. 6),

and (3) PCR reaction system: 10 × buffer (containing Mg)²⁺) 2.0 μ l, dNTP (2.5 mM) 1.6 μ l, bacterial liquid 2.0 μ l (OD)₆₀₀Between 0.4 and 0.6), TaXATFXba (10 uM) 0.1 μ l, TaXATRSac (10 uM) 0.1 μ l, rTaq 0.5 μ l, ddH₂Supplementing O to 20 mu l;

The electrophoresis results are shown in FIG. 3, lanes 1-3 are PCR amplification products of transformed plants, lane 4 is PCR product of control wild group, lane 5 is PCR product of recombinant plasmid pCAMBIA2301-CaMV35S-TaXAT, and lane 6 is PCR product of double distilled water.

2. Verification of transgene expression

Are respectively represented by T₁The cDNA of the arabidopsis transgenic and wild inflorescence is used as a template, and the relative expression quantity of the target gene is detected by using a Roche LightCycler96 real-time fluorescent quantitative PCR instrument. The gene specific primers are RTF (SEQ ID NO. 7) and RTR (SEQ ID NO. 8), the reference gene is Tubulin, and the amplification primers are TubF (SEQ ID NO. 9) and TubR (SEQ ID NO. 10).

The qPCR reaction system is as follows: SYBR Premix Ex Taq (2X) 10. mu.l, primer 1 (10. mu.M) 0.4. mu.l, primer 2 (10. mu.M) 0.4. mu.l, cDNA 2.0. mu.l, ddH₂O 7.2μl。

PCR procedure: a two-step PCR amplification standard procedure was used: 30 seconds at 95 ℃; 5 seconds at 95 ℃ and 30 seconds at 60 ℃ for 40 cycles. Each sample is repeated for three times, Ct value is obtained after reaction is finished, and 2 is adopted^-ΔΔCTThe method carries out relative quantitative expression analysis on the target gene.

The results are shown in FIG. 4, (1) is the wild type, (2) - (4) are the expression results of the selected three transgenic lines, and it can be seen that the target gene can be stably expressed in the transgenic line, and the expression level is improved by 9-18 times compared with the wild type.

3. Seed infection verification

Selecting T₁Transgenic arabidopsis and wild seeds are sown and then transplanted to a soil substrate, and the gibberellic disease can be inoculated when the seeds grow to the flowering phase and 2-3 siliques exist.

The inoculation method comprises the following steps: adjusting pathogenic bacteria spore number concentration to 1 × 10⁵And (2) inoculating the inflorescence by adopting a spraying method, spraying 0.5ml of conidium solution to each plant, and observing infected plants 2 days after inoculation.

The 2 mm position of the head end of the horn is cut off by sterile scissors, 5 mul of inoculated spore liquid drops are placed on the wound, and the disease condition is observed after 7 days of inoculation. Covering with a plastic film for moisture preservation after inoculation, performing dark culture at 22 ℃ for 2 days, and then transferring to a temperature chamber with the relative humidity of 80% at 25 ℃, and performing culture with the light-dark ratio of 16h to 8 h.

Fig. 5 is a picture of the identification result of the resistance of the inflorescences of two groups of plants to pathogenic bacteria, wherein fig. 5A is a graph of the identification of the resistance of the inflorescences of the plants in the transgenic group to the pathogenic bacteria, and fig. 5B is a graph of the identification of the resistance of the inflorescences of wild-type plants in the control group to the pathogenic bacteria.

Fig. 6 is a picture of identification of resistance of two groups of plant siliques to pathogenic bacteria, wherein fig. 6A is a picture of identification of resistance of transgenic plant siliques to pathogenic bacteria, fig. 6B is a picture of identification of resistance of wild plant siliques to pathogenic bacteria, as can be seen from fig. 6, the withered seeds are covered with hyphae to some extent, the wild arabidopsis thaliana siliques are withered and brown in whole, and the transgenic siliques are brownish green, which indicates that the resistance of transgenic plants to pathogenic bacteria is enhanced.

SEQUENCE LISTING

<110> agricultural science and academy of Jiangsu province

<120> wheat gene for improving plant gibberellic disease resistance and application thereof

<130> 10

<160> 10

<170> PatentIn version 3.3

<210> 1

<211> 2030

<212> DNA

<213> Artificial Synthesis

<400> 1

gtggtatcaa cgcagagtac tgggggaagc caagtcctac accgccacac ctcctcgcct 60

gctttctgtc cccgtctctc ttctatctcc atgtaaagtt gtgagccgtc gacgaagaaa 120

caggaggcgg aggaagaaac aaagaaagct cgaccgtgtc gccgggctcg tgcgcggtgc 180

cggaacaggt cgtcgtcggc gtcgcacgag aggattgcct gttgtgtacg cgtcgaggtg 240

ccggagggac agaggaggag gagatgggcg ccggcgaggg gaagcagggg aaggggcttg 300

tgaggagctg ggcgcagagg tacctcaacc tcggcttcgt cgccggcttc ctgctcgtgc 360

tcctcaccta catcgtcgtc acccagcagt tcgccatcac ctcccccgac gctgtcccca 420

cgatgacgcc gcaccagaag caggcgatca gcgcccccgg cgccggcgta acagggacgc 480

cggagggtcg agttgaggag aagtctgaga cgaaacctgt cgatgcttct tccggggatg 540

aaactcctaa ggaggagcag caacaacaac atgatgcgga gccggaatgg aagaagccgg 600

aagacacggt ggcgacggag gaagagcccc ccaaaagaga cgacaccgct gccgaaccgc 660

tcgacaacgg caaggtggtg tgtaccacgg agggcccctt ctccgacacg tgcgacgtct 720

tcggtgacgt ccggaccaac ggcacagctc acaccgtcac cctcgtccca gcgaccgaga 780

cggagagccg ggagtggaag attcagccct acgtccgccg tggcatgtct ggcatctcgg 840

aggtcacggt gacgcagctc gactcgacct ccgcggcatc cccggcgccg gcgtgcacgg 900

tgacgcaccg cgtcccggcc atcgtgttcg cgctcggtgg gctaacaggc aactacttcc 960

acgacttcag cgatgcgctg gtgccactat tcgtggcgtc acggcgatat ggcggcgagg 1020

tccagctatt ggccagcaac atccagccat ggtggctcgg caagtacgag gccgtggtga 1080

gacggctgtc caagtacgac gtcgtggacc tcgaccatga tgatcagatc cggtgctttc 1140

cgagcgtcac tgtcgggcta cgcatgcaca aggaattcga catcgtgccg gagttggtcc 1200

ccggcggcgc ccccctctcc atggtcgact tcaccgcctt cctccgcgag acctacaccc 1260

taccccgcgc cgcgcccatc agccttatga aggacatcag cccgccggag gaccaggaga 1320

agaggaaacc gcggctgatg ctgctccacc gtggccacta ccggaagttc gtgaacgtgc 1380

ctgagatcgt gaaggcggcg gagaaggccg ggttcgaggt gtccatcgcc gacccgcggt 1440

tcgacgtgcg ggtggaggag ctggcccggt cggtgaactc gttcgacgtg ctgctgggcg 1500

tgcacggcgc cgggctgacc aacgccgtgt tcatgcccac gggggcggtg gtgatccagg 1560

tggtgccgta cgggaaccta gagcacatgg cgaaggtgga cttcggcgac cccgtggcgg 1620

acatggggct ccggtacctc gagtacagca tcacggcgga ggagagcacg ctgctggaga 1680

tgctggggcc ggaccacccc gtgatcaagg accccgagtc ggtgcaccgg agtgggtggg 1740

acaaggtcgc cgagtactac ctcggcaagc aggacgtgcg cgttgacgtg gagcgcttcg 1800

cgcccacgct tgctctggcc atcgaacatc ttcgacagaa gtagaaaatt gcatgggcca 1860

ccgagaattc gttttttttt tgttggtaca gtttcggggc attctatggc ctctgaattt 1920

ggtggaacat acatacatag aagaattgct ggtatattgg ttgatttatt tacctcaagt 1980

gaatagaaaa attaacttgt ttcagcaaaa aaaaaaaaaa aaaaaaaaaa 2030

<210> 2

<211> 526

<212> PRT

<213> Artificial Synthesis

<400> 2

Met Gly Ala Gly Glu Gly Lys Gln Gly Lys Gly Leu Val Arg Ser Trp

1 5 10 15

Ala Gln Arg Tyr Leu Asn Leu Gly Phe Val Ala Gly Phe Leu Leu Val

20 25 30

Leu Leu Thr Tyr Ile Val Val Thr Gln Gln Phe Ala Ile Thr Ser Pro

35 40 45

Asp Ala Val Pro Thr Met Thr Pro His Arg Lys Gln Ala Ile Ser Ala

50 55 60

Pro Gly Ala Gly Val Thr Gly Thr Pro Glu Gly Arg Val Glu Glu Lys

65 70 75 80

Ala Glu Thr Lys Pro Val Asp Ala Ser Ser Gly Asp Glu Thr Pro Lys

85 90 95

Glu Glu Gln Gln Gln Gln His Asp Ala Glu Pro Glu Trp Lys Lys Pro

100 105 110

Glu Asp Thr Val Ala Thr Glu Glu Glu Pro Pro Lys Arg Asp Asp Thr

115 120 125

Ala Ala Glu Pro Leu Asp Asn Gly Lys Val Val Cys Thr Thr Glu Gly

130 135 140

Pro Phe Ser Asp Thr Cys Asp Val Phe Gly Asp Val Arg Thr Asn Gly

145 150 155 160

Thr Ala His Thr Val Thr Leu Val Pro Ala Thr Glu Thr Glu Ser Arg

165 170 175

Glu Trp Lys Ile Gln Pro Tyr Val Arg Arg Gly Met Ser Gly Ile Ser

180 185 190

Glu Val Thr Val Thr Gln Leu Asp Ser Thr Ser Ala Ala Ser Pro Ala

195 200 205

Pro Ala Cys Thr Val Thr His Arg Val Pro Ala Ile Val Phe Ala Leu

210 215 220

Gly Gly Leu Thr Gly Asn Tyr Phe His Asp Phe Ser Asp Ala Leu Val

225 230 235 240

Pro Leu Phe Val Ala Ser Arg Arg Tyr Gly Gly Glu Val Gln Leu Leu

245 250 255

Ala Ser Asn Ile Gln Pro Trp Trp Leu Gly Lys Tyr Glu Ala Val Val

260 265 270

Arg Arg Leu Ser Lys Tyr Asp Val Val Asp Leu Asp His Asp Asp Gln

275 280 285

Ile Arg Cys Phe Pro Ser Val Thr Val Gly Leu Arg Met His Lys Glu

290 295 300

Phe Asp Ile Val Pro Glu Leu Val Pro Gly Gly Ala Pro Leu Ser Met

305 310 315 320

Val Asp Phe Thr Ala Phe Leu Arg Glu Thr Tyr Thr Leu Pro Arg Ala

325 330 335

Ala Pro Ile Ser Leu Met Lys Asp Ile Ser Pro Pro Glu Asp Gln Glu

340 345 350

Lys Arg Lys Pro Arg Leu Met Leu Leu His Arg Gly His Tyr Arg Lys

355 360 365

Phe Val Asn Val Pro Glu Ile Val Lys Ala Ala Glu Lys Ala Gly Phe

370 375 380

Glu Val Ser Ile Ala Asp Pro Arg Phe Asp Val Arg Val Glu Glu Leu

385 390 395 400

Ala Arg Ser Val Asn Ser Phe Asp Val Leu Leu Gly Val His Gly Ala

405 410 415

Gly Leu Thr Asn Ala Val Phe Met Pro Thr Gly Ala Val Val Ile Gln

420 425 430

Val Val Pro Tyr Gly Asn Leu Glu His Met Ala Lys Val Asp Phe Gly

435 440 445

Asp Pro Val Ala Asp Met Gly Leu Arg Tyr Leu Glu Tyr Ser Ile Thr

450 455 460

Ala Glu Glu Ser Thr Leu Leu Glu Met Leu Gly Pro Asp His Pro Val

465 470 475 480

Ile Lys Asp Pro Glu Ser Val His Arg Ser Gly Trp Asp Lys Val Ala

485 490 495

Glu Tyr Tyr Leu Gly Lys Gln Asp Val Arg Val Asp Val Glu Arg Phe

500 505 510

Ala Pro Thr Leu Ala Leu Ala Ile Glu His Leu Arg Gln Lys

515 520 525

<210> 3

<211> 38

<212> DNA

<213> Artificial Synthesis

<400> 3

gattacgcca agcttgcagg tcctgcttgc cgaggtag 38

<210> 4

<211> 39

<212> DNA

<213> Artificial Synthesis

<400> 4

gattacgcca agcttgtggt gatccaggtg gtgccgtac 39

<210> 5

<211> 34

<212> DNA

<213> Artificial Synthesis

<400> 5

taatctctag aatgggcgcc ggcgagggga agcg 34

<210> 6

<211> 35

<212> DNA

<213> Artificial Synthesis

<400> 6

atgcgagctc ctacttctgt cgaagatgtt cgatg 35

<210> 7

<211> 18

<212> DNA

<213> Artificial Synthesis

<400> 7

ggtgggctaa caggcaac 18

<210> 8

<211> 18

<212> DNA

<213> Artificial Synthesis

<400> 8

gatctcaggc acgttcac 18

<210> 9

<211> 17

<212> DNA

<213> Artificial Synthesis

<400> 9

gtggaactgg ctctggc 17

<210> 10

<211> 17

<212> DNA

<213> Artificial Synthesis

<400> 10

cgctcaatgt caaggga 17

Claims

1. The application of wheat gene for raising plant's gibberellic disease resistance in strengthening plant's gibberellic disease resistance; the nucleotide sequence of the gene is shown in SEQ ID No. 1.

2. The use of claim 1, wherein the gene is transferred into a target plant via an expression vector to improve the disease resistance to head blight of the plant.

3. Use according to claim 2, wherein the plant is Arabidopsis thaliana.