CN110790831B

CN110790831B - Plant salt-tolerant drought-tolerant protein and coding gene and application thereof

Info

Publication number: CN110790831B
Application number: CN201911226533.0A
Authority: CN
Inventors: 蒋炳军; 武婷婷; 韩天富; 董萌; 侯文胜; 房裕东; 高友菲; 孙�石; 吴存祥; 袁珊
Original assignee: Institute of Crop Sciences of Chinese Academy of Agricultural Sciences
Current assignee: Institute of Crop Sciences of Chinese Academy of Agricultural Sciences
Priority date: 2019-12-04
Filing date: 2019-12-04
Publication date: 2021-08-24
Anticipated expiration: 2039-12-04
Also published as: CN110790831A

Abstract

The invention discloses a plant salt-tolerant drought-tolerant protein, a coding gene and application thereof. The invention provides application of a GmMYB173 protein and a coding gene thereof in regulation and control of salt tolerance and drought tolerance of plants. The invention creates conditions for cultivating salt-tolerant and drought-tolerant crops and cultivating widely-adapted salt-tolerant and drought-tolerant crop varieties by molecular means.

Description

Plant salt-tolerant drought-tolerant protein and coding gene and application thereof

Technical Field

The invention relates to the field of molecular genetic breeding, in particular to a plant salt-tolerant drought-tolerant protein, a coding gene and application thereof.

Background

Salt and drought are two major closely related abiotic stresses during plant growth and development. They seriously affect the growth and development of crops, cause great reduction of yield and even no harvest in severe cases. As a population large country in development, the living standard of people is continuously improved, which puts higher requirements on agricultural production. However, cultivated land resources in China are extremely limited, and grain safety and national safety face serious challenges. Meanwhile, China is also a big land for saline-alkali soil, and has over 5 hundred million acres of saline-alkali wasteland and saline-alkali soil influencing cultivated land, wherein the saline-alkali soil with the cultivated land development potential is not lacked. Therefore, the saline-alkali soil is scientifically utilized, the cultivated land area is effectively expanded, and the method has important significance for guaranteeing the stability and development of the society in China.

Soybean is the most important oil crop and high protein food crop in the world. In China, soybean is one of four major food crops, and plays an important role in ensuring national food safety, improving the life of people in urban and rural areas and increasing the income of farmers. However, the cultivated land area of China is limited, and the cultivated land area available for soybean planting is especially insufficient, so that the soybean yield of China can not meet the demand far, and a large amount of soybeans need to be imported every year. The soybean has nitrogen fixation capacity and is barren-resistant, can be used as a pioneer crop for scientific utilization of saline-alkali soil, can expand the planting area of the soybean, and can develop and utilize the saline-alkali soil.

Disclosure of Invention

The invention aims to provide a plant salt-tolerant drought-tolerant protein, a coding gene and application thereof.

The invention firstly protects the application of GmMYB173 protein or related biological materials thereof in regulating and controlling the stress tolerance of plants;

the related biological material is a nucleic acid molecule capable of expressing the GmMYB173 protein or an expression cassette, a recombinant vector, a recombinant bacterium or a transgenic cell line containing the nucleic acid molecule;

the GmMYB173 protein is any one of the following proteins:

(A1) a protein consisting of an amino acid sequence shown in a sequence 1 in a sequence table;

(A2) the protein which is derived from the sequence 1 and has the same function and 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;

(A3) a protein having 99% or more, 95% or more, 90% or more, 85% or more, or 80% or more homology to the amino acid sequence defined in any one of (A1) to (A2) and having the same function;

(A4) a fusion protein obtained by attaching a protein tag to the N-terminus and/or C-terminus of the protein defined in any one of (A1) to (A3).

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, homology means 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 homology can be obtained.

The "nucleic acid molecule capable of expressing the GmMYB173 protein" is a coding gene of the GmMYB173 protein.

The coding gene of the GmMYB173 protein is a DNA molecule as follows:

(D1) a DNA molecule shown in a sequence 2 of a sequence table;

(D2) a DNA molecule that hybridizes under stringent conditions to the DNA molecule defined in (D1) and encodes the GmMYB173 protein;

(D3) a DNA molecule which has more than 99%, more than 95%, more than 90%, more than 85% or more than 80% of identity with the DNA sequence limited by (D1) or (D2) and codes the GmMYB173 protein.

In the above genes, the stringent conditions may be as follows: 50 ℃ in 7% Sodium Dodecyl Sulfate (SDS), 0.5M Na₃PO₄Hybridization with 1mM EDTA, rinsing in 2 XSSC, 0.1% SDS at 50 ℃; also can be: 50 ℃ in 7% SDS, 0.5MNa₃PO₄Hybridization with 1mM EDTA, rinsing at 50 ℃ in 1 XSSC, 0.1% SDS; also can be: 50 ℃ in 7% SDS, 0.5M Na₃PO₄Hybridization with 1mM EDTA, rinsing in 0.5 XSSC, 0.1% SDS at 50 ℃; also can be: 50 ℃ in 7% SDS, 0.5M Na₃PO₄And 1Hybridization in a mixed solution of mM EDTA, rinsing at 50 ℃ in 0.1 XSSC, 0.1% SDS; also can be: 50 ℃ in 7% SDS, 0.5M Na₃PO₄Hybridization with 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 the above application, the stress tolerance is salt tolerance and/or drought tolerance.

The above application is embodied as follows: the activity and/or expression level of the GmMYB173 protein or the coding gene thereof in the plant is improved, and the salt tolerance of the plant is increased; the activity and/or expression level of the GmMYB173 protein or the coding gene thereof in the plant is improved, and the drought tolerance of the plant is increased.

The invention also protects a method for cultivating a plant variety with improved stress tolerance, which comprises the step of improving the expression level and/or activity of the GmMYB173 protein in a receptor plant.

The GmMYB173 protein is as described previously.

The stress tolerance is salt tolerance and/or drought tolerance.

The invention also protects a method for cultivating the transgenic plant, which comprises the following steps: introducing a nucleic acid molecule capable of expressing GmMYB173 protein into a receptor plant to obtain a transgenic plant; the stress tolerance of the transgenic plant is greater than that of a recipient plant.

The stress tolerance is salt tolerance and/or drought tolerance.

The nucleic acid molecule capable of expressing the GmMYB173 protein is as described hereinbefore.

The expression "introducing a nucleic acid molecule capable of expressing a GmMYB173 protein into a recipient plant" is realized by introducing a recombinant expression vector containing a coding gene of the GmMYB173 protein into the recipient plant.

The existing expression vector can be used for constructing a recombinant expression vector containing the coding gene of the GmMYB173 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 the gene coding the GmMYB173 protein is used for constructing a recombinant plant expression vector, any enhanced promoter or constitutive promoter (such as cauliflower mosaic virus (CAMV)35S promoter and maize Ubiquitin promoter (Ubiquitin)) or tissue-specific expression promoter (such as seed-specific expression promoter) can be added in front of the transcription initiation nucleotide, and the enhanced promoter or the constitutive promoter can be used alone or combined with other plant promoters. In addition, when a plant expression vector is constructed by using the gene encoding the GmMYB173 protein, enhancers including a translation enhancer or a transcription enhancer, and the enhancer region can be an ATG initiation codon or a initiation codon of a neighboring region and the like, but is required to be identical with the reading frame of the coding sequence so as to ensure the correct translation of the whole 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 XbaI and SacI enzyme cutting sites of a pTF101-GmFT1a vector with a DNA molecule shown in a sequence 2 in a sequence table.

In the above method, the introduction of the recombinant expression vector carrying the coding gene of the GmMYB173 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.

The invention also protects the GmMYB173 protein or related biological materials thereof, or the application of any one of the methods in plant breeding.

The breeding is aimed at breeding plants with high stress tolerance.

The stress tolerance is salt tolerance and/or drought tolerance.

In the above aspects, the plant is (C1) or (C2) or (C3):

(C1) a dicot or monocot;

(C2) leguminous plants;

(C3) and (4) soybeans.

The soybean is preferably a self-tribute winter bean.

The invention provides a plant salt-tolerant drought-tolerant protein, a coding gene and application thereof. The invention creates conditions for cultivating salt-tolerant and drought-tolerant crops and cultivating widely-adapted salt-tolerant and drought-tolerant crop varieties by molecular means.

Drawings

FIG. 1 shows a comparison of transgenic positive plants and wild type phenotype after glufosinate application.

FIG. 2 is a comparison of salt tolerance assay phenotypes.

FIG. 3 is a comparison of drought tolerance test phenotypes.

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.

Self-tribute winter beans: reference documents: hongbo Sun, Zhen Jia, Dong Cao, Bingjun Jiang, Cunxiang Wu, Wenshen gHou, Yike Liu, Zhihong Fei, Dazhong Zhuao, Tianfu Han GmFT2a, a sobean homolog of flowing motion LOCUS T, is invented in flowing transition and maintenance plos ONE,2011,6(12): e29238; the public is available from the institute of crop science, academy of agricultural sciences, china.

pTF101-GmFT1a vector: reference documents: wei Liu^#,Bingjun Jiang^#,Liming Ma,Shouwei Zhang,Hong Zhai,Xin Xu,Wensheng Hou,Zhengjun Xia,Cunxiang Wu,Shi Sun,Tingting Wu,Li Chen,Tianfu Han^*Functional differentiation of flying Lotus T-homologes in sobeans GmFT1a and GmFT2a/5a had positioning rolls in controlling flowing and mapping, New Photologist, 2018,217(3): 1335-1345; the public is available from the institute of crop science, academy of agricultural sciences, china.

Germination culture medium: 3.1G/L Gamborgs basic Salt mix (Phytotech G768), 20G/L sucrose, 1ml/L Gamborgs Vitamin Solution (Phytotech G219), 7G/L agar, pH 5.8.

Co-culture liquid medium: 2.0g/L of Murashige & Skoog basic Salt Mixture (Murashige & Skoog basic Salt Mixture), 3.9g/L of morpholine ethanesulfonic acid (MES), 30g/L of sucrose, 1ml/L of Gamboge vitamin solution, 150mg/L of dithiothreitol, 40mg/L of acetosyringone, 2mg/L of zeatin, pH 5.4.

Co-culture medium: 2.0g/L of the Muiragis Cuger's base salt mixture, 3.9g/L of morpholine ethanesulfonic acid, 30g/L of sucrose, 1ml/L of Gamboge vitamin solution, 150mg/L of dithiothreitol, 40mg/L of acetosyringone, 2mg/L of zeatin, 7g/L of agar, pH 5.4.

Recovering the culture medium: 3.1g/L Gamboge basic salt mixture, 0.98g/L morpholine ethanesulfonic acid, 30g/L sucrose, 1ml/L Gamboge vitamin solution, 150mg/L cefotaxime (cefotaxime), 450mg/L timentin, 1 mg/L6-Benzylaminopurine (6-benzamidopurines), 7g/L agar, pH 5.7.

Screening a culture medium: 3.1g/L Gamboge basic salt mixture, 0.98g/L morpholine ethanesulfonic acid, 30g/L sucrose, 1ml/L Gamboge vitamin solution, 150mg/L cefotaxime, 450mg/L timentin, 1 mg/L6-benzylaminopurine, 7g/L agar, 6mg/L glufosinate (glufosinate), pH 5.7.

Elongation culture medium: 4.0g/L MiluragisKuger base salt mixture, 0.6g/L morpholine ethanesulfonic acid, 30g/L sucrose, 1ml/L Gamboge vitamin solution, 150mg/L cefotaxime, 450mg/L timentin, 0.1mg/L indole acetic acid (3-Indoleacetic acid), 0.5mg/L gibberellin (Gibberellic acid), 1mg/L zeatin, 7g/L agar, 6mg/L glufosinate, pH 5.6.

Rooting culture medium: 2.0g/L of a mixture of muirag giskuge base salts, 0.6g/L of morpholine ethanesulfonic acid, 20g/L of sucrose, 1ml/L of a Gambog vitamin solution, 7g/L of agar, 3mg/L of glufosinate, pH 5.7.

The GmMYB173 protein disclosed by the embodiment of the invention is shown in a sequence 1 in a sequence table, and a coding gene (GmMYB173 gene) thereof is shown in a sequence 2 in the sequence table.

Example 1 transgenic overexpression of GmMYB173 improves salt tolerance

Obtaining of transgenic plant over-expressing GmMYB173

1. PCR amplification was carried out using the cDNA of the leaf of Hovenia dulcis Thunb as a template, and a primer pair consisting of a primer XbaI-MYB-F and a primer SacI-MYB-R, and the PCR amplification product (the objective fragment MYB173) was recovered using a common agarose gel recovery kit from Tiangen Bio Inc.

XbaI-MYB-F：5'-TCTAGAATGTCTCGCGCCTCCTCC-3'；

SacI-MYB-R：5'-GAGCTCTCAAGCAACACTAATGAT-3'。

In primer XbaI-MYB-F, the XbaI cleavage site is underlined.

In the primer SacI-MYB-R, the SacI cleavage site is underlined.

2. And (3) carrying out enzyme digestion on the PCR amplification product obtained in the step (1) by using restriction enzymes XbaI and SacI, and recovering an enzyme digestion product.

3. The pTF101-GmFT1a vector was digested with restriction enzymes XbaI and SacI, and an about 10kb vector backbone was recovered.

4. And (3) connecting the enzyme digestion product obtained in the step (2) with the vector skeleton obtained in the step (3) to obtain a recombinant expression vector pTF101-MYB 173. Based on the sequencing results, recombinant expression vector pTF101-MYB173 was described as follows: and (3) replacing a fragment between XbaI and SacI enzyme cutting sites of the pTF101-GmFT1a vector with a DNA molecule shown in a sequence 2 of a sequence table to obtain a recombinant expression vector (sequencing verification already).

5. And (3) introducing the recombinant expression vector pTF101-GmMYB173 obtained in the step (4) into Agrobacterium tumefaciens EHA105 to obtain the recombinant Agrobacterium tumefaciens.

6. Transforming the recombinant agrobacterium obtained in the step 5 into soybean to obtain T0 generation transgenic soybean, and the specific steps are as follows:

(1) selecting plump self-tribute winter bean seeds with uniform size, no scab, no crack, smooth surface and no wrinkle, and placing the seeds into a glass culture dish. Then put into a desiccator, and the culture dish was opened. A glass beaker was placed in the desiccator, and 100mL of sodium hypochlorite was added first, followed by dropwise addition of 4mL of concentrated hydrochloric acid to the beaker. The vaseline is coated on the periphery of the dryer cover, and then the dryer is covered to form a sealed state. The desiccator was then placed in a fume hood and the seeds were sterilized for 16-20 h.

(2) After the step (1) is finished, the sterilized seed hypocotyl is vertically and upwards inoculated into a germination culture medium, a culture dish is not sealed, and the seed hypocotyl is placed in a tissue culture room with the temperature of 25 ℃ and the illumination of 16 h/the darkness of 18 h for culture for 1 d.

(3) And (3) after the step (2) is completed, taking cotyledonary nodes of the germinated soybean seeds as explants. Firstly, peeling soybean seed coats, longitudinally cutting and separating two cotyledons, cutting a strip wound at the joint part (cotyledon node) of the cotyledons and an embryonic axis, putting the scratched explant into a co-culture liquid culture medium (bacterial liquid OD value is 0.6-0.8) containing recombinant agrobacterium heavy suspension, dip-dyeing for 30min at 28 ℃, after infection, downwards inoculating the explant (cotyledon) into the co-culture medium with the surface paved with filter paper, and culturing for 5 days under the conditions of 25 ℃, 16h illumination/18 h darkness.

(4) After completing step (3), the explants were transferred to recovery medium and cultured for 7d at 25 ℃ under 16h light/18 h dark conditions.

(5) After the step (4) is completed, the main bud generated by the explant is cut off and transferred into a screening culture medium, and the explant is cultured for 21d under the conditions of 25 ℃ and 16h of light/18 h of dark.

(6) After the step (5) is completed, the browned leaves are peeled off, and the produced adventitious bud is transferred into an elongation medium for elongation. And 4, subculturing once for 15d, and subculturing 2-3 times. During the period, the generated elongation seedlings are transferred into a rooting culture medium for rooting.

(7) And (6) finishing the step (6), taking out the seedlings from the culture medium after the roots grow out, and transplanting the seedlings into a small pot filled with the matrix for hardening the seedlings. After hardening off for 1 week, the seedlings are transferred to a big pot for growth. After three compound leaves grow on the regenerated seedling, 160mg/L glufosinate-butyl is smeared on the regenerated seedling, and an obvious screening effect can be seen 3-4 days after smearing. And selecting new three-leaf compound leaves to repeatedly smear for 2 times for detection. The detection result is shown in figure 1, the leaves of the wild type Yudongqidou become yellow and withered at the smearing part (marked with X), and the leaves of the transgenic positive plants have no obvious change at the smearing part.

T1 generations were obtained by selfing T0 transgenic positive soybeans, T2 generations were obtained by selfing T1 generations, and T3 generations were obtained by selfing T2 generations.

II, detecting salt tolerance of transgenic plants of overexpression GmMYB173

And (3) the plant to be detected: from Tribute winter beans and T3 generation GmMYB173 transgenic line T3-13-2.

The substrate and vermiculite were mixed at a ratio of 1:1 and placed in pots 30cm in diameter by 30cmx30cm and 30cm in height, 5 seeds were sown in each pot, 3 replicates were set, and all pots were placed in trays of 70cmx50cmx10cm and watered into the trays. The material was planted in a climatic chamber at 28 ℃ and humidity 60-80% for long days (LD, 16h light/8 h dark). On the 21 st day after sowing, 2L of NaCl saline solution with a concentration of 200mM was poured for salt stress treatment.

At 8 days of salt stress treatment, wild type ZGDD (fig. 2, right) showed obvious salt damage, and plants wilted and yellowed, while transgenic plants did not show obvious salt damage (fig. 2, left). By counting the plant damage degree at 3, 8 and 13 days of salt stress treatment (as shown in table 2), it can be seen that the wild type ZGDD already starts to generate salt damage at 3 days of salt stress treatment, while the transgenic plants are not damaged. Salt stress treatment for 8 and 13 days, the damage degree begins to deepen, but the transgenic plants in the same period perform better than the wild plants. The evaluation criteria of the damage rating are shown in table 1.

TABLE 1

Is free of	The plant is normally expressed
		Slight damage	The leaf edge of the 1 st to 3 rd section of the lower part of the plant is yellow, and the appearance above the 4 th section is normal
Moderate damage	Leaf of 1 st-3 rd section of the lower part of the plant is completely yellow and shed, and leaf of more than 4 th section is wilted
		Severe damage	The leaves at the 1 st to 3 rd nodes of the lower part of the plant are yellowed and fall off, and the edges of the leaves above the 4 th node are yellowed
Death was caused by death	The whole plant will wither, yellow and leave

TABLE 2

The above results indicate that GmMYB173 can improve salt tolerance.

Example 2 transgenic overexpression of GmMYB173 improves drought tolerance

Obtaining of transgenic plant over-expressing GmMYB173

The same procedure as in example 1.

Second, drought tolerance detection of transgenic plants with overexpression GmMYB173

Mixing the matrix and vermiculite at a ratio of 1:1, loading into flowerpots with a diameter of 30cm x30cm and a height of 30cm, sowing 5 seeds on each flowerpot for 29 days 11 months in 2018, setting 3 times of the sowing, placing all the flowerpots in trays with a length of 70cm x50cm x10cm, and watering all the flowerpots to the trays. The material is planted in a climatic chamber, the temperature is controlled at 28 ℃, and the humidity is maintained at 60-80%.

And (3) sowing and planting under a long-day condition (LD, 16h light/8 h dark), watering is not carried out until plant leaves wither 15 days after sowing, and watering is carried out again by 500 ml. The drought stress treatment was considered to have started on day 9 thereafter. Phenotypic observations 8 days after drought stress treatment as shown in fig. 3, it can be seen that transgenic plants (left) are significantly less damaged than wild type (right), indicating that GmMYB173 can improve drought tolerance.

Sequence listing

<110> institute of crop science of Chinese academy of agricultural sciences

<120> plant salt-tolerant drought-resistant protein and coding gene and application thereof

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Asn Lys Asp Ala Leu Ala Ala Gly Tyr Ala Ser Ala Asp Asp Ala Ala

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Pro Gln Asn Ser Gly Arg Leu Arg Glu Arg Glu Arg Lys Arg Gly Val

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Pro Trp Thr Glu Glu Glu His Lys Leu Phe Leu Val Gly Leu Gln Lys

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Val Gly Lys Gly Asp Trp Arg Gly Ile Ser Lys Asn Tyr Val Lys Thr

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Arg Thr Pro Thr Gln Val Ala Ser His Ala Gln Lys Tyr Phe Leu Arg

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Arg Ser Asn Leu Asn Arg Arg Arg Arg Arg Ser Ser Leu Phe Asp Ile

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Thr Thr Asp Thr Val Ser Ala Ile Pro Met Glu Gly Glu Gln Val Gln

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Asn Gln Asp Thr Leu Ser His Ser Gln Gln Gln Ser Pro Leu Phe Pro

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Ala Glu Thr Ser Lys Ile Asn Gly Phe Pro Met Met Pro Val Tyr Gln

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Phe Gly Phe Gly Ser Ser Gly Val Ile Ser Val Gln Gly Gly Asn Gly

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ccgtggacgg aggaagagca caagctgttt ttggttggat tgcagaaggt agggaaaggt 300

gattggagag gaatctccaa aaactacgtc aaaacgcgaa cgccaacgca ggttgcgagc 360

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aaaatcaatg ggtttccaat gatgccagtg tatcagtttg ggtttggttc ttctggagtg 600

atttcagtcc aaggtggcaa tggaaaccca atggaagaac tcactctggg acaaggaaac 660

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tcttctagtt ctgccgttga cccaccgaca ctgtccctgg ggctatcctt ttcatctgac 780

caaagacaga catcatcaag acattcagct ttacatgcca tacaatgttt cagcaatgga 840

gaaagcatca ttagtgttgc ttga 864

Claims

The application of GmMYB173 protein or related biological materials thereof in improving stress tolerance of soybeans;

the related biological material is a nucleic acid molecule capable of expressing the GmMYB173 protein or an expression cassette, a recombinant vector, a recombinant bacterium or a transgenic cell line containing the nucleic acid molecule;

the GmMYB173 protein is any one of the following proteins:

(A1) a protein consisting of an amino acid sequence shown in a sequence 1 in a sequence table;

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

the stress tolerance is drought tolerance;

the application is embodied as follows:

the expression level of the coding gene of the GmMYB173 protein in the soybeans is improved, and the drought tolerance of the soybeans is increased.
2. A method for breeding a soybean variety with improved stress tolerance, comprising the step of increasing the expression level of a gene encoding a GmMYB173 protein in recipient soybeans; the stress tolerance is drought tolerance;

the GmMYB173 protein is any one of the following proteins:

(A1) a protein consisting of an amino acid sequence shown in a sequence 1 in a sequence table;

(A2) and (b) a fusion protein obtained by attaching a protein tag to the N-terminus and/or C-terminus of the protein defined in (A1).
3. A method of breeding transgenic soybeans comprising the steps of: introducing a nucleic acid molecule capable of expressing GmMYB173 protein into receptor soybean to obtain transgenic soybean with improved expression level of a GmMYB173 protein coding gene; the stress tolerance of the transgenic soybean is greater than that of a receptor soybean; the stress tolerance is drought tolerance;

the GmMYB173 protein is any one of the following proteins:

(A1) a protein consisting of an amino acid sequence shown in a sequence 1 in a sequence table;

(A2) and (C) attaching a tag to the N-terminus and/or C-terminus of the protein defined in (A1).
4. The method of claim 3, wherein: the expression of introducing the nucleic acid molecule capable of expressing the GmMYB173 protein into the receptor soybean is realized by introducing a recombinant expression vector containing a coding gene of the GmMYB173 protein into the receptor soybean.
5. The method of claim 4, wherein: the coding gene of the GmMYB173 protein is a DNA molecule shown in a sequence 2 of a sequence table.