Soybean salt-tolerant related gene GmMATE85, and coding protein and application thereof
By cloning and regulating the soybean salt tolerance-related gene GmMATE85, the problem of insufficient research on the molecular mechanism of soybean salt tolerance has been solved, realizing the genetic engineering breeding of soybean salt tolerance and cultivating salt-tolerant or salt-sensitive soybean varieties.
Patent Information
- Application Number
- CN202410711234.0
- Authority / Receiving Office
- CN · China
- Patent Type
- Patents(China)
- Current Assignee / Owner
- Filing Date
- 2024-06-04
- Publication Date
- 2026-02-06
- Estimated Expiration
- 2044-06-04
AI Technical Summary
Existing technologies are insufficient to effectively elucidate the molecular mechanisms of soybean salt tolerance, and traditional breeding methods are insufficient to rapidly cultivate high-quality, high-yield salt-tolerant soybean varieties.
The soybean salt tolerance-related gene GmMATE85 and its encoded protein were cloned and expressed. The expression of GmMATE85 in soybean was regulated by recombinant expression vector and RNAi interference technology. The gene was inhibited or overexpressed to improve or reduce the salt tolerance of soybean.
By regulating the expression of the GmMATE85 gene, the salt tolerance or salt sensitivity of soybeans was significantly improved, thus achieving genetic engineering breeding for improved soybean salt tolerance.
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Figure HDA0004874088930000011 
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Abstract
Description
TECHNICAL FIELD
[0001] The application belongs to the field of genetic engineering, and particularly relates to a soybean salt-tolerant related gene GmMATE85, and a coding protein and application thereof. BACKGROUND
[0002] Soybean is an important economic oil crop in the world, and soil salinization is one of the important factors leading to serious yield reduction and quality decrease of soybean. Therefore, carrying out salt stress research on soybean, and analyzing its genetic and salt-tolerant molecular mechanism have important strategic significance for breeding salt-tolerant soybean varieties and rational utilization of saline-alkali land.
[0003] Salt stress can not only inhibit seed germination, but also reduce the emergence rate and seedling rate of soybean. NaCl stress reduces the emergence rate of wild and cultivated soybeans, and increases the time required from emergence to uniform emergence. Under NaCl stress, the seedling rate of seedlings is inhibited, and some emerged plants also die due to salt stress. Salt stress leads to decrease of normal plant rate, plant height, dry weight and water content, and increase of root / shoot ratio. Soil salinization seriously affects the agronomic traits of soybean, such as plant height, leaf size, biomass, main stem node number, branch number, nodule, pod number per plant, grain weight per plant and hundred-grain weight. The quality traits of soybean seeds, such as fatty acid, protein and fat content, are also affected by salt stress. It is generally believed that salt stress leads to significant increase of fat content and significant decrease of protein content in soybean seeds. Soybean yield is closely related to salt concentration and salt tolerance of varieties.
[0004] Salt-tolerant genetic engineering is the most promising way to breed salt-tolerant soybeans with high quality and high yield. Soybean has a complex salt stress response regulation network, and salt tolerance is a comprehensive performance of multiple physiological and biochemical reactions. It is difficult to improve the salt tolerance of soybean by traditional breeding methods. The use of molecular design breeding technology can accelerate the breeding process of salt-tolerant soybean varieties, but the research on the molecular mechanism of soybean salt tolerance is not deep enough, and the related work is slow. Therefore, it is of great significance to breed new salt-tolerant soybean varieties by mining salt-tolerant genes of soybean and analyzing their action mechanisms, so as to make full use of saline-alkali land resources. SUMMARY
[0005] To solve the above technical problems in the prior art, the purpose of the present application is to disclose a soybean salt-tolerant related gene GmMATE85, and a coding protein and application thereof.
[0006] The first purpose of the present application is to provide a salt-tolerant related gene GmMATE85, which is a DNA molecule as described in 1) or 2) or 3) below:
[0007] 1) a DNA molecule with a gene sequence as shown in SEQ ID NO. 1;
[0008] 2) a DNA molecule having a CDS sequence as shown in SEQ ID NO. 2;
[0009] 3) a DNA molecule hybridizing to the DNA sequence defined in 1) or 2) under stringent conditions and encoding the protein.
[0010] A second object of the present application is to provide the protein encoded by the aforementioned gene GmMATE85.
[0011] In particular, the protein provided by the present application is selected from any one of the following (a) or (b):
[0012] (a) a protein consisting of the amino acid sequence shown in SEQ ID NO. 3;
[0013] (b) a protein derived from the target sequence selected from SEQ ID NO. 2.
[0014] SEQ ID NO. 3 in the sequence listing consists of 535 amino acids.
[0015] A third object of the present application is to provide a recombinant expression vector, an expression cassette or a recombinant bacterium comprising the aforementioned gene GmMATE85.
[0016] Further, the recombinant expression vector or expression cassette is obtained by inserting the gene GmMATE85 into the recombination site of the vector pBA002 digested with XbaI; and the recombinant expression vector or expression cassette is transformed into an engineering bacterium to obtain the recombinant bacterium.
[0017] The recombinant expression vector containing any one of the aforementioned genes also falls within the protection scope of the present application.
[0018] The recombinant expression vector containing the aforementioned gene can be constructed using existing plant expression vectors.
[0019] The plant expression vector includes Agrobacterium binary vector and vector that can be used for plant microprojectile bombardment. The plant expression vector can further comprise the 3' untranslated region of the exogenous gene, i.e. comprising the polyadenylation signal and any other DNA fragment involved in mRNA processing or gene expression. The polyadenylation signal can guide the addition of polyadenylate to the 3' end of the mRNA precursor, such as the untranslated region of the 3' end of the Agrobacterium crown gall tumor-inducing (Ti) plasmid gene (such as the Nos gene of the nopaline synthase), the 3' end of the transcription of the untranslated region of the plant gene (such as the soybean storage protein gene) has similar functions.
[0020] When the gene construct is used to construct a recombinant plant expression vector, any one of the enhanced promoters or constitutive promoters, such as the Cauliflower Mosaic Virus (CAMV) 35S promoter, the Ubiquitin promoter of maize, can be added before the transcription initiation nucleotide, which can be used alone or in combination with other plant promoters; in addition, when the gene construct of the present application is used to construct a plant expression vector, enhancers, including translation enhancers or transcription enhancers, can also be used, and these enhancer regions can be ATG start codon or adjacent regions start codon, but must be the same reading frame as the coding sequence to ensure correct translation of the entire sequence. The source of the translation control signal and the start codon is wide, which can be natural or synthetic. The translation initiation region can be from the transcription initiation region or the structural gene.
[0021] In order to facilitate the identification and screening of transgenic plant cells or plants, the plant expression vector used can be processed, such as adding genes that can express enzymes or luminescent compounds that can produce color changes in plants (GUS genes, luciferase genes, etc.), antibiotic markers with resistance (gentamicin markers, kanamycin markers, etc.) or anti-chemical reagent marker genes (such as herbicide-resistant genes) and the like. For the safety of transgenic plants, no selective marker gene can be added, and the transformed plants can be directly screened under stress.
[0022] The recombinant expression vector can be a recombinant plasmid obtained by inserting the gene GmMATE85 into the recombination site of the restriction enzyme XbaI single enzyme digestion vector pBA002. The pBA002 containing GmMATE85 is named pBA002-GmMATE85.
[0023] The expression cassette containing any one of the above-mentioned genes GmMATE85, the transgenic cell line and the recombinant bacteria all belong to the protection scope of the present application.
[0024] The fourth object of the present application is to provide primers for amplifying the aforementioned gene GmMATE85, and the primer pair for amplifying the full length or any fragment of the gene GmMATE85 also belongs to the protection scope of the present application. In a specific example, the primers are shown as SEQ ID NO. 4 and SEQ ID NO. 5.
[0025] The fifth object of the present application is to provide an interfering RNA, an expression vector or a recombinant bacteria for silencing the aforementioned gene GmMATE85.
[0026] The silencing of the aforementioned gene GmMATE85 of the present application can be carried out according to the conventional method in the art, such as designing interfering RNA or knocking out the gene, etc. The present application provides an interfering RNA of the aforementioned gene GmMATE85, and the sequence is shown as SEQ ID NO. 6.
[0027] A sixth object of the present application provides a genetic engineering application of the aforementioned gene GmMATE85, the aforementioned protein, the aforementioned recombinant expression vector, expression cassette or recombinant bacteria, or the aforementioned primer, or the aforementioned interfering RNA, expression vector or recombinant bacteria in regulating salt tolerance of soybean.
[0028] Further, inhibiting expression of the aforementioned gene GmMATE85, for example, performing interfering silencing or knockout, can improve salt tolerance of soybean. Overexpressing the aforementioned gene GmMATE85 can improve salt sensitivity of soybean.
[0029] Preferably, the aforementioned interfering RNA, expression vector or recombinant bacteria is introduced into soybean to inhibit the aforementioned gene GmMATE85.
[0030] Preferably, the aforementioned recombinant expression vector, expression cassette or recombinant bacteria is introduced into soybean to overexpress the aforementioned gene GmMATE85.
[0031] A sixth object of the present application provides a method for regulating salt tolerance of soybean, which comprises inhibiting the aforementioned gene GmMATE85 in a soybean plant to improve salt tolerance of soybean; or overexpressing the aforementioned gene GmMATE85 in a soybean plant to improve salt sensitivity of soybean.
[0032] Inhibiting the aforementioned gene GmMATE85 in a soybean plant can be performed by using methods commonly used in the art, for example, silencing or knockout, and in particular, the gene is inhibited by using RNAi interference technology to close expression of the gene.
[0033] Overexpressing the aforementioned gene GmMATE85 in a soybean plant can be performed by introducing the aforementioned recombinant expression vector, expression cassette or recombinant bacteria into soybean to overexpress the aforementioned gene GmMATE85 in a soybean plant.
[0034] Any RNAi interference vector used in plants can be used to inhibit the gene encoding the protein to obtain a transgenic cell line and a transgenic plant. An expression vector carrying the gene can be transformed into plant cells or tissues by using a Ti plasmid, a Ri plasmid, a plant virus vector, direct DNA transformation, microinjection, electroporation, Agrobacterium-mediated transformation and other conventional biological methods, and the transformed plant tissue is cultivated into a plant.
[0035] A seventh object of the present application provides a method for cultivating a salt-tolerant or salt-sensitive soybean variety, which comprises inhibiting the gene GmMATE85 in a soybean plant to obtain a salt-tolerant soybean variety; or overexpressing the gene GmMATE85 in a soybean plant to obtain a salt-sensitive soybean variety.
[0036] Advantages:
[0037] The present application discloses a new plant salt tolerance related protein gene GmMATE85. The plant salt tolerance related protein of the present application affects the salt tolerance of plants. Inhibition of the expression of the protein coding gene can improve the salt tolerance of plants, so that salt-tolerant transgenic plants can be cultivated. Introduction of the protein coding gene into plants can cause the salt tolerance of plants to decrease, so that salt-sensitive transgenic plants can be cultivated. The protein and its coding gene can be applied to plant genetic improvement. BRIEF DESCRIPTION OF DRAWINGS
[0038] Figure 1 The expression amount of GmMATE85 gene in GmMATE85-RNAi and GmMATE85-OE soybean hairy root plants.
[0039] Figure 2 The phenotype (a) and the fresh weight of the first compound leaf (b) of GmMATE85-RNAi and GmMATE85-OE soybean hairy root plants under salt stress. DETAILED DESCRIPTION
[0040] The following examples facilitate a better understanding of the present application, but do not limit the present application. In the following examples, the experimental methods are conventional methods, unless otherwise specified. In the following examples, the test materials are commercially available, unless otherwise specified. The soybean variety Williams82 used in the transgenation is a publicly available variety.
[0041] Example 1, cloning of soybean gene GmMATE85
[0042] The following primers are designed:
[0043] Primer 1: 5'-ATGTCATGCATCAGCTTCACTG-3' (SEQ ID NO. 4);
[0044] Primer 2: 5'-ACTCTGAATCTTAGGTTCCTTC-3' (SEQ ID NO. 4).
[0045] PCR amplification was performed using primer 1 and primer 2 as primers and the root cDNA of Williams82 seedlings as template to obtain the target gene GmMATE85.
[0046] PCR amplification reaction was carried out in Bio-rad T100 PCR instrument, and the reaction system (50 μL) was as follows: 25 μL of 2x Phanta Max Buffer, 1 μL of dNTP Mix (10 mM), 1.5 μL of primer 1 (10 μM), 1.5 μL of primer 2 (10 μM), 2 μL of template cDNA (50 ng / uL), 1 μL of Phanta Max Super-Fidelity DNA Polymerase, and 18 μL of ddH2O; the program was as follows: 95 °C pre-denaturation for 3 min; 95 °C denaturation for 15 s, 58 °C annealing for 15 s, 72 °C extension for 2 min, 35 cycles; 72 °C extension for 5 min; 15 °C storage.
[0047] After the PCR product was recovered and purified, it was ligated to pEASY-Blunt (Beijing Zixingjin Biotechnology Co., Ltd.), and E. coli DH5α competent cells (CB101, Beijing Tiangen Co.) were transformed. After positive clones were selected, sequencing was performed.
[0048] The sequencing results showed that the GmMATE85 gene fragment obtained by PCR reaction had the nucleotide sequence shown in SEQ ID NO. 2, encoded a protein (SEQ ID NO. 3) composed of 535 amino acid residues, and the whole genome sequence was shown in SEQ ID NO. 1.
[0049] Example 2, Obtaining and Identifying of Soybean GmMATE85 Gene RNAi Rhizogenesis Combination Plant
[0050] I. Construction of GmMATE85 Gene RNAi Vector
[0051] The target sequence of GmMATE85 gene (SEQ ID NO. 2) was designed, the siRNA sequence was shown in SEQ ID NO. 6, the genomic cDNA of Williams82 root was used as a template, primer 3 and primer 4 were used for PCR amplification, and the sequence shown in SEQ ID NO. 6 was obtained.
[0052] Primer 3: 5'-ATGTCATGCATCAGCTTCAC-3';
[0053] Primer 4: 5'-GATCCATGTGGCGGAGAG-3'.
[0054] The amplification product was ligated to the pFGC5941 vector twice through NcoI and XbaI, and transformed into E. coli DH5α, and positive plasmids were extracted and sequenced. The sequencing results showed that a recombinant expression vector containing the sequence shown in SEQ ID NO. 6 was obtained, and was named pFGC5941-GmMATE85.
[0055] II. Obtaining of the recombinant Agrobacterium
[0056] The pFGC5941-GmMATE85 was transformed into the Agrobacterium K599 strain by heat shock method to obtain a recombinant strain, and the plasmid was extracted for PCR and enzyme digestion identification, and the recombinant strain with correct identification was named K5-pFGC5941-GmMATE85.
[0057] III. Obtaining of the transgenic plant
[0058] The K5-pFGC5941-GmMATE85 strain was transformed into soybean variety Williams82, and the specific method was as follows:
[0059] (1) Mature seeds with large size, fullness, and no disease spots were selected, and washed with deionized water. Then clean filter paper was sprayed with water and placed in a clean culture dish, and the selected soybean seeds were evenly spread on the filter paper, and germinated at 26°C in the dark.
[0060] (2) After the soybean seeds germinated, the seeds with consistent growth were selected and sown in a turnover box containing vermiculite, and a layer of vermiculite was covered on the surface of the seeds, and placed in a 26°C culture room for germination for 3-4 days. When the soybean cotyledons were about to open but had not yet opened (the color was light green, and the lateral roots were growing), the hairy roots could be infected.
[0061] (3) When the seeds germinated for 2-3 days, the K5-pFGC5941-GmMATE85 strain was cultured at 28°C and 200r overnight for about 12h; the bacterial liquid was inoculated into new LB medium at 1:100, and cultured at 28°C and 200r until the OD 600 = 0.6-0.8, and the bacterial cells were collected;
[0062] (4) The centrifuged bacterial cells were resuspended with an equal volume of prepared and sterilized co-culture medium (CCM).
[0063] (5) The germinated seeds without contamination and with intact cotyledons were selected, cut off the base with a scalpel, and the lower hypocotyls of about 2cm were reserved, and the soybean hypocotyls were soaked in the above resuspended bacterial liquid for 1h. Then the infected explants were transplanted in moist vermiculite and cultured in a 26°C culture room.
[0064] (6) After culturing for one week, when white callus grows at the hypocotyl wound of the soybean, the callus is cultured in 1 / 2 Hoagland nutrient solution, and after continuing to culture for one week, when soybean root hairs grow at the callus, the root hairs are obtained as transgenic soybean plants.
[0065] IV. Identification of transgenic plants
[0066] 1. PCR molecular identification
[0067] DNA of the root of the soybean hairy root combination plant is extracted as a template for PCR amplification, and a nucleotide fragment of an edited target point and both sides shown in SEQ ID NO. 6 is amplified by PCR. The PCR primers are as follows:
[0068] Primer 5: 5'-ATGTCATGCATCAGCTTCAC-3';
[0069] Primer 6: 5'-CTCAGGTTTTTTACAACGTGCAC-3'.
[0070] The primer 5 above is located in the target sequence of the GmMATE85 gene shown in SEQ ID NO. 6, and the primer 6 is located in the pFGC5941 vector sequence.
[0071] The PCR product is detected by 1% agarose gel electrophoresis, and the target band can be detected in the positive plants, but not in the negative plants.
[0072] 2. Detection of GmMATE85 gene expression amount
[0073] The root system sample of the GmMATE85 soybean hairy root combination plant is frozen and homogenized by liquid nitrogen, and RNA is extracted by the TRIzol method. An appropriate amount of RNA is used to obtain cDNA by using a reverse transcription kit as a template for fluorescence quantitative RT-PCR detection. An appropriate amount of template cDNA is used to detect the expression amount of the GmMATE85 gene by using the Hieff qPCR TM qPCR Green Master Mix (No RoxPlux) kit in a Bio-Rad fluorescence quantitative PCR instrument CFX96. The GmELF gene quantitative detection primers (primer 7 and primer 8) and the GmMATE85 gene quantitative detection primers (primer 9 and primer 10) used are as follows:
[0074] Primer 7: 5'-GTTGAAAAGCCAGGGGACA-3';
[0075] Primer 8: 5'-TCTTACCCCTTGAGCGTGG-3'.
[0076] Primer 9: 5'-ACTCTGCTGATTCCCGTCTC-3';
[0077] Primer 10: 5'-GGCGAGTTCGAGGTTTCC-3'.
[0078] The relative expression amount was quantitatively calculated by using 2 -△△CT The results are shown in Table 1. Figure 1 Figure 1 MATE85-RNAi-1, MATE85-RNAi-2, MATE85-RNAi-3) in Table 1.
[0079] Example 3, obtaining and identification of soybean GmMATE85 gene overexpression hairy root combination plants
[0080] I. Construction of GmMATE85 gene overexpression vector
[0081] The root genomic cDNA of Williams82 seedlings was used as a template, and primer 1 and primer 2 were used for PCR amplification to obtain a GmMATE85 gene full-length CDS sequence fragment (SEQ ID NO. 2).
[0082] Primer 1: 5'-ATGTCATGCATCAGCTTCACTG-3' (SEQ ID NO. 4);
[0083] Primer 2: 5'-ACTCTGAATCTTAGGTTCCTTC-3' (SEQ ID NO. 5).
[0084] The amplification product was single-enzyme digested by XbaI and connected to the pBA002 vector, and then transformed into E. coli DH5α, and the positive plasmid was extracted and sequenced. The sequencing results showed that a recombinant expression vector containing the sequence shown in SEQ ID NO. 2 was obtained, which was named pBA002-GmMATE85.
[0085] II. Obtaining of recombinant Agrobacterium
[0086] The pBA002-GmMATE85 was transformed into Agrobacterium K599 strain by heat shock method to obtain a recombinant strain, and the plasmid was extracted for PCR and enzyme digestion identification. The recombinant strain correctly identified was named K5-pBA002-GmMATE85.
[0087] III. Obtaining of transgenic plants
[0088] The K5-pBA002-GmMATE85 strain is transformed into soybean variety Williams82, and the specific method is as follows:
[0089] (1) Select mature seeds with large size, fullness, and no disease spots, and wash them with deionized water. Then take clean filter paper, spray it with water, and place it in a clean culture dish. Spread the selected soybean seeds evenly on the filter paper, and germinate them at 26°C in the dark.
[0090] (2) After the soybean seeds germinate, select seeds with consistent growth and sow them in a turnover box containing vermiculite. Cover the seeds with another layer of vermiculite and place them in a 26°C incubator for 3-4 days to germinate. When the soybean cotyledons are about to open but have not yet opened (the color is light green, and lateral roots are growing), the hairy roots can be infected.
[0091] (3) When the seeds germinate for 2-3 days, cultivate the K5-pBA002-GmMATE85 strain at 28°C and 200r overnight for about 12 hours. Inoculate the bacterial solution 1:100 into new LB medium and cultivate at 28°C and 200r until the OD 600 = 0.6-0.8, collect the bacterial cells;
[0092] (4) Resuspend the centrifuged bacterial cells with an equal volume of prepared and sterilized co-culture medium (CCM).
[0093] (5) Select germinated seeds without contamination and intact cotyledons, cut off the base with a scalpel, and reserve about 2cm of hypocotyls. Soak the soybean hypocotyls in the resuspended bacterial solution for 1 hour. Then transplant the infected explants into moist vermiculite and cultivate them in a 26°C incubator.
[0094] (6) After one week of cultivation, white callus tissue grows at the wound site of the soybean hypocotyls. Culture them in 1 / 2 Hoagland nutrient solution. After another week of cultivation, soybean root hairs grow at the callus site, and transgenic soybean plants with root hairs are obtained.
[0095] IV. Identification of transgenic plants
[0096] 1. PCR molecular identification
[0097] Extract DNA from the roots of the soybean hairy root combination plants, and use it as a template for PCR amplification. The PCR primers are as follows:
[0098] Primer 11: 5'-CTATCCTTCGCAAGACCCTTC-3';
[0099] Primer 12: 5'-CTTCTGTATGGGGAAGTTCAC-3'.
[0100] The primers described above, primer 11, are located in the pBA002 vector sequence, and primer 12 is located in the CDS sequence of the GmMATE85 gene shown in SEQ ID NO.2.
[0101] The PCR products were detected by 1% agarose gel electrophoresis. The target band could be detected in positive plants, but not in negative plants.
[0102] 2. Detection of GmMATE85 gene expression level
[0103] Root samples were collected from soybean rooting hybrid plants (GmMATE85), homogenized by liquid nitrogen freezing, and RNA was extracted using the TRIzol method. An appropriate amount of RNA was used to obtain cDNA using a transcription kit, which was then used as a template for quantitative RT-PCR detection. An appropriate amount of template cDNA was taken, and soybean GmELF was used as an internal control gene for analysis using Hieff. TM qPCR The Green Master Mix (No RoxPlux) kit was used to detect the expression level of the GmMATE85 gene in a Bio-Rad CFX96 real-time PCR instrument. The sequences of the primers used for quantitative detection of the GmELF gene (primer 7 and primer 8) and the GmMATE85 gene (primer 9 and primer 10) are as follows:
[0104] Primer7:5'-GTTGAAAAGCCAGGGGACA-3';
[0105] Primer8: 5'-TCTTACCCCTTGAGCGTGG-3'.
[0106] Primer9: 5'-ACTCTGCTGATTCCCGTCTC-3';
[0107] Primer10: 5'-GGCGAGTTCGAGGTTTCC-3'.
[0108] Relative expression level was 2 -△△CT The method is used for quantitative calculation, and the results are as follows: Figure 1 As shown ( Figure 1 MATE85-OE-1, MATE85-OE-2, and MATE85-OE-3 (among others).
[0109] Example 4: Salt tolerance identification of GmMATE85 gene RNAi and overexpression rooting combination plants
[0110] The identified GmMATE85-OE and GmMATE85-RNAi hairy root combination plants were further cultured with 1 / 2 Hoagland nutrient solution to the 2-3rd leaf expansion, wherein the soybean hairy root combination plants transformed with the empty strain K599 were used as a control, and then the soybean hairy root combination plants with relatively consistent growth were selected and transferred to 1 / 2 Hoagland nutrient solution and 1 / 2 Hoagland nutrient solution containing 100 mM NaCl, respectively, for further culture. After 7-10 days of further growth, the phenotypes of the soybean hairy root combination plants were observed by taking photos, and the related physiological indexes were determined.
[0111] Figure 2 The experimental results show that, compared with the soybean hairy root combination plants transformed with the empty strain, the GmMATE85-RNAi soybean hairy root plants exhibit lighter leaf chlorosis and wilting symptoms under salt stress, and the leaf fresh weight of the plants is higher than that of the control group.
[0112] Compared with the soybean hairy root combination plants transformed with the empty strain, the GmMATE85-OE soybean hairy root combination plants exhibit heavier leaf chlorosis and wilting symptoms under salt stress, and the leaf fresh weight of the plants is lower than that of the control group.
[0113] Therefore, according to these results, it can be determined that the GmMATE85 gene has an important negative regulatory effect on the salt tolerance of soybean, and inhibiting the expression of the protein coding gene can improve the salt tolerance of plants, and overexpression of the protein coding gene can increase the salt sensitivity of soybean.
Claims
1. Use of an interfering RNA silencing gene GmMATE85, a RNAi interference vector comprising the interfering RNA silencing gene GmMATE85 or a recombinant bacterium comprising the RNAi interference vector of the interfering RNA silencing gene GmMATE85 in improving salt tolerance of soybean, wherein the sequence of the gene GmMATE85 is shown as SEQ ID NO. 1 or SEQ ID NO.
2.
2. A method for improving salt tolerance in soybean, characterized by, The method is to inhibit the gene GmMATE85 in soybean plants, thereby improving salt tolerance of soybean, wherein the sequence of the gene GmMATE85 is shown as SEQ ID NO. 1 or SEQ ID NO.
2.
3. A method of breeding a salt tolerant soybean variety, characterized by, The method is to inhibit the gene GmMATE85 in soybean plants, thereby obtaining salt-tolerant soybean varieties, wherein the sequence of the gene GmMATE85 is shown as SEQ ID NO. 1 or SEQ ID NO. 2.