CN112250743B

CN112250743B - Wheat drought stress related protein TaWRKY-A and coding gene and application thereof

Info

Publication number: CN112250743B
Application number: CN201910599072.5A
Authority: CN
Inventors: 胡兆荣; 孙其信; 刘星贝; 倪中福; 彭惠茹; 姚颖垠; 辛明明; 郭伟龙
Original assignee: China Agricultural University
Current assignee: China Agricultural University
Priority date: 2019-07-04
Filing date: 2019-07-04
Publication date: 2022-05-06
Anticipated expiration: 2039-07-04
Also published as: CN112250743A

Abstract

The invention discloses a wheat drought stress related protein TaWRKY-A and a coding gene and application thereof. The invention obtains a gene which is from the wheat A subgenome and closely related to the wheat drought resistance by combining transcriptome analysis on the basis of identifying the wheat material seedling stage drought resistance, and deeply analyzes and identifies the function of the gene by a transgenic method, thereby further defining the function of the TaWRKY-A gene in the process of wheat responding to drought stress. The method has important significance for breeding the drought-resistant wheat.

Description

Wheat drought stress related protein TaWRKY-A and coding gene and application thereof

Technical Field

The invention relates to a wheat drought stress related protein TaWRKY-A and a coding gene and application thereof.

Background

In addition to the attack of diseases and pests, the growth and development of crops are often affected by adverse climatic and soil factors, which are called environmental stress or abiotic stress, and thus the yield and quality are affected. Along with global warming, water resource shortage, soil desertification and salinization, the abiotic stress suffered by crops is more and more frequent and serious.

Wheat is an important food crop in China, and the yield and the quality of the wheat are directly related to the life of people. Drought has a very important influence on the growth and development of wheat, and among various stress factors, the drought becomes an important factor for limiting wheat production. According to statistics, the yield reduction caused by drought and water shortage in China is nearly 1 hundred million tons in recent 40 years. Therefore, the molecular mechanism research of the drought tolerance of the wheat is developed, the drought-tolerant related gene is identified and applied to the molecular breeding of the wheat, the cultivation of the drought-tolerant variety of the wheat is facilitated, and the stable development of the wheat production in China is promoted.

Disclosure of Invention

The invention aims to provide a wheat drought stress related protein TaWRKY-A and a coding gene and application thereof.

In a first aspect, the protective protein of the invention (designated as TaWRKY-a) is a protein of (a1), (a2) or (A3) as follows:

(A1) the amino acid sequence is protein of sequence 1 in a sequence table;

(A2) a protein which is obtained by substituting and/or deleting and/or adding one or more amino acid residues in the amino acid sequence shown in the sequence 1 in the sequence table, has more than 90% of identity with the protein shown in (A1), and is related to the drought resistance of plants;

(A3) and (C) a fusion protein obtained by attaching a protein tag to the N-terminus or/and the C-terminus of (A1) or (A2).

The TaWRKY-A protein is derived from wheat.

The protein can be artificially synthesized, or can be obtained by synthesizing the coding gene and then performing biological expression.

In the above protein, the tag may be a Flag tag, a His tag, an MBP tag, an HA tag, a myc tag, a GST tag, and/or a SUMO tag, etc.

In a second aspect, the invention protects the biological material related to TaWRKY-A protein, which is (B1) or (B2) as follows:

(B1) a nucleic acid molecule encoding a TaWRKY-a protein;

(B2) an expression cassette, a recombinant vector, a recombinant microorganism or a transgenic plant cell line comprising (B1) the nucleic acid molecule.

(B1) The nucleic acid molecule is a coding gene of the protein shown in the following (b1), (b2) or (b 3):

(b1) DNA molecule shown in sequence 2 in the sequence table;

(b2) a DNA molecule which hybridizes under stringent conditions to the DNA sequence defined in (b1) and encodes said protein;

(b3) and (b) a DNA molecule which has more than 90% homology with the DNA sequence defined in (b1) or (b2) and encodes the protein.

In the above genes, the stringent conditions may be as follows: 50 ℃ in 7% Sodium Dodecyl Sulfate (SDS), 0.5M NaPO₄Hybridization with 1mM EDTA, rinsing in 2 XSSC, 0.1% SDS at 50 ℃; also can be: 50 ℃ in 7% SDS, 0.5M NaPO₄Hybridization with 1mM EDTA, rinsing at 50 ℃ in 1 XSSC, 0.1% SDS; also can be: 50 ℃ in 7% SDS, 0.5M NaPO₄Hybridization with 1mM EDTA, rinsing in 0.5 XSSC, 0.1% SDS at 50 ℃; also can be: 50 ℃ in 7% SDS, 0.5M NaPO₄Hybridization with 1mM EDTA, rinsing in 0.1 XSSC, 0.1% SDS at 50 ℃; also can be: 50 ℃ in 7% SDS, 0.5M NaPO₄Hybridization with a mixed solution of 1mM EDTA, rinsing in 0.1 XSSC, 0.1% SDS at 65 ℃; can also be: in a solution of 6 XSSC, 0.5% SDS at 65 ℃ and then washed once with each of 2 XSSC, 0.1% SDS and 1 XSSC, 0.1% SDS.

In a third aspect, the invention provides the protein, or the use of the biomaterial, as any one of (C1) to (C4) below:

(C1) regulating and controlling the drought resistance of the plant;

(C2) preparing a product for improving the drought resistance of plants;

(C3) cultivating drought-resistant plants;

(C4) and (5) plant breeding.

The above application is embodied as follows:

the activity and/or expression of the TaWRKY-A protein or the coding gene thereof in the plant are improved, and the drought resistance of the plant is improved;

the activity and/or expression of the TaWRKY-A protein or the coding gene thereof in the plant is reduced, and the drought resistance of the plant is reduced.

In a fourth aspect, the invention protects method a or method B.

The method A comprises the following steps: a method for breeding a plant variety with improved drought resistance, comprising the steps of: improving the expression quantity and/or activity of TaWRKY-A protein in a receptor plant to obtain a plant with improved drought resistance.

The method B comprises the following steps: a method of breeding a transgenic plant comprising the steps of: introducing a nucleic acid molecule for encoding TaWRKY-A protein into a receptor plant to obtain a transgenic plant; the drought resistance of the transgenic plant is greater than that of a receptor plant.

The 'introduction of a nucleic acid molecule encoding a TaWRKY-A protein into a receptor plant' is realized by introducing a recombinant expression vector containing an encoding gene of the TaWRKY-A protein into the receptor plant.

The existing expression vector can be used for constructing a recombinant expression vector containing the coding gene of the TaWRKY-A protein. The plant expression vector comprises a binary agrobacterium vector, a vector for plant microprojectile bombardment and the like. The plant expression vector may also comprise the 3' untranslated region of the foreign gene, i.e., a region comprising a polyadenylation signal and any other DNA segments involved in mRNA processing or gene expression. The poly A signal can lead poly A to be added to the 3 'end of mRNA precursor, and the untranslated regions transcribed at the 3' end of Agrobacterium crown gall inducible (Ti) plasmid genes (such as nopaline synthase gene Nos) and plant genes (such as soybean storage protein gene) have similar functions.

When a recombinant plant expression vector is constructed by using a coding gene of TaWRKY-A protein, any one of an enhanced promoter or a constitutive promoter (such as a cauliflower mosaic virus (CAMV)35S promoter and a maize Ubiquitin promoter (Ubiquitin)) or a tissue-specific expression promoter (such as a seed-specific expression promoter) can be added in front of a transcription initiation nucleotide, and the promoter can be used alone or combined with other plant promoters. In addition, when a plant expression vector is constructed by using a gene encoding the TaWRKY-a protein, enhancers, including translation enhancers or transcription enhancers, may be used, and these enhancer regions may be ATG initiation codons or initiation codons of adjacent regions, etc., but must be in the same reading frame as the coding sequence to ensure correct translation of the entire sequence. The translational control signals and initiation codons are widely derived, either naturally or synthetically. The translation initiation region may be derived from a transcription initiation region or a structural gene.

In order to facilitate the identification and screening of transgenic plant cells or plants, plant expression vectors to be used may be processed, for example, by adding a gene encoding an enzyme or a luminescent compound which can produce a color change (GUS gene, luciferase gene, etc.), an antibiotic marker having resistance (gentamicin marker, kanamycin marker, etc.), or a chemical-resistant marker gene (e.g., herbicide-resistant gene), etc., which can be expressed in plants.

In the invention, the recombinant expression vector can be specifically a recombinant expression vector obtained by replacing a fragment between BamHI and SpeI enzyme cutting sites of a pWMB110 vector with a DNA molecule shown in a sequence 2 in a sequence table.

In the above method, the introducing the recombinant expression vector carrying the coding gene of the TaWRKY-a protein into the recipient plant may specifically be: plant cells or tissues are transformed by conventional biological methods using Ti plasmids, Ri plasmids, plant viral vectors, direct DNA transformation, microinjection, conductance, agrobacterium mediation, etc., and the transformed plant tissues are grown into plants.

Transformed cells, tissues or plants are understood to comprise not only the end product of the transformation process, but also transgenic progeny thereof.

In a fifth aspect, the invention features method C or method D.

The method C comprises the following steps: a method of reducing drought resistance in a plant comprising the steps of: reducing the expression quantity and/or activity of TaWRKY-A protein in a receptor plant to obtain a plant with reduced drought resistance.

The method D comprises the following steps: a method of reducing drought resistance in a plant comprising the steps of: inhibiting the expression of a nucleic acid molecule encoding TaWRKY-A protein in a receptor plant to obtain a plant with reduced drought resistance.

The 'inhibiting the expression of a nucleic acid molecule encoding TaWRKY-A protein in a receptor plant' is realized by introducing an RNAi interference vector into the receptor plant.

The RNAi interference vector can be a recombinant expression vector obtained by replacing a fragment between the XhoI and SwaI enzyme cutting sites of the plasmid pFGC5941 with a DNA molecule shown in a sequence 3 of a sequence table, and replacing a fragment between the BamHI and XbaI enzyme cutting sites of the plasmid pFGC5941 with a DNA molecule shown in a sequence 4 of the sequence table.

In the above aspects, the plant is (D1) or (D2) or (D3):

(D1) a dicot or monocot;

(D2) a gramineous plant;

(D3) wheat (Triticum aestivum L.).

The wheat can be wheat farmer 199 or wheat CB 037.

In a sixth aspect, the invention protects the application of a substance for obtaining the full length or partial section of the coding gene promoter of the TaWRKY-A protein in the genome of the plant to be detected in the auxiliary identification of the drought resistance of the plant.

The substance can be a primer pair; the primer pair consists of a primer F and a primer R; the primer F is a single-stranded DNA molecule shown in a sequence 5 of the sequence table; the primer R is a single-stranded DNA molecule shown in a sequence 6 of a sequence table.

In a seventh aspect, the invention provides a method for assisting in identifying drought resistance of a plant, comprising the following steps:

(1) extracting the genome DNA of a plant to be detected;

(2) performing PCR amplification by using the genome DNA obtained in the step (1) as a template and adopting the primer pair, wherein if a specific strip of 677 +/-10 bp exists in an amplification product, the plant is or is a candidate drought-resistant plant; if a 1378 +/-10 bp specific band exists in the amplification product, the plant is or is selected as a drought-sensitive plant.

In the sixth and seventh aspects, the plant is (D1) or (D2) or (D3):

(D1) a dicot or monocot;

(D2) a gramineous plant;

(D3) wheat (Triticum aestivum L.).

The wheat may be Rumai No. 3, Weimai No. 8, Nongda 3338, Nongda 6554, Jin 50 or Zheng wheat 1817.

Drought, an abiotic stress, is an important limiting factor affecting wheat yield. The invention obtains a gene which is from the wheat A subgenome and closely related to the wheat drought resistance by combining transcriptome analysis on the basis of identifying the wheat material seedling stage drought resistance, and deeply analyzes and identifies the function of the gene by a transgenic method, thereby further defining the function of the TaWRKY-A gene in the process of wheat responding to drought stress. The method has important significance for breeding the drought-resistant wheat.

Drawings

FIG. 1 is the phenotypic observations of wheat from each group in example 2.

FIG. 2 is the weight and survival rate statistics of the upper part of each wheat group in example 2.

FIG. 3 is the phenotypic observations of wheat from each group of example 3.

FIG. 4 is the statistics of weight of upper part, water loss rate and survival rate of each group of wheat in example 3.

Fig. 5 is the survival rate statistics of wheat in each group in example 4.

FIG. 6 shows the statistical results of TaWRKY-A gene expression levels of wheat in each group in example 4.

FIG. 7 is a graph showing the results of electrophoresis of wheat groups in example 5.

Detailed Description

The following examples are given to facilitate a better understanding of the invention, but do not limit the invention. The experimental procedures in the following examples are conventional unless otherwise specified. The test materials used in the following examples were purchased from a conventional biochemical reagent store unless otherwise specified. The quantitative tests in the following examples, all set up three replicates and the results averaged.

Plasmid pFGC 5941: beijing Huayue ocean biology, cat # VECT 0360.

Escherichia coli Trans1-T1Phage Resistant: general gold, cat # goods: CD 501-03.

Wheat farmer 199, also known as wheat KN 199: reference documents: znwei, Wangjing, Jijun, et al, winter wheat new variety "Kenong 199" for breeding and promotion [ J ]. China ecological agriculture bulletin, 2011,19(5): 1215-; the public is available from the university of agriculture in china.

Wheat CB 037: reference: zhang Wei, Yi Mi Qi, Zhang pei, et al, China, part of the main thrust wheat variety tissue culture regeneration ability evaluation [ J ] crops, 2018.; the public is available from the university of agriculture in china.

Lumai No. 3: winter wheat new variety lumai No. 3 [ J ] Shandong agricultural science, 1984 (3); the public is available from the university of agriculture in china.

Weimai No. 8: chenxiang Zhi, Zhang Qilu, Zhang Lianjie, et al, Weimai No. 8 fertility characteristics and super wheat breeding target [ J ] Shandong agriculture, 2005(3): 14-16.; the public is available from the university of agriculture in china.

Nongda 3338: QTL analysis of the plant height and internode length of DH series 3338X Beijing winter 6 of common wheat (J) is carried out in the first, wax tiger, Weiqiang, Wangfei, et al, university of Chinese agriculture, 2014,19(1): 1-8.; the public is available from the university of agriculture in china.

"nongda" 6554: shanzhuang laboratory station of Chinese agricultural university.

Promoting 50: the experimental station for Shanzhuang of Chinese agriculture university.

Zheng wheat 1817: shanzhuang laboratory station of Chinese agricultural university.

Example 1, discovery of wheat drought stress related protein TaWRKY-A and coding gene thereof

A great deal of analysis is carried out on drought resistance and transcriptome data of the wheat material in the seedling stage, a gene related to wheat drought stress is found, and the gene is named as TaWRKY-A gene and is shown as a sequence 2 in a sequence table. The protein shown in the sequence 1 of the TaWRKY-A gene coding sequence table is named as TaWRKY-A protein.

Example 2 drought resistance analysis of TaWRKY-A RNAi transgenic wheat

1. The DNA molecule shown in the sequence 3 of the sequence table is replaced by the fragment between the XhoI and SwaI enzyme cutting sites of the plasmid pFGC5941, and the DNA molecule shown in the sequence 4 of the sequence table is replaced by the fragment between the BamHI and XbaI enzyme cutting sites of the plasmid pFGC5941, so that the WRKY-A-RNAi vector (which is sequenced and verified) is obtained.

2. And (2) introducing the WRKY-A-RNAi vector prepared in the step (1) into escherichia coli Trans1-T1Phage Resistant to obtain a recombinant bacterium.

3. Sending the recombinant bacteria obtained in the step 2 to genetic and developmental biology institute of Chinese academy of sciences to convert into a receptor wheat KN199 to obtain T₀And (5) plant generation.

4. For T obtained in step 3₀Extracting DNA of leaves from the generation plants, and performing PCR identification by using a primer pair consisting of a primer W RNAiF and a primer W RNAiR: selection of T identified as positive₀Selfing the plant to obtain T₁And (5) plant generation. The amplified band of the negative plant is 683bp, while the amplified band of the positive plant is 470bp and 683 bp.

W RNAiF：5’-CCATGACCTTGGACTTCACC-3’；

W RNAiR：5’-CACCGTGCTGCACTTGTAGT-3’。

5. For T obtained in step 4₁Identifying the generation plants according to the method in the step 4, and selecting the T which is identified as positive₁Selfing the plant to obtain T₂And (5) plant generation.

6. For T obtained in step 5₂Identifying the generation plants according to the method in the step 4, and selecting the T which is identified as positive₂Selfing the plant to obtain T₃And (4) generating plants, identifying the plants according to the method in the step (4), and screening out positive RNAi T₃And (5) plant generation. A total of 20 positive RNAi lines were obtained. Simultaneously screen out T₃Generation negative control RNAi (-) plants (transgenic plants with negative results detected by PCR).

Second, obtaining of empty vector line

And (3) replacing a WRKY-A-RNAi vector with a plasmid pFGC5941, and operating according to the steps 2-6 to obtain a transgenic empty vector strain.

Third, drought resistance identification of RNAi strain

The material to be tested: seed of wheat KN199, T of positive RNAi strains (RNAi-22 and RNAi-41)₃T of generation seed, empty carrier line₃Seed generation, control RNAi (-) T₃And (5) seed generation.

1. Disinfecting seeds to be detected (50 seeds are selected for each material) with 1% hydrogen peroxide for 10min, cleaning with distilled water for 3-4 times, placing in a culture dish, spreading two layers of filter paper in the culture dish, adding a small amount of distilled water, standing at room temperature for 48h, selecting seedlings with consistent germination, transplanting into a nutrition pot, and transplanting 8 seedlings in each pot.

2. After the step 1 is completed, dividing the test sample into a control group and an experimental group for treatment;

experimental groups: and (3) filling water once, then not watering again until the water content of the soil reaches about 4%, rehydrating, observing the phenotype 5 days after rehydration, photographing, and recording the survival rate and the weight of the overground part.

Control group: watering normally in the whole process.

The phenotypic observations are shown in figure 1. In fig. 1, a is the phenotypic observation of the control group, and B is the phenotypic observation of the experimental group. The above-ground weight statistics are shown in fig. 2A. The survival statistics are shown in fig. 2B.

The results show that under normal conditions, there was no significant difference between the different RNAi and control strains. After the drought treatment, obvious difference is shown, and the growth vigor and the recovery degree of RNAi strains RNAi-22 and RNAi-41 are obviously weaker than those of receptor KN199 and negative control strain RNAi (-). The survival rate of the RNAi strains is obviously lower than that of the KN199 and RNAi (-), wherein the survival rate of each RNAi strain is about 20-50%, and the survival rate of the KN199 and RNAi (-) is more than 70%. There was no significant difference in the dry weight of the upper part of the control group, while the dry weight of the upper part of each RNAi transgenic line in the treated group was significantly lower than those of the recipient and negative control lines. Therefore, the drought resistance of the RNAi strain is obviously weaker than that of the receptor and the negative control strain, which shows that the expression of TaWRKY-A is inhibited, and the drought resistance of the plant is reduced. The phenotype and statistical result of the empty vector transfer line have no significant difference from the wild type.

Example 3 drought resistance analysis of TaWRKY-A Gene-transferred wheat

Obtaining of wheat with TaWRKY-A gene

1. The fragment between the enzyme cutting sites of BamHI and speI of the vector pWMB110 (circular plasmid shown in sequence 8 of the sequence table) is replaced by a DNA molecule shown in sequence 2 of the sequence table to obtain an over-expression vector pWMB110-TaWRKY-A (sequencing verification is carried out).

2. And (3) introducing the over-expression vector pWMB110-TaWRKY-A prepared in the step 1 into Escherichia coli Trans1-T1Phage Resistant to obtain a recombinant bacterium.

3. Sending the recombinant bacteria obtained in the step 2 to a receptor wheat CB037 transformed by the institute of crop science of Chinese academy of agricultural sciences to obtain T₀And (5) plant generation.

4. For T obtained in step 3₀The generation plants are subjected to PCR identification by adopting a primer pair consisting of a primer Ubi F and a primer WrkyAR, and T which is identified as positive is selected₀Selfing the plant to obtain T₁And (5) plant generation. The positive plants can amplify bands, and the negative plants have no bands.

Ubi F：5’-TAGCCCTGCCTTCATACGCT-3’；

WrkyAR：5’-TCACCGACGACATGAAGGAT-3’。

6. For T obtained in step 5₂Identifying the generation plants according to the method in the step 4, and selecting the T which is identified as positive₂Selfing the plant to obtain T₃The generation plants are identified according to the method in the step 4, and positive transgenic T is screened out₃And (5) plant generation. A total of 12 positive transgenic lines were obtained.

Second, obtaining of empty vector line

And (3) replacing an overexpression vector pWMB110-TaWRKY-A with the vector pWMB110, and operating according to the steps 2-6 to obtain the empty-transferred vector strain.

Drought resistance identification of wheat with TaWRKY-A gene

The material to be tested: seeds of wheat CB037, T of positive transgenic lines (OE-3 and OE-1)₃T of generation seed, empty carrier line₃And (5) seed generation.

experimental groups: and (3) filling water once, then not watering again until the water content of the soil reaches about 4%, rehydrating, observing the phenotype 5 days after rehydration, photographing, and recording the survival rate and the fresh weight and dry weight of the overground part.

Control group: watering normally in the whole process.

The phenotypic observations are shown in figure 3. In fig. 3, a is the phenotypic observation of the control group, and B is the phenotypic observation of the experimental group. The results show that under normal conditions, there is no significant difference between the transgenic lines and the control lines. After drought treatment, transgenic lines OE-3 and OE-1 grew significantly more strongly than wild-type CB 037. The phenotype of the empty vector transferred strain has no significant difference with the wild type.

The above-ground weight statistics are shown in fig. 4A. The results show that after rehydration, the fresh weights of the strains are significantly different, the fresh weights of transgenic strains OE-3 and OE-1 are greater than CB037, wherein the fresh weight of OE-3 is significantly higher than that of the wild type. Furthermore, the dry weight of the transgenic lines after rehydration was slightly higher than the wild type. The statistical result of the empty vector-transferred strain has no significant difference with the wild type.

As can be seen from the statistical survival rate, the survival rate of the over-expression strains OE-1 and OE-3 is significantly higher than that of the wild type CB 037. The results are shown in FIG. 4C.

3. After the step 1 is completed, cutting leaves after the seedlings grow for about one month, recording the initial weight M1 of the leaves, paving the leaves in an environment with constant relative humidity (no requirement on specific relative humidity and temperature, only needing to be stable), recording the weight of the leaves every other hour, and calculating the water loss rate of the leaves in vitro by taking the weight of the leaves after water loss in different time periods as Mn.

The water loss rate (%) of the excised leaf was (M1-Mn)/M1X 100%

The results are shown graphically in FIG. 4B. The result shows that the water loss rate of the leaves in vitro of the transgenic lines OE-3 and OE-1 is obviously lower than that of wild CB037 under the same condition. The stomatal water loss rate of the transgenic line is less than that of the wild type.

Therefore, the drought resistance of plants can be remarkably enhanced by over-expressing the TaWRKY-A gene.

Example 4 analysis of correlation between expression level of TaWRKY-A Gene and drought resistance

Wheat to be tested: rumai No. 3, Weiwei mai No. 8, nong da 3338, nong da 6554, jin's 50 and zheng mai 1817.

First, identification of wheat drought resistance

Sowing seeds of wheat to be detected in black pots 9cm long, 7.5cm wide and 10cm high, wherein the cultivation soil consists of nutrient flower soil and vermiculite (3:1), fully stirring and uniformly mixing, adding nutrient soil (the weight of the soil is 300 g/pot) with the same weight into each pot, planting 9 wheat seeds in each pot, pouring excess water in the disc after each pot absorbs water fully for one night, and covering with a film to ensure the germination rate. After the seeds emerge, thinning the seedlings to the rest six seedlings, ensuring the growth vigor of the seedlings to be regular, and planting six pots of each material. After sufficient water absorption, no watering is needed. The position is moved randomly at regular intervals, so that the difference caused by the position effect is reduced. And (4) rehydrating when the water content of soil in each pot is reduced to 4%, and counting the survival rate of materials in each pot after five days of rehydration.

The results are shown in FIG. 5, where both the Rough 3 and Weichai No. 8 and the Nongda 3338 were drought-sensitive materials, with a 0% survival rate and a 0.4% survival rate for the Nongda 3338. The survival rates of the drought-resistant materials of agricultural crops 6554, Jin50 and Zheng wheat 1817 are all more than 0.8.

Detection of TaWRKY-A gene expression level under drought stress

1. Disinfecting seeds to be tested with 1% hydrogen peroxide for 10min, cleaning with distilled water for 3-4 times, placing in a culture dish, spreading two layers of filter paper in the culture dish, adding a small amount of distilled water, standing at room temperature for 48h, selecting seedlings with consistent germination, transplanting into a nutrition pot, and planting 8 seedlings in each pot for 6 times.

2. And (3) after the step 1 is finished and the plant grows for 5 days, taking out 3 repeated whole plants from the pot, cleaning the root system, paving the whole plants in an environment with constant relative humidity and temperature (no requirement on specific relative humidity and temperature, only needing to be stable), and continuously placing the other 3 repeated plants in the pot to grow to serve as a control group. After 4 hours, the treatment group and the control group are respectively sampled, RNA is extracted, cDNA is reversed, and qRT-PCR is carried out to detect the expression level of the TaWRKY-A gene (TaActin is used as an internal reference gene).

The qRT-PCR primer information is as follows:

TaWRKY-AF:5’-TGCTCATCGTGACCTACGAG-3’；

TaWRKY-AR:5’-ACATGCTTAGTGCGACTAAACC-3’；

TaActin-F：5’-GGAATCCATGAGACCACCTAC-3’；

TaActin-R：5’-GACCCAGACAACTCGCAAC-3’。

the results are shown in FIG. 6. The result shows that under normal conditions, the expression of the TaWRKY-A gene among different drought-resistant materials has no obvious difference, after drought stress treatment, the expression of the TaWRKY-A gene is obviously induced, and the expression level of the TaWRKY-A gene of the drought-resistant material is higher than that of a drought-sensitive material.

The up-regulation expression multiple and survival rate of the TaWRKY-A gene in different materials after induction are analyzed in a correlation mode, and the results are shown in table 1. The results show that the up-regulated expression multiple of the TaWRKY-A gene is in significant positive correlation with the survival rate.

TABLE 1 correlation analysis of survival rates with fold-up expression

Example 5 development of functional marker for drought resistance

Based on the results, the TaWRKY-A genes of various wheat materials are subjected to clone sequencing to discover that the TaWRKY-A genes have two allelic genotypes and length polymorphism in promoter regions thereof. Aiming at the discovery, a pair of primers for assisting in identifying the drought resistance of the plant is developed, and two sequences with different lengths can be amplified.

TaWRKY-AF: 5'-GGAGCATGCTTACAAAACCTATG-3' (sequence 5 of the sequence table);

TaWRKY-AR: 5'-GACCCACCTGTCAGAGACTTT-3' (SEQ ID NO: 6 of the sequence Listing).

The genomic DNA of the 6 wheat materials of example 4 was amplified using the primer set described above, and the results of electrophoresis are shown in FIG. 7. The size of the product band of the drought-resistant materials (Nongda 6554, Weida 50 and Zhengmai 1817) is 677bp (the sequencing result is shown as the sequence 7 in the sequence table), and the product band 1378bp of the drought-sensitive materials (Lumai No. 3, Weimai No. 8 and Nongda 3338) has obvious length polymorphism.

The results show that the molecular marker can be used as a functional marker for wheat molecular assisted breeding.

Sequence listing

<110> university of agriculture in China

<120> wheat drought stress related protein TaWRKY-A and coding gene and application thereof

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Asp Ala Gly Phe Ser Val Ser Ala Ala Ser Ser Ser Phe Met Ser Ser

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atgatgacca tggatctgat tggaggatac gggagggcgg acgagcaggt ggccatccag 60

gaggcggcgg cggcggggct gtgcgggatg gagcacctca tcctgcagct ctcccggaca 120

ggcaccagcg agagctcgcc ggttgggtcg tcggaggcgc cggagcagca ggtagactgc 180

cgggagatca ctgatatgac cgtgtccaag ttcaagaagg tgatttctat cctcaaccac 240

cgcactggcc acgccaggtt ccggcgcggg cctgtggtgg cgcagtccca gggccccgcc 300

gtgtccgagc cggcgccggt gagggcgtct tcgtcgaggt ccatgacctt ggacttcacc 360

aaggcgtctt ccgggtacgg aaacgacgcc gggttcagcg tctcggccgc gagctcatcc 420

ttcatgtcgt cggtgaccgg tgacgggagc gtgtccaacg acgcgggggc gggtcctcgc 480

tgatgctccc gccgccacct tcggccagct gcgggaaacc gccgctggcg tcctccgcgg 540

catccaccgg cgccggtgcc gggcagaagc gcaagtgcca cgaccacgcg cactcagaga 600

acgtcgccgg cggaaagtac ggcgcctccg gtggccgctg ccactgctcc aagcgcagga 660

aatcccgggt tcggcggatg actcgcgtgc cggcgatcag ctcgaaggcg gcggagatcc 720

ccgcggacga cttctcgtgg cgcaagtatg gccagaagcc tatcaagggc tccccctacc 780

cacgaggtta ctacaagtgc agcacggtgc gcgggtgccc ggcgcggaag cacgtggagc 840

gtgaccccag cgacccctcc atgctcatcg tgacctacga gggcgagcac cggcacaccc 900

ccgcggacca ggagccgctc gccccgctac cggagctctg a 941

<210> 3

<211> 469

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 3

ccatgacctt ggacttcacc aaggcgtctt ccgggtacgg aaacgacgcc gggttcagcg 60

tctcggccgc gagctcatcc ttcatgtcgt cggtgaccgg tgacgggagc gtgtccaacg 120

acgcgggggc gggtcctcgc tgatgctccc gccgccacct tcggccagct gcgggaaacc 180

gccgctggcg tcctccgcgg catccaccgg cgccggtgcc gggcagaagc gcaagtgcca 240

cgaccacgcg cactcagaga acgtcgccgg cggaaagtac ggcgcctccg gtggccgctg 300

ccactgctcc aagcgcagga aatcccgggt tcggcggatg actcgcgtgc cggcgatcag 360

ctcgaaggcg gcggagatcc ccgcggacga cttctcgtgg cgcaagtatg gccagaagcc 420

tatcaagggc tccccctacc cacgaggtta ctacaagtgc agcacggtg 469

<210> 4

<211> 469

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 4

ccatgacctt ggacttcacc aaggcgtctt ccgggtacgg aaacgacgcc gggttcagcg 60

tctcggccgc gagctcatcc ttcatgtcgt cggtgaccgg tgacgggagc gtgtccaacg 120

acgcgggggc gggtcctcgc tgatgctccc gccgccacct tcggccagct gcgggaaacc 180

gccgctggcg tcctccgcgg catccaccgg cgccggtgcc gggcagaagc gcaagtgcca 240

cgaccacgcg cactcagaga acgtcgccgg cggaaagtac ggcgcctccg gtggccgctg 300

ccactgctcc aagcgcagga aatcccgggt tcggcggatg actcgcgtgc cggcgatcag 360

ctcgaaggcg gcggagatcc ccgcggacga cttctcgtgg cgcaagtatg gccagaagcc 420

tatcaagggc tccccctacc cacgaggtta ctacaagtgc agcacggtg 469

<210> 5

<211> 23

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 5

ggagcatgct tacaaaacct atg 23

<210> 6

<211> 21

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 6

gacccacctg tcagagactt t 21

<210> 7

<211> 1378

<212> DNA

<213> wheat (Triticum aestivum L.)

<400> 7

ggagcatgct tacaaaacct atgtgatggg ggcgacctgc gccgaatgaa cctccggagg 60

ctatcgcatg agaaggccga acaccaagga ccccaggagg tgacgtgaac gggaccggcc 120

cgctagacat agctcggtcg cttggattta tcgacgctgc agggaagcac ccgagcatag 180

acgtcgtgcc cagtcttctc atgcaacaaa cccggggaag gacactcggc gcgctgctcc 240

tgcgacgaag aagcgccgaa ctgcggttgt atgaaccgag ccgtgcgccc tgactcaccc 300

actctcgatt agctcacgcg aggacaaggc accaaggacc ccgggaggtg acgtgcacgg 360

gagctggccc gcaagacaga gctcagtcgc ttggatttat caatgatata ctttttcctt 420

ttgcgagagg cacgaccatg ccttttgcga aagaaatgcg tgcctccacg agaaataaat 480

atgtgcctct catgaaagga aaaaaacacg tgtttttcta ttttacgagt ggcacgggcg 540

tgcctctcgc gtaagcaaat tcgtacctcc acaagaaata aaattgtgac tttcgcaggg 600

aaaaaaataa aatatacttt tttcatatat tttttgtcca aaacctaaga aaaaccgaag 660

acccaaaaaa attcatctaa aagccaaaaa agaccacaaa aaaataaaaa tgaaagacaa 720

aattcggagt gagcgtccat agcgcgacac gtggccgtgg ctgagagcgt caagtggcgc 780

gctccgagac cacctcaaat gaccttgtgg gggtttgtga aggagtactc tttaattact 840

cgtcttaatc actcgttggg tgaaattcaa ggcatcagtt gggttggtct aaggctggtc 900

atagtgggag taacataggt agtaacatag atgccacata agcaaaaatg atgatgtggc 960

aaatagttaa tgaggagaga agcaaataga gtaacataat atgttaccat cacatagcgg 1020

tttccaatgc ataatgagtc tacaaagtaa taaatgaagg caactatgtt accacaccta 1080

tgacactacc cactatgaag gtagtaacat agactagtaa catatgtatg ttactagtgt 1140

aagttactcc cactatgacc agcctaagca tttcctttgg agaccagaaa tggcaggacg 1200

cgcaaagaaa gtcagagctc atgattcttt cgttctcgaa ttgaatccac gaaccaacgg 1260

catttccatt caggcattaa agtccagcat tgacttttcg tcactccccg tcccacacga 1320

tatgccaaat cagacgtgaa tgaagaaaca acaaagcaaa gtctctgaca ggtgggtc 1378

<210> 8

<211> 11717

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 8

gcggccgcga taggccgacg cgaagcggcg gggcgtaggg agcgcagcga ccgaagggta 60

ggcgcttttt gcagctcttc ggctgtgcgc tggccagaca gttatgcaca ggccaggcgg 120

gttttaagag ttttaataag ttttaaagag ttttaggcgg aaaaatcgcc ttttttctct 180

tttatatcag tcacttacat gtgtgaccgg ttcccaatgt acggctttgg gttcccaatg 240

tacgggttcc ggttcccaat gtacggcttt gggttcccaa tgtacgtgct atccacagga 300

aagagacctt ttcgaccttt ttcccctgct agggcaattt gccctagcat ctgctccgta 360

cattaggaac cggcggatgc ttcgccctcg atcaggttgc ggtagcgcat gactaggatc 420

gggccagcct gccccgcctc ctccttcaaa tcgtactccg gcaggtcatt tgacccgatc 480

agcttgcgca cggtgaaaca gaacttcttg aactctccgg cgctgccact gcgttcgtag 540

atcgtcttga acaaccatct ggcttctgcc ttgcctgcgg cgcggcgtgc caggcggtag 600

agaaaacggc cgatgccggg atcgatcaaa aagtaatcgg ggtgaaccgt cagcacgtcc 660

gggttcttgc cttctgtgat ctcgcggtac atccaatcag ctagctcgat ctcgatgtac 720

tccggccgcc cggtttcgct ctttacgatc ttgtagcggc taatcaaggc ttcaccctcg 780

gataccgtca ccaggcggcc gttcttggcc ttcttcgtac gctgcatggc aacgtgcgtg 840

gtgtttaacc gaatgcaggt ttctaccagg tcgtctttct gctttccgcc atcggctcgc 900

cggcagaact tgagtacgtc cgcaacgtgt ggacggaaca cgcggccggg cttgtctccc 960

ttcccttccc ggtatcggtt catggattcg gttagatggg aaaccgccat cagtaccagg 1020

tcgtaatccc acacactggc catgccggcc ggccctgcgg aaacctctac gtgcccgtct 1080

ggaagctcgt agcggatcac ctcgccagct cgtcggtcac gcttcgacag acggaaaacg 1140

gccacgtcca tgatgctgcg actatcgcgg gtgcccacgt catagagcat cggaacgaaa 1200

aaatctggtt gctcgtcgcc cttgggcggc ttcctaatcg acggcgcacc ggctgccggc 1260

ggttgccggg attctttgcg gattcgatca gcggccgctt gccacgattc accggggcgt 1320

gcttctgcct cgatgcgttg ccgctgggcg gcctgcgcgg ccttcaactt ctccaccagg 1380

tcatcaccca gcgccgcgcc gatttgtacc gggccggatg gtttgcgacc gctcacgccg 1440

attcctcggg cttgggggtt ccagtgccat tgcagggccg gcaggcaacc cagccgctta 1500

cgcctggcca accgcccgtt cctccacaca tggggcattc cacggcgtcg gtgcctggtt 1560

gttcttgatt ttccatgccg cctcctttag ccgctaaaat tcatctactc atttattcat 1620

ttgctcattt actctggtag ctgcgcgatg tattcagata gcagctcggt aatggtcttg 1680

ccttggcgta ccgcgtacat cttcagcttg gtgtgatcct ccgccggcaa ctgaaagttg 1740

acccgcttca tggctggcgt gtctgccagg ctggccaacg ttgcagcctt gctgctgcgt 1800

gcgctcggac ggccggcact tagcgtgttt gtgcttttgc tcattttctc tttacctcat 1860

taactcaaat gagttttgat ttaatttcag cggccagcgc ctggacctcg cgggcagcgt 1920

cgccctcggg ttctgattca agaacggttg tgccggcggc ggcagtgcct gggtagctca 1980

cgcgctgcgt gatacgggac tcaagaatgg gcagctcgta cccggccagc gcctcggcaa 2040

cctcaccgcc gatgcgcgtg cctttgatcg cccgcgacac gacaaaggcc gcttgtagcc 2100

ttccatccgt gacctcaatg cgctgcttaa ccagctccac caggtcggcg gtggcccata 2160

tgtcgtaagg gcttggctgc accggaatca gcacgaagtc ggctgccttg atcgcggaca 2220

cagccaagtc cgccgcctgg ggcgctccgt cgatcactac gaagtcgcgc cggccgatgg 2280

ccttcacgtc gcggtcaatc gtcgggcggt cgatgccgac aacggttagc ggttgatctt 2340

cccgcacggc cgcccaatcg cgggcactgc cctggggatc ggaatcgact aacagaacat 2400

cggccccggc gagttgcagg gcgcgggcta gatgggttgc gatggtcgtc ttgcctgacc 2460

cgcctttctg gttaagtaca gcgataacct tcatgcgttc cccttgcgta tttgtttatt 2520

tactcatcgc atcatatacg cagcgaccgc atgacgcaag ctgttttact caaatacaca 2580

tcaccttttt agacggcggc gctcggtttc ttcagcggcc aagctggccg gccaggccgc 2640

cagcttggca tcagacaaac cggccaggat ttcatgcagc cgcacggttg agacgtgcgc 2700

gggcggctcg aacacgtacc cggccgcgat catctccgcc tcgatctctt cggtaatgaa 2760

aaacggttcg tcctggccgt cctggtgcgg tttcatgctt gttcctcttg gcgttcattc 2820

tcggcggccg ccagggcgtc ggcctcggtc aatgcgtcct cacggaaggc accgcgccgc 2880

ctggcctcgg tgggcgtcac ttcctcgctg cgctcaagtg cgcggtacag ggtcgagcga 2940

tgcacgccaa gcagtgcagc cgcctctttc acggtgcggc cttcctggtc gatcagctcg 3000

cgggcgtgcg cgatctgtgc cggggtgagg gtagggcggg ggccaaactt cacgcctcgg 3060

gccttggcgg cctcgcgccc gctccgggtg cggtcgatga ttagggaacg ctcgaactcg 3120

gcaatgccgg cgaacacggt caacaccatg cggccggccg gcgtggtggt gtcggcccac 3180

ggctctgcca ggctacgcag gcccgcgccg gcctcctgga tgcgctcggc aatgtccagt 3240

aggtcgcggg tgctgcgggc caggcggtct agcctggtca ctgtcacaac gtcgccaggg 3300

cgtaggtggt caagcatcct ggccagctcc gggcggtcgc gcctggtgcc ggtgatcttc 3360

tcggaaaaca gcttggtgca gccggccgcg tgcagttcgg cccgttggtt ggtcaagtcc 3420

tggtcgtcgg tgctgacgcg ggcatagccc agcaggccag cggcggcgct cttgttcatg 3480

gcgtaatgtc tccggttcta gtcgcaagta ttctacttta tgcgactaaa acacgcgaca 3540

agaaaacgcc aggaaaaggg cagggcggca gcctgtcgcg taacttagga cttgtgcgac 3600

atgtcgtttt cagaagacgg ctgcactgaa cgtcagaagc cgactgcact atagcagcgg 3660

aggggttgga tcaaagtact ttgatcccga ggggaaccct gtggttggca tgcacataca 3720

aatggacgaa cggataaacc ttttcacgcc cttttaaata tccgattatt ctaataaacg 3780

ctcttttctc ttaggtttac ccgccaatat atcctgtcaa acactgatag tttaaactga 3840

aggcgggaaa cgacaatctg atccaagctc aagctgctct agcattcgcc attcaggctg 3900

cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa 3960

gggggatgtg ctgcaaggcg attaagttgg gtaacgccag ggttttccca gtcacgacgt 4020

tgtaaaacga cggccagtgc caagcttgca tgcctgcaga gagatgcact gcaacatatg 4080

accaaaacgt catccatgat cgagtacaat ttcttagtgc ctttttcatc aaagttatat 4140

ttgaaacatt ttagaaaatt tgtcttaaat tttttcccat ggcgtttcga tcaaaaaaat 4200

gccttaaaac ctacctatat aaaggcaata agggaaaaca cttcaggata ccttcgaatc 4260

taaggtgtct ttctttagat ggtatgctaa actgttcttc atggcgcctc aaatcatcat 4320

tctgcaagta aagactaaaa ataacgctaa atatggagtt aaaggtttac aaacgacgag 4380

cggaaaagga gtcttatata attccagtag catttattcc attttgcttc acatagaaaa 4440

tgtagctgag gtggtcgatg tttattttga ctcgcgtgag gtcgccggtt cgatctcaga 4500

agcaacgctg gaatatccca tatagattta tttttcaggc tgagtgatgc tcgggtacct 4560

ccagcgtatg ggctgaaatt cgccccccct gcaaatcatg ggccctgtga cgttcgcacg 4620

ggttgcacat gccttggccc gggcctacta ggagtgtacc tggattatgt tggacgacgg 4680

gagatgaaag ggatgtatta attaacaaag ataatgaagc ttaattttct tatatgttgt 4740

taatattgac aagaaacaag ctgctaactc aaagttacgg ttacatagtc gcaacctttt 4800

atatctaaat aatatctctc tctcaacatg caaacatgcc acctcagcat gtagcatgca 4860

tggaaaattg tccacttcaa catgcaacca tgcatcaaaa tttccatttt actaggctat 4920

ttatttgata aaatttcaca aatatacaat aatcaaacac aatagatcat atgtgttttc 4980

agttttggtt ctcacattat tactccaaat ataaatgttt cgtataacca aatttcattc 5040

aaatatactg caaaacattt ccgtgaaaac atgtggggta catctagtta taaggaaata 5100

ttagtgatgt cctgcaagtg ataaggccaa ggagagaaga agtgcaccat ctacagaggc 5160

cagggaaaga caatggacat gcagagaggc gggggcgggg aagaaacaca tggagatcat 5220

agaagaacat aagaggttaa acataggagg aggatataat ggacaattaa atccacatta 5280

cctgaactca tttgggaagt ggaaaaatcc cctattctgg tgtaaatcaa actaattgac 5340

gcgagttttc tctgaagatt ctatgttaat tttagacatg aatgaccaaa ggtttcagtt 5400

agttgagttt tgtcatcgaa aggtgtttac ataagtccaa aaattctacc agcttttggt 5460

acggcgcgtc atagaacaga taaatgttgt gagtcattgg atagatatta tgagtcatag 5520

catggatttg tgttgcctgg aaatctaact atgacaagaa acaaaacata aatgggcttt 5580

tgaaagatga tttatcaact taccttatcc atgcaagcta ccttccacta gtcgacatgc 5640

ttagaagctt ttagtgaccg cagatttgca aaagcaatgg ctaacagaca cccaaacccc 5700

aagaagcata accacttctc ttagataaaa atagcagatc gatatacaaa cgtctacact 5760

tctgcaaaca atacccagaa gccagaatta ggattgaacc gattacgtgg ctttagcaga 5820

ccgtccaaaa atctgttttg caaagctcca attgctcctt gcttatccag cttcttttgt 5880

gttggcaaat tgttcttttc caaccgactt tattcttttc acatttcttc ttaggctgaa 5940

ctaacctcgt cgtgcacaca accattgtcc tgaaccttca ccacgtccct ataaaagccc 6000

aaccaatctc cacaatttca tcatcaccca caacaccgag caccacaaaa tagagatcaa 6060

ttcactgaca gtccaccgag ggatccccgg gtaccgagct cgaatttccc cgatcgttca 6120

aacatttggc aataaagttt cttaagattg aatcctgttg ccggtcttgc gatgattatc 6180

atataatttc tgttgaatta cgttaagcat gtaataatta acatgtaatg catgacgtta 6240

tttatgagat gggtttttat gattagagtc ccgcaattat acatttaata cgcgatagaa 6300

aacaaaatat agcgcgcaaa ctaggataaa ttatcgcgcg cggtgtcatc tatgttacta 6360

gatcgggaat tcgtaatcat gtcatagctg tttcctgtgt gaaattgtta tccgctcaca 6420

attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg 6480

agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg 6540

tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattggctag 6600

agcagcttgc caacatggtg gagcacgaca ctctcgtcta ctccaagaat atcaaagata 6660

cagtctcaga agaccaaagg gctattgaga cttttcaaca aagggtaata tcgggaaacc 6720

tcctcggatt ccattgccca gctatctgtc acttcatcaa aaggacagta gaaaaggaag 6780

gtggcaccta caaatgccat cattgcgata aaggaaaggc tatcgttcaa gatgcctctg 6840

ccgacagtgg tcccaaagat ggacccccac ccacgaggag catcgtggaa aaagaagacg 6900

ttccaaccac gtcttcaaag caagtggatt gatgtgatat tgccggtgaa cgacggtatg 6960

atccactctc ccaatatcct tcatacgatc tttcatcaat ataaggatat tgattttttt 7020

tggagaggac taatatcaaa taaacctcct ctcactccat tgccctctct ctcgagtctt 7080

cctcgagccc acaaacgacg cccggggtag cacctacgag tgaccgaggc ggacatgccg 7140

gcggtctgca ccatcgtcaa ccactacatc gagacaagca cggtcaactt ccgtaccgag 7200

ccgcaggaac cgcaggagtg gacggacgac ctcgtccgtc tgcgggagcg ctatccctgg 7260

ctcgtcgccg aggtggacgg cgaggtcgcc ggcatcgcct acgcgggccc ctggaaggca 7320

cgcaacgcct acgactggac ggccgagtcg accgtgtacg tctccccccg ccaccagcgg 7380

acgggactgg gctccacgct ctacacccac ctgctgaagt ccctggaggc acagggcttc 7440

aagagcgtgg tcgctgtcat cgggctgccc aacgacccga gcgtgcgcat gcacgaggcg 7500

ctcggatatg ccccccgcgg catgctgcgg gcggccggct tcaagcacgg gaactggcat 7560

gacgtgggtt tctggcagct ggacttcagc ctgccggtac cgccccgtcc ggtcctgccc 7620

gtcaccgaga tttgactcga gtttctccat aataatgtgt gagtagttcc cagataaggg 7680

aattagggtt cctatagggt ttcgctcatg tgttgagcat ataagaaacc cttagtatgt 7740

atttgtattt gtaaaatact tctatcaata aaatttctaa ttcctaaaac caaaatccag 7800

tactaaaatc cagatccccc gaattaattc ggcgttaatt cagtacatta aaaacgtccg 7860

caatgtgtta ttaagttgtc taagcgtcaa tttgtttaca ccacaatata tcctgccacc 7920

agccagccaa cagctccccg accggcagct cggcacaaaa tcaccactcg atacaggcag 7980

cccatcagtc cgggacggcg tcagcgggag agccgttgta aggcggcaga ctttgctcat 8040

gttaccgatg ctattcggaa gaacggcaac taagctgccg ggtttgaaac acggatgatc 8100

tcgcggaggg tagcatgttg attgtaacga tgacagagcg ttgctgcctg tgatcaccgc 8160

ggtttcaaaa tcggctccgt cgatactatg ttatacgcca actttgaaaa caactttgaa 8220

aaagctgttt tctggtattt aaggttttag aatgcaagga acagtgaatt ggagttcgtc 8280

ttgttataat tagcttcttg gggtatcttt aaatactgta gaaaagagga aggaaataat 8340

aaatggctaa aatgagaata tcaccggaat tgaaaaaact gatcgaaaaa taccgctgcg 8400

taaaagatac ggaaggaatg tctcctgcta aggtatataa gctggtggga gaaaatgaaa 8460

acctatattt aaaaatgacg gacagccggt ataaagggac cacctatgat gtggaacggg 8520

aaaaggacat gatgctatgg ctggaaggaa agctgcctgt tccaaaggtc ctgcactttg 8580

aacggcatga tggctggagc aatctgctca tgagtgaggc cgatggcgtc ctttgctcgg 8640

aagagtatga agatgaacaa agccctgaaa agattatcga gctgtatgcg gagtgcatca 8700

ggctctttca ctccatcgac atatcggatt gtccctatac gaatagctta gacagccgct 8760

tagccgaatt ggattactta ctgaataacg atctggccga tgtggattgc gaaaactggg 8820

aagaagacac tccatttaaa gatccgcgcg agctgtatga ttttttaaag acggaaaagc 8880

ccgaagagga acttgtcttt tcccacggcg acctgggaga cagcaacatc tttgtgaaag 8940

atggcaaagt aagtggcttt attgatcttg ggagaagcgg cagggcggac aagtggtatg 9000

acattgcctt ctgcgtccgg tcgatcaggg aggatatcgg ggaagaacag tatgtcgagc 9060

tattttttga cttactgggg atcaagcctg attgggagaa aataaaatat tatattttac 9120

tggatgaatt gttttagtac ctagaatgca tgaccaaaat cccttaacgc tgagagatcc 9180

cctcataatt tccccaaagc gtaaccatgt gtgaataaat tttgagctag tagggttgca 9240

gccacgagta agtcttccct tgttattgtg tagccagaat gccgcaaaac ttccatgcct 9300

aagcgaactg ttgagagtac gtttcgattt ctgactgtgt tagcctggaa gtgcttgtcc 9360

caaccttgtt tctgagcatg aacgcccgca agccaacatg ttagttgaag catcagggcg 9420

attagcagca tgatatcaaa acgctctgag ctgctcgttc ggctatggcg taggcctagt 9480

ccgtaggcag gacttttcaa gtctcggaag gtttcttcaa tctgcattcg cttcgaatag 9540

atattaacaa gttgtttggg tgttcgaatt tcaacaggta agttagttgc tagaacccat 9600

ggctcctttg ccgacgctga gtagatttta ggtgacgggt ggtgacaatg agtccgtgtc 9660

gagcgctgat tttttcggcc tttagagcga gatttataca atagaatttg gcatgagatt 9720

ggattgcttt tagtcagcct cttatagcct aaagtctttg agtgactaga tgacatatca 9780

tgtaagttgc tgataggttt ccagttttcc gctcctaggt ctgcatattg tacttttcct 9840

cttactcgac ttaaccagta ccaacccagc ttctcaacgg atttatacca tggcacttta 9900

aagccagcat cactgacaat gagcggtgtg gtgttactcg gtagaatgct cgcaaggtcg 9960

gctagaaatt ggtcatgagc tttctttgaa cattgctctg aaagcgggaa cgctttctca 10020

taaagagtaa cagaacgacc gtgtagtgcg actgaagctc gcaataccat aagtcgtttt 10080

tgctcacgaa tatcagacca gtcaacaagt acaatgggca tcgtattgcc cgaacagata 10140

aagctagcat gccaacggta tacagcgagt cgctctttgt ggaggtgacg attacctaac 10200

aatcggtcga ttcgtttgat gttatgtttt gttctcgctt tggttggcag gttacggcca 10260

agttcggtaa gagtgagagt tttacagtca agtaatgcgt ggcaagccaa cgttaagctg 10320

ttgagtcgtt ttaagtgtaa ttcggggcag aattggtaaa gagagtcgtg taaaatatcg 10380

agttcgcaca tcttgttgtc tgattattga tttttcgcga aaccatttga tcatatgaca 10440

agatgtgtat ccaccttaac ttaatgattt ttaccaaaat cattagggga ttcatcagcc 10500

cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt 10560

cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac 10620

cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct 10680

tcagcagagc gcagatacca aatactgtcc ttctagtgta gccgtagtta ggccaccact 10740

tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg 10800

ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata 10860

aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga 10920

cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag 10980

ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg 11040

agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac 11100

ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca 11160

acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg 11220

cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc 11280

gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcctga 11340

tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcatatgg tgcactctca 11400

gtacaatctg ctctgatgcc gcatagttaa gccagtatac actccgctat cgctacgtga 11460

ctgggtcatg gctgcgcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg 11520

tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca 11580

gaggttttca ccgtcatcac cgaaacgcgc gaggcagggt gccttgatgt gggcgccggc 11640

ggtcgagtgg cgacggcgcg gcttgtccgc gccctggtag attgcctggc cgtaggccag 11700

ccatttttga gcggcca 11717

Claims

1. The application of the protein or the related biological material thereof is any one of the following (C1) - (C3):

(C1) regulating and controlling the drought resistance of the plant;

(C2) preparing a product for improving the drought resistance of plants;

(C3) cultivating drought-resistant plants;

the amino acid sequence of the protein is shown as a sequence 1 in a sequence table;

the related biomaterial is (B1) or (B2) as follows:

(B1) a nucleic acid molecule encoding the protein;

(B2) an expression cassette, a recombinant vector, a recombinant microorganism or a transgenic plant cell line comprising the nucleic acid molecule of (B1);

the plant is wheat.

2. Use according to claim 1, characterized in that: (B1) the nucleic acid molecule is a DNA molecule shown in a sequence 2 in a sequence table.

3. Method A or method B;

the method A comprises the following steps: a method for breeding a plant variety with improved drought resistance, comprising the steps of: increasing the expression level and/or activity of the protein of claim 1 in a recipient plant to obtain a plant with increased drought resistance;

the method B comprises the following steps: a method of breeding a transgenic plant comprising the steps of: introducing into a recipient plant a nucleic acid molecule encoding the protein of claim 1 to produce a transgenic plant; the drought resistance of the transgenic plant is greater than that of a receptor plant;

the plant is wheat.

4. Method C or method D;

the method C comprises the following steps: a method of reducing drought resistance in a plant comprising the steps of: reducing the expression level and/or activity of the protein of claim 1 in a recipient plant to obtain a plant with reduced drought resistance;

the method D comprises the following steps: a method of reducing drought resistance in a plant comprising the steps of: inhibiting the expression in a recipient plant of a nucleic acid molecule encoding a protein according to claim 1, to obtain a plant with reduced drought resistance;

the plant is wheat.

5. The application of a substance for obtaining the full-length or partial section of a promoter of a coding gene of the protein in claim 1 in a plant genome to be tested in the auxiliary identification of the drought resistance of plants; the substance is a primer pair; the primer pair consists of a primer F and a primer R; the primer F is a single-stranded DNA molecule shown in a sequence 5 of the sequence table; the primer R is a single-stranded DNA molecule shown in a sequence 6 of a sequence table;

the plant is wheat.

6. A method for assisting in identifying drought resistance of a plant comprises the following steps:

(1) extracting the genome DNA of a plant to be detected;

(2) performing PCR amplification by using the genomic DNA obtained in the step (1) as a template and adopting the primer pair in claim 5, wherein if a specific band of 677 +/-10 bp exists in an amplification product, the plant is or is selected as a drought-resistant plant; if a specific band of 1378 +/-10 bp exists in the amplification product, the plant is or is selected as a drought-sensitive plant;

the plant is wheat.