Rice salt stress regulatory gene OsEra1 and its application

Abstract

The present invention relates to the field of plant gene technology, and in particular to a rice salt stress regulating gene OsEra1 and an application thereof. The present invention provides an OsEra1 gene for regulating rice salt tolerance, wherein the OsEra1 gene is any one of the following: A1) the coding sequence is the OsEra1 gene genome sequence of SEQ ID NO: 1 in the sequence list or the OsEra1 gene CDS sequence of SEQ ID NO: 2; A2) the nucleotide is the OsEra1 gene genome sequence of SEQ ID NO: 1 in the sequence list or the OsEra1 gene CDS sequence of SEQ ID NO: 2. The present invention found that after the OsEra1 gene was knocked out by CRISPR gene editing, two different strains of the osera1 knockout mutant showed a salt-sensitive phenotype after salt stress treatment compared with the recipient rice Nipponbare, proving that the OsEra1 gene plays an important role in mediating RNA m6A modification and thus responding to rice salt stress. Subsequent research on OsEra1 and m6A modification of its downstream regulatory genes will be of great significance to m6A modification regulating rice salt stress response.

Description

Technical Field

[0001] The invention relates to the field of plant gene technology, in particular to a rice salt stress regulating gene OsEra1 and an application thereof. Background Art

[0002] As one of the most important food crops in the world, rice production is of great significance to national food security. With the intensification of global warming, soil salinization has become more serious. The latest statistics show that the total area of ​​saline-alkali land in China has reached 99 million hectares. Soil salinization poses a serious threat to crop yields, especially for monocotyledonous rice.

[0003] From the perspective of the rice growth cycle, during the rice germination period, salt stress will inhibit the germination of rice seeds and reduce its germination rate; during the rice seedling period, the osmotic stress caused by salt stress will cause the growth of seedlings to slow down; during the rice heading period, salt stress will cause the heading period to be delayed, and the number of days of delay will increase with the increase of salt concentration. At present, the research on plant response to salt stress mainly focuses on ion stress, osmotic stress and oxidative stress (Park HJ, Kim WY, Yun DJ. A New Insight of Salt Stress Signaling in Plant. Mol Cells. 2016; 39(6): 447-459.). Plants under ion stress often suffer from the dual damage of salt ion poisoning and nutrient ion deficiency. Sodium ions and chloride ions are the main salt ions that are toxic to plants in saline soil. Under salt stress, the selectivity of plants for ion absorption decreases, and a large amount of Na + and Cl - After entering the plant cell, it makes Na + and Cl - Concentration is too high, K + , Ca 2+ and Mg 2+ The absorption of K is inhibited, which destroys the original ion balance in the plant and causes the plant to suffer from ion stress. In addition, salt stress also inhibits the plant's absorption of other nutrient ions, such as K + and Ca 2+etc., which results in insufficient intake of nutrients in the plant. Osmotic stress refers to a condition in which plants cannot get enough water due to changes in environmental factors. The osmotic pressure in the plant is lower than that in the soil solution, and the plant cannot absorb water or even loses water, which can seriously lead to death. In addition to causing plants to suffer from osmotic stress and ion stress, salt stress also produces secondary oxidative stress through the interaction of the two, which destroys the dynamic balance between the production and removal of reactive oxygen species (ROS) in the plant. When the accumulation of ROS exceeds its damage threshold, it will cause oxidative damage to the plant in two aspects: on the one hand, it will cause increased peroxidation and degreasing of cell membrane lipids, thereby destroying the cell membrane system of the plant; on the other hand, it will cause a decrease in the activity of specific enzymes responsible for the synthesis of photosynthetic pigments in the plant, and the chloroplast grana lamellae will swell and loosen, or even disintegrate, which will seriously inhibit the photosynthesis of the plant.

[0004] At present, the research on the molecular network regulating rice salt stress response is also focused on the above aspects. Recently, more and more evidence of epigenetic involvement in regulating rice salt stress response has been found, such as methylation and acetylation modifications on histones (Wei H, Wang X, He Y, Xu H, Wang L. Clock component OsPRR73 positively regulates rice salt tolerance by modulating OsHKT2;1-mediated sodium homeostasis. EMBO J. 2021;40(3):e105086.), DNA methylation modification (Wang J, Nan N, Li N, et al. A DNA Methylation Reader-chaperone regulator-transcription factor complex activates OsHKT1;5 expression during salinity stress. Plant Cell. 2020;32(11):3535-3558.), etc. However, there are no reports on the research on RNA epigenetic modification regulating rice salt stress. Summary of the invention

[0005] The present invention provides an OsEra1 gene for regulating rice salt tolerance, wherein the OsEra1 gene is any one of the following:

[0006] A1) The coding sequence is: the OsEra1 gene genomic sequence of SEQ ID NO: 1 or the OsEra1 gene CDS sequence of SEQ ID NO: 2 in the sequence listing;

[0007] A2) The nucleotide sequence is: the OsEra1 gene genomic sequence of SEQ ID NO: 1 or the OsEra1 gene CDS sequence of SEQ ID NO: 2 in the sequence listing.

[0008] The present invention provides an OsEra1 protein encoded by the gene, wherein the protein is OsEra1 is any one of the following:

[0009] B1): the amino acid sequence is the protein of SEQ ID NO: 3 in the sequence list;

[0010] B2): A protein having more than 90% identity with the protein shown in B1) and related to plant salt tolerance obtained by replacing and / or deleting and / or adding one or more amino acid residues of the amino acid sequence shown in SEQ ID NO: 3 in the sequence list;

[0011] B3): A fusion protein obtained by connecting a protein tag to the N-terminus or / and C-terminus of B1) or B2).

[0012] The present invention provides a biomaterial, characterized in that it contains at least one of the following:

[0013] C1) a nucleic acid molecule encoding the OsEra1 protein;

[0014] C2) an expression cassette containing the nucleic acid molecule described in C1);

[0015] C3) a recombinant vector containing the nucleic acid molecule described in C1), or a recombinant vector containing the expression cassette described in C2);

[0016] 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);

[0017] 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);

[0018] C6) transgenic plant tissue containing the nucleic acid molecule described in C1), or transgenic plant tissue containing the expression cassette described in C2);

[0019] 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);

[0020] C8) a nucleic acid molecule that reduces the expression of the OsEra1 protein;

[0021] C9) An expression cassette, a recombinant vector, a recombinant microorganism or a transgenic plant cell line containing the nucleic acid molecule described in C8).

[0022] The present invention provides a method for cultivating and regulating plant salt stress response, characterized in that the expression of a gene encoding a salt stress response-related protein in a target plant is regulated, wherein the salt stress response-related protein is any one of the OsEra1 proteins described in the protein.

[0023] The present invention provides a plant salt-tolerant substance, characterized in that: the plant salt-tolerant substance contains the OsEra1 gene or / and the OsEra1 protein or / and the biological material; and the plant is a monocotyledon or a dicotyledon.

[0024] The present invention provides an application of the rice salt stress regulatory gene OsEra1, characterized in that the application is based on any of the following:

[0025] Any one of the OsEra1 genes, and / or any one of the OsEra1 proteins, and / or any one of the biological materials;

[0026] The applications include one or more of the following:

[0027] D1) Application in regulating plant salt tolerance;

[0028] D2) Application in breeding salt-tolerant / intolerant plants;

[0029] D3) Use in the preparation of products for increasing / decreasing plant salt tolerance;

[0030] D4) Application in plant breeding.

[0031] Preferably, the plant is a monocotyledon or a dicotyledon.

[0032] The present invention also provides a method for cultivating a transgenic plant with reduced salt tolerance, which comprises reducing the expression of the coding gene of the OsEra1 protein in the target plant to obtain a transgenic plant with lower salt tolerance than the target plant.

[0033] Preferably, the reducing the expression of the gene encoding the OsEra1 protein in the target plant is achieved by inhibiting the content and / or activity of the OsEra1 protein in the target plant using the CRISPR / Cas9 gene editing system.

[0034] Preferably, the target plant is a monocotyledon or a dicotyledon.

[0035] The present invention provides a rice OsEra1 gene knockout mutant, characterized in that the rice mutant is a mutant knocking out a nucleotide sequence shown in SEQ ID NO:1.

[0036] Compared with the prior art, the present invention has the following significant improvements:

[0037] (1) The RNA m6A modification involved in the present invention belongs to RNA methylation modification, specifically, it refers to a very conservative chemical modification type formed after the hydrogen atom (H) at the N6 position of RNA is replaced by a methyl group (CH3) (Desrosiers RC, Friderici KH, Rottman FM. Characterization of Novikoff hepatoma mRNA methylation and heterogeneity in the methylated 5'terminus. Biochemistry. 1975; 14(20): 4367-4374.). The present invention found that after the OsEra1 gene was knocked out by CRISPR gene editing, compared with the recipient rice Nipponbare, two different strains of the osera1 knockout mutant showed a salt-sensitive phenotype after salt stress treatment, proving that the OsEra1 gene plays an important role in mediating RNA m6A modification and then responding to rice salt stress. Subsequent research on OsEra1 and the m6A modification of its downstream regulatory genes will be of great significance to the regulation of rice salt stress response by m6A modification.

[0038] (2) The present invention experimentally proves that knocking down the OsEra1 gene in wild-type rice can obtain transgenic rice with altered salt stress phenotype. Compared with the recipient rice, the expression of the OsEra1 gene in the transgenic rice is reduced, and there is no obvious change in the heading period under long-day and short-day conditions. Therefore, the OsEra1 gene is related to the response of rice to salt stress, laying a theoretical foundation for cultivating transgenic plants that positively regulate the response to salt stress.

[0039] Appendix Description

[0040] SEQ ID NO: 1 is the genomic nucleotide sequence of the OsEra1 gene,

[0041] SEQ ID NO: 2 is the CDS sequence of the OsEra1 gene,

[0042] SEQ ID NO: 3 is the amino acid sequence of OsEra1 protein. BRIEF DESCRIPTION OF THE DRAWINGS

[0043] Figure 1The phenotypes of wild-type NIP and mutants osera1-L1 and osera1-L2 after 180 mM NaCl salt stress treatment in Example 4 were first photographed before treatment, 180 mM NaCl was added to the nutrient solution during treatment, and photographed 14 days after treatment. Then, normal nutrient solution without salt was added during recovery, and photographed 14 days after recovery, and the survival rate was counted.

[0044] Figure 2 This is a statistical graph of the survival rate of the three-week-old seedlings of the homozygous mutants osera1-L1, osera1-L2 and wild-type NIP in Example 4 after recovery from salt stress.

[0045] Figure 3 These are the mutation types of the osera1 mutants in the NIP background obtained using CRISPR / Cas9 technology in Example 3, named osera1-L1 and osera1-L2, both of which are loss-of-function mutants. DETAILED DESCRIPTION

[0046] The present invention is further described in detail below in conjunction with specific embodiments, and the examples provided are only for illustrating the present invention, rather than for limiting the scope of the present invention. The examples provided below can be used as a guide for further improvements by those of ordinary skill in the art, and do not constitute a limitation of the present invention in any way.

[0047] The experimental methods in the following examples are conventional methods unless otherwise specified. The materials, reagents, etc. used in the following examples are commercially available unless otherwise specified.

[0048] The primary purpose of the present invention is to provide a rice salt stress regulatory gene, and its nucleotide sequence and a recombinant vector containing the gene. The inventor knocked out the OsEra1 gene in rice plants and found that compared with the recipient rice Nipponbare, two strains of the osera1 knockout mutant showed a salt-sensitive phenotype after salt stress treatment, that is, the salt stress phenotype of the target plant was successfully changed.

[0049] One object of the present invention is to provide a polypeptide, namely, protein OsEra1, whose amino acid sequence is shown in SEQ ID NO: 3. After the OsEra1 gene is knocked out, the expression level of the OsEra1 protein is greatly reduced, and the plant can exhibit a salt-sensitive phenotype.

[0050] Another object of the present invention is to provide an oligonucleotide encoding the aforementioned polypeptide, namely, the OsEra1 gene, whose nucleotide sequence is shown in SEQ ID NO: 1. After knocking out the gene in rice, its expression level is greatly reduced, and the plant can be made salt-sensitive phenotype.

[0051] The present invention provides the use of any one of the following substances (1)-(3) in regulating plant salt stress:

[0052] (1) Protein OsEra1;

[0053] (2) nucleic acid encoding protein OsEra1;

[0054] (3) a recombinant vector containing a nucleic acid encoding the protein OsEra1;

[0055] The protein OsEra1 is as follows (1) or (2):

[0056] (1) a protein consisting of the amino acid sequence shown in SEQ ID NO: 3;

[0057] (2) A protein derived from (1) having the same function as that of the amino acid sequence shown in SEQ ID NO: 3, wherein one or more amino acid residues are substituted and / or deleted and / or added.

[0058] The above-mentioned substitution and / or deletion and / or addition of one or several amino acid residues is a substitution and / or deletion and / or addition of no more than 10 amino acid residues.

[0059] In the above application, the DNA molecule encoding the protein OsEra1 is any one of the following DNA molecules 1) to 4):

[0060] (1) DNA molecules shown in SEQ ID NO: 1 and SEQ ID NO: 2;

[0061] (2) a DNA molecule that hybridizes with the DNA molecule defined in 1) or 2) under stringent conditions and encodes a protein consisting of the amino acid sequence shown in SEQ ID NO: 3;

[0062] (3) A DNA molecule having at least 70% homology with the DNA molecule defined in (1) or (2) and encoding a protein consisting of the amino acid sequence shown in SEQ ID NO: 3.

[0063] The present invention proves through experiments that knocking down the OsEra1 gene in wild-type rice can obtain transgenic rice with altered salt stress phenotype. Compared with the recipient rice, the expression of the OsEra1 gene in the transgenic rice is reduced, and there is no obvious change in the heading period under long-day and short-day conditions. Therefore, the OsEra1 gene is related to the rice response to salt stress, laying a theoretical foundation for cultivating transgenic plants that positively regulate the salt stress response.

[0064] The experimental material used in this study was the commonly used japonica rice variety Nipponbare (NIP).

[0065] The CRISPR / Cas9 vector system in the following embodiments is described in the following literature (Ma X, Zhang Q, Zhu Q, Liu W, Chen Y, Qiu R, Wang B, Yang Z, Li H, Lin Y, Xie Y, Shen R, Chen S, Wang Z, Chen Y, Guo J, Chen L, Zhao X, Dong Z, Liu YG (2015) A Robust CRISPR / Cas9 System for Convenient, High-Efficiency Multiplex Genome Editing in Monocot and Dicot Plants. Mol Plant 8: 1274-1284), which was kindly donated by Liu Yaoguang Laboratory of South China Agricultural University. The public can obtain it from South China Agricultural University to repeat the experiments of this application and cannot be used for other purposes.

[0066] The Agrobacterium EHA105 competent cells in the following examples were produced by our laboratory, and the same competent cells can also be purchased on the market.

[0067] Nutrient solution Kimura B formula: It is composed of A mother solution, B mother solution, EDTA-Fe mother solution and trace element mother solution, mixed in a ratio of 5:5:1:1 per liter of nutrient solution. Among them, 1L (200X) of mother liquor A contains 9.64g of (NH4)2SO4, 3.7g of KNO3, 4.96g of KH2PO4, 3.18g of K2SO4 and 29.965g of MgSO4·7H2O; 1L (200X) of mother liquor B contains 17.235g of Ca(NO3)2·4H2O; in 1L (1000X) of EDTA-Fe mother liquor: first dissolve 5.57g of FeSO4·7H2O in 200mL of distilled water, then heat to dissolve 7.45g of Na2EDTA in 200mL of distilled water, stir and mix the FeSO4·7H2O solution and the Na2EDTA solution continuously, and adjust the volume to 1L after cooling. 1L (1000X) of trace element mother solution contains: H3BO4 2.86g, CuSO4·H2O 0.08g, ZnSO4·7H2O 0.22g, MnCl2·4H2O 1.81g and NaMO4·H2O 0.09g. 300ng of sodium silicate is added to each liter of nutrient solution, and then the pH is adjusted to between 5.8 and 6.0 with concentrated hydrochloric acid.

[0068] The rice seedling material culture method used in the following embodiments of the present invention is as follows: select well-developed and plump seeds, soak them in distilled water in a 37°C incubator for 48 hours, change the water every 12 hours, and then continue to germinate for 24 hours in a humid environment. Select seeds with consistent germination status and place them in a bottomless 96-well plate, and grow them in a culture box filled with Kimura B rice nutrient solution. The greenhouse growth conditions are 12 hours of light / 12 hours of darkness, and the temperature is 28°C. During the culture process, fresh rice nutrient solution is replaced every 3 days.

[0069] Example 1

[0070] Cloning of the rice salt stress response gene OsEra1;

[0071] The inventors of the present invention isolated and cloned a salt stress-responsive rice gene OsEra1 from the rice variety Nipponbare, as shown in Sequence 1 of the sequence listing, and named the encoded protein OsEra1 protein, as shown in Sequence 2 of the sequence listing.

[0072] Total RNA of rice variety Nipponbare was extracted and reverse transcribed into cDNA, and PCR amplification was performed with primers F: ATGATCATACTGACCAAGGA and R: TTATTTTGCGGCATTCGGCT. The PCR reaction program was: 94℃ pre-denaturation for 2min; 94℃ denaturation for 30s, 56℃ annealing for 30s, 72℃ extension for 30s, 35 cycles; 72℃ extension for 5min. The PCR product was subjected to Sanger sequencing. The sequencing results showed that the nucleotide sequence of the PCR amplification product was sequence 1 in the sequence list, and its coding sequence was nucleotides 1-1113 of sequence 2 in the sequence list, encoding the protein OsEra1 shown in sequence 2 (amino acids 1-370 of sequence 3 in the sequence list); the DNA shown in sequence 1 in the sequence list was named OsEra1 gene.

[0073] Example 2

[0074] Construction of OsEra1-deficient rice mutant;

[0075] (1) Construction of vectors and recombinant bacteria

[0076] Using the DNA sequence shown in SEQ ID NO:1,

[0077] Screen suitable targets on the E-CRISPR website (http: / / www.e-crisp.org / E-CRISP / designcrispr.html).

[0078] Combined with the score and target location, select:

[0079] AGTGCATTGTCCGCCCCGAGAGG located in the first exon of OsEra1 is one of the CRISPR / Cas9 targets.

[0080] Synthetic primer sequences:

[0081] F:ggcaAGTGCATTGTCCGCCCCGAG;

[0082] and R:aaacCTCGGGGCGGACAATGCACT;

[0083] According to the literature (Ma X, Zhang Q, Zhu Q, Liu W, Chen Y, Qiu R, Wang B, Yang Z, Li H, Lin Y, Xie Y, Shen R, Chen S, Wang Z, Chen Y, Guo J, Chen L, Zhao X, Dong Z, Liu YG (2015) A Robust CRISPR / Cas9 System for Convenient, High-Efficiency Multiplex Genome Editing in Monocot and Dicot Plants. Mol Plant 8: 1274-1284), pCRISPR / Cas9 plasmid was constructed to finally obtain vector pCRISPR / Cas9-OsEra1. The plasmid pCRISPR / Cas9-OsEra1 was transformed into Agrobacterium EHA105 competent cells to obtain recombinant Agrobacterium pCRISPR / Cas9-OsEra1.

[0084] (2) pCRISPR / Cas9-OsEra1 transformation of rice

[0085] The genetic transformation of rice was completed by Wuhan Boyuan Biotechnology Co., Ltd., and the T0 generation transgenic strain of CRISPR / Cas9-OsEra1 under the NIP background, namely the T0 generation of rice osera1 mutant, was obtained.

[0086] Example 3

[0087] Identification of transgenic rice CRISPR / Cas9-OsEra1 T0 generation plants;

[0088] DNA was extracted from leaves of the T0 generation of the rice osera1 mutant, and PCR amplification was performed using the extracted DNA as a template and specific primers of the OsEra1 gene.

[0089] The primer sequences are:

[0090] F:ATGATCATACTGACCAAGGA;

[0091] R:TGCTTCTATGTGACAATTAG.

[0092] The PCR reaction program was: 94°C pre-denaturation for 2 min; 94°C denaturation for 30 s, 56°C annealing for 30 s, 72°C extension for 30 s, 35 cycles; 72°C extension for 5 min. The PCR product was subjected to Sanger sequencing to check whether the target site had mutations.

[0093] Among them, the extraction of leaf DNA was carried out according to the CTAB method: put the rice leaves into a 2.0mL centrifuge tube, add steel beads and grind them into powder with a grinder. Add 400μL CTAB extraction solution to the centrifuge tube and shake to mix. Add an equal volume of chloroform and phenol (1:1) mixture, shake to mix, and centrifuge at 12000rpm for 10min. Take 200μL supernatant, add an equal volume of chloroform, shake vigorously to mix, and centrifuge at 12000rpm for 10min. Take 100μL supernatant, add twice the volume of anhydrous ethanol and precipitate at 0℃ for 10min. Centrifuge at 4℃, 12000rpm for 10min. Remove the supernatant, add 500μL 75% ethanol to wash twice, blow dry and add 30μL ddH2O to dissolve.

[0094] The results showed that two independent transgenic plants (osera1-L1 and osera1-L2) with mutations in the OsEra1 gene were finally identified. The mutation sites were compared with the DNA sequences. Figure 3 As shown in Figure 2, osera1-L1 lacks 5 bp, causing a frameshift mutation in OsEra1, leading to loss of its function.

[0095] osera1-L2 inserted a base A 17 bp downstream of ATG, which also caused a frameshift mutation and led to its loss of function.

[0096] Example 4

[0097] Analysis of salt tolerance in osera1 mutant plants;

[0098] Similarly, the wild-type NIP and osera1-L1 and osera1-L2 mutant materials were grown for three weeks under the above culture conditions, and then subjected to 180 mM NaCl stress treatment. After 14 days of treatment and untreated materials, photos were taken, and then recovery treatment was started. After 14 days of recovery, photos were taken. Figure 1 As shown, Figure 1 osera1-L1 in the expression indicates OsEra1 gene mutant line 1. Figure 1osera1-L2 in the figure indicates OsEra1 gene mutant strain 2, and NIP indicates wild-type Nipponbare. After 14 days of recovery treatment, the survival rates of NIP, osera1-L1, and osera1-L2 were counted. Figure 2 As shown in the figure, the survival rate of NIP is about 40%, while the survival rate of osera1-L1 and osera1-L2 is only about 20%. This result shows that the functional loss of OsEra1 makes the plants more sensitive to salt stress. It can be seen that OsEra1 plays a positive regulatory role in the process of plant response to salt stress.

[0099] The present invention has been described in detail above. It will be apparent to those skilled in the art that the present invention may be implemented in a wide range under equivalent parameters, concentrations and conditions without departing from the spirit and scope of the present invention and without the need for unnecessary experimentation. Although the present invention provides specific embodiments, it should be understood that further improvements may be made to the present invention. In short, according to the principles of the present invention, this application intends to include any changes, uses or improvements to the present invention, including changes made by conventional techniques known in the art that depart from the scope disclosed in this application. Applications of some of the basic features may be made within the scope of the following appended claims.

[0100] It is easy to understand that those skilled in the art can combine, split, reorganize, etc. the embodiments of the present application to obtain other embodiments based on one or several embodiments provided in the present application, and these embodiments do not exceed the protection scope of the present application.

[0101] The present invention and its implementation methods are described schematically above, and the description is not restrictive. The embodiments shown in the embodiments are only part of the implementation methods of the present invention, and the actual structure is not limited thereto. Therefore, if a person skilled in the art is inspired by the embodiments and designs a structure and an implementation method similar to the technical solution without creativity without departing from the purpose of the invention, they should all fall within the protection scope of the present invention.

[0102] SEQ ID NO:1

[0103] OsEra1 gene genomic nucleotide sequence

[0104]

[0105] SEQ ID NO:2

[0106] OsEra1 gene CDS sequence

[0107]

[0108] SEQ ID NO:3

[0109] Amino acid sequence of OsEra1 protein

[0110] MIILTKDLKGNTAYQRVTLSALITRSALSAPRGSRFRKVNVVKGLKIYEDVFTTSELMKVADFINEIRQAGRNGELSGETFIFFNKQIKGNKREIIQLGVPLFQPTTEESNCHIEAIPLVLQAVIDHLVLWRLIPESRKPNSVIINFFDEDEHSQPYFKPPHLDNPISTLLLSETTMAFGRSLVTDSNGNYKGPLTLSLKQGSLLVMRGNSADMARHVVCPSSNRRVSITFVRVRPSTPVDLSPLPSPTKAMTLWQAPPPQQQQTIPAPGMQQKPQHGGAIIGYGPAPQAVLAPAWGMAVRAAPVMMVAAPAPARPMVMAPAGGNKRMSRSGTGVFLPWTVGPKRYNKHLPPRIQKRRFSAMMSPIEAQG

Claims

1. An application of reducing the expression of OsEra1 gene to reduce the salt tolerance of rice, characterized in that: The nucleotide sequence of the OsEra1 gene is: the OsEra1 gene genome sequence of SEQ ID NO: 1 or the OsEra1 gene CDS sequence of SEQ ID NO: 2 in the sequence list.

2. Use of reducing the expression of OsEra1 protein encoded by the OsEra1 gene as claimed in claim 1 to reduce the salt tolerance of rice, characterized in that: The OsEra1 protein is any one of the following: B1): the amino acid sequence is the protein of SEQ ID NO: 3 in the sequence list; B2): A fusion protein obtained by connecting a protein tag to the N-terminus or / and C-terminus of B1).

3. A method for cultivating transgenic rice with reduced salt tolerance, characterized in that: The method comprises reducing the expression of the gene encoding the protein according to claim 2 in the target rice to obtain a transgenic rice having lower salt tolerance than the target rice; the reducing the expression of the gene encoding the protein according to claim 2 in the target rice is achieved by inhibiting the content and / or activity of the protein according to claim 2 in the target rice using the CRISPR / Cas9 gene editing system.

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