CN114644699B

CN114644699B - Application of substance for regulating ZmARP1 gene expression in regulating and controlling plant drought resistance

Info

Publication number: CN114644699B
Application number: CN202011516801.5A
Authority: CN
Inventors: 巩志忠; 王瑜; 房婷婷
Original assignee: China Agricultural University
Current assignee: China Agricultural University
Priority date: 2020-12-21
Filing date: 2020-12-21
Publication date: 2023-03-28
Anticipated expiration: 2040-12-21
Also published as: CN114644699A

Abstract

The invention discloses application of a substance for regulating ZmARP1 gene expression in regulating and controlling plant drought resistance. The invention provides an application of a substance for regulating gene expression in regulating plant drought resistance, wherein the protein coded by the gene is ZmARP1 protein, and the amino acid sequence of the protein is protein of a sequence 1 in a sequence table or protein which is derived from the protein of the sequence 1 in the sequence table, has more than 80% of identity with the protein and has the same function. The invention leads ZmARP1 gene from corn 1145 into corn B73 to obtain ZmARP1 transgenic over-expression strain; overexpression of the ZmARP1 gene increased the susceptibility of transgenic maize to drought stress compared to untransformed B73 control plants. Thus indicating that ZmARP1 participates in the regulation and the adaptation of plants to drought-related adversity stress.

Description

Application of substance for regulating ZmARP1 gene expression in regulating and controlling plant drought resistance

Technical Field

The invention relates to the technical field of biology, in particular to application of a substance for regulating ZmARP1 gene expression in regulating plant drought resistance.

Background

Corn is one of three major food crops in the world, and the planting area of the corn is wide, but the corn is not uniformly distributed in each area, and different areas have different environments, different climatic conditions and different precipitation. Drought is an important factor influencing the yield of the corn, and can inhibit the growth rate of the corn in the seedling stage, so that the development stage is seriously shortened; it also inhibits the height of corn plants, causing leaf wilting, and thus reducing photosynthesis. During the corn fill stage, drought can result in under-filled kernels, which can lead to reduced corn yield.

ARP1 is a protein regulated by auxin and has an unknown functional domain, some researches about the ARP1 protein exist in plants at present, the homologous protein in Arabidopsis is SOSEKI, and previous reports show that the Arabidopsis homologous protein can participate in the polar growth of cells and can influence the direction of cell division. There have been few studies of ZmARP1 in corn.

Disclosure of Invention

The invention aims to solve the technical problem of determining the function and application of ZmARP1 protein and/or how to regulate and control the drought resistance of plants.

In order to solve the technical problem, the invention firstly provides any one of the following applications of a substance for regulating and controlling the ZmARP1 gene expression of corn:

f1, the application of the substance for regulating gene expression in regulating the drought resistance of plants,

f2, and the application of the substance for regulating gene expression in preparing the product for reducing the drought resistance of the plant,

f3, application of a substance for regulating gene expression in plant breeding;

in F1-F3, the gene encodes the following protein (named ZmARP 1) of A1), A2) or A3):

a1 Protein with the amino acid sequence of sequence 1 in the sequence table;

a2 Protein which is obtained by substituting and/or deleting and/or adding one or more amino acid residues to the amino acid sequence shown in the sequence 1 in the sequence table, is derived from A1) and has the same function, or has more than 80 percent of identity with the protein shown in A1) and has the same function;

a3 A fusion protein obtained by attaching a protein tag to the N-terminus or/and C-terminus of A1) or A2).

The plant may be corn.

In the protein, a sequence 1 in a sequence table consists of 421 amino acid residues.

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

In the above protein, the protein tag (protein-tag) refers to a polypeptide or protein that is expressed by fusion with a target protein using in vitro recombinant DNA technology, so as to facilitate expression, detection, tracking and/or purification of the target protein. The protein 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, among others.

In the above proteins, identity refers to the identity of amino acid sequences. The identity of the amino acid sequences can be determined using homology search sites on the Internet, such as the BLAST web pages of the NCBI home website. For example, in the advanced BLAST2.1, by using blastp as a program, setting the value of Expect to 10, setting all filters to OFF, using BLOSUM62 as a Matrix, setting Gap existence cost, per residual Gap cost, and Lambda ratio to 11,1, and 0.85 (default values), respectively, and performing a calculation by searching for the identity of a pair of amino acid sequences, a value (%) of identity can be obtained.

In the above protein, the 80% or more identity may be at least 81%, 82%, 85%, 86%, 88%, 90%, 91%, 92%, 95%, 96%, 98%, 99% or 100% identity.

In the above application, the substance for regulating gene expression may be a substance for regulating at least one of the following 6: 1) Regulation at the level of transcription of said gene; 2) Regulation after transcription of the gene (i.e., regulation of splicing or processing of a primary transcript of the gene); 3) Regulation of RNA transport of the gene (i.e., regulation of nuclear to cytoplasmic transport of mRNA of the gene); 4) Regulation of translation of the gene; 5) Regulation of mRNA degradation of the gene; 6) Post-translational regulation of the gene (i.e., regulation of the activity of a protein translated from the gene).

In the application, the drought resistance of the plant is regulated and controlled to improve the sensitivity of the plant to drought.

In the above application, the regulation of gene expression may be to enhance or increase the gene expression.

In the above application, the substance for regulating gene expression may be an agent for enhancing or increasing the gene expression.

The substance for regulating gene expression may be any of the following substances:

b1 Nucleic acid molecules encoding the above proteins;

b2 An expression cassette comprising the nucleic acid molecule according to B1);

b3 A recombinant vector containing the nucleic acid molecule according to B1) or a recombinant vector containing the expression cassette according to B2).

The protein also belongs to the protection scope of the invention.

In order to solve the technical problem, the invention also provides any one of the following applications of the protein:

p1, the application of the protein in regulating and controlling the drought resistance of plants,

p2 and the application of the protein in preparing products for reducing the drought resistance of plants,

p3 and application of the protein in plant breeding.

In order to solve the technical problems, the invention also provides any one of the following applications of the protein-related biological material:

q1, the application of the biological material in regulating and controlling the drought resistance of plants,

q2, application of the biological material in preparation of products for reducing drought resistance of plants,

q3 and application of the biological material in plant breeding.

The biological material is any one of the following B1) to B11):

b1 Nucleic acid molecules encoding the above proteins;

b3 A recombinant vector containing the nucleic acid molecule according to B1) or a recombinant vector containing the expression cassette according to B2);

b4 A recombinant microorganism containing the nucleic acid molecule according to B1), or a recombinant microorganism containing the expression cassette according to B2), or a recombinant microorganism containing the recombinant vector according to B3);

b5 A transgenic plant cell line containing the nucleic acid molecule according to B1) or a transgenic plant cell line containing the expression cassette according to B2);

b6 A transgenic plant tissue containing the nucleic acid molecule according to B1) or a transgenic plant tissue containing the expression cassette according to B2);

b7 A transgenic plant organ containing the nucleic acid molecule according to B1) or a transgenic plant organ containing the expression cassette according to B2);

b8 Nucleic acid molecules that promote or increase the gene expression of the above proteins;

b9 An expression cassette, a recombinant vector, a recombinant microorganism or a transgenic plant cell line containing the nucleic acid molecule according to B8);

b10 Nucleic acid molecules that inhibit or reduce gene expression of the above proteins;

b11 An expression cassette, a recombinant vector, a recombinant microorganism or a transgenic plant cell line containing the nucleic acid molecule according to B10).

In the above biological material, the nucleic acid molecule of B1) is a gene encoding the protein represented by the following B1), B2) or B3):

b1 ) the coding sequence of the coding strand is a cDNA molecule or a DNA molecule of the nucleotide of sequence 3 in the sequence table;

b2 Nucleotide is cDNA molecule or DNA molecule of sequence 3 in the sequence table;

b3 A cDNA or DNA molecule which hybridizes with a cDNA or DNA molecule defined in b 2) and encodes a protein having the same function.

In the above biological materials, the expression cassette containing a nucleic acid molecule described in B2) is a DNA capable of expressing the protein described in the above application in a host cell, and the DNA may include not only a promoter for initiating transcription of a gene encoding the protein but also a terminator for terminating transcription of a gene encoding the protein. Further, the expression cassette may also include an enhancer sequence. Promoters useful in the present invention include, but are not limited to: constitutive promoters, tissue, organ and development specific promoters, and inducible promoters. The existing plant expression vector can be used for constructing a recombinant expression vector containing the protein coding gene expression cassette. The plant expression vector comprises a binary agrobacterium vector, a vector for plant microprojectile bombardment and the like. Such as pAHC25, pWMB123, pBin438, pCAMBIA1302, pCAMBIA2301, pCAMBIA1301, pCAMBIA1300, pBI121, pCAMBIA1391-Xa or pCAMBIA1391-Xb (CAMBIA Corp.) and the like.

In the above biological material, the recombinant microorganism may be specifically yeast, bacteria, algae and fungi.

Any of the following products of the above gene-regulating substance and/or the above protein and/or the above biological material also fall within the scope of the present invention:

d1, regulating and controlling drought resistance of plants;

d2, reducing the drought resistance of the plants;

d3, improving the drought sensitivity of the plants.

As hereinbefore described, the product that reduces drought resistance in plants may be a plant stress resistance regulator. The plant stress resistance regulator can contain the protein or/and the biological material. In order to solve the technical problems, the invention also provides a method for cultivating the drought-sensitive plant, which comprises the step of increasing the biological expression quantity of the protein or/and the expression quantity of the nucleic acid molecule in the target plant to obtain the drought-sensitive plant. The drought-sensitive plant has higher drought sensitivity than the target plant. The plant of interest may be maize.

In the above method, the increase in the content of the ZmARP1 protein in the target plant or/and the expression level of the nucleic acid molecule may be achieved by introducing a gene encoding the ZmARP1 protein into the target plant.

In the above method, the stress-sensitive plant may be a transgenic plant or a plant obtained by conventional breeding techniques such as crossing.

In the above methods, the transgenic plant is understood to include not only the first to second generation transgenic plants but also the progeny thereof. For transgenic plants, the gene can be propagated in the species, and can also be transferred into other varieties of the same species, including particularly commercial varieties, using conventional breeding techniques. The transgenic plants include seeds, callus, whole plants and cells.

The drought resistance of the plant is regulated and controlled to improve the sensitivity of the plant to drought.

The plant described above may be maize.

The nucleic acid molecules described above may produce different transcripts and may be translated to different proteins, and it is within the scope of this patent that different transcripts be produced from the nucleic acid molecules and different proteins be translated.

The invention leads ZmARP1 gene from corn 1145 into corn B73 to obtain ZmARP1 transgenic over-expression strain; overexpression of the ZmARP1 gene increased the susceptibility of transgenic maize to drought stress compared to untransformed B73 control plants. Thus indicating that ZmARP1 participates in the regulation and the adaptation of plants to drought-related adversity stress.

Drawings

FIG. 1 is a control and ZmARP1 overexpression lines drought-treated phenotype.

Detailed Description

The present invention is described in further detail below with reference to specific embodiments, which are given for the purpose of illustration only and are not intended to limit the scope of the invention. The examples provided below serve as a guide for further modifications by a person skilled in the art and do not constitute a limitation of the invention in any way.

The experimental procedures in the following examples, unless otherwise indicated, are conventional and are carried out according to the techniques or conditions described in the literature in the field or according to the instructions of the products. Materials, reagents and the like used in the following examples are commercially available unless otherwise specified.

The agrobacterium strain is EHA105. The main reagents comprise: restriction enzymes, DNA polymerases, T4 ligases, etc. from biological companies such as NEB and Toyobo; reverse transcription kit from Thermo corporation; RNA extraction kit from magenta; quantitative PCR reagents from Taraka; the plasmid extraction kit and the DNA recovery kit are purchased from Tiangen corporation; MS culture medium, agar powder, agarose, ampicillin, kanamycin, gentamicin sulfate, rifampicin and other antibiotics are purchased from sigma; various other chemical reagents used in the examples are imported or domestic analytical reagents; primer synthesis and sequencing was done by invitro.

Example 1 construction of ZmARP1 Gene overexpression vector

The inventor of the application discovers a plant drought-enduring related gene in the maize ecotype 1145, wherein the nucleotide sequence of the genome gene of the plant drought-enduring related gene is shown as a sequence 2 in a sequence table, and the gene consists of 2879 basic groups. The reading frame of the gene T01 transcript is from the 1 st to the 2501 st bases of the 5' end. The transcript has 3 exons, wherein the coding exons are 2 and are respectively the 1 st to 971 st bases and the 2207 th to 2501 st bases of a reading frame, and the rest are intron sequences of the exons.

The protein coded by the gene is named ZmARP1, the amino acid sequence of the protein is shown as a sequence 1 in a sequence table, and the CDS of the cDNA gene of the ZmARP1 is shown as a sequence 3 in the sequence table.

Inserting a DNA molecule with a nucleotide sequence of sequence 3 in the sequence table into a pBCXUN vector to obtain a ZmARP1 gene expression recombinant vector, which is named as pBCXUN-ZmARP1. In pBCXUN-ZmARP1, a DNA molecule shown in a sequence 3 in a sequence table is started by a promoter of a maize ubiquitin gene Ubi and is terminated by a Nos terminator, and ZmARP1 protein can be expressed (an amino acid sequence is shown in a sequence 1 in the sequence table).

The pBCXUN vector is an expression vector obtained by replacing the HYG gene (hptII, hygromycin resistance gene) of pCXUN (GenBank: FJ905215.1, 06-JUL-2009) with a Bar gene (encoding phosphinothricin acetyltransferase) (GenBank: 284-835 th nucleotide in MG719235.1, 02-OCT-2018) and keeping the other nucleotides of pCXUN unchanged.

Example 2 acquisition of ZmARP1 Gene overexpressing plants

pBCXUN-ZmARP1 constructed in example 1 was transformed into competent Agrobacterium EHA105 strain by heat shock method, positive clones were identified by colony PCR, positive clones were selected for sequencing, and primers identified by colony PCR and sequencing were UbiP-seq (corresponding to in Ubi promoter) and NosR-seq (corresponding to in Nos terminator). The positive clone containing pBCXUN-ZmARP1 is the recombinant agrobacterium and is named as pBCXUN-ZmARP1/EHA105.

UbiP-seq:5′-TTTTAGCCCTGCCTTCATACGC-3′，

NosR-seq:5′-AGACCGGCAACAGGATTCAATC-3′。

Inoculating single pBCXUN-ZmARP1/EHA105 colony to 2-3mL liquid culture medium containing 100. Mu.g/mL kanamycin and 50. Mu.g/mL rifampicin, shake culturing at 28 deg.C overnight, inoculating to large amount of liquid culture medium containing kanamycin and rifampicin antibiotic, shake culturing the culture medium for the next day, collecting thallus after several times of inoculation, and re-suspending the thallus to OD ₆₀₀ Obtaining the recombinant agrobacterium tumefaciens suspension between 0.8 and 1.0. Using the obtained recombinant AgrobacteriumAfter the bacterial suspension infects B73 maize immature embryos scraped under the aseptic condition, screening and inducing callus to form seedlings by herbicide glufosinate, and screening by adopting a PCR identification method to obtain ZmARP1 transgenic plants (extracting genome DNA of plant leaves, and carrying out PCR amplification by adopting a primer pair consisting of Ubip-seq and NosR-seq to obtain the ZmARP1 transgenic plants of specific amplification products). The ZmARP1 transgenic plant is self-bred to obtain T3 generation stably inherited ZmARP1 gene over-expression plant for subsequent experiment.

Example 3 maize drought treatment phenotype detection with ZmARP1 Gene overexpression

The experiment contained three replicates. Each replicate contained 15 pots of control (B73) and 15 pots of ZmARP1 gene-overexpressing plants (T3 generation). Adding 140g of soil into each small pot, adding water into the tray, placing 4 seeds in each small pot, covering 50ml of soil, pouring the residual water in the tray after full water absorption, normally culturing in a culture room (the temperature is 25 ℃ and the humidity is 40 percent), removing one seedling with uneven growth after seedling emergence, adding 1L of water into the tray, pouring the water after full water absorption, and starting drought treatment without watering. The drought stress phenotype appeared at 3 weeks of treatment and the results are shown in FIG. 1, with ZmARP1 transgenic overexpressing plants (OE ZmARP1 in FIG. 1) growing worse than control B73 (WT in FIG. 1). The leaf wilting phenotype of the two material plants appears under the drought stress, the leaf wilting degree of the OE ZmARP1 plant is higher than that of the WT, the ZmARP1 transgenic plant is more sensitive to drought than a control B73 plant, and the ZmARP1 protein plays a negative regulation role in the resistance of the corn to the drought adversity stress.

The present invention has been described in detail above. It will be apparent to those skilled in the art that the invention can be practiced in a wide range of equivalent parameters, concentrations, and conditions without departing from the spirit and scope of the invention and without undue experimentation. While the invention has been described with reference to specific embodiments, it will be appreciated that the invention can be further modified. In general, this application is intended to cover any variations, uses, or adaptations of the invention following, in general, the principles of the invention and including such departures from the present disclosure as come within known or customary practice within the art to which the invention pertains. The use of some of the essential features is made possible within the scope of the claims attached below.

Sequence listing

<110> university of agriculture in China

<120> application of substance for regulating ZmARP1 gene expression in regulating and controlling plant drought resistance

<130> GNCSQ203217

<160> 3

<170> PatentIn version 3.5

<210> 1

<211> 421

<212> PRT

<213> corn (Zea mays)

<400> 1

Gly Pro Phe Tyr Pro Val Thr Val Ser Asn Gly Asn Gln Lys Gln Gln

1 5 10 15

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

20 25 30

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

35 40 45

Arg Ser Pro Ser Val Pro Ser Gln Asp Glu Glu Asp Asp Thr Pro Ser

50 55 60

Pro Cys Arg Asp Arg Ser Met Ser Gln Pro Gln Glu Leu Glu Pro Gln

65 70 75 80

Arg Asn Glu Arg Arg Thr Gln Pro Pro Ala Ser Gly Ser Ala Ser Pro

85 90 95

Val Glu Leu Arg Ala His Arg Pro Thr Thr Gly Cys Thr Asp Ala Ala

100 105 110

Thr Gln Thr Asp Asp Leu Gly Arg Arg Pro Ala Gly Arg Arg Ala Leu

115 120 125

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

130 135 140

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

145 150 155 160

Leu His Gly Ile Ser Arg Glu Leu Leu Arg Gln Cys Ile Thr Thr Pro

165 170 175

Leu Thr Val Pro Ser Thr Cys Ala Lys Ser Glu Ser Leu Glu Ser Leu

180 185 190

Ile Arg Ala Asp Asn Val Met Thr Asp Ser Phe Arg Ile Leu Glu Glu

195 200 205

Glu Asp Val Val Ala Arg Thr Cys Pro Lys Leu Arg Pro Ala Ser Val

210 215 220

Leu Met Gln Leu Val Thr Cys Gly Ser Leu Ser Val Lys Gly His Gly

225 230 235 240

Asp Ala Gly Val Val Arg Thr Cys Lys Pro Arg Phe Pro Asn Leu Arg

245 250 255

Phe Leu Pro Ser Pro Leu Ile Ser Arg Thr Met Met Met Gly Glu Leu

260 265 270

Asp Tyr Leu Ser Glu Asn Pro Arg Leu Met Gly Met Lys Leu Glu Glu

275 280 285

Lys Glu Tyr Phe Ser Gly Ser Leu Val Glu Thr Lys Lys Thr Gln Arg

290 295 300

Asp Gly Pro Ala Glu Arg Tyr Ser Ala Leu Lys Arg Ser Ser Ser Tyr

305 310 315 320

Asn Ala Glu Arg Ala Gly Glu Ala Leu Asp Cys Thr Arg Arg Glu Glu

325 330 335

Asp Lys Ala Asp Asp Val Ser Ser Arg Thr Arg Cys Leu Pro Arg Thr

340 345 350

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

355 360 365

Pro Val Ser Asp Cys Arg Arg Ser Ser Ser Ala Arg Arg Asp Tyr Asp

370 375 380

Ala Ala Ser Gly Asp Gly Ser Arg Arg Phe Ala Asp Ala Ser Val Ala

385 390 395 400

Ser Ala Thr Thr Gly Ala Glu Ser Phe Arg Lys Glu Glu Lys Leu Val

405 410 415

Lys Ile Glu Glu Ser

420

<210> 2

<211> 2879

<212> DNA

<213> corn (Zea mays)

<400> 2

ggtccatttt accctgtcac tgtcagcaac ggcaaccaga agcaacagag caggccgaaa 60

gagggcgcgc ggcagccact gccaagagac caatccgacc catcctcccc gcctagcgtg 120

acagtcagag aagccaaggc ccggcggtca ccgtccgtgc cttcgcaaga cgaggaggat 180

gacaccccgt ctccgtgcag ggatcgctcc atgtcacagc cacaggagct ggagccccag 240

cggaacgaga ggaggacgca accgccagcg agcgggtctg cgagcccagt ggagctcaga 300

gctcacaggc ccaccaccgg ctgcacggac gccgcaaccc agaccgacga tctcggcaga 360

aggccggcgg gccgcagggc gctcgagctg ccgcgcaaga agagtctgag cacggaccac 420

gacgccgtcg tccgtgagat cgcagagtac cggcagagcc accctcgccg gtcggcagac 480

ctccacggga tctcccggga gctcctgcgc cagtgcatca ctactccgct gaccgtaccc 540

tcgacctgcg ccaagtccga gagcctagag tcgctgatac gcgcggacaa cgtgatgacg 600

gacagcttca ggatcctcga agaggaggac gtcgtcgcgc gcacctgccc caagctgagg 660

ccggcgagcg tgctgatgca gctcgtcacc tgtggctcgc tctcggtgaa gggtcacggg 720

gacgctgggg tcgttcggac gtgcaagccg aggttcccca acctcaggtt cctcccctcg 780

ccgctgatct cgcgcaccat gatgatgggc gagctcgact acctgtcgga aaaccccagg 840

ttgatgggga tgaagctgga ggagaaggag tacttcagcg ggagccttgt tgagaccaag 900

aagacgcaaa gagatggtcc ggctgagagg tattcggcgc ttaaacggtc ttcttcctac 960

aatgcagaaa ggtaagcttg atcactgctt tgctatgttc ttgatatcct gcttaatcaa 1020

cttgatatct gggcacacat aaatatgtct aggctaactg ttgaaaccgt acatgattct 1080

ttgatgttat ttactggctg tcgattttga agaaaaaaat gttgcaaccc aagtgtcgag 1140

accttctaat ccatgacttc gtaccattat gcattgcaga gtttgcttgc cctcatgctt 1200

ttagcttacc aatttgattt ctgttagtta tgtttccttt tcgaaaacta tgtctcatta 1260

agagtagagc cgagccgatg acatgtgggc atttaatcca aaattttgta caaactgttg 1320

atagggacgc aaaccgttat tggttgggtt taggcttggg atcagagagt taagagttgg 1380

agaagagcca tgtactggag aatatttatg aacactagca ccattgcttt tgccgttgcc 1440

taaaaggcaa cactaacagg ttgataccag aaacactgta gggttgtcat atagcgccag 1500

catgcagctg gcatgtatct atcaaacaaa aataccaaag tacgggaatc tacctgccgg 1560

gagagaattt gaggtctgca gggtcccaaa tcggtaatga gagatttgta aatgaggcgt 1620

ggaacaacat ggcaaaacac tgccatctct ccgttttatc gatccaatat tttactttct 1680

tggaatcgta tttgaatgtt aggactgtcc ttagaacagg ggccatggct agctatggca 1740

aactgatgtg accagaaata tagatgggag ggtgatttct cctcctgtca ccaattcact 1800

tctgaaattc tggtgagatc ccctcaccct cacaacacga gccgcttgtc agttgtatag 1860

gaccacgctg ctggctgcca ggcaaggttc tcctgttgct ctactgaaat acaacagatg 1920

cgaccctgtt ggttcatcct catgaacagt cagttgtgtc cttccaagct gtcagtcagt 1980

tgtatccttg caatctgtca ttctattggc aggaccattt ttattttaaa aaaatgttgc 2040

cggctaaaaa tacagtgggt tccattccat ttatctgcgt ggctactgga tcatggtttc 2100

ttcagtatgt taaaagtcca aaatttacat ggaagaactg caacttatga atgcaatagc 2160

attttgatgg catatatgta tattttattt cataacaaat ccgcagggcc ggcgaggctc 2220

tggactgcac aagacgcgaa gaggataaag ccgacgacgt gtcgtcgcgc acgaggtgcc 2280

tcccgcggac gccgatcctg tcatccttcc tgcacccgaa gggcgactcg ctcaggtctc 2340

ccgtctcgga ctgccggcgg agctcctcgg cccggcggga ctacgacgcg gcctccgggg 2400

acgggagcag gaggttcgcc gacgcctccg tcgcgtccgc gaccacaggg gccgagtcgt 2460

tcaggaagga ggagaaactc gtcaagatcg aggaaagtta agtgccgccc gtggcctttc 2520

tttacctgaa cctactgtga actctttgtc agtgctcggt aatgcactcg atcgatgctt 2580

tcttcttgtt ttttgccccg tcgtcgtctt gacgcagtgg catcgagggc tggtattgag 2640

ggctgtgctt tgaacccttc gtgcaggctg tcgtccggag cccgtgtcgt gatccactac 2700

accgtccctt gcaacgacag cgacgaaggc tctgaacgct cggaggagtg acactgctgt 2760

tcagctagac gacccatttt ttcttcccgt tttttttttc actttctccc ttagaaacat 2820

taggccccct tatcattgta taagtgcctg ttataatgta tcattcatgt gtttcactt 2879

<210> 3

<211> 1266

<212> DNA

<213> corn (Artificial Sequence)

<400> 3

ggtccatttt accctgtcac tgtcagcaac ggcaaccaga agcaacagag caggccgaaa 60

gagggcgcgc ggcagccact gccaagagac caatccgacc catcctcccc gcctagcgtg 120

acagtcagag aagccaaggc ccggcggtca ccgtccgtgc cttcgcaaga cgaggaggat 180

gacaccccgt ctccgtgcag ggatcgctcc atgtcacagc cacaggagct ggagccccag 240

cggaacgaga ggaggacgca accgccagcg agcgggtctg cgagcccagt ggagctcaga 300

gctcacaggc ccaccaccgg ctgcacggac gccgcaaccc agaccgacga tctcggcaga 360

aggccggcgg gccgcagggc gctcgagctg ccgcgcaaga agagtctgag cacggaccac 420

gacgccgtcg tccgtgagat cgcagagtac cggcagagcc accctcgccg gtcggcagac 480

ctccacggga tctcccggga gctcctgcgc cagtgcatca ctactccgct gaccgtaccc 540

tcgacctgcg ccaagtccga gagcctagag tcgctgatac gcgcggacaa cgtgatgacg 600

gacagcttca ggatcctcga agaggaggac gtcgtcgcgc gcacctgccc caagctgagg 660

ccggcgagcg tgctgatgca gctcgtcacc tgtggctcgc tctcggtgaa gggtcacggg 720

gacgctgggg tcgttcggac gtgcaagccg aggttcccca acctcaggtt cctcccctcg 780

ccgctgatct cgcgcaccat gatgatgggc gagctcgact acctgtcgga aaaccccagg 840

ttgatgggga tgaagctgga ggagaaggag tacttcagcg ggagccttgt tgagaccaag 900

aagacgcaaa gagatggtcc ggctgagagg tattcggcgc ttaaacggtc ttcttcctac 960

aatgcagaaa gggccggcga ggctctggac tgcacaagac gcgaagagga taaagccgac 1020

gacgtgtcgt cgcgcacgag gtgcctcccg cggacgccga tcctgtcatc cttcctgcac 1080

ccgaagggcg actcgctcag gtctcccgtc tcggactgcc ggcggagctc ctcggcccgg 1140

cgggactacg acgcggcctc cggggacggg agcaggaggt tcgccgacgc ctccgtcgcg 1200

tccgcgacca caggggccga gtcgttcagg aaggaggaga aactcgtcaa gatcgaggaa 1260

agttaa 1266

Claims

1. The application of the protein in cultivating drought resistance-reduced plants;

the protein is the protein of the following A1) or A2):

a1 Protein of which the amino acid sequence is a sequence 1 in a sequence table;

a2 A fusion protein obtained by attaching a protein tag to the N-terminus or/and the C-terminus of A1);

the plant is a monocot.

2. Use according to claim 1, characterized in that: the plant is corn.

3. Use of a biological material related to a protein as defined in claim 1 or 2 for breeding drought-resistant reduced plants;

the biological material is any one of the following materials:

b1 A nucleic acid molecule encoding the protein of claim 1;

b2 An expression cassette containing the nucleic acid molecule according to B1);

the plant is a monocot.

4. Use according to claim 3, characterized in that: b1 The nucleotide sequence of the nucleic acid molecule is shown as a sequence 3 in a sequence table.

5. Use according to claim 3 or 4, characterized in that: the plant is corn.

6. A method for cultivating a drought-sensitive plant, comprising increasing the biological expression level of the protein of claim 1 or 2 and/or the expression level of the nucleic acid molecule of claim 3 or 4 in a plant of interest to obtain a drought-sensitive plant; the drought-sensitive plant has higher sensitivity to drought than the target plant; the plant is a monocot.

7. The method of claim 6, wherein: the plant is corn.