CN114480431A

CN114480431A - Application of corn ZmBES1/BZR1-10 gene in improving drought tolerance and yield of plants

Info

Publication number: CN114480431A
Application number: CN202210324341.9A
Authority: CN
Inventors: 于好强; 冯文奇; 刘媛; 付凤玲; 杨青青; 李晚忱
Original assignee: Sichuan Agricultural University
Current assignee: Sichuan Agricultural University
Priority date: 2022-03-30
Filing date: 2022-03-30
Publication date: 2022-05-13
Anticipated expiration: 2042-03-30
Also published as: CN114480431B

Abstract

The invention discloses application of a corn ZmBES1/BZR1-10 gene in improving plant drought tolerance and yield, defines the relationship between the corn ZmBES1/BZR1-10 gene and plant drought tolerance and grain development, and verifies the application effect of the ZmBES1/BZR1-10 gene in improving plant drought tolerance and grain size. The ZmBES1/BZR1-10 gene is separated from the leaves of maize seedlings, a 35S-ZmBES1/BZR1-10-eGFP vector is constructed, the vector is transformed into a plant by an agrobacterium-mediated method to be overexpressed, the drought tolerance and the seed yield of the transgenic plant are obviously improved, and the overexpression of the ZmBES1/BZR1-10 gene can obviously improve the drought tolerance and the yield of the transgenic plant, so that the application prospect in the field of cultivating drought-tolerant high-yield plants is wide.

Description

Application of corn ZmBES1/BZR1-10 gene in improving drought tolerance and yield of plants

Technical Field

The invention relates to the field of crop cultivation, in particular to application of a corn ZmBES1/BZR1-10 gene in improving plant drought tolerance and yield.

Background

Corn, rice and the like are important grain crops in China, and high yield and high resistance are important indexes considered by researchers in the breeding process. In recent years, the improvement of agronomic traits or resistance of crops by genetic engineering means is a common technical means, and a new solution is provided for the improvement of crop yield and resistance. Through a great deal of research for many years, people have accumulated abundant experience and data on adversity stress mechanisms such as crop drought resistance and the like in the aspects of physiology, biochemistry, metabolism, genetic evolution and the like, and along with the deepening of people on gene understanding, the combination of genes and traditional breeding for cultivating high-yield and high-drought-resistance varieties is an urgent problem to be solved, and although relevant reports on the research on gene level resistance of various crops exist, relevant reports on the effective separation and application of efficient drought-resistance yield-increasing genes on the cultivation and planting of crops such as corn, rice and the like do not exist.

Disclosure of Invention

The invention aims to provide application of a corn ZmBES1/BZR1-10 gene in improving the drought tolerance and yield of plants so as to solve the problems in the prior art, and the gene can obviously promote the drought tolerance and yield of transgenic plants.

In order to achieve the purpose, the invention provides the following scheme:

the invention provides a corn ZmBES1/BZR1-10 gene, wherein the nucleotide sequence of the corn ZmBES1/BZR1-10 gene is shown as SEQ ID NO. 1.

The invention also provides application of the corn ZmBES1/BZR1-10 gene in improving the drought tolerance and yield of plants, and the application of the corn ZmBES1/BZR1-10 gene in promoting the drought tolerance of plants to be improved and the grain length and thousand kernel weight of grains to be increased.

The invention also provides a method for cultivating drought-tolerant and high-yield plants by using the corn ZmBES1/BZR1-10 gene, which obtains plants with improved drought tolerance and grain size by up-regulating the expression of the ZmBES1/BZR1-10 gene in the plants.

Preferably, the up-regulation of the expression of the ZmBES1/BZR1-10 gene in plants specifically comprises: designing an amplification primer of the ZmBES1/BZR1-10 gene to carry out PCR amplification of the gene, constructing a recombinant plasmid containing the ZmBES1/BZR1-10 gene by using an amplification product and a vector, transforming the recombinant plasmid into a plant by using agrobacterium mediation, and carrying out over-expression of the ZmBES1/BZR1-10 gene to obtain the drought-resistant and high-yield plant.

Preferably, the nucleotide sequence of the amplification primer is shown as SEQ ID NO. 2-3.

Preferably, the recombinant plasmid is 35S-ZmBES1/BZR1-10-eGFP, and the nucleotide sequence is shown as SEQ ID NO:4, respectively.

Preferably, the plant comprises one of arabidopsis thaliana or rice.

The invention discloses the following technical effects:

the ZmBES1/BZR1-10 gene is separated from the leaves of corn seedlings, the gene is transferred into arabidopsis thaliana and rice to breed the transgenic plant, and experiments prove that the transgenic plant can obviously promote the increase of root length and fresh weight compared with wild type water control, and various physiological indexes of the plant after drought stress are obviously better than those of the wild type control, which shows that the ZmBES1/BZR1-10 gene can obviously increase the drought tolerance of the transgenic plant. Meanwhile, the gene can also obviously promote the grain length and thousand grain weight of transgenic plant grains, which shows that the gene can also improve the yield of transgenic plants.

Drawings

In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings needed in the embodiments will be briefly described below, and it is obvious that the drawings in the following description are only some embodiments of the present invention, and it is obvious for those skilled in the art to obtain other drawings without creative efforts.

FIG. 1 shows the growth of transgenic Arabidopsis in the medium;

FIG. 2 shows the root length measurement of transgenic Arabidopsis;

FIG. 3 shows the measurement results of fresh weight of transgenic Arabidopsis;

FIG. 4 shows the results of drought treatment of transgenic rice plants; a: before treatment; b: after drought treatment; c: after rehydration;

FIG. 5 is a graph showing the relative conductivity measurements of transgenic rice and wild-type NIP leaves after drought treatment;

FIG. 6 is the relative water content measurements of transgenic rice and wild-type NIP leaves after drought treatment;

FIG. 7 is a fluorescent observation of the root tips of homozygous transgenic Arabidopsis;

FIG. 8 is a homozygous transgenic Arabidopsis seed;

FIG. 9 shows the length and width of transgenic Arabidopsis seeds; and represents p <0.05, p <0.01, respectively;

FIG. 10 shows the thousand kernel weight of transgenic Arabidopsis seeds; and represents p <0.05, p <0.01, respectively;

FIG. 11 shows positive PCR detection of transgenic rice;

FIG. 12 is a phenotypic characterization of transgenic rice seeds;

FIG. 13 shows the grain length and width of transgenic rice seeds, respectively, indicating that p is <0.05 and p is < 0.01;

FIG. 14 shows the thousand kernel weight of transgenic rice seeds;

FIG. 15 is a map of pCAMBIA1300-35S-eGFP vector.

Detailed Description

Reference will now be made in detail to various exemplary embodiments of the invention, the detailed description should not be construed as limiting the invention but as a more detailed description of certain aspects, features and embodiments of the invention.

It is to be understood that the terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention. Further, for numerical ranges in this disclosure, it is understood that each intervening value, between the upper and lower limit of that range, is also specifically disclosed. Every smaller range between any stated value or intervening value in a stated range and any other stated or intervening value in a stated range is encompassed within the invention. The upper and lower limits of these smaller ranges may independently be included or excluded in the range.

Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Although only preferred methods and materials are described herein, any methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present invention. All documents mentioned in this specification are incorporated by reference herein for the purpose of disclosing and describing the methods and/or materials associated with the documents. In case of conflict with any incorporated document, the present specification will control.

It will be apparent to those skilled in the art that various modifications and variations can be made in the specific embodiments of the present disclosure without departing from the scope or spirit of the disclosure. Other embodiments will be apparent to those skilled in the art from consideration of the specification. The specification and examples are exemplary only.

As used herein, the terms "comprising," "including," "having," "containing," and the like are open-ended terms that mean including, but not limited to.

Example 1 application of maize ZmBES1/BZR1-10 Gene in improving drought tolerance and yield of plants

1. Gene cloning and vector construction

Taking five-leaf stage corn B73 seedling leaves, carrying out quick-freezing grinding by liquid nitrogen, and extracting total RNA by using a total RNA extraction kit Trizol (TaKaRa) according to the instruction. Using PrimeScript^TMRT regetant Kit (Takara) was used to synthesize cDNA using total RNA as a template. Cloning primers were designed using Primer 5.0, and specific PCR primers are shown in Table 1 using B73cDNA as template. Amplification was performed using high fidelity enzyme PrimerStar (Takara Inc.), PCR amplification reaction system as shown in Table 2, temperature cycling program: 94 ℃, 3 min; 98 ℃, 10s and the optimum annealing temperature of 10 s; 72 ℃ for 20 s;30 cycles; 5min at 72 ℃; keeping at 4 ℃. Sequencing the amplification product, wherein the sequence of the amplification product is shown as SEQ ID NO:1 is shown.

TABLE 1 ZmBES1/BZR1-10 cloning primer

TABLE 2 PCR amplification reaction System

1.1 transformation of Arabidopsis thaliana vector 35S-ZmBES1/BZR1-10-eGFP construction

According to the dicotyledonous plant expression vector pCAMBIA2300-35S-eGFP multiple cloning site, Sma I enzyme cutting site is introduced into an upstream primer, Spe I site is introduced into a downstream primer, and the specific sequence is as follows:

table 3 construction of pCAMBIA2300 vector primers

Samples were added to the reaction system shown in Table 2 and amplified. The temperature cycling program was: 3min at 94 ℃; 10s at 98 ℃, 10s at the optimum annealing temperature, 20s at 72 ℃ and 30 cycles; 5min at 72 ℃; keeping at 4 ℃.

And recovering the PCR product. The recovered product, pCAMBIA2300-35S-eGFP plasmid, was subjected to double digestion, and the sample was added according to the reaction system in Table 4. And (3) uniformly mixing the enzyme digestion system, placing the mixture in a water bath kettle at the temperature of 30 ℃, carrying out enzyme digestion for 6-8 h, adding the enzyme digestion product into a loading buffer solution, carrying out direct electrophoresis, recovering the enzyme digestion product, and carrying out enzyme digestion fragment connection according to the table 5.

TABLE 4 double digestion reaction System

TABLE 5 connection System

The ligation product was transformed into E.coli and tested by colony PCR. After the extracted recombinant plasmid is subjected to restriction enzyme digestion identification, the plasmid bacterium liquid is sent to a sequencing company for DNA sequence sequencing identification, and the plasmid with correct sequencing (the nucleotide sequence of the recombinant plasmid 35S-ZmBES1/BZR1-10-eGFP is shown in SEQ ID NO: 4) is transformed in the next step of Arabidopsis.

1.2 construction of transformed Rice vector 35S-ZmBES1/BZR1-10-eGFP

According to the multiple cloning site of the monocotyledon expression vector pCAMBIA1300-35S-eGFP (figure 15), the Hind III enzyme cutting site is added to the upstream primer, and the BamH I site is introduced to the downstream primer, and the specific sequence is shown in the table 6. Amplification was performed using high fidelity enzyme PrimerStar (Takara Co.) and the PCR amplification reaction system is shown in Table 2. The PCR product is then recovered.

TABLE 1 construction of pCAMBIA1300 vector primers

The expression vector pCAMBIA1300-35S-eGFP plasmid was subjected to double enzyme digestion, and the sample was added according to the reaction system in Table 4. Uniformly mixing the enzyme digestion system, placing the mixture in a water bath kettle at 30 ℃, carrying out enzyme digestion for 6-8 h, adding the enzyme digestion product into a loading buffer solution, carrying out direct electrophoresis, recovering the enzyme digestion product, using a Cloneexpress II One Step Cloning Kit (nunoprazan), wherein the reaction system is shown in Table 7, and the reaction conditions are as follows: 30mins at 37 ℃.

TABLE 2 homologous recombinase reaction System

The ligation product was transformed into E.coli and tested by colony PCR. After the extracted recombinant plasmid is subjected to restriction enzyme digestion identification, the plasmid bacterium liquid is sent to a sequencing company for DNA sequence sequencing identification, and the plasmid with correct sequencing (the nucleotide sequence of the recombinant plasmid 35S-ZmBES1/BZR1-10-eGFP is shown in SEQ ID NO: 4) is transformed into rice in the next step.

2. Arabidopsis and rice transformation and positive identification

2.1 transformation of Arabidopsis thaliana by Flowery infection

Exploring the function of maize ZmBES1/BZR1-10, we transformed the recombinant plasmid 35S-ZmBES1/BZR1-10-eGFP (nucleotide sequence shown in SEQ ID NO: 4) into Arabidopsis thaliana Columbia wild type Col (i.e., WT) using Agrobacterium-mediated catoptric floral dip method.

2.1.1 inflorescence Dip-dyeing transformation

(1) Taking agrobacterium containing the recombinant plasmid, streaking on a YEP plate containing Kana and Rif, and culturing for 2-3 d at 28 ℃.

(2) A single colony of Agrobacterium was picked and inoculated in 3mL of YEP liquid medium containing Kana and Rif, cultured at 28 ℃ at 200r/min, and shaken overnight.

(3) Inoculating 1mL of overnight-cultured starting Agrobacterium strain in 100mL of liquid YEP medium (containing Kana and Rif), and shake-culturing at 28 deg.C to OD₆₀₀The value is 1.2 to 1.5.

(4) Centrifuging at 4 deg.C for 10min at 5000r/min to collect cells, suspending thallus with 5% sucrose solution and adjusting OD₆₀₀When the value is 0.8-1.0, the surfactant silwet L-77 is added according to the proportion of 1-2%.

(5) And (3) carrying out pot culture on the arabidopsis wild type Col by using special nutrient soil for arabidopsis, pruning formed fruit pods and opened flowers after blooming, soaking inflorescences for 1.5-2.0 min by using the staining solution, and carrying out dark culture for 10 h.

(6) Culturing for 3-4 weeks under proper conditions, collecting seeds, and carrying out next screening treatment.

2.1.2 selection of resistant Medium

(1) Harvested infected seeds were aliquoted into 1.5mL EP tubes and soaked in 500. mu.L of 70% ethanol for 30 s.

(2) After the alcohol is sucked off, soaking the seeds in 500 mu L of 10% sodium hypochlorite for 5-10 min, and continuously shaking the centrifugal tube during the soaking process until the seeds are fully contacted with the sodium hypochlorite.

(3) Washing the seeds with sterilized water for 4-6 times, and abandoning the upper ddH layer after the seeds are settled₂O; 200 μ L of 0.1% agar water suspend seeds.

(4) The seeds were sown on 1/2MS medium containing 40mg/L kanamycin (Kana) and cultured.

(5) Repeating the steps until T3 homozygous positive seedlings are harvested.

Performing genome PCR identification on the obtained positive strain

(1) Taking 100mg of fresh plant tissue, adding liquid nitrogen, and fully grinding. 400 μ L of buffer FP1 and 6 μ L of RNaseA (10mg/mL) were added, vortexed for 1min, and allowed to stand at room temperature for 10 min.

(2) Add 130. mu.L of buffer FP2, mix well, vortex for 1min, centrifuge at 12000r/min (13400 Xg) for 5min, transfer the supernatant to a new centrifuge tube.

(3) Optional steps are as follows: the supernatant was centrifuged again at 12000r/min (13400 Xg) for 5min and transferred to a new centrifuge tube.

(4) 0.7 times volume of isopropanol is added into the supernatant, and the mixture is fully mixed, so that flocculent genomic DNA can appear. The mixture was centrifuged at 12000r/min (13400 Xg) for 2min, the supernatant was discarded, and the precipitate was retained.

(5) Add 600. mu.L 70% ethanol, vortex for 5s, centrifuge at 12000r/min (13400 Xg) for 2min, and discard the supernatant.

(6) And (5) repeating the step.

(7) And (5) opening the cover and inverting the cover, and completely airing the residual ethanol at room temperature for 5-10 min.

(8) Adding a proper amount of elution buffer TE, dissolving DNA in water bath at 65 ℃ for 10-60 min, and reversely and uniformly mixing for several times to aid dissolution to finally obtain a DNA solution.

(9) PCR detection was performed using PCR primers as shown in Table 8, using the extracted genomic DNA as template, at 94 ℃ for 3 min; 94 ℃ for 30s, the optimal annealing temperature for 30s, 72 ℃ for 60s/kb, 35 cycles; 5min at 72 ℃; storing at 4 ℃.

TABLE 3 transgenic Rice detection primers

(10) After harvesting and sterilizing transformed arabidopsis thaliana T2 generation seeds, sowing the seeds on 1/2MS solid culture medium containing kanamycin, and after 7-8 days, all green robust plants are homozygous transformants, while non-homozygous transformants show 3:1 segregation (proportion of normal growth and etiolated dead plants). Seeds of homozygous T3 generation were laid for further analysis.

Homozygous transgenic lines W10-13, W10-19 and W10-21 were selected for subsequent studies by kanamycin screening.

2.2 transformation of Rice

The recombinant plasmid 35S-ZmBES1/BZR1-10-eGFP (nucleotide sequence shown as SEQ ID NO: 4) vector is sent to Miami Bio Inc. (Nanjing) to transform the wild type "Nippon" (NIP) of rice.

3. Analysis of drought tolerance of transgenic lines

3.1 transgenic Arabidopsis thaliana simulated drought treatment

Homozygous transgenic Arabidopsis lines W10-13, W10-19, W10-21 and Wild Type (WT) were sterilized with 5% sodium hypochlorite solution, seeded on 1/2MS medium and 1/2MS medium containing 200mM mannitol, and cultured for 15d under normal conditions to observe changes in root length. And the root length and fresh weight were measured.

3.2 drought treatment of transgenic Rice

Positive plants R10-7, R10-9, R10-10 were identified by PCR, and R10-7, R10-9, R10-10 and wild type (NIP) were sown in water-containing soil, and after 20 days watering was stopped and phenotype was observed.

3. Analysis of pure and line yields

3.1 Arabidopsis seed phenotypic characterization

Homozygous transgenic arabidopsis lines W10-19 and W10-21 and Wild Type (WT) were disinfected with 5% sodium hypochlorite solution, sown on medium containing 1/2MS, and 15d later transplanted to nutrient-containing soil: in 50mm x 50mm pots with 3:1 soil of vermiculite, 4 plants were sown per pot, placed under the same conditions for direct harvest, and the seed grain length, grain width and thousand seed weight were measured.

3.2 phenotypic identification of Rice seeds

Homozygous transgenic rice lines R10-9, R10-10 and wild type (NIP) were sown in paddy fields, straight-harvested seeds were cultured under the same conditions, and seed grain length, grain width and thousand kernel weight were measured.

4. Analysis of results

4.1 transgenic Arabidopsis thaliana simulation of drought treatment and yield analysis on culture Medium

The growth conditions of the transgenic lines W10-13, W10-19, W10-19 and wild type WT did not change significantly on 1/2MS medium, but were significantly better than that of the wild type on 1/2MS medium containing 200mM mannitol (FIG. 1). Root length fresh weight measurements revealed that the transgenic lines W10-13, W10-19, W10-21 had significantly increased root length compared to wild-type WT (FIG. 2). Fresh weight measurements found that the fresh weights of transgenic lines W10-13, W10-19, and W10-21 were significantly increased compared to wild-type WT (FIG. 3). The results show that the target gene ZmBES1/BZR1-10 can obviously increase the drought tolerance of arabidopsis thaliana under simulated drought treatment.

Through resistance screening and root tip fluorescence observation, the stable expression of the target gene ZmBES1/BZR1-10 in Arabidopsis is determined (figure 7), and the seed grain length, grain width and thousand kernel weight of homozygous transgenic Arabidopsis are analyzed, and the result shows that the grain length of the seeds of over-expression strains W10-19 and W10-21 is remarkably increased, and the grain width of W10-21 is remarkably increased (figure 8 and figure 9). Thousand kernel weight measurements revealed a significant increase in thousand kernel weight for the overexpression lines W10-19 and W10-21 (FIG. 10).

4.2 drought treatment and yield analysis of transgenic Rice

The transgenic lines R10-7, R10-9, R10-10 and wild type (NIP) were sown in aqueous soil. Before treatment, the growth conditions of the transgenic lines and the NIP are not obviously changed (figure 4A), but after drought treatment, NIP leaves are completely wilted, while the leaves of the transgenic lines are still partially green (figure 4B), and after rehydration, the growth of the transgenic lines is obviously better than that of the NIP (figure 4C). The determination of physiological indexes of various drought-treated lines shows that the relative conductivity of the drought-treated transgenic lines is obviously smaller than that of wild-type NIP (figure 5), and the relative water content of the drought-treated transgenic lines is obviously higher than that of the wild-type NIP (figure 6). The results show that the target gene ZmBES1/BZR1-10 can obviously increase the drought tolerance of rice.

Positive lines were determined by PCR for detection of the gene of interest (FIG. 11). The analysis on the grain length, the grain width and the thousand grain weight of the positive strain shows that the grain length of over-expression strains R10-9 and R10-10 is remarkably increased, and the grain width has no obvious change; thousand kernel weight was significantly increased compared to control NIP (fig. 12-14).

The above-described embodiments are merely illustrative of the preferred embodiments of the present invention, and do not limit the scope of the present invention, and various modifications and improvements of the technical solutions of the present invention can be made by those skilled in the art without departing from the spirit of the present invention, and the technical solutions of the present invention are within the scope of the present invention defined by the claims.

Sequence listing

<110> Sichuan university of agriculture

Application of <120> corn ZmBES1/BZR1-10 gene in improving drought tolerance and yield of plants

<160> 4

<170> SIPOSequenceListing 1.0

<210> 1

<211> 954

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 1

atgacgtccg gggcggcggc ggcgggaggt ctggggcgga cgccgacgtg gaaggagcgg 60

gagaacaaca agcgccggga gcgccggcgg agggccatcg ccgccaagat cttcacgggc 120

ctccgcgccc tcggcaacta caagctgccc aagcactgcg acaacaacga ggtgctcaag 180

gcgctctgcc gcgaggcggg gtgggtcgtc gaggacgacg gcaccaccta ccggaagggt 240

tgcaggccgc cgccggggat gctgagcccg tgctcgtcgt cgcagctgct gagcgcgccg 300

tcctcgagct tcccgagccc ggtgccgtcc taccacgcca gcccggcgtc gtcgagcttc 360

ccgagcccga cgcgcctcga ccacagcagc ggcggcagca gcacccacaa ccccgccgcg 420

gcggccgcgg cggccgccgc ctccctgctc ccgttcctcc ggggcctgcc gaacctgccg 480

ccgctccgcg tgtccagcag cgcgcccgtc acgccgccgc tctcctcgcc cacggcggcg 540

gcggcgtcgc ggccgcccac caaggtccgc aggcccaact gggacgccgc cgccgccgtc 600

gtcgccgctg accccttccg gcaccccttg ttcgcggtct ccgcccccgc cagccccacc 660

cgcgcgcgcc ggcgcgagca cccggacacc atcccggagt gcgacgagtc cgacgtctgc 720

tccgcggtcg actccgcccg gtggatcagc ttccaggcca ccacggcgcc cgcgtcgccc 780

acgtacaacc tcgtccaccc ggcctccgac tccatggagc tggacgggac gacggcagcc 840

gtcgaggagt tcgagttcga caagggccgc gtcgtcacgc cgtgggaagg cgagcggatc 900

cacgaggtcg ccgccgagga gctcgagctc acgctcggcg tcggcgccaa gtga 954

<210> 2

<211> 18

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 2

atgacgtccg gggcggcg 18

<210> 3

<211> 22

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 3

tcacttggcg ccgacgccga gc 22

<210> 4

<211> 2483

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 4

aattcccatg gagtcaaaga ttcaaataga ggacctaaca gaactcgccg taaagactgg 60

cgaacagttc atacagagtc tcttacgact caatgacaag aagaaaatct tcgtcaacat 120

ggtggagcac gacacgcttg tctactccaa aaatatcaaa gatacagtct cagaagacca 180

aagggcaatt gagacttttc aacaaagggt aatatccgga aacctcctcg gattccattg 240

cccagctatc tgtcacttta ttgtgaagat agtggaaaag gaaggtggct cctacaaatg 300

ccatcattgc gataaaggaa aggccatcgt tgaagatgcc tctgccgaca gtggtcccaa 360

agatggaccc ccacccacga ggagcatcgt ggaaaaagaa gacgttccaa ccacgtcttc 420

aaagcaagtg gattgatgtg atatctccac tgacgtaagg gatgacgcac aatcccacta 480

tccttcgcaa gacccttcct ctatataagg aagttcattt catttggaga ggacagggta 540

cccggggatc ctctagagtc gacatgacgt ccggggcggc ggcggcggga ggtctggggc 600

ggacgccgac gtggaaggag cgggagaaca acaagcgccg ggagcgccgg cggagggcca 660

tcgccgccaa gatcttcacg ggcctccgcg ccctcggcaa ctacaagctg cccaagcact 720

gcgacaacaa cgaggtgctc aaggcgctct gccgcgaggc ggggtgggtc gtcgaggacg 780

acggcaccac ctaccggaag ggttgcaggc cgccgccggg gatgctgagc ccgtgctcgt 840

cgtcgcagct gctgagcgcg ccgtcctcga gcttcccgag cccggtgccg tcctaccacg 900

ccagcccggc gtcgtcgagc ttcccgagcc cgacgcgcct cgaccacagc agcggcggca 960

gcagcaccca caaccccgcc gcggcggccg cggcggccgc cgcctccctg ctcccgttcc 1020

tccggggcct gccgaacctg ccgccgctcc gcgtgtccag cagcgcgccc gtcacgccgc 1080

cgctctcctc gcccacggcg gcggcggcgt cgcggccgcc caccaaggtc cgcaggccca 1140

actgggacgc cgccgccgcc gtcgtcgccg ctgacccctt ccggcacccc ttgttcgcgg 1200

tctccgcccc cgccagcccc acccgcgcgc gccggcgcga gcacccggac accatcccgg 1260

agtgcgacga gtccgacgtc tgctccgcgg tcgactccgc ccggtggatc agcttccagg 1320

ccaccacggc gcccgcgtcg cccacgtaca acctcgtcca cccggcctcc gactccatgg 1380

agctggacgg gacgacggca gccgtcgagg agttcgagtt cgacaagggc cgcgtcgtca 1440

cgccgtggga aggcgagcgg atccacgagg tcgccgccga ggagctcgag ctcacgctcg 1500

gcgtcggcgc caagactagt accatggtga gcaagggcga ggagctgttc accggggtgg 1560

tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc gtgtccggcg 1620

agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gttcatctgc accaccggca 1680

agctgcccgt gccctggccc accctcgtga ccaccctgac ctacggcgtg cagtgcttca 1740

gccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg cccgaaggct 1800

acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg 1860

tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg 1920

aggacggcaa catcctgggg cacaagctgg agtacaacta caacagccac aacgtctata 1980

tcatggccga caagcagaag aacggcatca aggtgaactt caagatccgc cacaacatcg 2040

aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc 2100

ccgtgctgct gcccgacaac cactacctga gcacccagtc cgccctgagc aaagacccca 2160

acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg atcactctcg 2220

gcatggacga gctgtacaag taactgcagg catgccaggg ctctcaatgg agtttgaatc 2280

aaatcttcca gctgctttaa tgagatatgc gagacgccta tgatcgcatg atatttgctt 2340

tcaattctgt tgtgcacgtt gtaaaaaacc tgagcatgtg tagctcagat ccttaccgcc 2400

ggtttcggtt cattctaatg aatatatcac ccgttactat cgtattttta tgaataatat 2460

tctccgttca atttactgat tga 2483

Claims

1. The corn ZmBES1/BZR1-10 gene is characterized in that the nucleotide sequence of the corn ZmBES1/BZR1-10 gene is shown as SEQ ID NO: 1.

2. The use of the maize ZmBES1/BZR1-10 gene in improving drought tolerance and yield in plants of claim 1, wherein the application promotes the improvement of drought tolerance and increased grain length and thousand kernel weight of the grain.

3. A method of breeding drought tolerant and high yielding plants using the maize ZmBES1/BZR1-10 gene of claim 1, wherein plants with improved drought tolerance and grain size are obtained by up-regulating the expression of the ZmBES1/BZR1-10 gene in the plant.

4. The method of claim 3, wherein the upregulating expression of the ZmBES1/BZR1-10 gene in a plant specifically comprises: designing an amplification primer of the ZmBES1/BZR1-10 gene to carry out PCR amplification of the gene, constructing a recombinant plasmid containing the ZmBES1/BZR1-10 gene by using an amplification product and a vector, transforming the recombinant plasmid into a plant by using agrobacterium mediation, and carrying out over-expression of the ZmBES1/BZR1-10 gene to obtain the drought-resistant and high-yield plant.

5. The method of claim 3, wherein the nucleotide sequence of the amplification primer is shown in SEQ ID NO. 2-3.

6. The method of claim 3, wherein the recombinant plasmid is 35S-ZmBES1/BZR1-10-eGFP, and the nucleotide sequence is as set forth in SEQ ID NO:4, respectively.

7. The method of any one of claims 2 to 6, wherein the plant comprises one of Arabidopsis or rice.