CN1293095C - Drought resistant correlative protein and coded gene of plant and application - Google Patents

Abstract

The present invention discloses a protein relevant to plant drought resistance, a coded gene thereof and an application thereof. The protein relevant to plant drought resistance of the present invention is a protein with one of the following amino acid residue sequences: 1) SEQ ID No. 2 in a sequence list, and 2) a protein which is used for substituting, deleting or adding one or a plurality of amino acid residues of an amino acid residue sequence of SEQ ID No. 2 in the sequence list and is relevant to plant drought resistance. The coded gene of the protein relevant to plant drought resistance can be used for enhancing plant drought resistance; for example, the expression of the coded gene of the related protein of plant drought resistance in a plant is suppressed. The protein relevant to plant drought resistance and the coded gene of the present invention can be used for breeding new drought resisting plant varieties so as to have important meanings.

Description

A kind of plant drought-resistance related protein and its coding gene and application

technical field

The invention relates to a plant drought-resistance-related protein in the field of bioengineering and its coding gene and application, in particular to a method for enhancing plant drought-resistance by using the gene.

Background technique

The food problem is one of the major problems facing the world today. Increasing grain production by increasing unit yield or expanding planting area, increasing agricultural input to transform low-yielding fields, etc. will encounter the problem of overcoming adversity constraints or mitigating adversity hazards. Therefore, understanding the response mechanism of plants to adversity and improving the stress resistance of plants has become an important basic research for further increasing agricultural production. It has attracted the attention of governments and scientists around the world, and is also a hot spot in current life science research.

Drought is the main adversity that restricts the growth and development of plants. With the economic development of human beings, population expansion and the destruction of biological populations, the shortage of water resources is becoming more and more serious. It has seriously affected agricultural production and ecological environment, and has become a global concern. Therefore, it is very necessary to seek ways for plants, especially crops, to resist drought. To improve the drought resistance of crops, in addition to using traditional breeding methods, at present, the application of genetic engineering breeding has become one of the areas of concern to scientific and technological workers.

Arabidopsis is a typical model plant widely used in the research of plant genetics, developmental biology and molecular biology. Most genes in Arabidopsis can be found in other plants, and any discovery about Arabidopsis can be applied to other plant research. Therefore, research on the molecular biological mechanism of drought resistance in Arabidopsis will greatly help to find ways to improve crop drought resistance and increase yield.

Arabidopsis has a total of about 130 million base pairs and 29,000 genes. At present, the functions of most genes are still unclear, and the use of mutation technology to study gene functions has become an effective method. Through the study of mutants, the functions of some drought resistance genes, such as DREB, CBF, and ABRE, were known.

At present, no effective drought-resistant functional genes have been found. Facing the increasingly serious crop drought problem, it is of great theoretical and practical significance to find new drought-resistant functional genes and clarify their functions.

Contents of the invention

The purpose of the present invention is to provide a new plant drought resistance related protein and its coding gene.

The plant drought resistance-related protein provided by the present invention is named LEW2-1 (LEaf Wilt 2-1), which is derived from Arabidopsis thaliana of the Colombian ecotype, and is a protein with one of the following amino acid residue sequences:

1) SEQ ID №: 2 in the sequence listing;

2) A protein in which the amino acid residue sequence of SEQ ID No. 2 in the sequence listing has undergone one or several amino acid residue substitutions and/or deletions and/or additions and is related to plant drought resistance.

Sequence 2 in the sequence listing consists of 985 amino acid residues.

The substitution and/or deletion and/or addition of one or several amino acid residues refers to the substitution and/or deletion and/or addition of no more than 10 amino acid residues.

The coding gene of LEW2-1 (LEW2-1) also belongs to the protection scope of the present invention.

The cDNA gene of LEW2-1 may have one of the following nucleotide sequences:

1) The DNA sequence of SEQ ID №: 1 in the sequence listing;

2) A polynucleotide encoding the protein sequence of SEQ ID №: 2 in the sequence listing;

3) A nucleotide sequence that can hybridize to the DNA sequence defined by SEQ ID №: 1 in the sequence listing under high stringency conditions;

4) A DNA sequence that has more than 90% homology with the DNA sequence defined by SEQ ID №: 1 in the sequence listing and encodes the same functional protein.

The high stringency conditions can be 0.1×SSPE (or 0.1×SSC), 0.1% SDS solution, hybridization at 65° C. and membrane washing.

Sequence 1 in the sequence listing consists of 3217 bases, its open reading frame (ORF) is from the 68th to 3025th base at the 5' end, and encodes the protein of Sequence 2.

The expression vector, cell line and host bacteria containing LEW2-1 of the present invention all belong to the protection scope of the present invention. The primer pair for amplifying any fragment in LEW2-1 also belongs to the protection scope of the present invention.

The second object of the present invention is to provide a method for enhancing the drought resistance of plants by utilizing the gene.

The method for enhancing the drought resistance of plants provided by the present invention is to inhibit the expression of the genes encoding the above-mentioned plant drought resistance-related proteins in plants.

Inhibiting the expression of the above-mentioned plant drought resistance-related protein coding gene LEW2-1 in plants can be achieved by various methods, such as the method of gene silencing mediated by plant virus vectors, the method of silencing genes by antisense nucleic acid technology, and the gene silencing mediated by siRNA method etc. The method for inhibiting gene expression in the present invention is not limited to the above methods, as long as it can inhibit the expression of LEW2-1.

Utilize any kind of plant gene knockout technology, after this gene is knocked out (or silent), the plant shows drought resistance; Utilize any kind of vector that can guide exogenous gene to express in the plant, the LEW2- provided by the present invention 1 was transferred into plants, and the plants showed drought sensitivity.

When the LEW2-1 gene or its antisense nucleic acid of the present invention is constructed into a plant expression vector, any enhanced promoter or inducible promoter can be added before its transcription initiation nucleotide. In order to facilitate the identification and screening of transgenic plant cells or plants, the vectors used can be processed, such as adding plant selectable markers (GUS gene, luciferase gene, etc.) or antibiotic markers with resistance (qing Damycin, Kanamycin, etc.). The transformed plant host can be a monocotyledonous plant or a dicotyledonous plant, such as rice, wheat, corn, cucumber, tomato, poplar, lawn grass or alfalfa. Considering the safety of the transgenic plants, the transformed plants can be screened directly by the dehydration degree of leaves at the seedling stage without adding any selectable marker gene. The expression vector carrying the LEW2-1 gene of the present invention can transform plant cells or tissues by using conventional biological methods such as Ti plasmid, Ri plasmid, plant virus vector, direct DNA transformation, microinjection, conductance, Agrobacterium-mediated, and The transformed plants are tissue grown into plantlets.

The plant drought-resistance-related protein and its encoding gene of the invention provide genetic and technical support (guarantee) for crop drought-resistance breeding.

The technical solutions of the present invention will be further described below in conjunction with the embodiments.

Description of drawings

Fig. 1 is the comparative photo of drought resistance of lew2-1 and wild-type plants

Figure 2 is a comparison photo of lew2-1 and wild-type plant osmoregulation ability

Figure 3 shows the expression characteristics of stress-resistant genes of lew2-1 and wild-type plants under normal and adversity stress conditions

Detailed ways

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

Example 1, the acquisition of LEW2-1 and its coding gene

100-200 mg of wild-type Colombian ecotype Arabidopsis seedlings were used as materials, and total RNA was extracted from roots and stems with Trizol, and the integrity of RNA was detected by 1% agarose electrophoresis. Synthesis of ss cDNA Single-stranded (ss) cDNA was synthesized according to the method of SuperScript ^TM II RNase H-Reverse Transcriptase. Dilute the synthesized single-stranded cDNA 10 times and use it as a template for the following PCR reaction: 20 μl system containing 2 μl of 10×PCR buffer, 1.6 μl of 2.5 mM dNTP (dATP, dGTP, dCTP, dTTP) mixture, 5 μM primer 1 and 1.0 μl of each primer 2, 0.1 μl of Taq enzyme (15 U/μl). Among them, primer 1: 5'ATGATGGAGTCTAGGTCTCCCAT 3', primer 2: 5'ACTGTCTT GATCGTTATTGCTAA3'. Amplification on a PE 9700 instrument: 94°C pre-denaturation for 3 minutes; then 94°C for 30 sec, 55°C for 30 sec, 72°C for 1 min and 30 sec, a total of 35 cycles; finally, 72°C extension for 4 min, and the obtained PCR product was used as 1% agar Sugar gel electrophoresis. The cDNA fragment obtained by PCR was connected to the pGEM-T vector to obtain the vector pT-LEW2-1 containing the target fragment, which was sequenced and identified. The results showed that the cDNA fragment obtained by PCR had the DNA sequence of sequence 1 in the sequence table, which was LEW2-1 The cDNA gene consists of 3217 bases, and its coding sequence is from the 68th base to the 3025th base at the 5' end, encoding a protein with the amino acid residue sequence of Sequence 2 in the sequence listing.

Embodiment 2, cultivate the Arabidopsis thaliana that drought resistance enhances

1. Obtaining the homozygous mutant lew2-1 whose LEW2-1 gene has been knocked out

According to the specific fragments in the LEW2-1 gene sequence, primers were designed: RNAi-F: 5'ATGATGGAGTCTAGGTCTCCCAT 3' and RNAi-R: 5'ACTGTCTTGATCGTTATTGCTAA 3', using the Arabidopsis thaliana genomic DNA as a template for PCR amplification. The target fragment with a length of about 350bp was obtained, and the 350bp target fragment was integrated into the plasmid pJawoh-8 (purchased from INVITROGEN) using the gateway system (introduction vector construction transformation system, purchased from INVITROGEN) to form an inverted repeat sequence After sequencing, it was detected that the integrated fragment and the insertion direction were correct, transformed into Agrobacterium (selection markers were kanamycin, ampicillin, and gentamycin), and transformed Arabidopsis thaliana using the Agrobacterium infection method, and the obtained F1 generation plants were treated with The herbicide basta (basta, purchased from INVITROGEN) was used to screen the transformed seedlings and isolate and purify them. Total RNA was extracted from Trizol, a strain that had no segregation traits after being screened by glufosinate, and transferred to a nylon membrane after agarose electrophoresis for Northern-blot detection. Among them, the probe used is a DNA fragment obtained by PCR amplification using the wild-type Arabidopsis DNA as a template, using primer 1 and primer 2, wherein the 20μl PCR reaction system contains 2μl of 10×PCR buffer, 2.5mM dNTP (dATP, dGTP, dCTP, dTTP) mixture 1.6 μl, 5 μM primer 1 and primer 2 each 1.0 μl, Taq enzyme (15 U/μl) 0.1 μl. Amplification on PE 9700 instrument: 94°C pre-denaturation for 3min; 94°C for 30sec, 55°C for 30sec, 72°C for 1min and 30sec, a total of 35 cycles; 72°C for 4min, and the obtained PCR product was made into 1% agarose gel Electrophoresis. After the probe P32 was labeled and hybridized with the total RNA, no hybridization band was obtained, which proved that the RNA expressed by the LEW2-1 gene had been degraded, and a homozygous mutant lew2-1 was obtained.

2. Comparison of drought resistance of lew2-1 and wild-type plants

The lew2-1 plants and wild-type plants grown in the soil for 2 weeks were stopped from watering for 10-14 days, and the other culture conditions were the same. The survival rate of the plants and the wilting of the leaves were observed and compared to determine their drought resistance. The results are shown in Figure 1, showing that the survival rate of wild-type plants (WT) is 2%, and 1/3 of the leaves of surviving plants are withered and yellow; the lew2-1 plants whose LEW2-1 gene is knocked out are all survived, and there is no wilting and withered leaves ; It shows that the deletion of LEW2-1 gene can make the plants resistant to drought,

3. Comparison of osmoregulation ability of lew2-1 and wild-type plants

The seeds of lew2-1 and wild-type plants were sowed on ordinary (MS) medium, vernalized at 4°C for 2-3 days, then cultured for 5-6 days at 22°C with 16 hours of light and 8 hours of darkness, and then transferred Cultivate on medium containing different concentrations of mannose (0, 420, 520mmol/L) at 22°C, 16 hours of light and 8 hours of darkness for 7-10 days, observe and compare the growth of the root system and the whole plant to determine their osmotic regulation ability. The results are shown in Figure 2, showing that with the increase of mannose concentration, the root growth of wild-type plants (WT) slowed down, and the change was obvious; when the mannose concentration reached 420mmol/L, three-quarters of the leaves of all plants When the mannose concentration reached 520mmol/L, all the leaves of all plants turned yellow; although the root growth of the lew2-1 plants whose LEW2-1 gene was knocked out also slowed down, the change was not as obvious as that of the wild-type plants. When the mannose concentration reached 520mmol/L, only one-third of the leaves of one-fifth of the plants turned yellow. This indicated that after the deletion of the LEW2-1 gene, the osmotic adjustment ability of the plants was enhanced, which was beneficial to drought resistance.

Example 3, detection of stress-resistant gene expression in lew2-1 and wild-type plants

Select lew2-1 grown for 2 weeks in ordinary (MS) medium, and wild-type plants are subjected to the following treatments:

Hydroponic treatment: grow in ordinary liquid (MS) medium for 2 weeks;

Drought treatment: exposure at room temperature for 3 hours;

High light treatment: grow for 2 weeks in ordinary (MS) medium under high light intensity (260 micromol/s·cm²);

Extract the plant RNA of above-mentioned treatment respectively, take rRNA as control (the probe used is the DNA fragment that utilizes plant genome DNA as template, utilizes primers ATGATGGAGTCTAGGTCTCCCAT and ACTGTCTTGATCGTTATTGCTAA to carry out PCR to obtain), utilize Northern blot hybridization technique to detect stress resistance gene: Arabidopsis The expression of drought, high-salt and low-temperature response gene RD29A, salt-drought tolerance gene P5CS and ABA (abscisic acid) synthesis gene SDR1 in lew2-1 and wild-type plants to determine their resistance to corresponding stress size. Wherein, the probe used for detecting RD29A is the DNA fragment obtained by PCR using the primers 5'GACGAG TCAGGA GCTGAGCTG3' and 5'CGATGCTGCCTTCTCGGTAGAG3' using the plant genomic DNA as a template; the probe used for detecting P5CS is using the plant genomic DNA as a template, The DNA fragment obtained by PCR using primers 5'AGCCTTGGCACAGGAGCAACG3' and 5'TGAGAC CAGTGACAGCATCAAACA3; the probe used for SDR1 is a DNA fragment obtained by PCR using the primers 5'TGATCACTGGAGGAGCCACAGG 3' and 5'CTCCGCTTATGTACC GCGAGTC3' using the plant genomic DNA as a template .

The results are shown in Figure 3, indicating that lew2-1 plants had significantly higher expression levels of RD29A, P5CS and SDR1 than wild-type plants under normal conditions (control), hydroponics, drought and high light treatment conditions. This shows that the deletion of the LEW2-1 gene can make the plants resistant to drought. In Figure 3, W represents the wild-type plant, and M represents the lew2-1 plant.

Sequence Listing

<160>2

<210>1

<211>3217

<212>DNA

<213> Arabidopsis thaliana

<400>1

atccatccaa atctcaatcc ctaattaggg ttcatttctc tgtttctcca aacaggggaa 60

ttcgaagatg atggagtcta ggtctcccat ctgcaacact tgtggtgaag agattggtgt 120

aaaatcaaac ggagagttct ttgtggcttg tcatgagtgt agtttcccga tctgcaaagc 180

ttgtcttgag tatgaattca aagaaggtcg aagaatttgc ttgcgttgcg gcaatcctta 240

cgatgagaat gtgtttgatg atgttgagac aaagacatct aaaactcaat ccattgttcc 300

aacacagacc aataacactt ctcaggattc agggattcat gctagacata taagtacagt 360

ctcaacaata gacagtgaac tgaatgatga atatggcaat ccaatttgga agaacagagt 420

ggagagctgg aaagacaaga aagacaagaa gagcaagaag aagaagaaag atccaaaagc 480

aacaaaagct gaacaacatg aggctcagat tcctacccaa cagcacatgg aagatacgcc 540

accgaacaca gaatctggtg ctacagatgt gctttcggtt gtgattccta tcccaaggac 600

aaaaatcact tcatatagga ttgtcatcat catgcggttg atcatcttgg ctctgttttt 660

taactaccgt atcacgcatc ctgtcgatag cgcttacggt ttatggctaa catctgtgat 720

atgtgagatt tggtttgctg tttcttgggt gttggatcag ttccctaaat ggtctcctat 780

taaccgagaa acttacatcg accggttatc cgcaagattc gaaagagaag gcgagcaatc 840

acagcttgca gctgtagatt tctttgttag tacggtagat ccattaaagg agccaccttt 900

gataactgca aacacggttc tttcgatcct cgcgcttgat tatccggtgg ataaagtctc 960

ttgctatgta tctgatgatg gtgctgcaat gctttcgttt gagtctttgg ttgagacagc 1020

agattttgct aggaaatggg tacctttctg caaaaagtac tccatcgagc cacgagctcc 1080

cgagttttac ttctcgctta aaatcgatta cttgagggat aaagttcaac cttcttttgt 1140

gaaagaacgt agagccatga aaagagatta tgaagagttt aaaataagaa tgaatgcttt 1200

agtcgccaag gctcaaaaga caccagaaga aggatggaca atgcaagatg gaacatcttg 1260

gcccgggaac aacactcgtg accatcccgg gatgattcag gtttttcttg gatatagcgg 1320

tgctcgcgac attgaaggaa atgaacttcc aagattagtt tacgtctcta gagagaagag 1380

acctggttat cagcatcaca aaaaggccgg ggcagagaac gcattggtga gggtgtctgc 1440

ggttttaacg aatgctccat tcattcttaa ccttgattgt gatcactacg tcaacaatag 1500

caaagccgtg cgtgaagcaa tgtgcttttt aatggatcct gttgttggtc aagacgtttg 1560

ctttgttcag ttcccacaga gatttgatgg aatcgacaag agtgatcgat atgctaaccg 1620

caacattgtt ttcttcgatg ttaatatgag agggcttgat gggattcaag gtccagttta 1680

tgttggtaca ggtactgtct ttagaagaca agcactttac ggatacagtc caccttcaaa 1740

accgaggatt ttaccgcaat cttcatcatc gtcgtgttgc tgtctaacca agaagaaaca 1800

acctcaagat ccttccgaga tttacaaaga tgcaaagcga gaagaacttg atgctgcaat 1860

ctttaatctt ggggacctag acaactacga tgagtacgac agatcgatgc tgatttcaca 1920

aacaagcttt gagaaaacgt ttggtctctc tacggttttc atcgagtcta ctcttatgga 1980

gaatggcggt gttcccgact ctgtaaaccc gtcaacgctc atcaaagaag ctattcatgt 2040

cattagctgt ggatacgaag agaaaactga atggggaaaa gaaataggat ggatttacgg 2100

gtcgatcacc gaagacattt tgacgggttt caagatgcat tgtcgtggat ggaggtcgat 2160

ttactgtatg ccattaagac cagcatttaa aggatctgct ccaatcaatc tatcagatag 2220

gcttcaccag gttctacgtt gggctcttgg ctcggttgag atcttcctta gccgacattg 2280

tcctttgtgg tacggttgca gcggaggccg cctcaagttg ctccagagat tagcttatat 2340

aaacactatt gtctacccat tcacttcttt gcctcttgtt gcttactgta ctcttccagc 2400

tatttgcctt cttaccggca aatttatcat cccaacgcta tcaaacctag caagcatgct 2460

gtttctaggt ctctttatat caatcatctt aacgagtgtc ctcgagcttc gatggagcgg 2520

agtcagtatc gaagacttat ggagaaacga acagttttgg gttattggag gtgtctcagc 2580

tcatctcttt gccgttttcc aaggattcct caaaatgctc gctggtctcg acacaaattt 2640

cacagtcaca tcgaaaaccg cagatgattt agaattcggt gagctttaca ttgtcaaatg 2700

gacaactctc ttgatccctc cgacgtcact tcttataatc aacttggtcg gagttgttgc 2760

tggattctct gacgctctta acaaaggtta tgaagcttgg ggacctttgt ttgggaaggt 2820

atttttcgcc ttttgggtga ttcttcatct ttatccattc ctcaaaggtc ttatgggaag 2880

acaaaacaga acacctacta ttgttattct ctggtctatc ttgcttgctt ctgtgttttc 2940

acttgtttgg gttcgtatca atcctttcgt ctccaaaacc gatacgactt ccctttctct 3000

gaactgtctt ttgatcgatt gctaagagaa gatacgttat gtttgtattt tgaaagattg 3060

atcatatgtg ttttggttgt tttataaatt ttcatatggt tgcttgagcc acaagttaag 3120

taatgttctt attttagcaa caagtcttgg cgggttccgc aagttaggtt tctattgttt 3180

caacatcaac atgtttttaa aaagcaaaat attttat

<210>2

<211>985

<212>PRT

<213> Arabidopsis thaliana

<400>2

Met Met Glu Ser Arg Ser Pro Ile Cys Asn Thr Cys Gly Glu Glu Ile

1 5 10 15

Gly Val Lys Ser Asn Gly Glu Phe Phe Val Ala Cys His Glu Cys Ser

20 25 30

Phe Pro Ile Cys Lys Ala Cys Leu Glu Tyr Glu Phe Lys Glu Gly Arg

35 40 45

Arg Ile Cys Leu Arg Cys Gly Asn Pro Tyr Asp Glu Asn Val Phe Asp

50 55 60

Asp Val Glu Thr Lys Thr Ser Lys Thr Gln Ser Ile Val Pro Thr Gln

65 70 75 80

Thr Asn Asn Thr Ser Gln Asp Ser Gly Ile His Ala Arg His Ile Ser

85 90 95

Thr Val Ser Thr Ile Asp Ser Glu Leu Asn Asp Glu Tyr Gly Asn Pro

100 105 110

Ile Trp Lys Asn Arg Val Glu Ser Trp Lys Asp Lys Lys Asp Lys Lys

115 120 125

Ser Lys Lys Lys Lys Lys Asp Pro Lys Ala Thr Lys Ala Glu Gln His

130 135 140

Glu Ala Gln Ile Pro Thr Gln Gln His Met Glu Asp Thr Pro Pro Asn

145 150 155 160

Thr Glu Ser Gly Ala Thr Asp Val Leu Ser Val Val Ile Pro Ile Pro

165 170 175

Arg Thr Lys Ile Thr Ser Tyr Arg Ile Val Ile Ile Met Arg Leu Ile

180 185 190

Ile Leu Ala Leu Phe Phe Asn Tyr Arg Ile Thr His Pro Val Asp Ser

195 200 205

Ala Tyr Gly Leu Trp Leu Thr Ser Val Ile Cys Glu Ile Trp Phe Ala

210 215 220

Val Ser Trp Val Leu Asp Gln Phe Pro Lys Trp Ser Pro Ile Asn Arg

225 230 235 240

Glu Thr Tyr Ile Asp Arg Leu Ser Ala Arg Phe Glu Arg Glu Gly Glu

245 250 255

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

260 265 270

Leu Lys Glu Pro Pro Leu Ile Thr Ala Asn Thr Val Leu Ser Ile Leu

275 280 285

Ala Leu Asp Tyr Pro Val Asp Lys Val Ser Cys Tyr Val Ser Asp Asp

290 295 300

Gly Ala Ala Met Leu Ser Phe Glu Ser Leu Val Glu Thr Ala Asp Phe

305 310 315 320

Ala Arg Lys Trp Val Pro Phe Cys Lys Lys Tyr Ser Ile Glu Pro Arg

325 330 335

Ala Pro Glu Phe Tyr Phe Ser Leu Lys Ile Asp Tyr Leu Arg Asp Lys

340 345 350

Val Gln Pro Ser Phe Val Lys Glu Arg Arg Ala Met Lys Arg Asp Tyr

355 360 365

Glu Glu Phe Lys Ile Arg Met Asn Ala Leu Val Ala Lys Ala Gln Lys

370 375 380

Thr Pro Glu Glu Gly Trp Thr Met Gln Asp Gly Thr Ser Trp Pro Gly

385 390 395 400

Asn Asn Thr Arg Asp His Pro Gly Met Ile Gln Val Phe Leu Gly Tyr

405 410 415

Ser Gly Ala Arg Asp Ile Glu Gly Asn Glu Leu Pro Arg Leu Val Tyr

420 425 430

Val Ser Arg Glu Lys Arg Pro Gly Tyr Gln His His Lys Lys Ala Gly

435 440 445

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

450 455 460

Phe Ile Leu Asn Leu Asp Cys Asp His Tyr Val Asn Asn Ser Lys Ala

465 470 475 480

Val Arg Glu Ala Met Cys Phe Leu Met Asp Pro Val Val Gly Gln Asp

485 490 495

Val Cys Phe Val Gln Phe Pro Gln Arg Phe Asp Gly Ile Asp Lys Ser

500 505 510

Asp Arg Tyr Ala Asn Arg Asn Ile Val Phe Phe Asp Val Asn Met Arg

515 520 525

Gly Leu Asp Gly Ile Gln Gly Pro Val Tyr Val Gly Thr Gly Thr Val

530 535 540

Phe Arg Arg Gln Ala Leu Tyr Gly Tyr Ser Pro Pro Ser Lys Pro Arg

545 550 555 560

Ile Leu Pro Gln Ser Ser Ser Ser Ser Ser Cys Cys Cys Cys Leu Thr Lys Lys

565 570 575

Lys Gln Pro Gln Asp Pro Ser Glu Ile Tyr Lys Asp Ala Lys Arg Glu

580 585 590

Glu Leu Asp Ala Ala Ile Phe Asn Leu Gly Asp Leu Asp Asn Tyr Asp

595 600 605

Glu Tyr Asp Arg Ser Met Leu Ile Ser Gln Thr Ser Phe Glu Lys Thr

610 615 620

Phe Gly Leu Ser Thr Val Phe Ile Glu Ser Thr Leu Met Glu Asn Gly

625 630 635 640

Gly Val Pro Asp Ser Val Asn Pro Ser Thr Leu Ile Lys Glu Ala Ile

645 650 655

His Val Ile Ser Cys Gly Tyr Glu Glu Lys Thr Glu Trp Gly Lys Glu

660 665 670

Ile Gly Trp Ile Tyr Gly Ser Ile Thr Glu Asp Ile Leu Thr Gly Phe

675 680 685

Lys Met His Cys Arg Gly Trp Arg Ser Ile Tyr Cys Met Pro Leu Arg

690 695 700

Pro Ala Phe Lys Gly Ser Ala Pro Ile Asn Leu Ser Asp Arg Leu His

705 710 715 720

Gln Val Leu Arg Trp Ala Leu Gly Ser Val Glu Ile Phe Leu Ser Arg

725 730 735

His Cys Pro Leu Trp Tyr Gly Cys Ser Gly Gly Arg Leu Lys Leu Leu

740 745 750

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

755 760 765

Pro Leu Val Ala Tyr Cys Thr Leu Pro Ala Ile Cys Leu Leu Thr Gly

770 775 780

Lys Phe Ile Ile Pro Thr Leu Ser Asn Leu Ala Ser Met Leu Phe Leu

785 790 795 800

Gly Leu Phe Ile Ser Ile Ile Leu Thr Ser Val Leu Glu Leu Arg Trp

805 810 815

Ser Gly Val Ser Ile Glu Asp Leu Trp Arg Asn Glu Gln Phe Trp Val

820 825 830

Ile Gly Gly Val Ser Ala His Leu Phe Ala Val Phe Gln Gly Phe Leu

835 840 845

Lys Met Leu Ala Gly Leu Asp Thr Asn Phe Thr Val Thr Ser Lys Thr

850 855 860

Ala Asp Asp Leu Glu Phe Gly Glu Leu Tyr Ile Val Lys Trp Thr Thr

865 870 875 880

Leu Leu Ile Pro Pro Thr Ser Leu Leu Ile Ile Asn Leu Val Gly Val

885 890 895

Val Ala Gly Phe Ser Asp Ala Leu Asn Lys Gly Tyr Glu Ala Trp Gly

900 905 910

Pro Leu Phe Gly Lys Val Phe Phe Ala Phe Trp Val Ile Leu His Leu

915 920 925

Tyr Pro Phe Leu Lys Gly Leu Met Gly Arg Gln Asn Arg Thr Pro Thr

930 935 940

Ile Val Ile Leu Trp Ser Ile Leu Leu Ala Ser Val Phe Ser Leu Val

945 950 955 960

Trp Val Arg Ile Asn Pro Phe Val Ser Lys Thr Asp Thr Thr Ser Leu

965 970 975

Ser Leu Asn Cys Leu Leu Ile Asp Cys

980 985

Claims (9)

1. A plant drought resistance-related protein, the amino acid residue sequence of which is shown in SEQ ID №:2. 2. The gene encoding the plant drought resistance-related protein of claim 1. 3. The gene according to claim 2, characterized in that: the cDNA gene of the plant drought resistance-related protein is selected from one of the following nucleotide sequences: 1) The DNA sequence of SEQ ID №: 1 in the sequence listing; 2) A polynucleotide encoding the protein sequence of SEQ ID №: 2 in the sequence listing. 4. The expression vector containing the gene encoding the plant drought resistance-related protein according to claim 2 or 3. 5. A cell line containing the gene encoding the plant drought resistance-related protein as claimed in claim 2 or 3. 6. A host bacterium containing the gene encoding the plant drought resistance-related protein as claimed in claim 2 or 3. 7. A primer for amplifying the gene encoding the plant drought resistance-related protein as claimed in claim 2 or 3. 8. A method for enhancing plant drought resistance by using the gene encoding the plant drought resistance-related protein according to claim 2 or 3, comprising inhibiting the expression of the gene encoding the plant drought resistance-related protein in the plant. 9. The method according to claim 8, characterized in that: the method of inhibiting the expression of the gene encoding the plant drought resistance-related protein in the plant includes the method of plant virus vector-mediated gene silencing, the method of antisense nucleic acid technology silencing gene and siRNA-mediated gene silencing methods.

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