CN120665885A - MADS22 and application of coding gene thereof in regulation and control of saline-alkali tolerance of plants - Google Patents

Abstract

The invention discloses MADS22 and application of a coding gene thereof in regulating and controlling saline-alkali 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 regulation and control of saline-alkali tolerance of plants. MADS22 protein can be applied to 1) application in regulating and controlling the saline-alkali tolerance of plants, 2) application in preparing products for regulating and controlling the saline-alkali tolerance of plants, 3) application in cultivating plants with changed saline-alkali tolerance, 4) application in preparing products for cultivating plants with changed saline-alkali tolerance, and 5) application in plant breeding. The over-expression MADS22 protein can improve the stress tolerance of plants to high saline and alkaline and is used for corn breeding.

Description

MADS22 and application of coding gene thereof in regulation and control of saline-alkali tolerance of plants

Technical Field

The invention belongs to the technical field of biology, and particularly relates to application of MADS22 and a coding gene thereof in regulation and control of saline-alkali tolerance of plants.

Background

The salinization trend of soil is continuously expanded worldwide, the growth of crops is severely limited, and the soil salinization trend becomes an important factor for restricting agricultural production. The damage caused by saline-alkali stress to plants is high concentration of Na ⁺ and excessively high pH in saline-alkali soil. Along with the accumulation of salt ions in plant cells, ion toxic effects are generated on the cells, so that the normal functions of the cells are influenced in various aspects. In alkaline earth environment, most of metal elements except alkali metal elements form insoluble salts, and Na ⁺ is used as the alkali metal element with the highest content in nature, and is enriched in alkaline earth to cause salinization of soil.

With the rapid development of molecular biology, genomics, genetics, biochemistry and gene editing technology, the molecular mechanism of plant resistance to saline-alkali stress is continuously and deeply studied, and a plurality of new genes or proteins are involved in the regulation and control of the saline-alkali stress response process. Previous researches find that MADS22 is mainly involved in the disease resistance response process of plants, and our researches find that MADS22 is also involved in the saline-alkali stress response process, and the saline-alkali resistance of plants is regulated.

Disclosure of Invention

The invention solves the technical problem of how to regulate and control the saline-alkali tolerance of plants.

In order to solve the above problems, the present invention provides related applications of a protein, a substance regulating the expression of a gene encoding the protein, or a substance regulating the activity or content of the protein.

The invention provides the application of the protein, the substance for regulating the expression of the encoding gene of the protein or the substance for regulating the activity or the content of the protein in any one of the following:

1) The application in regulating and controlling the saline-alkali tolerance of plants;

2) The application in preparing the product for regulating and controlling the saline-alkali tolerance of plants;

3) The application in cultivating plants with modified saline-alkali tolerance;

4) The application in preparing a product for cultivating plants with modified saline-alkali tolerance;

5) The application in plant breeding.

The protein is any one of the following proteins:

a1 Protein with the amino acid sequence of SEQ ID No. 2;

a2 A protein which is obtained by substituting and/or deleting and/or adding one or more amino acid residues in the amino acid sequence shown in SEQ ID No.2 and has the same function;

a3 A protein having 80% or more identity and the same function as the amino acid sequence defined in a 1) or (a 2);

a4 A fusion protein obtained by ligating a tag to the end of the protein defined in any one of a 1) to a 3).

Among the above proteins, the protein tag (protein-tag) refers to a polypeptide or protein that is fusion expressed together with a target protein by using a DNA in vitro recombination technique, so as to facilitate the expression, detection, tracing and/or purification of the target protein. The protein tag may be a Flag tag, his tag, MBP tag, HA tag, myc tag, GST tag, and/or SUMO tag, etc.

In the above proteins, the identity refers to the identity of amino acid sequences. The identity of amino acid sequences can be determined using homology search sites on the internet, such as BLAST web pages of the NCBI homepage website. For example, in advanced BLAST2.1, by using blastp as a program, expect values are set to 10, all filters are set to OFF, BLOSUM62 is used as Matrix, gap existence cost, per residue gap cost and Lambda ratio are set to 11,1 and 0.85 (default values), respectively, and identity of a pair of amino acid sequences is searched for and calculated, and then the value (%) of identity can be obtained.

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

In the above protein, SEQ ID No.2 consists of 74 amino acid residues. This was designated MADS22 protein and its coding gene was MADS22 gene.

In the above application, the protein is derived from corn (Zea mays l.).

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

In the above, the substance regulating the expression of a gene may be a substance which performs at least one of 1) regulation at the level of transcription of the gene, 2) regulation after transcription of the gene (i.e., regulation of splicing or processing of the primary transcript of the gene), 3) regulation of RNA transport of the gene (i.e., regulation of nuclear to cytoplasmic transport of mRNA of the gene), 4) regulation of translation of the gene, 5) regulation of mRNA degradation of the gene, and 6) regulation after translation of the gene (i.e., regulation of activity of protein translated by the gene).

In the present invention, the regulation may be up-regulation or enhancement or improvement, or the regulation may be down-regulation or attenuation or reduction.

The enhancement, increase or up-regulation of the expression level of the gene encoding the aforementioned protein in the recipient plant, or/and the enhancement, increase or up-regulation of the activity and/or content of the gene encoding the aforementioned protein, is achieved by introducing the gene encoding the aforementioned protein into the recipient plant.

Herein, the modulating the expression of the encoding gene of the protein may be inhibiting or reducing or down-regulating the expression of the encoding gene. Inhibition or reduction or downregulation of expression of the coding gene may be achieved by gene knockout or gene silencing.

In the above application, the substance regulating the expression of the gene encoding the protein or the substance regulating the activity or content of the protein may be a biological material related to the protein as described above, and the biological material may be any of the following:

c1 A nucleic acid molecule encoding a protein as described above;

c2 An expression cassette comprising c 1) said nucleic acid molecule;

c3 A recombinant vector comprising c 1) said nucleic acid molecule, or a recombinant vector comprising c 2) said expression cassette;

c4 A recombinant microorganism comprising c 1) said nucleic acid molecule, or a recombinant microorganism comprising c 2) said expression cassette, or a recombinant microorganism comprising c 3) said recombinant vector;

c5 A transgenic plant cell line comprising c 1) said nucleic acid molecule, or a transgenic plant cell line comprising c 2) said expression cassette;

c6 A transgenic plant tissue comprising c 1) said nucleic acid molecule, or a transgenic plant tissue comprising c 2) said expression cassette;

c7 A transgenic plant organ comprising c 1) said nucleic acid molecule, or a transgenic plant organ comprising c 2) said expression cassette;

e1 A nucleic acid molecule that inhibits or reduces or silences the expression of a gene encoding a protein as described above;

e2 An expression cassette comprising e 1) said nucleic acid molecule;

e3 A recombinant vector comprising e 1) said nucleic acid molecule, or a recombinant vector comprising e 2) said expression cassette;

e4 A recombinant microorganism comprising e 1) said nucleic acid molecule, or a recombinant microorganism comprising e 2) said expression cassette, or a recombinant microorganism comprising e 3) said recombinant vector;

e5 A transgenic plant cell line comprising e 1) said nucleic acid molecule, or a transgenic plant cell line comprising e 2) said expression cassette;

e6 A transgenic plant tissue comprising e 1) said nucleic acid molecule, or a transgenic plant tissue comprising e 2) said expression cassette;

e7 A transgenic plant organ containing e 1) said nucleic acid molecule, or a transgenic plant organ containing e 2) said expression cassette.

In the above application, the nucleic acid molecule of c 1) may be a DNA molecule as shown in any one of the following,

D1 A DNA molecule with a nucleotide sequence shown as SEQ ID No. 3;

d2 A DNA molecule with a coding sequence shown in SEQ ID No. 1;

d3 A DNA molecule which has 90% or more identity to the nucleotide sequence defined in d 1) or d 2) and which encodes a protein as described above;

d4 A DNA molecule which hybridizes under stringent conditions to the nucleotide sequence defined under d 1) or d 2) and which codes for a protein as described above.

The nucleic acid molecule described herein may be DNA, such as cDNA, genomic DNA or recombinant DNA, or RNA, such as gRNA, mRNA, siRNA, shRNA, sgRNA, miRNA or antisense RNA.

The vectors described herein are well known to those of skill in the art and include, but are not limited to, plasmids, phages (e.g., lambda phage or M13 filamentous phage, etc.), cosmids (i.e., cosmids), ti plasmids, or viral vectors. Specifically, the maize transformation vector pXUE C-BG can be used.

The recombinant expression vector containing the MADS22 gene may be constructed using existing plant expression vectors. Such plant expression vectors include, but are not limited to, vectors such as binary Agrobacterium vectors and vectors useful for microprojectile bombardment of plants, and the like. The plant expression vector may also comprise the 3' -untranslated region of a foreign gene, i.e., comprising a polyadenylation signal and any other DNA segments involved in mRNA processing or gene expression. The polyadenylation signal may direct the addition of polyadenylation to the 3 'end of the mRNA precursor and may function similarly to untranslated regions transcribed from the 3' end of plant genes including, but not limited to, agrobacterium tumefaciens induction (Ti) plasmid genes (e.g., nopaline synthase Nos genes), plant genes (e.g., soybean storage protein genes).

When the MADS22 gene is used to construct a recombinant plant expression vector, any one of an enhanced promoter or a constitutive promoter including, but not limited to, a 35S promoter such as cauliflower mosaic virus (CAMV), a ubiquitin promoter (ubiquitin) of maize, which can be used alone or in combination with other plant promoters, may be added before the transcription initiation nucleotide thereof, and in addition, when the gene of the present invention is used to construct a plant expression vector, enhancers including a translation enhancer or a transcription enhancer may be used, and these enhancer regions may be ATG initiation codons or adjacent region initiation codons, etc., but are necessarily identical to the reading frame of the coding sequence to ensure proper translation of the entire sequence. The sources of the translational control signals and initiation codons are broad, and can be either natural or synthetic. The translation initiation region may be derived from a transcription initiation region or a structural gene.

In order to facilitate the identification and selection of transgenic plant cells or plants, the plant expression vectors used may be processed, such as by adding genes encoding enzymes or luminescent compounds that produce a color change (GUS gene, luciferase gene, etc.), antibiotic markers with resistance (gentamicin markers, kanamycin markers, etc.), or anti-chemical marker genes (e.g., anti-herbicide genes), etc., which may be expressed in plants. From the safety of transgenic plants, transformed plants can be screened directly in stress without adding any selectable marker gene.

The invention also provides a method for changing the saline-alkali tolerance of plants, which comprises the following steps of M or P:

The step M is to enhance, increase or up-regulate the activity and/or content of the protein in the target plant, or/and enhance, increase or up-regulate the expression level of the encoding gene of the protein to improve the saline-alkali tolerance of the plant;

the method comprises a step P, wherein the step P is used for inhibiting or reducing or silencing the activity and/or content of the protein in the target plant, or/and inhibiting or reducing or silencing the expression level of the encoding gene of the protein so as to reduce the saline-alkali tolerance of the plant.

The invention also provides a method for cultivating the plant with strong salt and alkali resistance, which comprises the steps of up-regulating or enhancing or improving the expression quantity of the coding gene of the protein in the target plant, and/or obtaining the plant with strong salt and alkali resistance by the activity and/or the content of the protein, wherein the salt and alkali resistance of the plant with strong salt and alkali resistance is higher than that of the target plant.

In a specific embodiment, said up-regulating or enhancing or increasing expression in a plant of a gene encoding a protein as described above comprises introducing into said plant of interest a nucleic acid molecule, expression cassette or recombinant vector as described above, resulting in a plant that is resistant to saline and alkaline.

The invention also provides a method for cultivating a plant with weak salt and alkali resistance, which comprises the steps of inhibiting or reducing or silencing the expression quantity of the coding gene of the protein in a target plant, and/or obtaining the plant with weak salt and alkali resistance by the activity and/or the content of the protein, wherein the salt and alkali resistance of the plant with weak salt and alkali resistance is weaker than that of the target plant.

The breeding purpose comprises the cultivation of plants with strong saline-alkali tolerance, and the breeding purpose also comprises the cultivation of plants with weak saline-alkali tolerance.

Herein, the saline-alkali tolerance may be an improvement in plant height, and an improvement in the yellowing degree of the leaf.

Herein, the corn may be corn B73.

The proteins and/or the biological materials described above are also within the scope of the claimed invention.

In the above application or method, the plant may be any of the following:

n1) monocotyledonous plants;

N2) gramineae plants;

n3) a gramineous plant;

n4) zea plants;

N5) corn.

Experiments prove that compared with wild type B73-329, the MADS22 over-expression strain shows remarkable saline-alkali stress tolerance, and the MADS22 protein can improve the stress tolerance of plants to high saline-alkali and is used for corn breeding.

Drawings

FIG. 1 shows the phenotype of maize wild-type B73-329 and MADS22 overexpressing lines in soil treated with NaHCO ₃. Wherein CAUB1815 is MADS22 overexpressing strain.

Detailed Description

The following detailed description of the invention is provided in connection with the accompanying drawings that are presented to illustrate the invention and not to limit the scope thereof. The examples provided below are intended as guidelines for further modifications by one of ordinary skill in the art and are not to be construed as limiting the invention in any way.

The experimental methods in the following examples, unless otherwise specified, are conventional methods, and are carried out according to techniques or conditions described in the literature in the field or according to the product specifications. Materials, reagents and the like used in the examples described below are commercially available unless otherwise specified.

Unless otherwise indicated, the quantitative tests in the examples below were all performed in triplicate, and the results averaged.

Maize B73-329 in the examples described below 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. public availability of this biomaterial from the applicant, which was used only for repeated experiments of the invention and was not used for other purposes.

Example 1, use of MADS22 Gene for modulating salt and alkaline tolerance in plants

The coding sequence (CDS) of MADS22 gene in maize variety B73-329 is SEQ ID No. 1 and the coding amino acid sequence is MADS22 protein of SEQ ID No. 2. In the genome DNA of the maize variety B73-329, the genome gene for encoding MADS22 protein is shown as SEQ ID No. 3 of the sequence table.

1. Obtaining MADS22 overexpressing lines

1.1 Preparation of Medium

LB medium (1000 mL) 10g tryptone, 5g yeast extract, 10g NaCl, 8g agarose (LB liquid medium does not add agarose).

1.2 Amplified fragments, gel recovery, restriction ligation

The target gene and the matched promoter and terminator are inserted into T-DNA, and plant expression vectors are constructed according to the conventional molecular biology method, and the plant expression vectors are named pXUE C-BG-OverExp-ZmMADS22 respectively. And positive clones were identified for maize transformation by enzymatic cleavage and two-way sequencing.

The sequence of the upstream and downstream primers was designed based on the sequence of MADS22 gene, wherein F:5'-CTATAAGTTACTCACGTTCTCGTATC-3', R:5'-CACCAATTCTATTCATGTAACGAAA-3', the target fragment was obtained about 678bp (CDS sequence of MADS22 having the nucleotide sequence of SEQ ID No. 1) after amplification, and then the target fragment was ligated to the starting vector pXUE C-BG to obtain recombinant vector pXUE C-BG-OverExp-ZmMADS22.

The structure of the over-expression vector pXUE C-BG-OverExp-ZmMADS22 is described as follows, namely a recombinant vector is obtained by inserting a DNA fragment with the sequence of SEQ ID No. 1 between EcoR I and KPn I of the starting vector pXUE C-BG and keeping other sequences of the vector pXUE C-BG unchanged. The vector is expressed by a strong promoter Ubi, pXUE C-BG-OverExp-ZmMADS22 vector can overexpress MADS22, and the amino acid sequence of the vector is SEQ ID No. 2.

1.3 E.coli transformation culture

50 Μl of E.coli competent cells were thawed on ice, added with recombinant vector pXUE C-BG-OverExp-ZmMADS22, gently mixed, and ice-bathed for 30min. The tube was quickly transferred to an ice bath for 2min by heat shock in a 42 ℃ water bath for 30 s. mu.L of sterile LB medium (without antibiotics) was added to each centrifuge tube, mixed well, incubated at 37℃for 1h at 200rpm, and strains were recovered. Centrifuge at 5000-6000rpm for 5min, aspirate part of the supernatant and shake. And (5) coating a plate. (antibiotic-containing kana concentration of 50. Mu.g/ml) 37 ℃. And 12-16 h. The monoclonal was picked according to colony growth. Colony PCR was performed with reference to Gel Extraction Kit kit (Omega, cat# QYM 10016).

And (3) carrying out electrophoresis verification on the PCR product to obtain a positive clone, selecting to shake bacteria, extracting plasmids by using a kit, and carrying out sequencing to successfully obtain the recombinant vector pXUE411C-BG-OverExp-ZmMADS22.

1.4 Agrobacterium transformation (2-3 d)

1) 50. Mu.L of EHA105 Agrobacterium competent cells were thawed on ice, 1-2. Mu.L of plasmid DNA was added and gently mixed, and ice-bathed for 30min.

2) The tube was quickly transferred to an ice bath for 2min by freezing 1min with liquid nitrogen and heat-shock in a 37 ℃ water bath for 5 min.

3) 1ML of sterile LB medium (without antibiotics) was added to each centrifuge tube, mixed well, cultured at 28℃for 3h at 180rpm, and strains were recovered.

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

5) And (5) coating a plate. (containing kana 50 ug/mL+rib 40 ug/mL) at 28 ℃.2-3d.

1.5 Agrobacterium infection and transgenic lines obtained

Transgenic plants are obtained by a method of infecting maize immature embryos by agrobacterium. Transforming the ZmMADS22 gene plant transformation vector into agrobacterium EHA105 to obtain EHA105/pXUE411 CBG-OverExp-ZmMADS 22, infecting maize young embryo with the agrobacterium EHA105/pXUE C-BG-OverExp-ZmMADS22 containing the target gene, and screening by herbicide dipropylamine phosphorus to obtain a transgenic plant, wherein the specific transgenic method is as follows:

The receptor used in the transgenic process was inbred line B73. Firstly, planting an inbred line B73 in the field, bagging when the inbred line is scattered, then preparing pollination, 9-11 days after pollination, taking immature embryo on the seed grain of the pollination cluster, then carrying out agrobacterium infection indoors, placing the young embryo invaded by agrobacterium on a selection medium for multiple screening to obtain a resistant callus, and regenerating the resistant callus into seedlings to obtain a transgenic T0 generation plant. After the transgenic T0 generation is obtained, pollen of the T0 generation transgenic plant is used for selfing, and the T2 generation MADS22 overexpression strain is regenerated.

2. Application of MADS22 gene in plant salt and alkali tolerance

B73-329 and MADS22 over-expressed line corn seeds were taken as controls, and B73-329 was used as a control. Planting in pure vermiculite without nutrient soil, irrigating with half corn nutrient solution (：0.75mM K₂SO₄、0.65mM MgSO₄.7H₂O、2mM Ca(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)6Mo₇O₂₄.4H₂O、0.001mM H₃BO₃), for preparation), irrigating with half corn nutrient solution containing 100mM NaHCO ₃ (saline-alkali stress) after about 7-10 days of corn growth (three leaf stage), irrigating every 7-10 days, and growing for about 30-40 days every two liters of each disk, screening, and observing corn growth and leaf yellowing.

As a result, as shown in FIG. 1, under the condition of 100mM NaHCO ₃ treatment, MADS22 over-expression strain shows stronger salt-alkali resistance than wild type B73-329, leaves are greener, and the plant height is higher. The results show that MADS22 can positively regulate the salt-alkali resistance of plants.

The present application is described in detail above. It will be apparent to those skilled in the art that the present application can be practiced in a wide range of equivalent parameters, concentrations, and conditions without departing from the spirit and scope of the application and without undue experimentation. While the application has been described with respect to specific embodiments, it will be appreciated that the application may be further modified. In general, this application is intended to cover any variations, uses, or adaptations of the application following, in general, the principles of the application and including such departures from the present disclosure as come within known or customary practice within the art to which the application pertains.

Claims (10)

1. Use of a protein or a substance regulating the expression of a gene encoding said protein or a substance regulating the activity or content of said protein in any of the following;

1) The application in regulating and controlling the saline-alkali tolerance of plants;

2) The application in preparing the product for regulating and controlling the saline-alkali tolerance of plants;

3) The application in cultivating plants with modified saline-alkali tolerance;

4) The application in preparing a product for cultivating plants with modified saline-alkali tolerance;

5) Application in plant breeding;

The protein is any one of the following proteins:

a1 Protein with the amino acid sequence of SEQ ID No. 2;

a2 A protein which is obtained by substituting and/or deleting and/or adding one or more amino acid residues in the amino acid sequence shown in SEQ ID No.2 and has the same function;

a3 A protein having 80% or more identity to the amino acid sequence defined in a 1) or a 2) and having the same function;

a4 A fusion protein obtained after the tag has been attached to the end of the protein defined in any one of a 1) to a 3).

2. The use according to claim 1, wherein the protein is derived from maize.

3. The use according to claim 1 or 2, wherein the substance regulating the expression of a gene or the substance regulating the activity or content of the protein is a biological material related to the protein in the use according to claim 1 or 2, said biological material being any of the following:

c1 A nucleic acid molecule encoding said protein;

c2 An expression cassette comprising c 1) said nucleic acid molecule;

c3 A recombinant vector comprising c 1) said nucleic acid molecule, or a recombinant vector comprising c 2) said expression cassette;

c4 A recombinant microorganism comprising c 1) said nucleic acid molecule, or a recombinant microorganism comprising c 2) said expression cassette, or a recombinant microorganism comprising c 3) said recombinant vector;

c5 A transgenic plant cell line comprising c 1) said nucleic acid molecule, or a transgenic plant cell line comprising c 2) said expression cassette;

c6 A transgenic plant tissue comprising c 1) said nucleic acid molecule, or a transgenic plant tissue comprising c 2) said expression cassette;

c7 A transgenic plant organ comprising c 1) said nucleic acid molecule, or a transgenic plant organ comprising c 2) said expression cassette.

4. The use according to claim 3, wherein c 1) the nucleic acid molecule is a DNA molecule as shown in any one of the following,

D1 A DNA molecule with a nucleotide sequence shown as SEQ ID No. 3;

d2 A DNA molecule with a coding sequence shown in SEQ ID No. 1;

d3 A DNA molecule which has 90% or more identity to the nucleotide sequence defined in d 1) or d 2) and which encodes a protein as described in claim 1;

d4 A DNA molecule which hybridizes under stringent conditions to the nucleotide sequence defined in d 1) or d 2) and which codes for a protein according to claim 1.

5. A method for altering the saline-alkali tolerance of a plant, which comprises the step P of enhancing, increasing or up-regulating the activity and/or content of the protein of claim 1 or 2 in the plant of interest, or/and enhancing, increasing or up-regulating the expression level of the gene encoding the protein of claim 1 or 2 to enhance the saline-alkali tolerance of a plant, or the step M of inhibiting or reducing or silencing the activity and/or content of the protein of claim 1 or 2 in the plant of interest, or/and inhibiting or reducing or down-regulating the expression level of the gene encoding the protein of claim 1 or 2 to reduce the saline-alkali tolerance of a plant.

6. A method for growing a plant having high salt and alkali tolerance, comprising enhancing, increasing or upregulating the expression level of a gene encoding a protein according to claim 1 or 2 in a plant of interest, and/or the activity and/or content of said protein, to obtain a plant having high salt and alkali tolerance, said plant having high salt and alkali tolerance being stronger than said plant of interest.

7. The method of claim 6, wherein said enhancing, increasing or up-regulating the expression of a gene encoding the protein of claim 1 or 2 in a plant comprises introducing the nucleic acid molecule of claim 4 c 1), the expression cassette of c 2) or the recombinant vector of c 3) into said plant of interest to obtain a plant with enhanced saline-alkali tolerance.

8. A method for growing a plant with weak salt and alkali tolerance, comprising inhibiting or reducing or silencing the expression level of a gene encoding the protein of claim 1 or 2 in a plant of interest, and/or the activity and/or content of the protein, to obtain a plant with weak salt and alkali tolerance, which is weaker than the plant of interest.

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

10. The method of any one of claims 5-8, wherein the plant is any one of the following:

n1) monocotyledonous plants;

N2) gramineae plants;

n3) a gramineous plant;

n4) zea plants;

N5) corn.