Application of rice histone H1 gene in improving salt stress resistance of rice

Abstract

The application relates to an application of a rice histone H1 gene in improving the salt stress resistance of rice. The application discloses an application of a rice histone H1 gene or a gene with a nucleotide sequence as shown in SEQ ID NO. 9 in improving the salt stress resistance of rice. The DNA sequence of the rice in the application is 837 bp, and the rice has a good effect on enhancing the salt tolerance.

Description

Technical Field

[0001] This invention relates to the application of the rice histone H1 gene in improving the rice's ability to cope with salt stress, and belongs to the field of genetics. Background Technology

[0002] Currently, soil salinization is a major factor limiting sustainable agricultural development. High concentrations of sodium ions in the soil cause osmotic and ion stress, hindering the normal absorption of water and nutrients by plants. On the one hand, high-salt environments rapidly trigger osmotic stress in plants, leading to reduced water absorption by cells, rapid stomatal closure, and thus reducing the plant's ability to absorb carbon dioxide and inhibiting photosynthesis. Sustained salt stress reduces turgor pressure in plant cells, thereby inhibiting cell growth. On the other hand, plants have evolved various strategies to adapt to high-salt environments, possessing a certain degree of salt stress resistance. Plants regulate their adaptive responses to salt stress by inducing the expression of stress-response genes, and the expression products of these response genes directly participate in the repair of various aspects of primary and secondary stress induced by salt stress. Therefore, identifying salt-tolerance-related genes and applying them to improve plant salt tolerance provides stronger support for cultivating salt-tolerant crops on genetic engineering platforms. Summary of the Invention

[0003] Purpose of the invention: The technical problem to be solved by the present invention is to provide an application of the rice histone H1 gene in improving the ability of rice to cope with salt stress. It is the first time that this gene has been found to have a good effect on enhancing the salt tolerance of rice.

[0004] Technical solution: To solve the above-mentioned technical problems, the present invention provides the application of the rice histone H1 gene with nucleotides as shown in SEQ ID NO.1 or the gene with nucleotides as shown in SEQ ID NO.9 in improving the ability of rice to cope with salt stress.

[0005] Among them, SEQ ID NO.1:

[0006] ATGGCGACTGAGGAGGTTGTCCCTGAGGTTCCGGTGACCGAGGTGGAGGCTGCCGCGGCGGAGGAGGCCGTCGAGGAGACGACGGCGGCGGAGGAGAAGGCCGCCAAGCCGGCGAAGGAGAAGAAGAAGGCCGGCAGGCCGCCGAAGGAGAAGAAGGAGGCCAAGCCGGCGAAGGAGAAGAAGGTGAAGGAGGCCAAGGCCAAGAAGCCCCGCGTCGCCGCCGCCCACCCGCCGTACGCCGAGATGATCATGGAGGCGATCGTGGCGCTGAAGGAGAGGACTGGATCGAGCTCCCAGGCCATCGGCAAGCACATCCATGCCAACCATGGCGCCAACCTGCCGCCCAACTTCCGCAAGCTCCTCTCCGGCAACCTCAAGAAGCTCACCGCCGCCGGCAAGCTGGCCAAGGTCAAGAACTCCTTCAAGCTCTCCTCCACCCGCCCTGCCGCTCCGGCCGCCGCCGACGCCAAGCCCAAGGCCGCCCCCGCCACGAAGCCCAAGGTCAAGACCACCAAGGCCGCGAAGCCGGCTGCCAAGGCGAAGGCTCCTGCTACCACCAAGGCCGCGAAGCCGGCGACCAAGACCAAGATCAAGGTCGCCGCCGCGCCGGCGGCGAAGCCCAAGGCGTCTCCCAAGGCGAAGGCCAAGACCGCCACCTCGCCGGTGAAGCCCCGCGGCCGCCCTGCCAAGTCCGCCAAGACCTCTGCCAAGGACTCGCCTGCCAAGAAGGCGGCGCCGGTGGCTGCCAAGAAGAAGGCGGCGGCGACCAAGAAGAAGGCGTCGGTGGCTGCGGCGCCGGCTGCTCGCAAGGGTGCGGCGAGGAAGAGCATGAAGTAG

[0007] The amino acid sequence encoded by the DNA sequence is shown in SEQ ID NO. 2.

[0008] SEQ ID NO. 2:

[0009] MATEEVVPEVPVTEVEAAAAEEAVEETTAAEEKAAKPAKEKKKAGRPPKEKKEAKPAKEKKVKEAKAKKPRVAAAHPPYAEMIMEAIVALKERTGSSSQAIGKHIHANHGANLPPNFRKLLSGNLKKLTAAGKLAKVKN SFKLSSTRPAAPAAADAKPKAAPATKPKVKTTKAAKPAAKAKAPATTKAAKPATKTKIKVAAAPAAKPKASPKAKAKTATSPVKPRGRPAKSAKTSAKDSPAKKAAPVAAKKKAAATKKKASVAAAPAARKGAARKSMK

[0010] The process includes the following steps: linking the gene to the L177 vector after double enzyme digestion, transforming it into a strain, infecting rice, screening, and obtaining rice with high salt stress resistance.

[0011] The double digestion system consists of NEB buffer, SacⅠ, SalⅠ, plasmid, and ddH2O.

[0012] The nucleotides, such as those shown in SEQ ID NO.9, were obtained by PCR amplification using the genomic DNA of Nipponbare japonica rice as a template.

[0013] Furthermore, the rice mentioned is Nipponbare, a japonica rice species belonging to the genus Oryza of the family Poaceae.

[0014] Furthermore, using the genomic DNA of Nipponbare japonica rice (Oryza sativa) as a template, an 837bp amplification product (SEQ ID NO.1) was obtained.

[0015] Furthermore, the primers used for amplification have the following sequences:

[0016] Forward F: SEQ ID NO.3:5'-ATGGCGACTGAGGAGGTTGT-3';

[0017] Reverse R:SEQ ID NO.4:5'-CTACTTCATGCTCTTCCTCG-3'.

[0018] The method for identifying rice salt tolerance enhanced by the rice coding gene is as follows, including the following steps:

[0019] (1) Detection of LOC_Os07g08710 gene transcription level

[0020] Five tissues of Nipponbare were collected: seeds, roots, stems, leaves, and anthers. The transcriptional levels of the LOC_Os07g08710 gene in different tissues were analyzed and compared.

[0021] The amplification primers are:

[0022] Forward F: SEQ ID NO.5:5'-GAAGGTGAAGGAGGCCAAGG-3';

[0023] Reverse R:SEQ ID NO.6:5'-ATGGATGTGCTTGCCGATG-3'.

[0024] (2) Detection of LOC_Os07g08710 gene expression pattern under salt stress conditions

[0025] Salt stress was applied to Nipponbare samples under the following conditions: Nipponbare samples were hydroponically grown in 1 / 2 MS nutrient solution for 14 days, followed by treatment with 150 mM NaCl solution. Samples were collected at 0 h, 6 h, 12 h, and 24 h after treatment. RNA was extracted from the collected materials, reverse transcribed into cDNA, and subjected to real-time PCR to detect the expression pattern of the LOC_Os07g08710 gene under salt stress.

[0026] The amplification primers are:

[0027] Forward F: SEQ ID NO.5:5'-GAAGGTGAAGGAGGCCAAGG-3';

[0028] Reverse R:SEQ ID NO.6:5'-ATGGATGTGCTTGCCGATG-3'.

[0029] (3) Constructing transgenic rice LOC_Os07g08710 gene lines

[0030] The specific steps for constructing transgenic material of rice LOC_Os07g08710 gene are as follows: amplify the promoter sequence and full-length sequence of rice LOC_Os07g08710 gene (SEQ ID NO.9), and ligate it with the linear vector p1300U1-FLAG(L177) to form a transformation plasmid, that is, the promoter of rice LOC_Os07g08710 gene itself initiates its gene expression, and at the same time, a Flag tag is fused to the C-terminus of its gene.

[0031] The amplification primers are:

[0032] Forward F: SEQ ID NO.7:5'-GATATTTGCTCGCAAAGTCG-3';

[0033] Reverse F:SEQ ID NO.8:5'-CTTCATGCTCTTCCTCGCCG-3'.

[0034] The constructed plasmid was transformed into Agrobacterium EHA105 strain, and the callus tissue of Nipponbare was infected by Agrobacterium-mediated genetic transformation. Transgenic positive plants were obtained through further screening.

[0035] (4) Phenotypic identification of LOC_Os07g08710 transgenic material in response to salt stress

[0036] The LOC_Os07g08710 transgenic material and the control Nipponbare were grown in 1 / 2 MS hydroponics for 14 days, followed by salt stress treatment with 150 mM NaCl solution. The growth status and phenotypic changes of the two materials were observed for 3 consecutive days. Then, a rehydration recovery experiment was conducted to continue to observe phenotypic changes and record the survival rate.

[0037] Identification principle:

[0038] LOC_Os07g08710 is a rice histone H1 gene, whose coding region contains 837 deoxyribonucleotides. Analysis of this gene expression in different rice tissues revealed tissue specificity, with relatively high expression levels in leaves. Treatment of Nipponbare rice with 150 mM NaCl solution to induce salt stress, and comparison of gene transcription levels at 0 h, 6 h, 12 h, and 24 h, showed a gradual increase in gene expression from 0 to 24 h after salt stress, reaching its maximum at 24 h, indicating that the gene expression is induced by salt stress. Transgenic materials of the rice LOC_Os07g08710 gene were constructed. These transgenic materials, along with the control Nipponbare rice, were treated with 150 mM NaCl salt stress. After 3 days, the transgenic material showed greater salt tolerance than Nipponbare. Subsequent recovery experiments showed that the transgenic material recovered growth more quickly, with a survival rate of 61%, significantly higher than Nipponbare. In summary, the expression level of the LOC_Os07g08710 gene gradually increased in Nipponbare rice during the first 24 hours after salt stress treatment. Increasing the expression of this gene in transgenic materials promotes the plant's tolerance to salt stress, and the experimental results suggest that this gene plays an important role in the salt tolerance of rice.

[0039] This invention identifies that the LOC_Os07g08710 gene is induced by salt stress in the rice cultivar Nipponbare, and that increasing the expression of this gene in transgenic materials promotes the plant's tolerance to salt stress.

[0040] The present invention also provides a method for constructing rice with high salt stress capacity, comprising the following steps: linking the gene to the L177 vector after double enzyme digestion, transforming it into a strain, infecting rice, screening, and obtaining rice with high salt stress capacity.

[0041] The double digestion system consists of NEB buffer, SacⅠ, SalⅠ, plasmid, and ddH2O.

[0042] The nucleotides, such as those shown in SEQ ID NO.9, were obtained by PCR amplification using the genomic DNA of Nipponbare japonica rice as a template.

[0043] The present invention also provides a method for identifying rice with high salt stress capacity. When the rice to be tested contains the gene, it is rice with high salt stress capacity; otherwise, it is not rice with high salt stress capacity.

[0044] Beneficial effects: Compared with the prior art, the present invention has the following significant advantages: 1. The rice coding DNA sequence of the present invention is 837bp, which has a good effect on enhancing the salt tolerance of rice; 2. In particular, the transgenic rice plants constructed using the SEQ ID NO.9 sequence were found to have better tolerance to salt stress; The SEQ ID NO.9 sequence has the advantage of providing regulatory information to guide the spatiotemporal expression of the above-mentioned coding DNA. Attached Figure Description

[0045] Figure 1 The expression levels of the rice LOC_Os07g08710 gene in different tissues;

[0046] Figure 2 The expression level of the rice LOC_Os07g08710 gene after salt stress;

[0047] Figure 3 This is a schematic diagram of the LOC_Os07g08710 transgenic vector structure.

[0048] Figure 4 Salt stress phenotypes of OsH1 (LOC_Os07g08710 gene) transgenic material: A: Phenotype of LOC_Os07g08710 transgenic material (labeled T2-OsH1) and control Nipponbare after 14 days of growth under 1 / 2 MS hydroponic conditions; B: Phenotype after 72 hours of salt stress; C: Phenotype after 3 days of recovery from salt stress.

[0049] Figure 5 The survival rate of OsH1 transgenic material after salt stress. Detailed Implementation

[0050] The technical solution of the present invention will be further described below with reference to the accompanying drawings.

[0051] Example 1: Rice coding DNA sequence and its identification for enhancing rice salt tolerance

[0052] The rice coding DNA sequence and its identification method for enhancing rice salt tolerance are as follows:

[0053] 1. Rice histone H1 gene

[0054] LOC_Os07g08710 is a rice histone H1 gene. Its genetic information is available in the Rice Genome Access Database (RGAP, http: / / rice.plantbiology.msu.edu / ). The coding region of this gene consists of 837 deoxyribonucleotides, and the protein itself is composed of 278 amino acid residues.

[0055] Sequence-specific primers were designed and prepared by Sangon Biotech (Shanghai) Co., Ltd. as follows:

[0056] Forward F: SEQ ID NO.3:5'-ATGGCGACTGAGGAGGTTGT-3';

[0057] Reverse R:SEQ ID NO.4:5'-CTACTTCATGCTCTTCCTCG-3'.

[0058] Using the above primers and the genomic DNA of Nipponbare japonica rice as a template, the target DNA fragment of 837 bp was amplified by PCR using the above-mentioned primers and procedures.

[0059] The PCR amplification system is as follows (using Phanta Max Super-Fidelity DNA Polymerase, catalog number P505, from Nanjing Novizan Biotechnology Co., Ltd.):

[0060] 50 μL reaction system: 25 μL 2×Phanta Max Buffer; 1 μL dNTP mix (10 mmol / L); 1 μL PhantaMax Super-Fidelity DNA Polymerase (100 U); 2 μL each of upstream and downstream primers (10 μmol / L); 1 μL template (30 ng); ddH2O to bring the total volume to 50 μL.

[0061] PCR reaction conditions: 95℃ pre-denaturation for 3 min; 95℃ denaturation for 15 s, 58℃ annealing for 15 s, 72℃ extension for 45 s, 30 cycles; final extension at 72℃ for 5 min.

[0062] After sequencing, the nucleotide sequence of the amplification product is shown as positions 1 to 837 from the 5' end of SEQ ID NO.1.

[0063] SEQ ID NO.1:

[0064] ATGGCGACTGAGGAGGTTGTCCCTGAGGTTCCGGTGACCGAGGTGGAGGCTGCCGCGGCGGAGGAGGCCGTCGAGGAGACGACGGCGGCGGAGGAGAAGGCCGCCAAGCCGGCGAAGGAGAAGAAGAAGGCCGGCAGGCCGCCGAAGGAGAAGAAGGAGGCCAAGCCGGCGAAGGAGAAGAAGGTGAAGGAGGCCAAGGCCAAGAAGCCCCGCGTCGCCGCCGCCCACCCGCCGTACGCCGAGATGATCATGGAGGCGATCGTGGCGCTGAAGGAGAGGACTGGATCGAGCTCCCAGGCCATCGGCAAGCACATCCATGCCAACCATGGCGCCAACCTGCCGCCCAACTTCCGCAAGCTCCTCTCCGGCAACCTCAAGAAGCTCACCGCCGCCGGCAAGCTGGCCAAGGTCAAGAACTCCTTCAAGCTCTCCTCCACCCGCCCTGCCGCTCCGGCCGCCGCCGACGCCAAGCCCAAGGCCGCCCCCGCCACGAAGCCCAAGGTCAAGACCACCAAGGCCGCGAAGCCGGCTGCCAAGGCGAAGGCTCCTGCTACCACCAAGGCCGCGAAGCCGGCGACCAAGACCAAGATCAAGGTCGCCGCCGCGCCGGCGGCGAAGCCCAAGGCGTCTCCCAAGGCGAAGGCCAAGACCGCCACCTCGCCGGTGAAGCCCCGCGGCCGCCCTGCCAAGTCCGCCAAGACCTCTGCCAAGGACTCGCCTGCCAAGAAGGCGGCGCCGGTGGCTGCCAAGAAGAAGGCGGCGGCGACCAAGAAGAAGGCGTCGGTGGCTGCGGCGCCGGCTGCTCGCAAGGGTGCGGCGAGGAAGAGCATGAAGTAG

[0065] Using the online bioinformatics tool software https: / / www.novopro.cn / tools / translate.html, the protein sequence of SEQ ID NO.1 was obtained by translating from the first base of the 5' sequence.

[0066] SEQ ID NO.2:

[0067] MATEEVVPEVPVTEVEAAAAEEAVEETTAAEEKAAKPAKEKKKAGRPPKEKKEAKPAKEKKVKEAKAKKPRVAAAHPPYAEMIMEAIVALKERTGSSSQAIGKHIHANHGANLPPNFRKLLSGNLKKLTAAGKLAKVKNSFKLSSTRPAAP AAADAKPKAAPATKPKVKTTKAAAKPAAKAKAPATTKAAKPATKTKIKVAAAPAAKPKASPKAKAKTATSPVKPRGRPAKSAKTSAKDSPAKKAAPVAAKKKAAATKKKASVAAAPAARKGAARKSMK Example 2 LOC_Os07g08710 Transcription Level Detection

[0068] Five tissues were collected from Nipponbare: seeds, roots, stems, leaves, and anthers. A kit was used to analyze these tissues. RNA was extracted using the UniversalPlant Total RNA Isolation Kit (RC411-01, Nanjing Novizan Biotechnology Co., Ltd.). The RNA was reverse transcribed into cDNA using the HiScript III RT SuperMix for qPCR (+gDNA wiper) (see R323-01, Nanjing Novizan Biotechnology Co., Ltd. for detailed steps). Using the cDNA as a template, the expression level of the LOC_Os07g08710 gene was detected using the ChamQ SYBR qPCR Master Mix (Q311-02, Nanjing Novizan Biotechnology Co., Ltd.) and gene-specific primers SEQ ID NO. 5 and SEQ ID NO. 6. The amplified sequence is shown in SEQ ID NO. 10, with a length of 139 bp.

[0069] SEQ ID NO.10:

[0070] GAAGGTGAAGGAGGCCAAGGCCAAGAAGCCCCGCGTCGCCCGCCGCCCACCCGCC

[0071] GTACGCCGAGATGATCATGGAGGCGATCGTGGCGCTGAAGGAGAGGACTGGATCG

[0072] AGCTCCCAGGCCATCGGCAAGCACATCCAT

[0073] The primers used were prepared by Sangon Biotech (Shanghai) Co., Ltd.:

[0074] Forward F: SEQ ID NO.5:5'-GAAGGTGAAGGAGGCCAAGG-3';

[0075] Reverse R:SEQ ID NO.6:5'-ATGGATGTGCTTGCCGATG-3'.

[0076] The results are as follows Figure 1 As shown, the rice LOC_Os07g08710 gene exhibits tissue-specific expression, with significant differences in transcriptional levels across different tissues, and higher expression levels observed in leaves.

[0077] Example 3 Salt stress induces LOC_Os07g08710 gene expression

[0078] Salt stress treatment was applied to Nipponbare samples under the following conditions: Nipponbare samples were hydroponically grown in 1 / 2 MS nutrient solution (phytotechlab, catalog number M519) for 14 days, followed by treatment with 150 mM NaCl solution. Samples were taken at 0 h, 6 h, 12 h and 24 h after treatment, and the collected materials were used for RNA extraction.

[0079] RNA was extracted and reverse transcribed into cDNA according to the method in Example 2. Real-time quantitative PCR was performed using gene-specific primers SEQ ID NO. 5 and SEQ ID NO. 6 to detect the expression changes of the LOC_Os07g08710 gene at different time points under salt stress. The results showed that the gene expression trend gradually increased during the 0-24 h period after salt stress. Figure 2 As shown in the figure, the expression level reached its maximum at 24 hours, indicating that the gene was induced to express under salt stress.

[0080] Example 4: Construction of transgenic rice line with LOC_Os07g08710 gene

[0081] The expression vector used was p1300U1-FLAG (L177), provided by Professor Chen Chen's research group at Yangzhou University. For details, please refer to the reference: Niu, B., Xu, J., Zhiguo, E., Zhang, Z., Lu, X., & Chen, C. (2023). Ectopic expression of OsNF-YA8, an endosperm-specific nuclear factor Y transcription-factor gene, causes vegetative and reproductive development defects in rice. The Crop Journal, 11(6), 1719-1730. The L177 vector was double-digested with SacⅠ-HF and SalⅠ-HF (NEB), and the vector fraction was recovered. The digestion system was as follows: 50 μL digestion system: 10×NEB buffer 5 μL; SacⅠ 1 μL; SalⅠ 1 μL; plasmid 1 μg; ddH2O to 50 μL. The digestion conditions were 37℃ for 15 minutes.

[0082] Specific primers SEQ ID NO.7 and SEQ ID NO.8 were designed to amplify the target fragment. The amplified sequence is shown in SEQ ID NO.9. The SEQ ID NO.9 sequence includes the promoter sequence of the LOC_Os07g08710 gene and its full-length gene sequence.

[0083] The following primers were prepared by Sangon Biotech (Shanghai) Co., Ltd.:

[0084] Forward F: SEQ ID NO.7:5'-GATATTTGCTCGCAAAGTCG-3';

[0085] Reverse F:SEQ ID NO.8:5'-CTTCATGCTCTTCCTCGCCG-3'.

[0086] Using the above primers and the genomic DNA of Nipponbare japonica rice as a template, a 1349bp target DNA fragment was amplified using the PCR system and steps.

[0087] The PCR amplification system is as follows (using Novizan Phanta Max Super-Fidelity DNA Polymerase, catalog number P505):

[0088] 50 μL reaction system: 25 μL 2×Phanta Max Buffer; 1 μL dNTP mix (10 mmol / L); 1 μL PhantaMax Super-Fidelity DNA Polymerase (100 U); 2 μL each of upstream and downstream primers (10 μmol / L); 1 μL template (30 ng); ddH2O to bring the total volume to 50 μL.

[0089] PCR reaction conditions: 95℃ pre-denaturation for 3 min; 95℃ denaturation for 15 s, 58℃ annealing for 15 s, 72℃ extension for 60 s, 30 cycles; final extension at 72℃ for 5 min.

[0090] Sequencing revealed that the nucleotide sequence of the amplified product is shown from position 1 to position 1349 of SEQ ID NO.9.

[0091] SEQ ID NO.9:

[0092]

[0093] The amplified fragment of SEQ ID NO.9 was ligated with the enzyme-digested L177 vector, as shown in the schematic diagram below. Figure 3 As shown, the rice LOC_Os07g08710 gene's own promoter initiates its gene expression, and a Flag tag is fused to the C-terminus of the gene. The constructed plasmid was transformed into Agrobacterium EHA105 strain (Shanghai Weidi Biotechnology Co., Ltd., catalog number AC1010). Agrobacterium-mediated genetic transformation was used to infect callus tissue of Nipponbare rice. Transgenic positive plants were obtained through further screening (detailed steps were performed according to the following method: Chen Taiyu, Wu Hao, Lin Yongjun, Chen Hao. (2018). General steps for the cultivation of transgenic rice materials. Bio-101:e1010177.DOI:10.21769 / BioProtoc.1010177).

[0094] Example 5: Phenotypic identification of LOC_Os07g08710 transgenic material in response to salt stress

[0095] The LOC_Os07g08710 transgenic material and the control Nipponbare were grown in 1 / 2 MS hydroponic conditions for 14 days (phenotype as shown). Figure 4 As shown in Figure A), the materials were then subjected to salt stress treatment with 150 mM NaCl solution, during which the growth status and phenotypic changes of the two materials were observed. Three days after treatment with 150 mM NaCl, the leaves of the LOC_Os07g08710 transgenic material (labeled T2-OsH1 in the image) remained green, while the leaves of the Nipponbare (labeled Nip in the image) had partially turned yellow. The phenotypic changes after salt stress were as follows: Figure 4 As shown in Figure B, the LOC_Os07g08710 transgenic material exhibits significant salt tolerance. Next, a recovery experiment was conducted, in which both salt-stressed materials were transferred to hydroponics to resume growth, and their phenotypic changes were further observed. During the recovery process, the salt-stressed LOC_Os07g08710 transgenic material recovered growth relatively quickly, as shown in Figure B. Figure 4 As shown in C. The survival rate of the transgenic material was 61%, significantly higher than that of Nipponbare (28%). Figure 5 As shown. In summary, the expression level of the LOC_Os07g08710 gene gradually increased in Nipponbare rice during the first 24 hours after salt stress treatment. Increasing the expression of this gene in transgenic materials promotes the plant's tolerance to salt stress. The experimental results suggest that this gene plays an important role in the salt tolerance of rice.

Claims

1. Application of rice histone H1 gene with nucleotides as shown in SEQ ID NO.1 or gene with nucleotides as shown in SEQ ID NO.9 in improving the ability of rice to cope with salt stress.

2. The application according to claim 1, characterized in that, The process includes the following steps: linking the gene described in claim 1 to the L177 vector after double enzyme digestion, transforming it into a strain, infecting rice, screening, and obtaining rice with high salt stress resistance.

3. The application according to claim 2, characterized in that, The double digestion system consisted of NEB buffer, SacⅠ, SalⅠ, plasmid, and ddH2O.

4. The application according to claim 1, characterized in that, The nucleotides, as shown in SEQ ID NO.9, were obtained by PCR amplification using the genomic DNA of Nipponbare japonica rice as a template.

5. A method for constructing rice with high salt stress resistance, characterized in that, The process includes the following steps: linking the gene described in claim 1 to the L177 vector after double enzyme digestion, transforming it into a strain, infecting rice, screening, and obtaining rice with high salt stress resistance.

6. The method according to claim 5, characterized in that, The double digestion system consisted of NEB buffer, SacⅠ, SalⅠ, plasmid, and ddH2O.

7. The method according to claim 5, characterized in that, The nucleotides, as shown in SEQ ID NO.9, were obtained by PCR amplification using the genomic DNA of Nipponbare japonica rice as a template.

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