Application of Glyma03G. 201100 gene in improving salt tolerance of soybean or yeast

Abstract

The invention discloses application of a Glyma03G.201100 gene in improving the salt tolerance of soybeans or yeast, and belongs to the technical field of biology. The invention aims to provide a method for improving the salt tolerance of soybeans. The invention provides an application of a Glyma03G.201100 gene in improving the salt tolerance of soybeans or yeasts. The transgenic plant and the microorganism provided by the invention can provide theoretical guidance and gene resources for genetic improvement of crop salt tolerance, and have important significance in coping with global soil salinization.

Description

Technical Field

[0001] The present invention belongs to the field of biotechnology, and specifically relates to the application of the Glyma03G.201100 gene in improving the salt tolerance of soybeans or yeast. Background Art

[0002] As an oilseed and cash crop, soybeans have become one of the most widely planted and utilized cash crops and food crops in the world, accounting for approximately 25% of the global edible oil market and two-thirds of the world's protein demand. Soil salinization is an environmental stress on a global scale, seriously threatening the sustainable development of crop production. According to statistics, approximately 7% of the world's land area, that is, about 9.5 million square kilometers of soil, is affected by salinization. Salt stress is different from drought stress. It starts to stress from the germination stage of plants and continues until the senescence of plants. The germination stage, as a sensitive period of stress, is likely to affect the later growth of plants, resulting in a significant reduction in yield and causing great economic losses.

[0003] Currently, the food problem faced by humanity is intensifying day by day. It is necessary to make full use of this reserve resource of saline-alkali land, use modern biotechnology to explore the salt tolerance mechanism of soybeans, and screen its salt tolerance genes, which can provide theoretical guidance and gene resources for the genetic improvement of crop salt tolerance and is of great significance in dealing with global soil salinization. Summary of the Invention

[0004] The purpose of the present invention is to provide a method for improving the salt tolerance of soybeans.

[0005] The present invention provides the application of the Glyma03G.201100 gene in improving the salt tolerance of soybeans or yeast.

[0006] The present invention provides the application of the Glyma03G.201100 protein in improving the salt tolerance of soybeans or yeast.

[0007] The present invention provides the application of a recombinant vector containing the Glyma03G.201100 gene in improving the salt tolerance of soybeans or yeast.

[0008] Further defined, the starting vector of the recombinant vector is pYES2.

[0009] The present invention provides the application of a recombinant microbial cell containing the Glyma03G.201100 gene in improving the salt tolerance of soybeans or yeast.

[0010] Further defined, the microbial cell is a eukaryotic microbial cell or a prokaryotic microbial cell.

[0011] Further defined, the concentration of the salt is 70 - 100 mM of NaCl.

[0012] The present invention provides a breeding method for improving the salt tolerance of soybeans. The steps of the method are as follows:

[0013] Step 1: Connect the gene shown in SEQ ID NO.1 to the vector pHair to obtain a recombinant vector;

[0014] Step 2: Transform the recombinant vector obtained in Step 1 into Agrobacterium to obtain recombinant Agrobacterium;

[0015] Step 3: Transfer the recombinant Agrobacterium obtained in Step 2 into soybeans to obtain transgenic soybean plants, and obtain positive transgenic soybean plants after identification.

[0016] The present invention provides a method for improving the salt tolerance of yeast. The steps of the method are as follows:

[0017] Step 1: Connect the gene shown in SEQ ID NO.1 to the vector pYES2 to obtain a recombinant vector;

[0018] Step 2: Transform the recombinant vector obtained in Step 1 into yeast to obtain recombinant yeast.

[0019] Further defined, the primers for amplifying the sequence shown in SEQ ID NO.1 are SEQ ID NO.3 and SEQ ID NO.4.

[0020] Beneficial effects: Overexpress the Glyma03G.201100 gene in soybeans to obtain transgenic soybean hairy roots of soybeans. Treat with 70 mM sodium chloride solution for salt stress for 5 days, and observe the development of roots and leaves. There are significant differences in the MDA content in the roots. Under salt stress, the MDA content increases significantly, while the MDA content in the transgenic overexpression plants is significantly lower than that of the empty vector plants ( Figure 7 ). Compared with the control plants, there are significant differences in the activities of POD, SOD, and CAT in the roots. Under salt stress, the activities of POD, SOD, and CAT increase significantly, while the activities of POD, SOD, and CAT in the transgenic overexpression plants are significantly higher than those of the empty vector plants ( Figures 8 - 10 ). This shows that the transgenic soybean hairy roots also have salt tolerance.

[0021] Overexpress the Glyma03G.201100 gene in yeast. The transgenic yeast has obvious differences in growth compared with the empty vector yeast in the YPDA medium under salt stress. In the normal YPDA medium, there is no difference in the growth of yeast. This shows that the transgenic yeast has salt tolerance. Description of the Drawings

[0022] Figure 1PCR amplification product for constructing the expression vector of Glyma03G.201100 in soybean

[0023] Figure 2 PCR product for transforming yeast with the soybean expression vector

[0024] Figure 3 Results of the transgenic yeast verification experiment for Glyma03G.201100; among them, CK is the normal culture medium; S is the salt stress culture medium

[0025] Figure 4 Results of the identification of the recombinant vector for Glyma03G.201100

[0026] Figure 5 Results of the overexpression of genes in transgenic soybean hairy roots of Glyma03G.201100 verified by qpcr in the transgenic soybean hairy root verification experiment; note: 1 is the gene expression level of transgenic plants, and 2 is the gene expression level of empty vector plants

[0027] Figure 6 Results of the PCR product of the overexpression vector construction in the transgenic soybean hairy root verification experiment for Glyma03G.201100

[0028] Figure 7 Graph of the MDA detection results of transgenic soybean hairy roots of Glyma03G.201100

[0029] Figure 8 Graph of the SOD enzyme activity detection results of transgenic soybean hairy roots of Glyma03G.201100

[0030] Figure 9 Graph of the POD enzyme activity detection results of transgenic soybean hairy roots of Glyma03G.201100

[0031] Figure 10 Graph of the CAT enzyme activity detection results of transgenic soybean hairy roots of Glyma03G.201100

[0032] Figure 11 Results of the successful transfection of transgenic soybean hairy roots of Glyma03G.201100 verified by qpcr in the transgenic soybean hairy root verification experiment and determination of gene overexpression results; note: 1 is the gene expression level of transgenic plants, and 2 is the gene expression level of empty vector plants Specific implementation method

[0033] The present invention will be further described in detail below in conjunction with specific embodiments and the accompanying drawings. The following embodiments facilitate a better understanding of the present invention, but do not limit the present invention. The experimental methods in the following embodiments are conventional experimental methods in the art unless otherwise specified. The drugs, reagents and materials used in the following embodiments can be obtained through commercial channels unless otherwise specified.

[0034] Example 1. Construction of a recombinant vector

[0035] Nucleotide sequence of the Glyma03G.201100 gene (SEQ ID NO.1):

[0036] ATGGCCGATAAGCAACCCCACTTGAATGGTGCTTATTACGGTCCCGCCATTCCCCCGGCGGAGCAACCACACTACCGCCCTAGCCGCGAAAGAAGCTGCTGTTGCTGCCTCTTCGGAATCTTGTGGAAGATTCTGGTTGCACTCATTGTCCTCGTTGGCCTCGCGGTCCTCATCTTCTGGCTGGTGGTTCAACCCCGTTCCTTCAAGTTCCACGTCACGAAAGCCAACCTAACACAATTTGATTACTATACCAACAACAACACCCTTCACTACAACATGGTCCTCAACTTCACTGCACGCAACCCCAACAAAAAACTCAGCATATACTACGACAAAGTAGAGGCATTAGCATTCTACGAGGATGTCAGGTTCGCCAATTACAGTGTGATAACGCACATGAACTCCTTCCGCCAGTACAAGAAGACCACCAGCCACATGAGCGCCGTTTTCTCGGGGCAGCAAGTGTTGCCGCTCGACAACGACCTAGTCTCAGAGTTGAACCAAGACAAGAGTGGTGGGGTGTACGAGATCGATGTGAAGCTCTACTTCAGGATTAGGTTCAGGCTCGGGGATGTCAAAACCCGTCGCTTCAAGCCCGAGGTCAAATGTGATATCAGGGTTCCCTTGAGGACCAATGGCAGCGTAACTTTGTTTCAGACCACCAAGTGTGATGTCGATTACTAG;

[0037] Protein sequence: (SEQ ID NO.2)

[0038] MADKQPHLNGAYYGPAIPPAEQPHYRPSRERSCCCCLFGILWKILVALIVLVGLAVLIFWLVVQPRSFKFHVTKANLTQFDYYTNNNTLHYNMVLNFTARNPNKKLSIYYDKVEALAFYEDVRFANYSVITHMNSFRQYKKTTSHMSAVFSGQQVLPLDNDLVSELNQDKSGGVYEIDVKLYFRIRFRLGDVKTRRFKPEVKCDIRVPLRTNGSVTLFQTTKCDVDY。

[0039] 1. Molecular cloning of the Glyma03G.201100 gene

[0040] Synthesis of the first strand of cDNA: Calculate the volume according to the RNA sample concentration. Take 1 μg of total RNA as the template and perform RT-PCR reverse transcription into cDNA with reference to the reverse transcription kit instructions.

[0041] 2. Cloning of the Glyma03G.201100 gene and recovery and purification of the PCR product

[0042] Primer design: First, use Phytozome to find the CDS sequence, design primers using Snapgene, and send them to Sangon Biotech (Shanghai) Co., Ltd. for synthesis. The primer sequences are shown in Table 3:

[0043] Table 3 Primer sequences

[0044]

[0045] 3. Amplification of the Glyma03G.201100 gene fragment and recovery and purification of the PCR product

[0046] The PCR amplification system is shown in Table 4:

[0047] Table 4 PCR system

[0048] Composition Volume 10×Buffer for KOD-Fx 25 μL 2 mM dNTPs 10 μL Primer 3 μL Template 1 μL Nuclease-free Water 10 μL KOD-Fx 1 μL

[0049] The PCR reaction program is as follows: Pre-denaturation at 94 °C for 2 min; denaturation at 98 °C for 10 s, annealing at 55 °C for 45 s; extension at 68 °C for 2 min, set 35 cycles; extension at 68 °C for 5 min. Verify the length of the PCR product by 1.0% agarose gel electrophoresis. Under ultraviolet light, use a newly opened blade to excise the single target DNA band as much as possible. Put the excised gel containing the DNA fragment into a pre-weighed 1.5 mL centrifuge tube, weigh it, calculate the gel weight, use the recovery kit for product recovery, perform the experiment according to the instructions, and collect the DNA solution for storage at -20 °C.

[0050] 4. Acquisition of pYES2 Plasmid

[0051] The yeast expression vector used in this experiment was pYES2. A small amount of purchased pYES2 plasmid was inoculated on an LB solid medium (with 100 mg / mL ampicillin as the antibiotic) by streak plating and cultured overnight. Subsequently, a single colony was picked and cultured in a liquid medium by shaking overnight, and the pYES2 plasmid was extracted with reference to the instructions of the plasmid extraction kit.

[0052] To ligate the amplified target fragment to the vector, pYES2 plasmid and gene fragment with the same sticky ends need to be prepared. First, the pYES2 plasmid was digested with EcorRⅠ and HindⅢ double enzymes. After agarose gel electrophoresis, the corresponding fragment was purified and recovered.

[0053] The digestion system is shown in Table 5:

[0054] Table 5 Digestion System

[0055] Composition Volume Restriction Enzyme 1 μL each DNA 1 μg 10×NEBuffer 5 μL Nuclease-free Water Make up to 50 μL

[0056] The components of the system were added to a 200 μL centrifuge tube, gently flicked to mix evenly, briefly centrifuged for 5 s, then incubated at 37 °C for 30 min and heat inactivated at 65 °C for 20 min.

[0057] Transgenic identification result: The recombinant vector was transformed into Escherichia coli to obtain recombinant Agrobacterium. A single colony was picked, and with the primers shown in Table 3, pre-denaturation was carried out at 95 °C for 3 min; denaturation was carried out at 95 °C for 15 s, annealing was carried out at 55 °C for 15 s; extension was carried out at 72 °C for 15 s, and 30 cycles were set; extension was carried out at 72 °C for 5 min. Colony PCR was performed. The length of the PCR product was verified by 1.0% agarose gel electrophoresis. The results showed that the recombinant plasmid vector contained the target gene. ( Figure 1 )

[0058] Example 2. Construction of Recombinant Microbial Cells

[0059] 1. Ligation and Transformation of Target Fragment and Yeast Expression Vector

[0060] The concentration of the corresponding fragment after recovery and purification was measured, and the target fragment was ligated to the vector fragment using homologous recombinase to construct a recombinant plasmid vector. The ligation system is shown in Table 6:

[0061] Table 6 Ligation System

[0062] Composition Volume 2×SoSoo Mix 5 μL Linear Vector 4 μL Target Fragment 1 μL

[0063] The components of the system were added to a 200 μL centrifuge tube, gently flicked to mix evenly, briefly centrifuged for 5 s, and then placed at 50 °C for 30 min.

[0064] The recombinant plasmid vector was transferred into DH5α competent cells, and the antibiotic in the medium was 100 mg / mL ampicillin. After overnight culture, the colonies growing on the plate were ampicillin-resistant bacteria after transformation. Single colonies were picked and inoculated into a liquid medium containing ampicillin, and cultured overnight at 37 °C with 200 rpm. Using the overnight cultured bacterial solution as a template, bacterial solution PCR was performed. Positive bacterial solutions identified by PCR were selected, and the recombinant plasmid vector was extracted using a plasmid extraction kit.

[0065] 2. Yeast transformation

[0066] Take freshly prepared yeast competent cells and perform yeast transformation at room temperature according to the transformation section in the instruction manual. Finally, spread 50 μL of the transformation mixture evenly on an SD-Ura (glucose) solid medium and incubate at 30 °C for 3 d until white colonies grow.

[0067] Pick the growing white single colonies into an SD-Ura liquid medium and culture overnight at 30 °C with 200 rpm. Take the bacterial solution and perform cell lysis treatment at 100 °C for 5 min. Use 5 μL of the lysed bacterial solution as a template for PCR detection to determine whether the target gene has been accurately integrated into the yeast expression vector.

[0068] 3. Phenotypic identification of recombinant yeast

[0069] The recombinant yeast was renamed INVScⅠ(pYES2-Glyma03G.201100), and INVScⅠ(pYES2-Glyma03G.201100) was the Glyma03G.201100 gene introduced in Example 1; to verify the effect of salt stress on transgenic yeast, a salt stress method with 100 mmol NaCl was used to observe its growth status, and it was repeated three times.

[0070] (1) In a laminar flow hood, pick the verified recombinant yeast and yeast transfected with the empty vector into a YPDA liquid medium (glucose) and culture overnight at 30 °C with 200 rpm shaking;

[0071] (2) Measure the OD600 of the overnight bacterial solution, calculate the required volume of the bacterial solution to make the OD value of 5 mL of the induction medium supplemented with galactose (SD-Ura) be 0.4;

[0072] (3) Take the required volume of the bacterial solution and centrifuge at 8000 rpm for 1 min;

[0073] (4) First resuspend the cells with 1 mL of SD-Ura, and then make up to 5 mL;

[0074] (5) Culture overnight at 30 °C with 200 rpm shaking;

[0075] (6) Measure the OD600 of the overnight bacterial solution, and after calculation, uniformly adjust the OD600 to 2.0;

[0076] (7) Uniformly dilute the bacterial solution by 10-3 times, 10-4 times, and 10-5 times respectively;

[0077] (8) Using INVScⅠ (pYES2) of yeast transfected with the empty vector as a control, take 5 μL of the undiluted bacterial solution and the diluted bacterial solution respectively, and inoculate them on the normal YPDA solid medium and the YPDA solid medium under salt stress;

[0078] (9) Incubate at 30 °C for 5 days;

[0079] (10) Observe and record the growth of the plates, and analyze and compare the differences in yeast growth.

[0080] Process of salt stress resistance treatment: Take 5 μL of the diluted bacterial solution respectively with the yeast with the empty vector as a control and inoculate them on the normal YPDA solid medium and the YPDA solid medium under 100 mM sodium chloride salt stress. Incubate at 30 °C for 5 days. Observe and compare the growth of transgenic yeast.

[0081] Results of positive identification: Dip the transgenic yeast with the primers shown in Table 3, pre-denature at 95 °C for 3 min; denature at 95 °C for 15 s, anneal at 55 °C for 15 s; extend at 72 °C for 15 s, set 30 cycles; extend at 72 °C for 5 min. Perform colony PCR. Verify the length of the PCR product by 1.0% agarose gel electrophoresis. ( Figure 2 )

[0082] In this example, a transgenic verification experiment was carried out in yeast to verify the function of Glyma03G.201100. The results showed that there were obvious differences in the growth of transgenic yeast compared with that of yeast with the empty vector on the YPDA medium under salt stress. However, there was no difference in the growth of yeast on the normal YPDA medium. It shows that the transgenic yeast has acquired salt tolerance ( Figure 3 ).

[0083] Example 3. Genetic transformation of soybean hairy roots

[0084] Obtaining of the pHair plasmid: The expression vector used in this experiment is pHair. A small amount of purchased pHair plasmid was streaked on the LB solid medium (antibiotic: 50 mg / mL kanamycin) and cultured overnight. Subsequently, a single colony was picked and cultured in the liquid medium overnight with shaking, and the pHair plasmid was extracted with reference to the plasmid extraction kit instructions.

[0085] To ligate the amplified target fragment to the vector, it is necessary to prepare the pHair plasmid and the gene fragment with the same sticky ends. First, double digest the pHair plasmid with ascⅠ and xbaⅠ. After agarose gel electrophoresis, purify and recover the corresponding fragment.

[0086] The digestion system is shown in Table 7:

[0087] Table 7 Digestion System

[0088] Composition Volume Restriction Enzyme 1 μL each DNA 1 μg 10×NEBuffer 5 μL Nuclease-free Water Make up to 50 μL

[0089] Add the components of the system to a 200 μL centrifuge tube, flick to mix well, briefly centrifuge for 5 s, then incubate at 37 °C for 30 min and inactivate at 65 °C for 20 min.

[0090] Measure the concentration of the corresponding fragment after recovery and purification, and use homologous recombinase to ligate the target fragment and the vector fragment to construct the recombinant plasmid vector pHair Glyma03G.201100. The ligation system is shown in Table 8:

[0091] Table 8 Ligation System

[0092] Composition Volume 2×SoSoo Mix 5 μL Linear Vector 4 μL Target Fragment 1 μL

[0093] Add the components of the system to a 200 μL centrifuge tube, flick to mix well, briefly centrifuge for 5 s, then place at 50 °C for 30 min.

[0094] Transfer the recombinant plasmid vector into DH5α competent cells, and the medium antibiotic is 50 mg / mL kanamycin. After overnight culture, the colonies growing on the plate are kanamycin-resistant bacteria after transformation. Pick a single colony and inoculate it into a liquid medium containing kanamycin, and culture it overnight at 37 °C with 200 rpm. Use the overnight cultured bacterial solution as a template for bacterial solution PCR. Select the bacterial solution with positive PCR identification, and extract the recombinant plasmid vector using a plasmid extraction kit.

[0095] Results of successful identification of the recombinant vector construction: Pick a single colony of Escherichia coli, pre-denature at 95 °C for 3 min with the primers shown in Table 3; denature at 95 °C for 15 s, anneal at 55 °C for 15 s; extend at 72 °C for 15 s, set 30 cycles; extend at 72 °C for 5 min. Perform colony PCR. Verify the length of the PCR product by 1.0% agarose gel electrophoresis. The results show that the recombinant plasmid vector contains the target gene.( Figure 4 )。

[0096] Agrobacterium strain K599 transformation: Take 5 μL of the pHair Glyma03G.201100 vector plasmid, add it to 100 μL of K599 competent cells, incubate on ice for 30 min, freeze in liquid nitrogen for 5 min, incubate at 37 °C for 5 min, add 800 μY T liquid medium, pre-culture at 28 °C with 200 rpm for 3 h, spread the bacterial solution on a YT (50 μg / mL str + 50 μg / mL kana) solid medium, incubate in the dark at 28 °C for 2 days, select positive monoclonal colonies, and shake the bacteria at 28 °C with 200 rpm for 16 - 18 h for soybean transformation.

[0097] Infection solution preparation: Shake the bacteria 3 - 4 days in advance. Select the empty vector bacterial solution and the overexpression bacterial solution of Agrobacterium rhizogenes k599. After culturing the Agrobacterium on a TY medium solid (containing kana + str antibiotics) for 2 d, pick single colonies and put them into a 2 mL centrifuge tube containing 1 mL of TY liquid medium, and culture overnight at 28 °C with 200 r / min. Pipette 500 μL of the well-shaken bacterial solution into 100 mL of TY medium and culture overnight at 28 °C with 200 r / min until it becomes turbid. Centrifuge the well-shaken bacterial solution at 28 °C with 4000 r / min for 10 min. After centrifuging out the bacterial pellet, resuspend it with distilled water until OD600 is about 0.8. Centrifuge at 28 °C and 4000×g for 5 min, discard the supernatant, collect the bacterial cells, and resuspend the bacterial cells in an infection solution containing 45 μL of acetosyringone (AS) with a final concentration of 100 μmol / L, 6 mL of MES with a concentration of 0.5 mol / L, 3 mL of MgCl2 with a concentration of 1 mol / L, and 500 mL of distilled water for infecting soybeans.

[0098] Seed germination: Select plump soybean seeds, soak them in the diluted sodium hypochlorite solution for 3 min (constantly mix the solution and screen the soybean seeds), and wash the soaked soybean seeds three times with distilled water. After the treatment, sow them in vermiculite and germinate them in a growth chamber.

[0099] Infection treatment: Use the hypocotyl seeds of about 2 mm, make wounds at the cotyledon node, and use them as explants for Agrobacterium rhizogenes-mediated hairy root transformation. Immerse the prepared explants in the suspension of Agrobacterium rhizogenes K599 containing the target construct. After infection, briefly air-dry them on sterile filter paper. Then place the explants on a filter paper containing a small amount of liquid co-culture medium and co-culture them in the dark at 23 °C for 1 day and in the light for 3 days.

[0100] Transgenic identification results: Verify the successful transfection of soybeans and determine the gene overexpression results by qrt-pcr ( Figure 11) The expression levels of the Glyma03G.201100 gene in the transfected soybean and the empty vector plants were measured by qRT-PCR (primer F: TTCTCGGGGCAGCAAGTG, SEQ ID NO.5); primer R: TGACATCCCCGAGCCTGA, SEQ ID NO.6). The expression level increased by about 14-fold.

[0101] Process of salt stress resistance treatment: Salt stress treatment was carried out with 70 mM sodium chloride solution for 5 days, and the root and leaf development were observed ( Figure 6 ). The following data were all detected in the root parts of the transgenic plants and the empty vector plants treated with water for 5 days and 70 mM sodium chloride solution.

[0102] Results of transgenic identification: After the above-mentioned stress, the transgenic materials were verified by qRT-PCR for the successful transfection of soybean and the determination of gene overexpression results ( Figure 5 ) The expression levels of the Glyma03G.201100 gene in the transfected soybean and the empty vector plants were measured by qRT-PCR (primer F: TTCTCGGGGCAGCAAGTG, SEQ ID NO.5); primer R: TGACATCCCCGAGCCTGA, SEQ ID NO.6). The expression level increased by about 16-fold.

[0103] In this example, during the genetic transformation experiment of soybean hairy roots, the physiological indexes of the function of Glyma03G.201100 were measured. The results showed that after 5 days of salt stress treatment, there were significant differences in the MDA content in the roots between the transgenic soybean plants and the control plants. Under salt stress, the MDA content increased significantly, while the MDA content in the transgenic overexpression plants was significantly lower than that in the empty vector plants ( Figure 7 ). There were significant differences in the activities of POD, SOD and CAT in the roots between the transgenic overexpression plants and the control plants. Under salt stress, the activities of POD, SOD and CAT increased significantly, while the activities of POD, SOD and CAT in the transgenic overexpression plants were significantly higher than those in the empty vector plants ( Figures 8 - 10 ). This indicates that the transgenic soybean hairy roots also developed salt tolerance.

[0104] The above-described embodiments are only descriptions of the preferred embodiments of the present invention, and do not limit the scope of the present invention. Without departing from the design spirit of the present invention, various deformations and improvements made by those of ordinary skill in the art to the technical solutions of the present invention should all fall within the protection scope determined by the claims of the present invention.

Claims

1. Application of Glyma03G.201100 gene in improving the salt tolerance of soybean or yeast.

2. Application of Glyma03G.201100 protein in improving the salt tolerance of soybeans or yeast.

3. Application of recombinant vector containing Glyma03G.201100 gene in improving the salt tolerance of soybean or yeast.

4. The use according to claim 3, characterized in that The starting vector of the recombinant vector is pYES2.

5. Application of recombinant microbial cells containing the Glyma03G.201100 gene in improving the salt tolerance of soybeans or yeast.

6. The use according to claim 5, characterized in that: The microbial cell is a eukaryotic microbial cell or a prokaryotic microbial cell.

7. The use according to any one of claims 1 to 6, characterized in that: The salt concentration is 70-100 mM NaCl.

8. A breeding method for improving the salt tolerance of soybean, characterized in that: The steps of the method are as follows: Step 1: connect the gene shown in SEQ ID NO.1 with the vector pHair to obtain a recombinant vector; Step 2: transforming the recombinant vector described in step 1 into Agrobacterium to obtain recombinant Agrobacterium; Step 3: The recombinant Agrobacterium described in step 2 is transferred into soybean to obtain transgenic soybean plants, and positive transgenic soybean plants are obtained after identification.

9. A method for improving the salt tolerance of yeast, characterized in that: The steps of the method are as follows: Step 1: connect the gene shown in SEQ ID NO.1 to vector pYES2 to obtain a recombinant vector; Step 2: Transform the recombinant vector described in step 1 into yeast to obtain recombinant yeast.

10. The method according to claim 8 or 9, characterized in that: The primers for amplifying the sequence shown in SEQ ID NO.1 are SEQ ID NO.3 and SEQ ID NO.4.

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