CN114716521B - Maize drought-resistant related protein and application thereof in plant drought resistance - Google Patents

Abstract

The invention discloses a corn drought resistance-related protein and its application in plant drought resistance. The protein is GRMZM2G121851. After the GRMZM2G121851 protein is overexpressed in plants, the growth of the overexpressed plants under drought treatment conditions is significantly better than that of the wild type, indicating that this Gene overexpression can significantly improve plant drought resistance. In the embodiments of the present invention, transgenic overexpression technology is used to obtain drought-resistant plants, which have good growth conditions under drought conditions, and their leaf wilting degree is significantly lower than that of the wild type. Compared with traditional breeding methods, the time is shorter and the purpose is stronger. It provides genetic resources for cultivating and improving new drought-resistant plant varieties, and provides a theoretical basis for elucidating the molecular mechanism of this gene in plant drought stress signal response.

Description

Corn drought resistance-related proteins and their application in plant drought resistance

Technical field

The invention belongs to the field of biotechnology, and specifically relates to a corn drought resistance-related protein and its application in plant drought resistance.

Background technique

More than half of my country's corn is grown on dry land in the northwest, southwest, north China and northeastern regions that rely on natural precipitation. Water loss is rapid and precipitation variability is also large, seriously affecting the growth of corn. Corn is an important food crop in my country, and research on its drought resistance is of great significance to solving the problem of corn yield in my country. We aimed to find drought-related genes in corn overexpression lines and study their gene functions and signal transduction pathways. Through research on drought-related genes, corn can be improved to improve its drought resistance and increase its yield.

Drought can cause osmotic stress in plants. The osmotic pressure in the plant is lower than the osmotic pressure of the environment (such as soil solution), and the plant cannot absorb water or even lose water. There are two pathways for osmotic stress: ABA-dependent and ABA-independent pathways. When subjected to environmental stress, ABA is produced depending on the ABA pathway. PYL, PYR, and RCAR are ABA receptors that can bind to ABA. The protein phosphatase activity of PP2C is inhibited, allowing SnRK2 to have kinase activity, which can phosphorylate downstream transcription factors, etc., thereby regulating the expression of downstream stress-responsive genes, inhibiting stomatal opening, regulating ABA sensitivity, and producing other responses. . In an ABA-independent pathway, DREB2A is phosphorylated, thereby regulating the expression of downstream stress-responsive genes. Drought stress can induce the production of abscisic acid (ABA) in plants, which can promptly guide stomata to close and reduce water loss in plants. At the same time, it can also activate the expression of a large number of drought-related genes to cause various drought stress responses in plants. As an extremely important plant hormone, ABA plays a key role in stomatal closure and expression of stress-responsive genes.

Contents of the invention

The purpose of the present invention is to provide a corn GRMZM2G121851 protein and its application in regulating plant drought resistance.

The present invention provides a protein that is primarily related to corn drought resistance. The protein is GRMZM2G121851 protein, and the GRMZM2G121851 protein is any one of the following proteins b1) to b3):

b1) The amino acid sequence is the protein of SEQ ID No. 2 in the sequence listing;

b2) A protein with the same biological function obtained by substituting and/or deleting and/or adding one or several amino acid residues to the amino acid sequence shown in SEQ ID No. 2 in the sequence listing;

b3) A protein that has 80% or more identity with the amino acid sequence defined by SEQ ID No. 2 in the sequence listing, originates from corn and has the same biological function.

The present invention also provides related biological materials of the GRMZM2G121851 protein, and the related biological materials are any one of the following B1) to B9):

B1) Nucleic acid molecule encoding the GRMZM2G121851 protein of claim 1;

B2) An expression cassette containing the nucleic acid molecule described in B1);

B3) A recombinant vector containing the nucleic acid molecule described in B1), or a recombinant vector containing the expression cassette described in B2);

B4) A recombinant microorganism containing the nucleic acid molecule described in B1), or a recombinant microorganism containing the expression cassette described in B2), or a recombinant microorganism containing the recombinant vector described in B3);

B5) A transgenic plant cell line containing the nucleic acid molecule described in B1), or a transgenic plant cell line containing the expression cassette described in B2);

B6) Transgenic plant tissue containing the nucleic acid molecule described in B1), or transgenic plant tissue containing the expression cassette described in B2);

B7) Transgenic plant organs containing the nucleic acid molecule described in B1), or transgenic plant organs containing the expression cassette described in B2);

B8) Nucleic acid molecules that increase or promote the expression of the GRMZM2G121851 protein;

B9) Expression cassette, recombinant vector, recombinant microorganism or transgenic plant cell line containing the nucleic acid molecule described in B8).

The above-mentioned protein can be synthesized artificially, or its encoding gene can be first synthesized and then biologically expressed.

In the above-mentioned proteins, identity refers to the identity of the amino acid sequence. The identity of the amino acid sequence can be determined using homology search sites on the Internet, such as the BLAST web page of the NCBI homepage. For example, in advanced BLAST2.1, you can use blastp as the program, set the Expect value to 10, set all Filters to OFF, use BLOSUM62 as the Matrix, and set the Gap existence cost, Per residue gap cost, and Lambda ratio to respectively 11, 1 and 0.85 (default value) and search for the identity of a pair of amino acid sequences to calculate, and then the identity value (%) can be obtained.

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

The related biological materials of the GRMZM2G121851 protein in any one of the above B1) to B9) also fall within the protection scope of the present invention. Wherein, the nucleic acid molecule can be DNA, such as cDNA, genomic DNA or recombinant DNA; the nucleic acid molecule can also be RNA, such as mRNA or hnRNA, etc.

The plant expression vector can specifically be a pBCXUN vector.

The nucleic acid molecule of B1) is any one of the following a1)-a3):

a1) DNA molecule with coding region as shown in SEQ ID No.3;

a2) The nucleotide sequence is the DNA molecule shown in SEQ ID No. 1 in the sequence listing;

a3) A DNA molecule that has 90% or more identity with the nucleotide sequence defined by a1) or a2), originates from corn and encodes the GRMZM2G121851 protein described in claim 1.

The above-mentioned "identity" refers to sequence similarity with natural nucleic acid sequences. "Identity" includes nucleotide sequences that are 90% or higher, or 95% or higher identical to the nucleotide sequence encoding the GRMZM2G121851 protein of the invention.

Alternatively, the maize GRMZM2G121851 gene consists of 4945 nucleotides, and the reading frame of the T01 transcript is from 5' end 745th to 3970th nucleotide. The gene consists of 7 exons, of which the T01 transcript encodes 7 exons, with the reading frame ranging from nucleotides 1 to 158, 265 to 340, and 825 to 340. Nucleotide 877, Nucleotide 968 to 1458, Nucleotide 2039 to 2199, Nucleotide 2314 to 2514, Nucleotide 2636 to 3226 , and the rest are intronic sequences. The gene is derived from B73-329 maize, and its number in the maize genome database is GRMZM2G121851. Since the same DNA sequence in corn can produce different transcripts and translate into different proteins, the different transcripts produced by this sequence and the different proteins translated into it are within the scope of this patent protection.

The application of the protein or the related biological material in regulating plant drought resistance should also be within the protection scope of the present invention.

The application of the protein or the related biological material in breeding drought-resistant plant varieties should also be within the protection scope of the present invention.

The present invention also provides a method for cultivating highly drought-resistant plants, which includes promoting or improving the expression of the nucleic acid of the GRMZM2G121851 protein in a target plant to obtain a highly drought-resistant plant, and the target plant is a plant containing the nucleic acid of the GRMZM2G121851 protein. , the drought resistance of the highly drought-resistant plant is higher than the drought resistance of the target plant.

The present invention also provides a plant breeding method, which includes the following steps: increasing the content and/or activity of the protein according to claim 1 in the target plant, thereby increasing the drought resistance of the plant.

In the above, the plant is a monocotyledonous plant or a dicotyledonous plant.

The plant is a gramineous plant.

The plant is corn.

After the GRMZM2G121851 gene provided by the invention is overexpressed in plants, the growth of the overexpressed plants under drought treatment conditions is significantly better than that of the wild type, indicating that overexpression of this gene can significantly improve plant drought resistance. In the embodiments of the present invention, transgenic overexpression technology is used to obtain drought-resistant plants, which have good growth conditions under drought conditions, and their leaf wilting degree is significantly lower than that of the wild type. Compared with traditional breeding methods, the time is shorter and the purpose is stronger. It provides genetic resources for cultivating and improving new drought-resistant plant varieties, and provides a theoretical basis for elucidating the molecular mechanism of this gene in plant drought stress signal response.

Description of the drawings

Figure 1 shows the drought treatment phenotypes of the control and GRMZM2G121851 overexpression lines.

Detailed ways

The present invention will be described in further detail below in conjunction with specific embodiments. The examples given are only for illustrating the present invention and are not intended to limit the scope of the present invention. The examples provided below can serve as a guide for those of ordinary skill in the art to make further improvements, and do not limit the present invention in any way.

The experimental methods in the following examples, unless otherwise specified, are all conventional methods and are carried out in accordance with the techniques or conditions described in literature in the field or in accordance with product instructions. Materials, reagents, etc. used in the following examples can all be obtained from commercial sources unless otherwise specified.

The corn ecotype is B73; the Agrobacterium strain is EHA105. The main reagents include: restriction endonucleases, DNA polymerase, T4 ligase, etc. from biological companies such as NEB and Toyobo; reverse transcription kits from Thermo; RNA extraction kits from Magen; quantitative PCR reagents from Taraka; Plasmid extraction kit and DNA recovery kit were purchased from Tiangen Company; MS culture medium, agar powder, agarose, ampicillin, kanamycin, gentamicin sulfate, rifampicin antibiotics and other reagents were purchased from sigma; implementation Various other chemical reagents used in the examples are all imported or domestic analytical pure reagents; primer synthesis and sequencing were completed by Yingjun Company.

Example 1

1. Discovery of corn GRMZM2G121851 protein

A new protein related to plant drought tolerance was discovered in corn B73-329, named GRMZM2G121851. Its amino acid sequence is shown in SEQ ID No. 2 in the sequence listing.

In the maize B73-329 genomic DNA, the corresponding maize GRMZM2G121851 gene consists of 4945 nucleotides as shown in SEQ ID No. 1 in the list, and the reading frame of the transcript is from the 5' end of position 745 to position 3970 Nucleotides. The gene consists of 7 exons, of which the transcript encodes 7 exons. The reading frame includes nucleotides 1 to 158, 265 to 340, and 825 to 340. Nucleotide 877, nucleotide 968 to 1458, nucleotide 2039 to 2199, nucleotide 2314 to 2514, nucleotide 2636 to 3226, The rest are intronic sequences.

2. Construction and detection of GRMZM2G121851 gene overexpression vector

2.1 Construction of GRMZM2G121851 gene overexpression vector

The sequences of the seven exons in the corn GRMZM2G121851 gene are connected sequentially to form a DNA molecule shown in Sequence 3, which can encode a protein with an amino acid sequence shown in Sequence 2.

The DNA molecule shown in sequence 3 was inserted into the pBCXUN vector to obtain the recombinant plasmid pBCXUN-GRMZM2G121851. In the recombinant plasmid pBCXUN-GRMZM2G121851, the DNA molecule shown in Sequence 3 of the sequence list is started by the Ubi promoter and terminated by the Nos terminator, thereby expressing the GRMZM2G121851 protein.

The pBCXUN vector replaces the HYG gene (hptII, hygromycin resistance gene) of the pCXUN vector (GenBank: FJ905215.1, 06-JUL-2009) with the Bar gene (encoding phosphinothricin acetyltransferase) (GenBank: MG719235 .1, 02-OCT-2018), keeping other nucleotides of pCXUN unchanged.

2.2 Verification of recombinant vector

Transform the recombinant vector pBCXUN-GRMZM2G121851 into E. coli competent cells. Screen on LB plates containing 50 μg/mL kanamycin. Colony PCR was used to identify single clones, and positive clones were selected for sequencing. The obtained recombinant expression vector with correct sequencing was named pBCXUN-GRMZM2G121851. Common primers for colony PCR and sequencing are as follows:

UbiP-seq:TTTTAGCCCTGCCTTCATACGC

NosR-seq:AGACCGGCAACAGGATTCAATC.

3. Construction of GRMZM2G121851 gene overexpression plants

3.1 Transform the pBCXUN-GRMZM2G121851 overexpression plasmid constructed in step 2 into the competent Agrobacterium EHA105 strain using the heat shock method. Colony PCR identifies positive clones to obtain recombinant Agrobacterium.

3.2 Inoculate a single colony of recombinant Agrobacterium into 2-3 mL of liquid culture medium containing 100 μg/mL kanamycin and 50 μg/mL rifampicin, culture it overnight at 28°C with shaking, and transfer to a large amount of liquid culture containing antibiotics the next day. Culture in the medium with shaking, transfer the cells several times, collect the cells, and resuspend until the OD ₆₀₀ is between 0.8-1.0. The obtained recombinant Agrobacterium suspension was used to infect the young B73 corn embryos picked out under aseptic conditions, and the callus was induced to grow into seedlings to obtain T ₀ generation regenerated plants.

3.3 The transgenic plants were self-propagated to obtain the T3 generation for subsequent experiments. The cultivation method of T3 generation plants is as follows: conduct PCR identification on T ₀ generation regenerated plants, and screen to obtain transgenic plants; self-cross the T ₀ generation transgenic plants, and the seeds obtained are T ₁ generation seeds, and the seeds grown from T ₁ generation seeds are The plant is the T ₁ generation plant; by selfing the T ₁ generation plant, the seeds obtained are the T ₂ generation seeds, and the plants grown from the T ₂ generation seeds are the T ₂ generation plants; by selfing the T ₂ generation plants, we get The seeds are T ₃ generation seeds, and the plants grown from T ₃ generation seeds are T ₃ generation plants. The screening method for transgenic plants is as follows: extract the genomic DNA from plant leaves, and perform PCR amplification using the primer pair composed of UbiP-seq and NosR-seq in step 2.2. If a specific amplification product is obtained, the plant is a transgenic plant.

4. Phenotypic detection of drought treatment in corn overexpressed with GRMZM2G121851 gene

The seeds of the transgenic lines OE1 and OE2 in the T3 generation plants were selected, and the seeds of corn B73 were used as wild-type controls to conduct phenotypic detection of corn drought treatment. The control and transgenic plants were replicated in three pots each, and the experiment was repeated three times.

The specific steps are:

1. Add 100g of soil to each small pot, add water to the tray, put 4 seeds in each small pot, cover with 50ml of soil, drain out the remaining water in the tray after it is full of water, and remove the uneven growth after the seedlings emerge. Remove the seedlings, add 1L of water to the tray, pour out the water when it is full, and start the drought treatment. Observe the drought treatment phenotypes of the control and transgenic plants. The results after one week are shown in Figure 1. Figure 1 shows that the growth status of transgenic plants overexpressing GRMZM2G121851 is better than that of the control, and the degree of leaf wilting is lower than that of the control, indicating that the transgenic plants are more drought-resistant than the control.

2. On the basis of step 1, continue without watering for 20 days, resume normal watering and culture for 7 days, and then calculate the survival rate. The survival rate is the percentage of surviving plants to the total number of plants. The results showed that the survival rate of the transgenic line OE1 in the T3 generation plants was 68.95%, the survival rate of the transgenic line OE2 in the T3 generation plants was 70.11%, and the survival rate of the wild type was 36.25%.

The present invention has been described in detail above. For those skilled in the art, the present invention can be implemented in a wider range under equivalent parameters, concentrations and conditions without departing from the spirit and scope of the invention and without performing unnecessary experiments. Although specific embodiments of the present invention have been shown, it should be understood that further modifications can be made to the invention. In short, based on the principles of the present invention, this application is intended to include any changes, uses, or improvements to the present invention, including changes that depart from the scope disclosed in this application and are made using conventional techniques known in the art. Some essential features may be applied within the scope of the appended claims below.

sequence list

<110> China Agricultural University

<120> Corn drought resistance-related proteins and their application in plant drought resistance

<160> 3

<170> SIPOSequenceListing 1.0

<210> 1

<211> 4945

<212> DNA

<213> Artificial Sequence

<400> 1

ggatcagttc cacgcctgat ccttcgttaa attattctcg ctttgacgcc acagcctttt 60

gcactggcat gtgcttaccc tctgagggaa cgtgccatgt ctacccttcc tgcagtgaca 120

tctttattat ctctacaccc ctttcctgtt atgagtgtat ttactgttcc tctacttggc 180

ctggtctcga tctgatcgaa ctcttggctg ttgttacggc tgcgaaaaaa gagttgcctg 240

caggctgcag tccgccaatt catcgacgcc tgcaagttgt ccaggttcaa tgcaccgata 300

ttctattgca cgtggttgca aagtttatgt aatttattta tgttgttttg gttgatgggt 360

tgcattgttg caagattatt ttagcttggt caccggtcag gaattgtggc tggtatctta 420

aaacggaaca aagcagacca cttgctgtta ctaggctctt aattaaaaga atgctggcct 480

ggggccgacc cggcctgcct ccgagtggac cggtcgtgcc gtaccgccca tctgcccgcc 540

cccaacggcc caccagagga acccgaggtc attttgtgga cgggaaagga aaagaaaagt 600

aatgtatttc catggaaccg aacgcgtgct gcgcctcctg tacggcgcgg aggtaggtca 660

agccgcggtt gggacgcggt gtggcgtggg cgagccgaac caagcggcga tcgatcgaaa 720

cgaggcgtcg cgtggcagcc ggagatgtcg cagaagaggc agccggagga gggcgacgtc 780

ccgcggcgcg ccgccgatgc cggcggcggc ggcggaggag acgagcccgg tgggagctcg 840

tcgcgctcct ctctgcccca gcgccacggc gagccgaagc ggcagaggat cgcccttcgc 900

gagtgagttt gtcctccccc gcgcgccctg gattgggtcg cgccgattcg tgcgccgcgc 960

tctctgattc gatgcgccgt ggtgacttgg attgttcccc ccttgcagtg tgatcacgga 1020

ggtgatgcgg aacaccagca tcgagaagtt tctaatcgcg ctcgagcccc tcatcaggag 1080

agtggtgaga ccgccttttt tatcttttac tatctgcccc atttctcact tcagtccttc 1140

ttgaatggaa attcatagga ttggatagtg atctgatgtg agccctctag gttgtttgcc 1200

tttcctcttg ttggatacat aacgatacat agtgctgttg taatcccatc tgcatgtgtg 1260

gctgattggc actctgatat tggttttggc aagagcaagt tcttctttcc ctcccaaact 1320

actatctgag aagctaaggc atttatctgt atcgtattaa tagctctata ggctttgctg 1380

tcatgattgt atccataatg attccagcct tacaaaataa ggcttggctt cactgtttcc 1440

gatgtgttca gccattccaa ttcagtgata attttacagt gggctacatc atatgttctc 1500

cttgaaccat ttgcttattt tagccatcta ctgctaccag tggggtttga tagtctattc 1560

atttctaggt aaaagaagaa attgagtcgg cttttgcaaa ccatgcctct atgatggcaa 1620

ggtatggagt ttaaacactg cttttgacaa ccaccatgat ttgttatattgg tcgctgtatc 1680

tctaataagt ttatttctta tttttcatta ggagtgtcac agacagtgtt ccatctgtgt 1740

caaagaattt gcagctgcag ttcatgacca gactttctct tccaatattt actggatcca 1800

agattgaagg agagggctcc ttaagtataa ctattgctct agtcgacgct ttgacaagac 1860

aaattgtagc accaggcaaa gagttccaga taaaggtcga gattgtagtt ctggaggggg 1920

attttgaaag tggagaagat gatgactgga cagctcagga gtttaacaat aacattgtta 1980

aagaaagaga aggcaaaagg cccttgattt ctggggatgc attcattgcc ctcgtcgatg 2040

gcattggaac agtaggggaa ctttcattca cagataactc cagctggaca cggagccgaa 2100

agttcaggct aggagcaaga acagaggatg gttctttcaa tggtgtaaga gtacgggaag 2160

caaaaactga atcatttgtg gttaaggacc atcgaggaga atgtgagttt attattggat 2220

ttttaaaatt ttaatgccca agctatcctt ttactttgtt gcctttttgc tctgctagac 2280

tcattctagg ttcaagaacg atttggcgta gtggtactta attcaacaat ttcatgaaaa 2340

ataaatgaat tctttcttgt cctctactgt ctagacttag ttctagttga ggaagactag 2400

ttgaggtgga aagtatgatc atgcacccaa tcttcaccga ggtttgtgtc ctcttgaact 2460

taaatttggg tgcctatttt cttcttaggg ctagtttggg aaccacattt tcccaagggt 2520

ttttcgtttt cccaaggtaa attagttcat tttcccttgg gaaattgaaa accccatgga 2580

aaaatgcggt tgccaaacta gcccttaatg aaaaaaccta gtttttccta gtatgttctc 2640

aactttcccc tctagactta gttttgtagt atgtttcctt aaatatctac tgatctacat 2700

ctccaattga atgatgagac atgctatttc cctgaatact tctaaaatgg tgtaatttaa 2760

ttgattacct cctattgacc agtgtacaag aagcaccacc caccatttct tgaagatgaa 2820

gtctggcgcc tagagaaaat tggcaaggaa ggtgcttttc acaagcgttt gaataggggag 2880

aacatttgca ctgtcaaaga ttttctcacc ttgttaaatc ttgatgcttc taggcttcga 2940

aaggtattgt tcttgtcttc tattgttttt ctgatgttct gtatattgtt gtctatggta 3000

gttgcagttc agcaaagatg tgtattacta accctgcttt atgtccaaat cgtgcagata 3060

ttaggtggtg gcatgtcaac aaagatgtgg gaggccactg tggaacatgc aaaaacatgt 3120

gttctaactg acaaagtgca tcactactat cctgatggtc taaacaaagc tggtgttgta 3180

ttcaatgtag ttggagaagt aagagggtta atatctgata aatatgtttt tgttgatgac 3240

tttactgaaa aggagaaggt aaccatgttg cattgctttc caagttttct ttaaacaagt 3300

ccatccattt tccttatatg cccacctttt ctgacccgga ccatgtgttt ttacatgttt 3360

gctgtttgcc tattttcagg ccgaagcacg tgcagcagtg aagcaagcat atgaacactg 3420

gaaggatgtc catacttgtg acaatgaaac gcttgtggaa aacccttcac atccattcaa 3480

tttggggatct ccatctttgc atgaaaatca gtataaccag ttgcccacac aagtttctac 3540

tgatggtttt agtttgagca attcggccat actatcaccc aacattttct caatggagcc 3600

atcgagtgct ttagaccctt gtagtatatt ggagactgaa gaaagcagtg ctaatcaatt 3660

tcagtcggtg ttgcccccag ttggtggcca tgaagtgccc cgagaacctc agacgctgga 3720

taagttctcc aactcgttgg tatatgatga ctgcagcgcc catccctcat tcagtgaaag 3780

ctattacagc actgtagatc caagcatgtc cttcgacaca caagatcttg gagctgcact 3840

gaaaggcttc attgcaacca tatcgaagcc taaggcggca tatagaggat ggagaacatt 3900

gtcttatgtg ctaggatgga ttttctatac caagagaatt gttgcaagga gaaagaaaca 3960

tgggaaataa gtgaacacaa gtctttgtgt agttaaatgc gaatataagc ttagagaatg 4020

agagattatg tacattagtt tgtttcttgt cacagctggt aagctctgct ggtttcttca 4080

gctgtacatg ggctgtcagc tatggttcgc tatgcggctg taaataatca aatgaagttt 4140

gtacagggtg agggatgcgc atagcatttc tgatgttttg gatggatgcc ggagcttgct 4200

ttttgatgcg gcccgtgtat tatctatgta gtagtatgca gtatgctgct taattctgca 4260

gtattttcga catttggaat ctatatgtcg catggtaaca agtttaattc tgcttatttt 4320

cgtacacaga taatactgca gaattaccat gccgcaaact gatttttatg tagacagtaa 4380

tgtgtttatg gatatggttg acacttaaat aaaaagctgg gcagcgtgca catgccattt 4440

acaatagaga tttgccttgc gcgggaaatt gtattccatt tacagtagag tcttgccttg 4500

aacaacaatg ttcgatgtgg tgggataact gacttgatcc tacaactacc atagtagcta 4560

atctccccgt tgctcgtatg actagtaaca cacttgagga gcacagataa acatattgct 4620

gctactctgg atatacgggt atgcgtactt cttgttgtgc ttttcttcta ctatgctgtg 4680

tgcctagtga taataattta tgatcctcta ctcacacgtt atcttgcttg gtatggtaaa 4740

tatgctagtg catagtgtaa ttaaaaatgg ttgttttacg agaataatat attgatttcc 4800

gttttaatgc atgcactttc atttaaaatt gagttgagta gaacttgaag taatctataa 4860

aaaatgctgt acctctagaa caattggcac aaggtcctgg actgtttttt gtatggttct 4920

tatattgaat agcacaaaaa aattt 4945

<210> 2

<211> 576

<212> PRT

<213> Artificial Sequence

<400> 2

Met Ser Gln Lys Arg Gln Pro Glu Glu Gly Asp Val Pro Arg Arg Ala

1 5 10 15

Ala Asp Ala Gly Gly Gly Gly Gly Gly Gly Asp Glu Pro Gly Gly Ser Ser

20 25 30

Ser Arg Ser Ser Leu Pro Gln Arg His Gly Glu Pro Lys Arg Gln Arg

35 40 45

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

50 55 60

Glu Lys Phe Leu Ile Ala Leu Glu Pro Leu Ile Arg Arg Val Val Lys

65 70 75 80

Glu Glu Ile Glu Ser Ala Phe Ala Asn His Ala Ser Met Met Ala Arg

85 90 95

Ser Val Thr Asp Ser Val Pro Ser Val Ser Lys Asn Leu Gln Leu Gln

100 105 110

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

115 120 125

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

130 135 140

Arg Gln Ile Val Ala Pro Gly Lys Glu Phe Gln Ile Lys Val Glu Ile

145 150 155 160

Val Val Leu Glu Gly Asp Phe Glu Ser Gly Glu Asp Asp Asp Trp Thr

165 170 175

Ala Gln Glu Phe Asn Asn Asn Ile Val Lys Glu Arg Glu Gly Lys Arg

180 185 190

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

195 200 205

Thr Val Gly Glu Leu Ser Phe Thr Asp Asn Ser Ser Trp Thr Arg Ser

210 215 220

Arg Lys Phe Arg Leu Gly Ala Arg Thr Glu Asp Gly Ser Phe Asn Gly

225 230 235 240

Val Arg Val Arg Glu Ala Lys Thr Glu Ser Phe Val Val Lys Asp His

245 250 255

Arg Gly Glu Leu Tyr Lys Lys His Pro Pro Phe Leu Glu Asp Glu

260 265 270

Val Trp Arg Leu Glu Lys Ile Gly Lys Glu Gly Ala Phe His Lys Arg

275 280 285

Leu Asn Arg Glu Asn Ile Cys Thr Val Lys Asp Phe Leu Thr Leu Leu

290 295 300

Asn Leu Asp Ala Ser Arg Leu Arg Lys Ile Leu Gly Gly Gly Met Ser

305 310 315 320

Thr Lys Met Trp Glu Ala Thr Val Glu His Ala Lys Thr Cys Val Leu

325 330 335

Thr Asp Lys Val His His Tyr Tyr Pro Asp Gly Leu Asn Lys Ala Gly

340 345 350

Val Val Phe Asn Val Val Gly Glu Val Arg Gly Leu Ile Ser Asp Lys

355 360 365

Tyr Val Phe Val Asp Asp Phe Thr Glu Lys Glu Lys Ala Glu Ala Arg

370 375 380

Ala Ala Val Lys Gln Ala Tyr Glu His Trp Lys Asp Val His Thr Cys

385 390 395 400

Asp Asn Glu Thr Leu Val Glu Asn Pro Ser His Pro Phe Asn Leu Gly

405 410 415

Ser Pro Ser Leu His Glu Asn Gln Tyr Asn Gln Leu Pro Thr Gln Val

420 425 430

Ser Thr Asp Gly Phe Ser Leu Ser Asn Ser Ala Ile Leu Ser Pro Asn

435 440 445

Ile Phe Ser Met Glu Pro Ser Ser Ala Leu Asp Pro Cys Ser Ile Leu

450 455 460

Glu Thr Glu Glu Ser Ser Ala Asn Gln Phe Gln Ser Val Leu Pro Pro

465 470 475 480

Val Gly Gly His Glu Val Pro Arg Glu Pro Gln Thr Leu Asp Lys Phe

485 490 495

Ser Asn Ser Leu Val Tyr Asp Asp Cys Ser Ala His Pro Ser Phe Ser

500 505 510

Glu Ser Tyr Tyr Ser Thr Val Asp Pro Ser Met Ser Phe Asp Thr Gln

515 520 525

Asp Leu Gly Ala Ala Leu Lys Gly Phe Ile Ala Thr Ile Ser Lys Pro

530 535 540

Lys Ala Ala Tyr Arg Gly Trp Arg Thr Leu Ser Tyr Val Leu Gly Trp

545 550 555 560

Ile Phe Tyr Thr Lys Arg Ile Val Ala Arg Arg Lys Lys His Gly Lys

565 570 575

<210> 3

<211> 1731

<212> DNA

<213> Artificial Sequence

<400> 3

atgtcgcaga agaggcagcc ggaggagggc gacgtcccgc ggcgcgccgc cgatgccggc 60

ggcggcggcg gaggagacga gcccggtggg agctcgtcgc gctcctctct gccccagcgc 120

cacggcgagc cgaagcggca gaggatcgcc cttcgcgatg tgatcacgga ggtgatgcgg 180

aacaccagca tcgagaagtt tctaatcgcg ctcgagcccc tcatcaggag agtggtaaaa 240

gaagaaattg agtcggcttt tgcaaaccat gcctctatga tggcaaggag tgtcacagac 300

agtgttccat ctgtgtcaaa gaatttgcag ctgcagttca tgaccagact ttctcttcca 360

atatttactg gatccaagat tgaaggagag ggctccttaa gtataactat tgctctagtc 420

gacgctttga caagacaaat tgtagcacca ggcaaagagt tccagataaa ggtcgagatt 480

gtagttctgg agggggattt tgaaagtgga gaagatgatg actggacagc tcaggagttt 540

aacaataaca ttgttaaaga aagagaaggc aaaaggccct tgatttctgg ggatgcattc 600

attgccctcg tcgatggcat tggaacagta ggggaacttt cattcacaga taactccagc 660

tggacacgga gccgaaagtt caggctagga gcaagaacag aggatggttc tttcaatggt 720

gtaagagtac gggaagcaaa aactgaatca tttgtggtta aggaccatcg aggagaattg 780

tacaagaagc accacccacc atttcttgaa gatgaagtct ggcgcctaga gaaaattggc 840

aaggaaggtg cttttcacaa gcgtttgaat agggagaaca tttgcactgt caaagatttt 900

ctcaccttgt taaatcttga tgcttctagg cttcgaaaga tattaggtgg tggcatgtca 960

acaaagatgt gggaggccac tgtggaacat gcaaaaacat gtgttctaac tgacaaagtg 1020

catcactact atcctgatgg tctaaacaaa gctggtgttg tattcaatgt agttggagaa 1080

gtaagagggt taatatctga taaatatgtt tttgttgatg actttactga aaaggagaag 1140

gccgaagcac gtgcagcagt gaagcaagca tatgaacact ggaaggatgt ccatacttgt 1200

gacaatgaaa cgcttgtgga aaacccttca catccattca atttggggatc tccatctttg 1260

catgaaaatc agtataacca gttgcccaca caagtttcta ctgatggttt tagtttgagc 1320

aattcggcca tactatcacc caacattttc tcaatggagc catcgagtgc tttagaccct 1380

tgtagtatat tggagactga agaaagcagt gctaatcaat ttcagtcggt gttgccccca 1440

gttggtggcc atgaagtgcc ccgagaacct cagacgctgg ataagttctc caactcgttg 1500

gtatatgatg actgcagcgc ccatccctca ttcagtgaaa gctattacag cactgtagat 1560

ccaagcatgt ccttcgacac acaagatctt ggagctgcac tgaaaggctt cattgcaacc 1620

atatcgaagc ctaaggcggc atatagagga tggagaacat tgtctttatgt gctaggatgg 1680

attttctata ccaagagaat tgttgcaagg agaaagaaac atgggaaata a 1731

Claims (4)

1. Application of proteins related to corn drought resistance in improving corn drought resistance; the protein is GRMZM2G121851 protein, and the GRMZM2G121851 protein is a protein whose amino acid sequence is shown in SEQ ID No. 2. 2. Application of biological materials related to the GRMZM2G121851 protein described in claim 1 in improving the drought resistance of corn; The relevant biological material is any one of the following B1) to B4): B1) DNA molecule with coding region as shown in SEQ ID No.3; B2) An expression cassette containing the nucleic acid molecule described in B1); B3) A recombinant vector containing the nucleic acid molecule described in B1), or a recombinant vector containing the expression cassette described in B2); B4) A recombinant microorganism containing the nucleic acid molecule described in B1), or a recombinant microorganism containing the expression cassette described in B2), or a recombinant microorganism containing the recombinant vector described in B3). 3. A method for cultivating highly drought-resistant corn, which is characterized by: overexpressing the GRMZM2G121851 protein described in claim 1 in the target corn to obtain highly drought-resistant corn, and the target corn contains the GRMZM2G121851 protein described in claim 1 The corn with nucleic acid, the drought resistance of the high drought resistance corn is higher than the drought resistance of the target corn. 4. A corn breeding method, comprising the following steps: increasing the protein content of claim 1 in target corn, thereby obtaining corn with increased drought resistance.