Application of MADS22 and coding gene thereof in regulation and control of saline-alkaline tolerance of plants

Abstract

The invention discloses application of MADS22 and a coding gene thereof in regulation and control of saline-alkaline tolerance of plants. The invention belongs to the technical field of biology, and particularly relates to application of MADS22 and a coding gene thereof in regulating and controlling saline-alkaline tolerance of plants. The MADS22 protein can be applied to the following aspects: 1) application to regulation and control of saline-alkaline tolerance of plants; 2) application in preparation of products for regulating and controlling saline-alkaline tolerance of plants; 3) application in cultivation of plants with changed saline-alkaline tolerance; 4) application in preparation of products for cultivating plants with changed saline-alkaline tolerance; and 5) application in plant breeding. The overexpressed MADS22 protein can improve the stress tolerance of plants to high salinity and alkalinity, and is used in corn breeding.

Description

Technical Field

[0001] The invention belongs to the field of biotechnology, and particularly relates to application of MADS22 and its encoding gene in regulating plant salt and alkali tolerance. Background Art

[0002] The trend of soil salinization is expanding worldwide, severely restricting the growth of crops and becoming an important factor restricting agricultural production. The harm caused by salinity stress to plants is that the high concentration of Na in salinized soil + and too high pH. As salt ions accumulate in plant cells, they will produce ion toxicity to the cells, thereby affecting the normal function of the cells in many ways. In an alkaline soil environment, most metal elements except alkali metal elements will form insoluble salts, such as Na + As the most abundant alkali metal element in nature, it accumulates in alkaline soil and causes soil salinization.

[0003] With the rapid development of molecular biology, genomics, genetics, biochemistry, and gene editing technologies, research on the molecular mechanisms of plant resistance to saline-alkali stress has continued to deepen, and many new genes and proteins have been identified as being involved in regulating saline-alkali stress responses. Previous studies have found that MADS22 is primarily involved in plant disease resistance. Our research has discovered that MADS22 also participates in saline-alkali stress responses, regulating plant tolerance to saline-alkali stress. Summary of the Invention

[0004] The technical problem solved by the present invention is how to regulate the salt-alkali tolerance of plants.

[0005] In order to solve the above problems, the present invention provides related applications of proteins, substances that regulate the expression of genes encoding the proteins, or substances that regulate the activity or content of the proteins.

[0006] Use of the protein provided by the present invention, a substance for regulating the expression of a gene encoding the protein, or a substance for regulating the activity or content of the protein in any of the following:

[0007] 1) Application in regulating plant salt-alkali tolerance;

[0008] 2) Application in the preparation of products for regulating plant salt and alkali tolerance;

[0009] 3) Application in cultivating plants with altered salt and alkali tolerance;

[0010] 4) Application in the preparation of products for cultivating plants with altered salt and alkali tolerance;

[0011] 5) Application in plant breeding.

[0012] The protein is any one of the following proteins:

[0013] a1) a protein having the amino acid sequence of SEQ ID No. 2;

[0014] a2) a protein having the same function as the amino acid sequence shown in SEQ ID No. 2, wherein one or more amino acid residues are substituted and / or deleted and / or added;

[0015] a3) a protein having an amino acid sequence of at least 80% identity with the amino acid sequence specified in a1) or (a2) and having the same function;

[0016] a4) A fusion protein obtained by ligating a tag to the end of the protein defined in any one of a1) to (a3).

[0017] In the above proteins, the protein tag refers to a polypeptide or protein that is fused and expressed with the target protein using in vitro DNA recombination technology to facilitate the expression, detection, tracing, and / or purification of the target protein. The protein tag can be a Flag tag, His tag, MBP tag, HA tag, myc tag, GST tag, and / or SUMO tag, etc.

[0018] In the above-mentioned proteins, identity refers to amino acid sequence identity. Amino acid sequence identity can be determined using homology search sites on the Internet, such as the BLAST page on the NCBI homepage. For example, using Advanced BLAST 2.1, blastp can be used as the program, with the Expect value set to 10, all filters set to OFF, BLOSUM62 as the matrix, and the Gap existence cost, Per residue gap cost, and Lambda ratio set to 11, 1, and 0.85 (default values), respectively. The identity of a pair of amino acid sequences can be calculated and the identity value (%) can be obtained.

[0019] In the above proteins, the above 80% or greater identity may be at least 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 95%, 96%, 98%, 99% or 100% identity.

[0020] Among the above proteins, SEQ ID No. 2 consists of 74 amino acid residues and is named MADS22 protein, and its encoding gene is MADS22 gene.

[0021] In the above application, the protein is derived from corn (Zea mays L.).

[0022] Herein, the substance that regulates the activity and / or content of the protein may be a substance that regulates the expression of a gene encoding the protein MADS22.

[0023] In the above, the substance that regulates gene expression may be a substance that performs at least one of the following six types of regulation: 1) regulation at the transcription level of the gene; 2) regulation after transcription of the gene (that is, regulation of the splicing or processing of the primary transcript of the gene); 3) regulation of RNA transport of the gene (that is, regulation of the transport of the mRNA of the gene from the cell nucleus to the cytoplasm); 4) regulation of the translation of the gene; 5) regulation of the degradation of the mRNA of the gene; 6) post-translational regulation of the gene (that is, regulation of the activity of the protein translated from the gene).

[0024] In the present invention, the regulation may be up-regulation, enhancement or increase; the regulation may also be down-regulation, attenuation or reduction.

[0025] Herein, the enhancement, increase or upregulation of the expression level of the gene encoding the protein mentioned above in the recipient plant, or / and the enhancement, increase or upregulation of the activity and / or content of the gene encoding the above protein is achieved by introducing the gene encoding the above protein into the recipient plant.

[0026] Herein, the regulation of the expression of the gene encoding the protein may be inhibition, reduction or down-regulation of the expression of the gene encoding the protein. Inhibition, reduction or down-regulation of the expression of the gene encoding the protein may be achieved by gene knockout or gene silencing.

[0027] In the above application, the substance that regulates the expression of the gene encoding the protein or the substance that regulates the activity or content of the protein may be a biological material related to the protein mentioned above, and the biological material may be any of the following:

[0028] c1) a nucleic acid molecule encoding the protein described above;

[0029] c2) an expression cassette containing the nucleic acid molecule described in c1);

[0030] c3) a recombinant vector containing the nucleic acid molecule described in c1), or a recombinant vector containing the expression cassette described in c2);

[0031] c4) a recombinant microorganism containing the nucleic acid molecule described in c1), or a recombinant microorganism containing the expression cassette described in c2), or a recombinant microorganism containing the recombinant vector described in c3);

[0032] c5) a transgenic plant cell line containing the nucleic acid molecule described in c1) or a transgenic plant cell line containing the expression cassette described in c2);

[0033] c6) transgenic plant tissue containing the nucleic acid molecule described in c1), or transgenic plant tissue containing the expression cassette described in c2);

[0034] c7) a transgenic plant organ containing the nucleic acid molecule described in c1), or a transgenic plant organ containing the expression cassette described in c2);

[0035] e1) a nucleic acid molecule that inhibits, reduces or silences the expression of the protein encoding gene mentioned above;

[0036] e2) an expression cassette containing the nucleic acid molecule described in e1);

[0037] e3) a recombinant vector containing the nucleic acid molecule described in e1), or a recombinant vector containing the expression cassette described in e2);

[0038] e4) a recombinant microorganism containing the nucleic acid molecule described in e1), or a recombinant microorganism containing the expression cassette described in e2), or a recombinant microorganism containing the recombinant vector described in e3);

[0039] e5) a transgenic plant cell line containing the nucleic acid molecule described in e1), or a transgenic plant cell line containing the expression cassette described in e2);

[0040] e6) transgenic plant tissue containing the nucleic acid molecule described in e1), or transgenic plant tissue containing the expression cassette described in e2);

[0041] e7) A transgenic plant organ containing the nucleic acid molecule described in e1), or a transgenic plant organ containing the expression cassette described in e2).

[0042] In the above application, the nucleic acid molecule in c1) can be any of the following DNA molecules,

[0043] d1) the nucleotide sequence is the DNA molecule shown in SEQ ID No. 3;

[0044] d2) the coding sequence is the DNA molecule shown in SEQ ID No. 1;

[0045] d3) a DNA molecule that has 90% or more identity with the nucleotide sequence defined in d1) or d2) and encodes the protein described above;

[0046] d4) A DNA molecule that hybridizes under stringent conditions to the nucleotide sequence defined in d1) or d2) and encodes the protein described above.

[0047] The nucleic acid molecule described herein can be DNA, such as cDNA, genomic DNA or recombinant DNA; the nucleic acid molecule can also be RNA, such as gRNA, mRNA, siRNA, shRNA, sgRNA, miRNA or antisense RNA.

[0048] The vectors described herein are well known to those skilled in the art, and include, but are not limited to, plasmids, phages (such as lambda phage or M13 filamentous phage), cosmids (i.e., cosmids), Ti plasmids, or viral vectors. Specifically, the maize transformation vector pXUE411C-BG may be used.

[0049] Existing plant expression vectors can be used to construct a recombinant expression vector containing the MADS22 gene. Such plant expression vectors include, but are not limited to, binary Agrobacterium vectors and vectors useful for plant microprojectile bombardment. Such plant expression vectors may also contain the 3' untranslated region of the exogenous gene, i.e., a polyadenylation signal and any other DNA fragments involved in mRNA processing or gene expression. The polyadenylation signal can direct the addition of polyadenylic acid to the 3' end of the mRNA precursor. Examples include, but are not limited to, Agrobacterium crown gall-inducing (Ti) plasmid genes (e.g., the rouge synthase Nos gene) and the 3' transcribed untranslated region of plant genes (e.g., the soybean storage protein gene), all of which have similar functions.

[0050] When using the MADS22 gene to construct a recombinant plant expression vector, any enhanced promoter or constitutive promoter can be added before its transcription initiation nucleotide, including but not limited to the cauliflower mosaic virus (CAMV) 35S promoter and the ubiquitin promoter of corn. These promoters can be used alone or in combination with other plant promoters. In addition, when using the gene construction plant expression vector of the present invention, enhancers can also be used, including translation enhancers or transcription enhancers. These enhancer regions can be ATG start codons or adjacent region start codons, but must be identical to the reading frame of the coding sequence to ensure correct translation of the entire sequence. The source of the translation control signal and the start codon is extensive and can be natural or synthetic. The translation initiation region can be from a transcription initiation region or a structural gene.

[0051] To facilitate identification and screening of transgenic plant cells or plants, the plant expression vectors used can be modified to include, but are not limited to, genes encoding enzymes or luminescent compounds that can be expressed in plants (such as the GUS gene or luciferase gene), antibiotic resistance markers (such as gentamicin and kanamycin), or chemical resistance marker genes (such as herbicide resistance genes). For safety reasons, it is possible to omit any selectable marker genes and directly screen for transformed plants using stress.

[0052] The present invention also provides a method for changing the salt-alkali tolerance of plants, the method comprising the following steps M or P:

[0053] The step M is to enhance, increase or upregulate the activity and / or content of the protein mentioned above in the target plant, or / and enhance, increase or upregulate the expression of the gene encoding the protein mentioned above, so as to improve the salt and alkali tolerance of the plant;

[0054] The method includes step P, which is to inhibit, reduce or silence the activity and / or content of the protein mentioned above in the target plant, or / and inhibit, reduce or silence the expression level of the gene encoding the protein mentioned above, so as to reduce the plant's salt and alkali tolerance.

[0055] The present invention also provides a method for cultivating plants with strong salt-alkali tolerance, comprising upregulating, enhancing or increasing the expression level of the gene encoding the protein described above in the target plant, and / or the activity and / or content of the protein to obtain plants with strong salt-alkali tolerance, wherein the salt-alkali tolerance of the plants with strong salt-alkali tolerance is stronger than that of the target plant.

[0056] In a specific embodiment, the upregulation, enhancement or improvement of the expression of the gene encoding the protein described above in the plant comprises introducing the nucleic acid molecule, expression cassette or recombinant vector described above into the target plant to obtain a plant with strong salt and alkali tolerance.

[0057] The present invention also provides a method for cultivating plants with weak salt-alkali tolerance, comprising inhibiting, reducing or silencing the expression level of the gene encoding the protein described above in the target plant, and / or the activity and / or content of the protein, to obtain salt-alkali tolerant plants, wherein the salt-alkali tolerant plants have weaker salt-alkali tolerance than the target plant.

[0058] Herein, the purpose of the breeding includes cultivating plants with strong salt-alkali tolerance; the purpose of the breeding also includes cultivating plants with weak salt-alkali tolerance.

[0059] Herein, the salt-alkali tolerance can be to increase plant height and improve the degree of leaf yellowing.

[0060] Herein, the corn may be corn B73.

[0061] The above-mentioned proteins and / or biological materials also fall within the scope of protection claimed by the present invention.

[0062] In the above application or method, the plant may be any one of the following:

[0063] N1) Monocots;

[0064] N2) Gramineae;

[0065] N3) Grasses;

[0066] N4) Zea mays;

[0067] N5) Corn.

[0068] The experiments of the present invention show that the MADS22 overexpression strain shows significant salt-alkali stress tolerance compared with the wild type B73-329. The MADS22 protein can improve the plant's resistance to high salt and alkali stress and is used for corn breeding. BRIEF DESCRIPTION OF THE DRAWINGS

[0069] Figure 1 Figure 2 shows the phenotypes of wild-type maize B73-329 and MADS22 overexpressing strains in soil treated with NaHCO3. CAUB1815 is a MADS22 overexpressing strain. DETAILED DESCRIPTION

[0070] The present invention will be further described in detail below in conjunction with specific embodiments. The examples provided are only for illustrating the present invention and are not intended to limit the scope of the present invention. The examples provided below can serve as a guide for further improvements by those skilled in the art and are not intended to limit the present invention in any way.

[0071] Unless otherwise specified, the experimental methods in the following examples are conventional methods and were performed according to the techniques or conditions described in the literature in the field or according to the product instructions. The materials and reagents used in the following examples, unless otherwise specified, were all commercially available.

[0072] Unless otherwise specified, the quantitative tests in the following examples were performed three times, and the results were averaged.

[0073] The maize B73-329 used in the following examples has been described in Liu M, Zhang S, Li W, Zhao X, Wang XQ. Identifying yield-related genes in maize based on ear trait plasticity. Genome Biol. 2023; 24(1): 94. The public can obtain the biological material from the applicant for use only in repeating the experiments of the present invention and cannot be used for other purposes.

[0074] Example 1: Application of MADS22 gene in regulating plant salt-alkali tolerance

[0075] The coding sequence (CDS) of the MADS22 gene in corn variety B73-329 is SEQ ID No: 1, and the encoded amino acid sequence of the MADS22 protein is SEQ ID No: 2. In the genomic DNA of corn variety B73-329, the genomic gene encoding the MADS22 protein is shown in SEQ ID No: 3 in the sequence listing.

[0076] 1. Obtaining MADS22 overexpression lines

[0077] 1.1 Culture medium preparation

[0078] LB medium (1000 mL): 10 g of tryptone, 5 g of yeast extract, 10 g of NaCl, and 8 g of agarose (LB liquid medium without agarose).

[0079] 1.2 Amplification of fragments, gel recovery, enzyme digestion and ligation

[0080] The target gene and its associated promoter and terminator were inserted into the T-DNA, and a plant expression vector, named pXUE411C-BG-OverExp-ZmMADS22, was constructed using conventional molecular biology methods. Positive clones were identified using enzyme digestion and bidirectional sequencing for use in maize transformation.

[0081] According to the sequence of the MADS22 gene, upstream and downstream primer sequences were designed, wherein F: 5'-CTATAAGTTACTCACGTTCTCGTATC-3', R: 5'-CACCAATTCTATTCATGTAACGAAA-3'. After amplification, the target fragment of approximately 678 bp was obtained (containing the CDS sequence of MADS22 with the nucleotide sequence of SEQ ID No: 1). The target fragment was then ligated to the starting vector pXUE411C-BG to obtain the recombinant vector pXUE411C-BG-OverExp-ZmMADS22.

[0082] The structure of the overexpression vector pXUE411C-BG-OverExp-ZmMADS22 is described as follows: The DNA fragment (SEQ ID No: 1) is inserted between the EcoRI and KPnI restriction sites of the starting vector pXUE411C-BG, while the other sequences of the pXUE411C-BG vector remain unchanged. This recombinant vector is driven by the strong Ubi promoter and can overexpress MADS22, whose amino acid sequence is shown in SEQ ID No: 2.

[0083] 1.3 E. coli transformation culture

[0084] Melt 50 μL of E. coli competent cells on ice, add the recombinant vector pXUE411C-BG-OverExp-ZmMADS22, mix gently, and place on ice for 30 minutes. Heat shock in a 42°C water bath for 30 seconds, and quickly transfer the centrifuge tube to an ice bath for 2 minutes. Add 700 μL of sterile LB medium (without antibiotics) to each centrifuge tube, mix well, and culture at 37°C, 200 rpm for 1 hour to revive the strain. Centrifuge at 5000-6000 rpm for 5 minutes, aspirate part of the supernatant, and shake. Spread the plate. (Contains antibiotic kana at a concentration of 50 μg / ml) and culture at 37°C for 12 hours to 16 hours. Pick single clones based on colony growth. Colony PCR was performed with reference to the Gel Extraction Kit (Omega, product number QYM10016).

[0085] The PCR products were verified by electrophoresis, and positive clones were obtained. The clones were selected for shaking, and the plasmids were extracted with a kit and sent for sequencing. The recombinant vector pXUE411C-BG-OverExp-ZmMADS22 was successfully obtained.

[0086] 1.4 Agrobacterium transformation (2-3 days)

[0087] 1) Thaw 50 μL EHA105 Agrobacterium competent cells on ice, add 1-2 μL plasmid DNA, mix gently, and incubate on ice for 30 minutes.

[0088] 2) Freeze in liquid nitrogen for 1 min, heat shock in a 37°C water bath for 5 min, and quickly transfer the centrifuge tube to an ice bath for 2 min.

[0089] 3) Add 1 mL of sterile LB medium (without antibiotics) to each centrifuge tube, mix well, and incubate at 28°C, 180 rpm for 3 h to resuscitate the strain.

[0090] 4) Centrifuge at 5000-6000 rpm for 5 min, aspirate part of the supernatant and shake.

[0091] 5) Spread the plate (containing 50 μg / mL of Kana and 40 μg / mL of Rif) and incubate at 28°C in the dark for 2-3 days.

[0092] 1.5 Agrobacterium infection and transgenic strain acquisition

[0093] Transgenic plants were obtained by Agrobacterium infection of maize embryos. The ZmMADS22 gene plant transformation vector was transformed into Agrobacterium EHA105 to generate EHA105 / pXUE411C-BG-OverExp-ZmMADS22. Maize embryos were then infected with Agrobacterium EHA105 / pXUE411C-BG-OverExp-ZmMADS22 containing the target gene. Transgenic plants were obtained after selection with the herbicide bialaphos. The specific transgenic method is as follows:

[0094] The recipient strain used in the transgenic process is the inbred line B73. First, the inbred line B73 is planted in the field and bagged when it begins to shed pollen. Then, preparations for pollination are made. Nine to eleven days after pollination, immature embryos are harvested from the kernels of the pollinated ears. These are then infected with Agrobacterium indoors. These Agrobacterium-infected embryos are placed on a selective medium and screened multiple times to obtain resistant calli. These calli are then regenerated into seedlings, resulting in the T0 generation of transgenic plants. Once the T0 generation of transgenic plants is obtained, pollen from these T0 plants is self-pollinated to regenerate the T2 generation of MADS22-overexpressing strains.

[0095] 2. Application of MADS22 gene in plant salt-alkali tolerance

[0096] Corn seeds of B73-329 and MADS22 overexpressing strains were taken, with B73-329 as the control. They were planted in pure vermiculite without nutrient soil and nutrient solution (composition of 1 / 2 corn nutrient solution as follows: 0.75mM K2SO4, 0.65mM MgSO4.7H2O, 2mMCa(NO3)2.4H2O, 0.1mM EDTA-Fe, 0.1mM KCl, 0.25mM KH2PO4, trace elements including 0.001mM MnSO4.H2O, 0.001mM ZnSO4.7H2O, 0.0001mM CuSO4.5H2O, 0.00005mM (NH4)6Mo7O) was used. 24 After about 7-10 days of growth (three-leaf stage), corn was irrigated with a nutrient solution containing 1 / 2 corn nutrient solution containing 100mM NaHCO3 (for saline-alkali stress). Watering was done every 7-10 days, using two liters per tray. Phenotypes were observed after about 30-40 days of growth. Screening criteria: corn growth and leaf yellowing were observed.

[0097] The results are as follows Figure 1 As shown in the results, under 100 mM NaHCO₃ treatment, the MADS22 overexpressing line showed stronger salt-alkali tolerance than the wild-type B73-329, with greener leaves and taller plants. These results indicate that MADS22 can positively regulate plant salt-alkali resistance.

[0098] The present invention has been described in detail above. For those skilled in the art, without departing from the purpose and scope of the present invention, and without the need to carry out unnecessary experimental conditions, the present invention can be implemented in a wide range under equivalent parameters, concentrations and conditions. Although the present invention provides specific embodiments, it should be understood that further improvements can be made to the present invention. In short, according to the principles of the present invention, this application is intended to include any changes, uses or improvements to the present invention, including changes that depart from the disclosed scope in this application and are made using conventional techniques known in the art.

Claims

1. Use of a protein or a substance that regulates the expression of a gene encoding the protein or a substance that regulates the activity or content of the protein in any of the following; 1) Application in regulating plant salt-alkali tolerance; 2) Application in the preparation of products for regulating plant salt and alkali tolerance; 3) Application in cultivating plants with altered salt and alkali tolerance; 4) Application in the preparation of products for cultivating plants with altered salt and alkali tolerance; 5) Application in plant breeding; The protein is any one of the following proteins: a1) a protein having an amino acid sequence of SEQ ID No. 2; a2) a protein having the same function as the amino acid sequence shown in SEQ ID No. 2 after one or more amino acid residues are substituted and / or deleted and / or added; a3) a protein having an amino acid sequence identity of at least 80% with that specified in a1) or a2) and having the same function; a4) A fusion protein obtained by ligating a tag to the end of the protein defined in any one of a1) to a3).

2. The use according to claim 1, characterized in that The protein is derived from corn.

3. The use according to claim 1 or 2, characterized in that The substance that regulates gene expression or the substance that regulates the activity or content of the protein is a biological material related to the protein in the application according to claim 1 or 2, and the biological material is any one of the following: c1) a nucleic acid molecule encoding the protein; c2) an expression cassette containing the nucleic acid molecule described in c1); c3) a recombinant vector containing the nucleic acid molecule described in c1), or a recombinant vector containing the expression cassette described in c2); c4) a recombinant microorganism containing the nucleic acid molecule described in c1), or a recombinant microorganism containing the expression cassette described in c2), or a recombinant microorganism containing the recombinant vector described in c3); c5) a transgenic plant cell line containing the nucleic acid molecule described in c1) or a transgenic plant cell line containing the expression cassette described in c2); c6) transgenic plant tissue containing the nucleic acid molecule described in c1), or transgenic plant tissue containing the expression cassette described in c2); c7) A transgenic plant organ containing the nucleic acid molecule described in c1) or a transgenic plant organ containing the expression cassette described in c2).

4. The use according to claim 3, characterized in that c1) The nucleic acid molecule is a DNA molecule as shown below, d1) the nucleotide sequence is the DNA molecule shown in SEQ ID No. 3; d2) the coding sequence is the DNA molecule shown in SEQ ID No. 1; d3) a DNA molecule that has 90% or more identity with the nucleotide sequence defined in d1) or d2) and encodes the protein of claim 1; d4) A DNA molecule that hybridizes under stringent conditions with the nucleotide sequence defined in d1) or d2) and encodes the protein of claim 1.

5. A method for changing the salt-alkali tolerance of plants, characterized by: The method includes steps P or M, wherein step P is to enhance, increase or upregulate the activity and / or content of the protein according to claim 1 or 2 in the target plant, or / and, enhance, increase or upregulate the expression level of the gene encoding the protein according to claim 1 or 2, so as to improve the salt-alkali tolerance of the plant; and step M is to inhibit, reduce or silence the activity and / or content of the protein according to claim 1 or 2 in the target plant, or / and, inhibit, reduce or downregulate the expression level of the gene encoding the protein according to claim 1 or 2, so as to reduce the salt-alkali tolerance of the plant.

6. A method for cultivating plants with high salt-alkali tolerance, characterized in that: The method comprises enhancing, increasing or upregulating the expression level of the gene encoding the protein according to claim 1 or 2 in the target plant, and / or the activity and / or content of the protein, to obtain a plant with strong salt-alkali tolerance, wherein the salt-alkali tolerance of the plant with strong salt-alkali tolerance is stronger than that of the target plant.

7. The method according to claim 6, characterized in that The enhancing, increasing or upregulating the expression of the gene encoding the protein of claim 1 or 2 in the plant comprises introducing the nucleic acid molecule described in c1) of claim 4, the expression cassette described in c2) or the recombinant vector described in c3) of claim 4 into the target plant to obtain a plant with enhanced salt and alkali tolerance.

8. A method for cultivating plants with weak salt and alkali tolerance, characterized in that: The method comprises inhibiting, reducing or silencing the expression level of the gene encoding the protein according to claim 1 or 2 in the target plant, and / or the activity and / or content of the protein, thereby obtaining a plant with weak salt-alkali tolerance, wherein the salt-alkali tolerance of the plant with weak salt-alkali tolerance is weaker than that of the target plant.

9. The protein according to claim 1 or 2 and / or the biomaterial according to claim 3 or 4.

10. The method according to any one of claims 5 to 8, characterized in that The plant is any one of the following: N1) Monocotyledons; N2) Gramineae; N3) Grasses; N4) Zea mays; N5) Corn.

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