Salt stress regulation gene osdnaj of rice and application thereof

Abstract

The application discloses a salt stress regulation gene of rice OsDNAJ and application thereof. The application belongs to the technical field of biotechnology, and particularly relates to a salt stress regulation gene of rice OsDNAJ and application thereof. The protein OsDNAJ is any one of the following proteins: A1) an amino acid sequence shown in SEQ ID No: 3; A2) a protein obtained by substitution, deletion and / or addition of amino acid residues of the protein of A1) and having more than 75% identity with the protein shown in A1) and having the same function; and A3) a fusion protein obtained by connecting a protein tag to the N terminal or / and C terminal of A1) or A2). After CRISPR gene editing knockout of the gene, OsDNAJ after salt stress treatment, osdnaj the knockout mutant presents a salt tolerance phenotype, indicating that OsDNAJ the gene plays an important role in regulating salt tolerance of rice.

Description

Technical Field

[0001] This invention belongs to the field of biotechnology, specifically relating to rice salt stress regulatory genes. OsDNAJ And its applications. Background Technology

[0002] As one of the world's most important food crops, ensuring rice production is of great significance to national food security. With the intensification of global warming, soil salinization is becoming increasingly serious, posing a particularly severe threat to the yield of monocotyledonous rice.

[0003] Current research on plant responses to salt stress mainly focuses on ion stress, osmotic stress, and oxidative stress. When the osmotic pressure within a plant is lower than that of the soil solution, the plant cannot absorb water and may even lose water, potentially leading to death. Besides osmotic and ion stress, salt stress also generates secondary oxidative stress through their interaction, disrupting the dynamic balance between the production and scavenging of reactive oxygen species (ROS) within the plant. When the accumulation of ROS exceeds its damage threshold, it causes oxidative damage to the plant in two ways: firstly, it exacerbates cell membrane lipid peroxidation and defatting, thereby damaging the plant's cell membrane system; secondly, it severely reduces the activity of specific enzymes responsible for photosynthetic pigment synthesis, impairs chloroplast function, and severely inhibits photosynthesis. Ion stress mainly involves Na+ in the roots. + and Cl - Plants prevent sodium from being transported to cells by opening ion transport channels. + and Cl - Excessive accumulation of osmotic stress. High-salt environments activate osmotic stress signaling pathways, accelerating the synthesis and accumulation of osmotic regulators. In addition, they induce the initiation of MAPK cascade reactions, thereby participating in the scavenging of ROS and the maintenance of ion homeostasis.

[0004] DNAJ family proteins are a class of molecular chaperones that interact with heat shock protein 70 (HSP70), playing crucial roles in the proper folding, assembly, and transport of proteins. Recent research advances have shown that DNAJ family proteins play important roles in plant growth, development, and stress responses. Studies have found that the heat shock protein NAL11, containing a DnaJ domain, affects rice plant architecture by regulating gibberellin (GA) homeostasis. DnaJ proteins participate in the folding, assembly, and transport of nascent peptides by regulating the ATPase activity of HSP70. The functional diversity of DnaJ proteins makes them extremely important in plant morphogenesis, growth, development, and stress responses. However, whether DNAJ proteins are involved in regulating rice salt tolerance remains unclear. Summary of the Invention

[0005] The technical problem to be solved by this invention is how to regulate the salt tolerance of plants.

[0006] To address the problems existing in the prior art, the present invention provides a protein.

[0007] The protein provided by this invention may be any of the following: A1) A protein with the amino acid sequence shown in SEQ ID No:3; A2) A protein obtained by substituting and / or deleting and / or adding amino acid residues of the protein in A1) that has more than 75% identity with the protein shown in A1) and has the ability to regulate plant salt tolerance; for example, those skilled in the art can, based on the amino acid sequence shown in SEQ ID No:3 and conventional techniques such as the conserved substitution of amino acids, obtain a protein mutant with the same function as the amino acid sequence shown in SEQ ID No:3 by substituting, deleting and / or adding one or more amino acids without affecting its activity. A3) A fusion protein obtained by attaching a protein tag to the N-terminus and / or C-terminus of A1) or A2).

[0008] The protein described in A1 above is named OsDNAJ. The OsDNAJ protein consists of 668 amino acids.

[0009] To facilitate the purification or detection of the protein in A1), a tag protein can be attached to the amino or carboxyl terminus of the protein, which consists of the amino acid sequence shown in SEQ ID No:3 in the sequence listing.

[0010] The proteins mentioned above can be synthesized artificially, or their encoding genes can be synthesized first and then expressed biologically.

[0011] The tag proteins include, but are not limited to: GST (glutathione thiotransferase) tag protein, His6 tag protein (His-tag), MBP (maltose-binding protein) tag protein, Flag tag protein, SUMO tag protein, HA tag protein, Myc tag protein, eGFP (enhanced green fluorescent protein), eCFP (enhanced cyan fluorescent protein), eYFP (enhanced yellow-green fluorescent protein), mCherry (monomer red fluorescent protein), or AviTag tag protein.

[0012] Those skilled in the art can readily mutate the nucleotide sequence encoding the protein OsDNAJ of this invention using known methods, such as directed evolution or point mutation. Any artificially modified nucleotides that possess 75% or more of the nucleotide sequence identity with the protein OsDNAJ isolated in this invention, provided they encode and function as protein OsDNAJ, are derived from and equivalent to the nucleotide sequence of this invention.

[0013] The aforementioned 75% or higher degree of identity can be 80%, 85%, 90%, or 95% or higher degree of identity.

[0014] In this article, identity refers to the similarity of amino acid or nucleotide sequences. The identity of amino acid or nucleotide sequences can be determined using homology search sites on the Internet, such as the BLAST page on the NCBI homepage. For example, in Advanced BLAST 2.1, using blastp as the procedure, setting the Expect value to 10, setting all filters to OFF, using BLOSUM62 as the matrix, setting the Gap existence cost, Per residue gap cost, and Lambda ratio to 11, 1, and 0.85 (default values) respectively, and performing a search to calculate the identity of a pair of amino acid sequences or nucleotide sequences, then the identity value (%) can be obtained.

[0015] In this document, the 80% or more of identity can be at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity.

[0016] In this document, the above 90% identity can be at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity.

[0017] The protein mentioned above is derived from rice ( Oryza sativa L.).

[0018] The present invention also provides biomaterials related to the above-mentioned proteins, said biomaterials may be any of the following: B1) Nucleic acid molecules that encode the proteins described above; B2) An expression cassette containing the nucleic acid molecule described in B1); B3) A recombinant vector containing the nucleic acid molecule described in B1), or a recombinant vector containing the expression cassette described in B2); B4) Recombinant microorganisms containing the nucleic acid molecules described in B1), or recombinant microorganisms containing the expression cassette described in B2), or recombinant microorganisms containing the recombinant vector described in B3); B5) A transgenic plant cell line containing the nucleic acid molecule described in B1), or a transgenic plant cell line containing the expression cassette described in B2); B6) Transgenic plant tissue containing the nucleic acid molecules described in B1), or transgenic plant tissue containing the expression cassette described in B2); B7) Transgenic plant organs containing the nucleic acid molecules described in B1), or transgenic plant organs containing the expression cassette described in B2); C1) Nucleic acid molecules that inhibit, reduce, or silence the expression of the genes encoding the proteins described above; C2) expresses the gene encoding the nucleic acid molecule described in C1); C3) contains an expression cassette containing the gene encoding described in C2); C4) A recombinant vector containing the encoding gene described in C2), or a recombinant vector containing the expression cassette described in C3); C5) A recombinant microorganism containing the encoding gene described in C2), or a recombinant microorganism containing the expression cassette described in C3), or a recombinant microorganism containing the recombinant vector described in C4); C6) A transgenic plant cell line containing the encoding gene described in C2), or a transgenic plant cell line containing the expression cassette described in C3), or a transgenic plant cell line containing the recombinant vector described in C4); C7) Transgenic plant tissue containing the encoding gene described in C2), or transgenic plant tissue containing the expression cassette described in C3), or transgenic plant tissue containing the recombinant vector described in C4); C8) A transgenic plant organ containing the encoding gene described in C2), or a transgenic plant organ containing the expression cassette described in C3), or a transgenic plant organ containing the recombinant vector described in C4).

[0019] In the above-mentioned biological materials, the nucleic acid molecule described in B1) may be a gene as shown in E1) or E2) below: E1) The coding sequence is a cDNA molecule or DNA molecule of SEQ ID No:2; E2) The nucleotide sequence is the cDNA molecule or DNA molecule of SEQ ID No:1.

[0020] The DNA molecule shown in SEQ ID No:2 (which regulates plant salt tolerance) OsDNAJ The gene encodes the protein OsDNAJ with the amino acid sequence SEQ ID No:3.

[0021] The nucleotide sequence shown in SEQ ID No:2 is the nucleotide sequence of the protein OsDNAJ encoding gene (CDS).

[0022] The present invention OsDNAJ Genes can be any nucleotide sequence that encodes the protein OsDNAJ. Considering codon degeneracy and the codon preferences of different species, those skilled in the art can use codons suitable for expression in a specific species as needed.

[0023] B1) The nucleic acid molecule may also include a nucleic acid molecule obtained by codon preference modification based on the nucleotide sequence shown in SEQ ID No:2.

[0024] B1) The nucleic acid molecule may also include nucleic acid molecules that have a nucleotide sequence identity of more than 95% with the nucleotide sequence shown in SEQ ID No:2 and originate from the same species.

[0025] The nucleic acid molecules mentioned in this article can be DNA, such as cDNA, genomic DNA, or recombinant DNA; the nucleic acid molecules can also be RNA, such as gRNA, mRNA, siRNA, shRNA, sgRNA, miRNA, or antisense RNA.

[0026] The vectors described herein are known to those skilled in the art and include, but are not limited to: plasmids, bacteriophages (such as λ phage or M13 filamentous phage), granules (i.e., Cos plasmids), Ti plasmids, or viral vectors.

[0027] To facilitate the identification and screening of transgenic plant cells or plants, the plant expression vectors used can be processed, such as by adding genes that can be expressed in plants, encoding enzymes or luminescent compounds that produce color changes (GUS genes, luciferase genes, etc.), antibiotic resistance markers (gentamicin markers, kanamycin markers, etc.), or chemical reagent resistance marker genes (such as herbicide resistance genes). From a safety perspective, transgenic plants can be screened directly under stress without adding any selective marker genes.

[0028] The microorganisms mentioned in this article may be yeast, bacteria, algae, or fungi. Among them, bacteria may originate from the genus *Escherichia* (…). Escherichia Erwinia ( Erwinia Agrobacterium tumefaciens ( ), Agrobacterium tumefaciens Agrobacterium Flavobacterium ( Flavobacterium Alcaligenes ( ) Alcaligenes ), Pseudomonas ( Pseudomonas ), Bacillus spp. ( Bacillus (e.g., Agrobacterium tumefaciens EHA105).

[0029] The present invention also provides the use of the protein OsDNAJ described above, or the expression substance of the gene regulating it, or the substance regulating the activity or content of the protein, in any of the following: Application of U1 in regulating plant salt tolerance; Application of U2 in the preparation of products that regulate plant salt tolerance; U3) Applications in cultivating plants with salt tolerance; U4) Applications in the preparation of products that cultivate salt-tolerant plants; U5) Applications in plant breeding.

[0030] In this article, the substance that regulates the activity and / or content of the protein may be a substance that regulates gene expression, wherein the gene encodes the protein OsDNAJ.

[0031] In the above applications, the substance regulating gene expression or the substance regulating protein activity or content can be a biological material related to the protein, and the biological material can be any of the following: B1) Nucleic acid molecules that encode the proteins described above; B2) An expression cassette containing the nucleic acid molecule described in B1); B3) A recombinant vector containing the nucleic acid molecule described in B1), or a recombinant vector containing the expression cassette described in B2); B4) Recombinant microorganisms containing the nucleic acid molecules described in B1), or recombinant microorganisms containing the expression cassette described in B2), or recombinant microorganisms containing the recombinant vector described in B3); B5) A transgenic plant cell line containing the nucleic acid molecule described in B1), or a transgenic plant cell line containing the expression cassette described in B2); B6) Transgenic plant tissue containing the nucleic acid molecules described in B1), or transgenic plant tissue containing the expression cassette described in B2); B7) Transgenic plant organs containing the nucleic acid molecules described in B1), or transgenic plant organs containing the expression cassette described in B2); C1) Nucleic acid molecules that inhibit, reduce, or silence the expression of the genes encoding the proteins described above; C2) expresses the gene encoding the nucleic acid molecule described in C1); C3) contains an expression cassette containing the gene encoding described in C2); C4) A recombinant vector containing the encoding gene described in C2), or a recombinant vector containing the expression cassette described in C3); C5) A recombinant microorganism containing the encoding gene described in C2), or a recombinant microorganism containing the expression cassette described in C3), or a recombinant microorganism containing the recombinant vector described in C4); C6) A transgenic plant cell line containing the encoding gene described in C2), or a transgenic plant cell line containing the expression cassette described in C3), or a transgenic plant cell line containing the recombinant vector described in C4); C7) Transgenic plant tissue containing the encoding gene described in C2), or transgenic plant tissue containing the expression cassette described in C3), or transgenic plant tissue containing the recombinant vector described in C4); C8) A transgenic plant organ containing the encoding gene described in C2), or a transgenic plant organ containing the expression cassette described in C3), or a transgenic plant organ containing the recombinant vector described in C4).

[0032] The present invention also provides a method for regulating the salt tolerance of plants, including regulating the activity and / or content of the proteins described above in the target plant, and / or the expression level of the genes encoding the proteins, to regulate the salt tolerance of plants.

[0033] In the above method, regulating the activity and / or content of the protein OsDNAJ in the target plant, and / or the expression level of the gene encoding the protein, includes introducing a gene encoding the protein that inhibits its expression into the recipient plant. OsDNAJ The expressed substance was used to obtain a target plant with altered salt tolerance; OsDNAJ The gene encodes the protein OsDNAJ.

[0034] In the above applications and methods, the regulation can be to increase, enhance, or upregulate.

[0035] In the above applications and methods, the regulation can be suppression, reduction, or silencing.

[0036] The present invention also provides a method for cultivating plants with altered salt tolerance, comprising: 1) inhibiting, reducing or silencing the expression level of the coding gene of the protein described above in the target plant, and / or inhibiting, reducing or silencing the activity and / or content of the coding gene of the protein described above, to obtain a plant with improved salt tolerance. 2) Increase, enhance, or upregulate the expression level of the coding genes of the proteins mentioned above in the target plant, or / and increase, enhance, or upregulate the activity and / or content of the coding genes of the proteins mentioned above, to obtain plants with reduced salt tolerance.

[0037] In one specific embodiment, a method for cultivating plants with enhanced salt tolerance includes the following steps: inhibiting the expression of nucleic acid molecules encoding OsDNAJ protein in the target plant to obtain transgenic plants with enhanced salt tolerance. Specifically, inhibiting the expression of nucleic acid molecules encoding OsDNAJ protein in the target plant can be achieved by introducing an interference vector or a knockout vector targeting the nucleic acid molecule encoding OsDNAJ protein into the target plant.

[0038] In this invention, the purpose of plant breeding includes cultivating plants with increased / decreased salt tolerance.

[0039] In the above applications or methods, the plant is any one of the following: N1) Monocotyledonous or dicotyledonous plants; N2) Plants of the order Poales; N3) Gramineae plants; N4) Rice plants; N5) rice.

[0040] The rice salt stress regulator OsDNAJ involved in this invention belongs to the molecular chaperone class that interacts with heat shock protein 70 (HSP70) and participates in regulating the correct folding and assembly of proteins. This study found that... OsDNAJ After CRISPR gene editing and knockout, compared with the recipient rice Zhonghua 11 (ZH11), the salt stress treatment resulted in... osdnaj The knockout mutant exhibits a salt-tolerant phenotype, indicating that... OsDNAJ Genes play a crucial role in regulating salt tolerance in rice, and further detailed analysis is needed. OsDNAJ Understanding the molecular mechanisms involved in regulating rice salt tolerance is of great significance for improving rice salt tolerance. Attached Figure Description

[0041] Figure 1 Wild-type ZH11 and mutant after treatment with 180 mM NaCl salt stress osdnaj Phenotypic images were obtained. First, photos were taken before treatment. During treatment, 180 mM NaCl was added to the nutrient solution. Photos were taken 21 days after treatment. Then, during recovery, normal nutrient solution without salt was added, and photos were taken 14 days after recovery.

[0042] Figure 2 homozygous mutant osdnaj A statistical chart showing the survival rate of wild-type ZH11 three-week-old seedlings after salt stress recovery.

[0043] Figure 3 To obtain ZH11 background using CRISPR / Cas9 technology osdnaj Mutation type of mutant (abbreviation) osdnaj This mutant is a loss-of-function mutant. Detailed Implementation

[0044] The present invention will now be described in further detail with reference to specific embodiments. The given embodiments are merely illustrative of the invention and not intended to limit its scope. The embodiments provided below can serve as a guide for further improvements by those skilled in the art and do not constitute a limitation on the invention in any way.

[0045] Unless otherwise specified, the experimental methods used in the following examples are conventional methods, performed according to the techniques or conditions described in the literature in this field or according to the product instructions. Unless otherwise specified, the materials and reagents used in the following examples are commercially available.

[0046] Unless otherwise specified, the quantitative experiments in the following examples are all repeated three times, and the results are averaged.

[0047] CRISPR / Cas9 mutants in the following examples - osdnaj The mutant was purchased from Biogle Gene Technology (Jiangsu) Co., Ltd., catalog number BG110816G09, and can be purchased from: http: / / biogle.cn / geo / index / geo / val / BG110816G09.

[0048] The japonica rice variety Zhonghua 11 (ZH11, abbreviated as ZH11) in the following examples was purchased from Baige Gene Technology (Jiangsu) Co., Ltd. Zhonghua 11 is a CRISPR / Cas9 mutant. osdnaj Receptor control material for mutants.

[0049] The nutrient solution B formula in the following examples is as follows: It is prepared by mixing A mother liquor, B mother liquor, EDTA-Fe mother liquor and trace element mother liquor in a ratio of 5:5:1:1 per liter of nutrient solution.

[0050] Among them, 1 L (200×) of mother liquor A contains 9.64 g of (NH4)2SO4, 3.7 g of KNO3, 4.96 g of KH2PO4, 3.18 g of K2SO4 and 29.965 g of MgSO4·7H2O; 1 L (200×) of mother liquor B contains 17.235 g of Ca(NO3)2·4H2O; In 1 L (1000×) of EDTA-Fe mother liquor: first, dissolve 5.57 g FeSO4·7H2O in 200 mL of distilled water, then heat and dissolve 7.45 g Na2EDTA in 200 mL of distilled water. Stir the FeSO4·7H2O solution and Na2EDTA solution continuously, cool, and then bring the volume to 1 L. 1 L (1000×) of trace element mother liquor contains: 2.86 g H3BO4, 0.08 g CuSO4·H2O, 0.22 g ZnSO4·7H2O, 1.81 g MnCl2·4H2O and 0.09 g NaMO4·H2O.

[0051] Add 300 ng sodium silicate per liter of nutrient solution, then adjust the pH to between 5.8 and 6.0 with concentrated hydrochloric acid.

[0052] The following examples use SPSS statistical software to process the data. The experimental results are expressed as mean ± standard deviation. One-way ANOVA test was used. P < 0.05 (*) indicates a significant difference, P < 0.01 (**) indicates a highly significant difference, and P < 0.001 (***) indicates a highly significant difference.

[0053] Example 1, CRISPR / Cas9- osdnaj Identification of homozygous mutants OsDNAJ The coding sequence (CDS) of the gene in Nipponbare rice is SEQ ID No:2, encoding the OsDNAJ protein with the amino acid sequence SEQ ID No:3. The genomic gene encoding the OsDNAJ protein in the genomic DNA of Nipponbare rice is shown in SEQ ID No:1 of the sequence listing.

[0054] Rice purchased from Baige Gene Technology (Jiangsu) Co., Ltd. osdnaj mutants only OsDNAJ The gene has mutated. Since the purchased mutant is not homozygous, it is necessary to identify a homozygous mutant.

[0055] Pick osdnaj DNA was extracted from the leaves of the mutant. Leaf DNA extraction was performed according to the CTAB method: Rice leaves were placed in 2.0 mL centrifuge tubes, and steel balls were added to grind them into powder using a grinder. 400 µL of CTAB extraction buffer was added to the centrifuge tube and vortexed to mix. An equal volume of a 1:1 mixture of chloroform and phenol was added, vortexed to mix, and centrifuged at 12000 rpm for 10 min. 200 µL of the supernatant was collected, and an equal volume of chloroform was added. The mixture was vigorously vortexed to mix, and centrifuged at 12000 rpm for 10 min. 100 µL of the supernatant was collected, and twice the volume of anhydrous ethanol was added to precipitate at 0℃ for 10 min. The mixture was then centrifuged at 12000 rpm for 10 min at 4℃. The supernatant was discarded, and the mixture was washed twice with 500 µL of 75% ethanol, dried, and dissolved in 30 µL of ddH2O.

[0056] Using the extracted DNA as a template, OsDNAJGene-specific primers were used for PCR amplification. The primer sequences were F: 5'-GCGCCTCTAGGATCTTGGAG -3'; R: 5'-GTGTGCCTGTCGGAGGAC -3'. The PCR reaction program was: 94℃ pre-denaturation for 2 min; 94℃ denaturation for 30 s, 56℃ annealing for 30 s, 72℃ extension for 30 s, 35 cycles; 72℃ extension for 5 min. The PCR products were then subjected to Sanger sequencing to check for mutations at the target site.

[0057] The final identification will be obtained OsDNAJ Transgenic plants with gene mutations are named osdnaj Comparison OsDNAJ The DNA sequence of the gene (nucleotide sequence is SEQ ID No:1), and its mutation sites are as follows: Figure 3 As shown.

[0058] Sequencing analysis revealed that, compared to the genomic DNA of rice Nipponbare, osdnaj In both homologous chromosomes of the plant, the gene encoding the OsDNAJ protein underwent the following mutation: "5'-CCTTCAGGGAACACCACCAAATC-3' (corresponding to positions 1679-1701 of SEQ ID No:1 and positions 700-722 of SEQ ID No:2)" was mutated to "5'-CCTTCA(A)GAACACCACCAAATC-3'", where "GG" was mutated to "A". This caused the protein coding sequence of SEQ ID No:3 to mutate from arginine to glycine at position 236 and terminate prematurely at position 292, resulting in loss of function and thus knocking out the gene encoding the OsDNAJ protein.

[0059] Example 2 osdnaj Salt tolerance analysis of mutants Samples to be tested: wild-type ZH11 and osdnaj mutant materials Rice seedlings were cultured as follows: Well-developed and plump seeds were selected and soaked in distilled water at 37°C for 48 hours, with the water changed every 12 hours. Germination was then continued for 24 hours under a humid environment. Seeds with uniform germination were selected and placed in bottomless 96-well plates, then placed in culture boxes containing Kimura B rice nutrient solution. Greenhouse growing conditions were 12 hours light / 12 hours darkness at 28°C. The rice nutrient solution was replaced with fresh solution every 3 days during the cultivation process.

[0060] wild-type ZH11 and osdnajAfter three weeks of growth, the mutant material was subjected to 180 mM NaCl stress treatment. After 21 days of treatment, photos were taken of the treated and untreated materials. Then, a recovery treatment was initiated, and photos were taken 14 days later. Figure 1 As shown, where Figure 1 In osdnaj express OsDNAJ The mutant strain, ZH11, represents the wild-type ZH11. After 14 days of recovery treatment, the genetic mutations of ZH11 and... osdnaj Survival rate, such as Figure 2 As shown.

[0061] The results showed that the survival rate of ZH11 was approximately 27.8%, while osdnaj The survival rate is approximately 86.3%, indicating that... OsDNAJ The loss of this function enhances the plant's tolerance to salt stress. Therefore, it can be seen that... OsDNAJ It plays a negative regulatory role in the plant's response to salt stress.

[0062] The present invention has been described in detail above. Those skilled in the art will recognize that the invention can be practiced in a wide range of ways with equivalent parameters, concentrations, and conditions without departing from its spirit and scope, and without requiring unnecessary experiments. While specific embodiments have been provided, it should be understood that further modifications can be made to the invention. In summary, according to the principles of the invention, this application is intended to include any changes, uses, or improvements to the invention, including changes made using conventional techniques known in the art that depart from the scope disclosed herein.

Claims

1. A protein, wherein the protein is any of the following: A1) A protein with the amino acid sequence shown in SEQ ID No:3; A2) A protein obtained by substituting and / or deleting and / or adding amino acid residues of the protein in A1) that has more than 75% identity with the protein shown in A1) and has the same function. A3) A fusion protein obtained by attaching a protein tag to the N-terminus and / or C-terminus of A1) or A2).

2. The protein according to claim 1, characterized in that: The protein is derived from rice ( Oryza sativa L.).

3. A biomaterial relating to the protein of claim 1 or 2, wherein the biomaterial is any one of the following: B1) A nucleic acid molecule encoding the protein described in claim 1 or 2; B2) An expression cassette containing the nucleic acid molecule described in B1); B3) A recombinant vector containing the nucleic acid molecule described in B1), or a recombinant vector containing the expression cassette described in B2); B4) Recombinant microorganisms containing the nucleic acid molecules described in B1), or recombinant microorganisms containing the expression cassette described in B2), or recombinant microorganisms containing the recombinant vector described in B3); B5) A transgenic plant cell line containing the nucleic acid molecule described in B1), or a transgenic plant cell line containing the expression cassette described in B2); B6) Transgenic plant tissue containing the nucleic acid molecules described in B1), or transgenic plant tissue containing the expression cassette described in B2); B7) Transgenic plant organs containing the nucleic acid molecules described in B1), or transgenic plant organs containing the expression cassette described in B2); C1) A nucleic acid molecule that inhibits, reduces, or silences the expression of the gene encoding the protein described in claim 1 or 2; C2) expresses the gene encoding the nucleic acid molecule described in C1); C3) contains an expression cassette containing the gene encoding described in C2); C4) A recombinant vector containing the encoding gene described in C2), or a recombinant vector containing the expression cassette described in C3); C5) A recombinant microorganism containing the encoding gene described in C2), or a recombinant microorganism containing the expression cassette described in C3), or a recombinant microorganism containing the recombinant vector described in C4); C6) A transgenic plant cell line containing the encoding gene described in C2), or a transgenic plant cell line containing the expression cassette described in C3), or a transgenic plant cell line containing the recombinant vector described in C4); C7) Transgenic plant tissue containing the encoding gene described in C2), or transgenic plant tissue containing the expression cassette described in C3), or transgenic plant tissue containing the recombinant vector described in C4); C8) A transgenic plant organ containing the encoding gene described in C2), or a transgenic plant organ containing the expression cassette described in C3), or a transgenic plant organ containing the recombinant vector described in C4).

4. The biomaterial according to claim 3, characterized in that: B1) The nucleic acid molecule described is a gene as shown in E1) or E2) below: E1) The coding sequence is a cDNA molecule or DNA molecule of SEQ ID No:2; E2) The nucleotide is a cDNA molecule or DNA molecule of SEQ ID No:

1.

5. The use of the protein or gene expression regulator or substance regulating the activity or content of said protein as described in claim 1 or 2 in any of the following: U1) Application in regulating plant salt tolerance; U2) Application in the preparation of products that regulate plant salt tolerance; U3) Applications in cultivating plants with salt tolerance; U4) Applications in the preparation of products that cultivate salt-tolerant plants; U5) Applications in plant breeding.

6. The application according to claim 5, characterized in that: The substance that regulates the expression of the gene or the substance that regulates the activity or content of the protein is a biological material related to the protein, and the biological material is the biological material according to claim 3 or 4.

7. A method for regulating the salt tolerance of plants, characterized in that: This includes regulating the activity and / or content of the protein described in claim 1 or 2 in the target plant, or / and the expression level of the gene encoding the protein described in claim 1 or 2, to regulate the plant's salt tolerance.

8. The method according to claim 7, characterized in that: The regulation of the activity and / or content of the protein described in claim 1 or 2 in the target plant, or / and the expression level of the gene encoding the protein described in claim 1 or 2 includes the introduction of a substance that inhibits the expression of the gene encoding the protein into the recipient plant, resulting in a target plant with a salt tolerance higher than that of the recipient plant; the gene encoding the protein described in claim 1 or 2.

9. Methods for cultivating plants with altered salt tolerance, including: 1) Inhibit or reduce or silence the expression level of the gene encoding the protein of claim 1 in the receptor plant, and / or inhibit or reduce or silence the activity and / or content of the gene encoding the protein of claim 1, to obtain a plant with improved salt tolerance. 2) Increase, enhance and / or upregulate the expression level of the gene encoding the protein described in claim 1 in the recipient plant, or / and increase, enhance and / or upregulate the activity and / or content of the gene encoding the protein described in claim 1, to obtain a plant with reduced salt tolerance.

10. The application according to claim 5 or 6, or the method according to any one of claims 7-9, characterized in that: The plant is any one of the following: N1) Monocotyledonous or dicotyledonous plants; N2) Plants of the order Poales; N3) Gramineae plants; N4) Rice plants; N5) rice.

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