Application of wheat TaSSRP1 protein and coding gene thereof in regulation and control of plant salt tolerance

Abstract

The invention discloses application of a wheat TaSSRP1 protein and a coding gene thereof in regulation and control of plant salt tolerance. According to the invention, DNA molecules for coding TaSSRP1 protein are respectively introduced into wild type arabidopsis thaliana and wild type wheat, so that TaSSRP1 overexpressed arabidopsis thaliana and TaSSRP1 overexpressed wheat are respectively obtained. Experiments prove that under salt stress, compared with wild type arabidopsis thaliana and wild type wheat, the salt tolerance of TaSSRP1 overexpression arabidopsis thaliana and the salt tolerance of TaSSRP1 overexpression wheat are both improved, and it is indicated that the salt stress tolerance of plants is improved through TaSSRP1 overexpression. The invention discovers that the TaSSRP1 protein and the coding gene thereof have the function of regulating and controlling the salt tolerance of the plant for the first time, and the TaSSRP1 protein and the coding gene thereof have an important value for cultivating salt-tolerant plant varieties.

Description

Technical Field

[0001] The present invention relates to the field of biotechnology, and in particular to application of wheat TaSSRP1 protein and a coding gene thereof in regulating plant salt tolerance. Background Art

[0002] Saline-alkali land is widely distributed around the world and covers a large area. Due to the influence of climate, hydrology and human activities, the trend of land salinization has worsened. Soil salinization seriously affects agricultural production and crop yields, threatening global food security. There are about 1.5 billion mu of saline-alkali land in my country, of which at least 1 / 3 has the potential for development and utilization. Cultivating salt-alkali tolerant crop varieties, making full use of and transforming saline-alkali land, increasing the effective arable land area, and thus increasing the total grain output are of great significance to safeguarding China's granary and keeping China's rice bowl.

[0003] Wheat is an important food crop, and salt stress is the main factor restricting wheat yield. At present, the key to ensuring wheat production lies in improving wheat's own salt tolerance. Under abiotic stress conditions, plants activate transcription factors through a variety of signal transduction pathways. These transcription factors can regulate the expression of a variety of stress response-related genes, thereby enhancing the plant's stress resistance. Therefore, cloning and transforming genes related to transcription factors and further regulating the expression of a variety of functional genes have become important strategies and means to improve crop stress resistance, and have broad application prospects. In plants, ERF transcription factors are a large family of transcription factors. At present, research on the role of the ERF transcription factor TaSSRP1 is insufficient, and no relevant research on TaSSRP1 in regulating salt tolerance has been found. Summary of the invention

[0004] The technical problem to be solved by the present invention is how to regulate plant stress tolerance.

[0005] In order to solve the above technical problems, the present invention first provides a new use of TaSSRP1 protein.

[0006] The present invention provides the use of TaSSRP1 protein in any of the following A1)-A4): A1) Regulate plant stress tolerance; A2) Cultivate plants resistant to reverse gene; A3) Preparation of products for regulating plant stress tolerance; A4) Preparation of products for breeding plants resistant to reverse gene mutation; The TaSSRP1 protein is derived from wheat and is any one of the following B1)-B4): B1) The amino acid sequence is the protein shown in SEQ ID NO:2; B2) A fusion protein having the same function obtained by connecting a tag to the N-terminus and / or C-terminus of the amino acid sequence shown in SEQ ID NO: 2; B3) A protein having the same function obtained by replacing and / or deleting and / or adding one or more amino acid residues in the amino acid sequence shown in SEQ ID NO: 2; B4) A protein that has 80% or more identity with the amino acid sequence shown in SEQ ID NO: 2 and has the same function.

[0007] In the protein described in B2) above, the tag refers to a polypeptide or protein that is fused and expressed with the target protein using DNA in vitro recombination technology to facilitate the expression, detection, tracing and / or purification of the target protein. The tag includes but is not limited to: GST (glutathione sulfhydryl transferase) 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 (monomeric red fluorescent protein) or AviTag tag protein.

[0008] In the protein described in B3) above, the substitution and / or deletion and / or addition of one or several amino acid residues is substitution and / or deletion and / or addition of no more than 10 amino acid residues, or substitution and / or deletion and / or addition of no more than 9 amino acid residues, or substitution and / or deletion and / or addition of no more than 8 amino acid residues, or substitution and / or deletion and / or addition of no more than 7 amino acid residues, or substitution and / or deletion and / or addition of no more than 6 amino acid residues, or substitution and / or deletion and / or addition of no more than 5 amino acid residues, or substitution and / or deletion and / or addition of no more than 4 amino acid residues, or substitution and / or deletion and / or addition of no more than 3 amino acid residues, or substitution and / or deletion and / or addition of no more than 2 amino acid residues, or substitution and / or deletion and / or addition of no more than 1 amino acid residue.

[0009] In the protein described in B4) above, the identity refers to the identity of the amino acid sequence. The identity of the amino acid sequence can be determined using a homology search site on the Internet, such as the BLAST page on the NCBI homepage website. For example, in Advanced BLAST2.1, by using blastp as a program, setting the Expect value to 10, setting all filters to OFF, using BLOSUM62 as a matrix, setting the Gap existence cost, Per residue gap cost and Lambda ratio to 11, 1 and 0.85 (default value) respectively, and searching for the identity of a pair of amino acid sequences for calculation, the value of the identity (%) can be obtained. The identity includes amino acid sequences having 80% or more, or 85% or more, or 90% or more, or 91% or more, or 92% or more, or 93% or more, or 94% or more, or 95% or more, or 96% or more, or 97% or more, or 98% or more, or 99% or more homology with the amino acid sequence shown in Sequence 2 of the present invention.

[0010] The protein described in B1) or B2) or B3) or B4) above can be synthesized artificially, or its encoding gene can be synthesized first and then expressed biologically.

[0011] In order to solve the above technical problems, the present invention further provides new uses of biomaterials related to the above TaSSRP1 protein.

[0012] The present invention provides the use of the biomaterial related to the above-mentioned TaSSRP1 protein in any of the following A1)-A4): A1) Regulate plant stress tolerance; A2) Cultivate plants resistant to reverse gene; A3) Preparation of products for regulating plant stress tolerance; A4) Preparation of products for breeding plants resistant to reverse gene mutation; The biological material is any one of the following E1) to E5): E1) a nucleic acid molecule encoding the above-mentioned TaSSRP1 protein; E2) an expression cassette containing the nucleic acid molecule described in E1); E3) a recombinant vector containing the nucleic acid molecule described in E1), or a recombinant vector containing the expression cassette described in E2); E4) a recombinant microorganism containing the nucleic acid molecule described in E1), or a recombinant microorganism containing the expression cassette described in E2), or a recombinant microorganism containing the recombinant vector described in E3); E5) A recombinant host cell containing the nucleic acid molecule of E1), or a recombinant host cell containing the expression cassette of E2), or a recombinant host cell containing the recombinant vector of E3).

[0013] In the above application, the nucleic acid molecule E1) is any one of the following: F1) the DNA molecule shown in SEQ ID NO: 21-839; F2) A DNA molecule having 75% or more identity with the nucleotide sequence defined in F1) and encoding the above-mentioned TaSSRP1 protein.

[0014] A person skilled in the art can easily mutate the nucleotide sequence encoding the TaSSRP1 protein of the present invention by using known methods, such as directed evolution and point mutation. Those artificially modified nucleotides having 80% or higher identity with the TaSSRP1 nucleotide sequence isolated from the present invention are derived from the nucleotide sequence of the present invention and are equivalent to the sequence of the present invention as long as they encode the TaSSRP1 protein and have the same function. The identity refers to the sequence similarity with the natural nucleic acid sequence, including nucleotide sequences with 75% or higher, or 80% or higher, or 85% or higher, or 90% or higher, or 95% or higher identity with the nucleotide sequence of the protein composed of the amino acid sequence shown in the coding sequence 2 of the present invention. The identity can be evaluated by the naked eye or computer software. Using computer software, the identity between two or more sequences can be expressed as a percentage (%), which can be used to evaluate the identity between related sequences.

[0015] The above 75% or more identity may be 75%, 80%, 85%, 90% or more identity. The above 75% identity may be at least 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity. The above 80% identity may 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. The 85% or greater identity may be at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity. The 90% or greater identity may be at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity. The 95% or greater identity may be at least 95%, 96%, 97%, 98%, or 99% identity.

[0016] In the above application, the expression cassette refers to a DNA capable of expressing the above TaSSRP1 protein in a host cell. The expression cassette may also include a single-stranded or double-stranded nucleic acid molecule of all regulatory sequences necessary for expressing the above TaSSRP1 protein nucleic acid molecule. The regulatory sequence can guide the coding sequence to express the above TaSSRP1 protein in a suitable host cell under compatible conditions. The regulatory sequence includes, but is not limited to, a leader sequence, a polyadenylation sequence, a propeptide sequence, a promoter, a signal sequence and a transcription terminator. At a minimum, the regulatory sequence should include a promoter and termination signals for transcription and translation. In order to introduce specific restriction enzyme sites into the vector so as to connect the regulatory sequence to the coding region of the nucleic acid sequence encoding the protein, a regulatory sequence with a linker can be provided. The regulatory sequence may be a suitable promoter sequence, i.e., a nucleic acid sequence that can be recognized by the host cell expressing the nucleic acid sequence. The promoter sequence contains a transcriptional regulatory sequence that mediates protein expression. The promoter may be any nucleic acid sequence that is transcriptionally active in the selected host cell, including mutant, truncated and hybrid promoters, and may be obtained from a gene encoding an extracellular or intracellular protein that is homologous or heterologous to the host cell. The regulatory sequence can also be a suitable transcription termination sequence, i.e., a sequence that can be recognized by the host cell to terminate transcription. The termination sequence can be operably connected to the 3' end of the nucleic acid sequence encoding the protein. Any terminator that can function in the selected host cell can be used in the present invention. The regulatory sequence can also be a suitable leader sequence, i.e., an mRNA non-translated region that is very important for the translation of the host cell. The leader sequence can be operably connected to the 5' end of the nucleic acid sequence encoding the protein. Any leader sequence that can function in the selected host cell can be used in the present invention. The regulatory sequence can also be a signal peptide coding region, which encodes an amino acid sequence connected to the amino terminus of the protein and can guide the encoded protein into the cell secretory pathway. The signal peptide coding region that can guide the expressed protein into the secretory pathway of the host cell used can be used in the present invention. It may also be necessary to add a regulatory sequence that can regulate protein expression according to the growth of the host cell.

[0017] In the above application, the carrier refers to a TaSSRP1 A vector for carrying genes into host cells for amplification and expression, which may be a cloning vector or an expression vector, including but not limited to: a plasmid, a phage (such as lambda phage or M13 filamentous phage, etc.), a cosmid (i.e., a cosmid), a Ti plasmid, a viral vector (such as a retrovirus (including a lentivirus), adenovirus, adeno-associated virus, etc.).

[0018] The recombinant vector refers to TaSSRP1 The recombinant DNA molecule is constructed by connecting the gene with the vector in vitro. Existing plant expression vectors can be used to construct a recombinant DNA molecule containing the gene. TaSSRP1 Gene or TaSSRP1Recombinant vector of gene expression cassette. The plant expression vector includes binary Agrobacterium vector and vector that can be used for plant microprojectile bombardment, etc. Such as pAHC25, pBin438, pCAMBIA1302, pCAMBIA2300, pCAMBIA2301, pCAMBIA1301, pCAMBIA1300, pBI121, pCAMBIA1391-Xa or pCAMBIA1391-Xb (CAMBIA Company), etc. The plant expression vector may also contain the 3′ non-translated region of the foreign gene, i.e., a polyadenylic acid signal and any other DNA fragments involved in mRNA processing or gene expression. The polyadenylic acid signal can guide the addition of polyadenylic acid to the 3′ end of the mRNA precursor, such as the Agrobacterium crown gall induction (Ti) plasmid gene (such as the nopaline synthase gene Nos), the 3′ non-translated region of the plant gene (such as the soybean storage protein gene) has similar functions. When using the gene of the present invention to construct a plant expression vector, enhancers may also be used, including translation enhancers or transcription enhancers. These enhancer regions may be ATG start codons or adjacent region start codons, etc., but must be the same as the reading frame of the coding sequence to ensure the correct translation of the entire sequence. The sources of the translation control signals and start codons are wide-ranging and may be natural or synthetic. The translation initiation region may come from a transcription initiation region or a structural gene. In order to facilitate the identification and screening of transgenic plant cells or plants, the plant expression vector used may be processed, such as adding genes that can be expressed in plants and encode enzymes or luminescent compounds that can produce color changes (GUS genes, luciferase genes, etc.), antibiotic marker genes (such as genes that confer resistance to kanamycin and related antibiotics) and nptII Gene that confers resistance to the herbicide phosphinothricin bar Gene that confers resistance to the antibiotic hygromycin hph genes, and those conferring resistance to methotrexate dhfr Gene, conferring resistance to glyphosate EPSPS Gene) or chemical resistance marker gene (such as herbicide resistance gene), mannose-6-phosphate isomerase gene that provides the ability to metabolize mannose. Considering the safety of transgenic plants, no selective marker gene can be added, and transformed plants can be directly screened by adversity.

[0019] In the above application, the microorganism may be bacteria, fungi, actinomycetes, protozoa, algae or viruses. Among them, the bacteria may be from the genus Escherichia ( Escherichia sp. ), Erwinia ( Erwinia sp. ), Agrobacterium ( Agrobacterium sp. ), Flavobacterium ( Flavobacterium sp. )、Alcaligenes spp. Alcaligenes sp. ), Pseudomonas ( Pseudomonas sp. ), Bacillus (Bacillus sp. ) etc. but not limited thereto, for example, the bacteria may be Escherichia coli ( Escherichia coli )、Bacillus subtilis( Bacillus subtilis ) or Bacillus pumilus ( Bacillus pumilus The fungus may be a yeast, and the yeast may be from the genus Saccharomyces (e.g., Saccharomyces cerevisiae Saccharomyces cerevisiae ), Kluyveromyces (such as Kluyveromyces lactis Kluyveromyces lactis ), Pichia (such as Pichia pastoris Pichia pastoris ), Schizosaccharomyces spp. (e.g. Schizosaccharomyces pombe Schizosaccharomyces pombe ), Hansenula (such as Hansenula polymorpha Hansenula polymorpha ) etc. but not limited thereto. The fungus may also be from the genus Fusarium ( Fusarium sp. ), Rhizoctonia spp. Rhizoctonia sp. ), Verticillium ( Verticillium sp. )、Penicillium( Penicillium sp. )、Aspergillus( Aspergillus sp. )、Cephalosporium( Cephalosporium sp. ) etc. but not limited thereto. The actinomycetes may be from the genus Streptomyces ( Streptomyces sp. ), Nocardia ( Nocardia sp. ), Micromonospora ( Micromonospora sp. ), Neurospora ( Streptosporangium sp. )、Actinomyces spp.( Actinoplanes sp. )、Thermoactinomycetes( Thermoactinomyces sp. ) etc. but not limited thereto. The algae may be from the genus Fucus ( Fucus sp. ), the genus A. Achnanthes sp. ), Cocoon algae ( Amphiprora sp. ), Diplocoriaceae ( Amphora sp. ), Fibrocystis spp. Ankistrodesmus sp. ), Astrophytum ( Asteromonas sp. )、Golden Chromophytes( Boekelovia sp. ) etc. but not limited thereto. The virus may be rotavirus, herpes virus, influenza virus, adenovirus etc. but not limited thereto.

[0020] In the above applications, the host cell (also referred to as a recipient cell) may be a plant cell or an animal cell. The host cell may be understood to refer not only to a specific recipient cell, but also to the offspring of such a cell, and due to natural, accidental or intentional mutations and / or changes, the offspring may not necessarily be completely identical to the original parent cell, but is still included in the scope of the host cell. Suitable host cells are known in the art, among which: the plant cell may be Arabidopsis thaliana ( Arabidopsis thaliana ),tobacco( Nicotiana tabacum ),corn( Zea mays )、Rice( Oryza sativa ),wheat( Triticum aestivum) and other plant cells, but not limited thereto; the animal cells may be mammalian cells (e.g., Chinese hamster ovary cells (CHO cells), African green monkey kidney cells (Vero cells), baby hamster kidney cells (BHK cells), mouse breast cancer cells (C127 cells), human embryonic kidney cells (HEK293 cells), human HeLa cells, fibroblasts, bone marrow cell lines, T cells or NK cells, etc.), avian cells (e.g., chicken or duck cells), amphibian cells (e.g., African clawed frog (Xenopus laevis) cells or giant salamander (Andrias davidianus) cells), fish cells (e.g., grass carp, carp, rainbow trout or catfish cells), insect cells (e.g., Sf21 cells or Sf-9 cells), etc., but not limited thereto.

[0021] In the present invention, the recombinant microorganism (or recombinant host cell) refers to a recombinant microorganism (or recombinant host cell) with a changed function obtained by operating and modifying the genes of the target microorganism (or target host cell). TaSSRP1 The recombinant microorganism (or recombinant host cell) obtained after the gene or the above-mentioned recombinant vector is introduced into the target microorganism (or target host cell). The recombinant microorganism (or recombinant host cell) can be understood to refer not only to the specific recombinant microorganism (or recombinant host cell), but also to the offspring of such a cell, and due to natural, accidental or intentional mutations and / or changes, the offspring may not be completely identical to the original parent cell, but is still included in the scope of the recombinant microorganism (or recombinant host cell).

[0022] In the above application, the regulating plant stress tolerance is to improve plant stress tolerance.

[0023] Furthermore, the stress resistance is salt resistance.

[0024] Furthermore, the salt tolerance is NaCl tolerance.

[0025] In one embodiment of the present invention, the regulation of plant stress tolerance is to improve plant salt tolerance, and the improvement of plant salt tolerance is specifically embodied in that under salt stress, when the content and / or activity of TaSSRP1 protein in the plant is increased, the salt tolerance of the plant is enhanced; further embodied in that under salt stress, TaSSRP1 The gene expression level increases and the chlorophyll content of the plant increases.

[0026] In another embodiment of the present invention, the regulation of plant stress tolerance is to improve plant salt tolerance, and the improvement of plant salt tolerance is specifically embodied in that under salt stress, when the content and / or activity of TaSSRP1 protein in the plant is increased, the salt tolerance of the plant is enhanced; further embodied in that under salt stress, TaSSRP1The gene expression level increased, the MDA content in the plant leaves decreased, and the proline content increased.

[0027] In order to solve the above technical problems, the present invention also provides a method for cultivating plants resistant to gene reversal.

[0028] The method for cultivating a gene-reversal resistant plant provided by the present invention comprises the following steps: increasing the content and / or activity of TaSSRP1 protein in a target plant to obtain a transgenic plant; the transgenic plant has higher stress tolerance than the target plant.

[0029] In the above method, the stress resistance is salt resistance, specifically NaCl resistance.

[0030] In the above method, the method for increasing the content and / or activity of TaSSRP1 protein in the target plant is to overexpress the TaSSRP1 protein in the target plant. The overexpression method can be to introduce the coding gene of the TaSSRP1 protein into the target plant. The introduction includes but is not limited to: transfecting plant cells or tissues by conventional biological methods such as Ti plasmid, Ri plasmid, plant virus vector, direct DNA transformation, microinjection, electroporation, Agrobacterium-mediated, and cultivating the transfected plant cells or tissues into plants.

[0031] In one embodiment of the present invention, the gene encoding the TaSSRP1 protein can be introduced into the target plant via a recombinant vector.

[0032] In a specific embodiment of the present invention, the gene encoding the TaSSRP1 protein is introduced into the target plant via the recombinant vector pBI121-TaSSRP1. The recombinant vector pBI121-TaSSRP1 is a vector obtained by inserting the DNA molecule shown in positions 21-839 of SEQ ID NO: 1 between the BamHI and SaCl restriction sites of the pBI121 vector.

[0033] In the above method, the salt tolerance of the transgenic plant is higher than that of the target plant, which is embodied in any one of the following X1)-X3): X1) Under salt stress, the chlorophyll content of the transgenic plant is higher than that of the target plant; X2) under salt stress, the proline content of the transgenic plant is greater than that of the target plant; X3) Under salt stress, the MDA content of the transgenic plant is lower than that of the target plant.

[0034] Furthermore, the salt stress is NaCl stress.

[0035] Furthermore, the NaCl stress is 300 mM NaCl.

[0036] In any of the above applications or methods, the transgenic plant not only comprises TaSSRP1 The first generation of transgenic plants obtained by gene transformation of the recipient plant also includes its progeny. For transgenic plants, the gene can be propagated in the species, or the gene can be transferred into other varieties of the same species using conventional breeding techniques, especially including commercial varieties. The transgenic plants include seeds, callus, complete plants and cells.

[0037] In any of the above applications or methods, the plant is a dicotyledon or a monocotyledon; further, the monocotyledon is a Poaceae plant; further, the Poaceae plant is rice (such as Kongyu 131).

[0038] The present invention introduces DNA molecules encoding TaSSRP1 protein into Arabidopsis and wheat, respectively, to obtain TaSSRP1 Overexpression of Arabidopsis and TaSSRP1 Overexpression of wheat. Experiments have shown that under salt stress, compared with the recipient plants, TaSSRP1 Overexpression of Arabidopsis and TaSSRP1 The salt tolerance of overexpressed wheat was improved. TaSSRP1 Overexpression improves the tolerance of plants to salt stress. The present invention first discovered that TaSSRP1 protein and its encoding gene have the function of regulating plant salt tolerance, which is of great value for breeding salt-tolerant plant varieties. BRIEF DESCRIPTION OF THE DRAWINGS

[0039] Figure 1 For wheat TaSSRP1 qPCR expression analysis of genes and TaSSRP1 Identification of salt tolerance of overexpressed Arabidopsis. A shows the expression of wheat root, stem, leaf, ear and grain tissues. B shows the response expression to 200 mM NaCl salt stress. C shows the schematic diagram of pBI121 plant overexpression vector. D shows TaSSRP1 RT-PCR analysis of overexpressed Arabidopsis (before salt stress treatment, wild-type Arabidopsis and TaSSRP1 The leaves of the overexpressing Arabidopsis strains were analyzed). E is the growth status of Arabidopsis seedlings after 300 mM NaCl salt stress treatment. Among them, WT: wild-type Arabidopsis; OE1, OE2, OE3: TaSSRP1 Overexpression Arabidopsis lines. F: WT and TaSSRP1 Chlorophyll content in overexpressed Arabidopsis thaliana.

[0040] Figure 2 For wheat TaSSRP1 qPCR expression analysis of genes and TaSSRP1 Identification of salt tolerance in overexpressed wheat. A is wheat TaSSRP1 Gene in wild type and TaSSRP1Expression levels in overexpressed wheat (before salt stress treatment, wild type and TaSSRP1 The leaves of the overexpressing wheat lines were analyzed. B shows the growth of wheat seedlings after 300 mM NaCl salt stress. C shows the growth of WT and TaSSRP1 MDA content in overexpressed wheat. D is the difference between WT and TaSSRP1 Overexpression of proline content in wheat.

[0041] Figure 3 Figure 1 shows the activation effect of TaSSRP1 on the promoters of stress and antioxidant enzyme genes. A shows the analysis of cis-acting elements in the promoters of stress and antioxidant enzyme genes. B shows the structure of the reporter and effector vectors used in the dual luciferase system. C shows the dual luciferase test of TaSSRP1 activating stress and antioxidant enzyme genes. ** indicates the difference reaches 1% level.

[0042] Figure 4 The subcellular localization, transcriptional activity analysis and yeast two-hybrid screening of TaSSRP1. A is the subcellular localization of wheat TaSSRP1 (the subcellular localization of wheat TaSSRP1 protein was observed under a laser confocal fluorescence microscope, and the wavelengths of 488nm and 633nm were used for GFP and chloroplast autofluorescence, respectively, and the scale bar is 50 μm). B is the transcriptional activity analysis of wheat TaSSRP1 (yeast cells carrying the bait vectors pGBKT7-TaSSRP1 (1-272 aa, 1-127aa and 128-272 aa) were cultured in SD / -Trp and SD / -Trp / -His / -Ade medium, respectively. DETAILED DESCRIPTION

[0043] 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.

[0044] The experimental methods in the following examples, unless otherwise specified, are all conventional methods, and are performed according to the techniques or conditions described in the literature in the field or according to the product instructions. The materials, reagents, etc. used in the following examples, unless otherwise specified, can all be obtained from commercial channels.

[0045] The wheat variety Fielder in the following examples is recorded in the document “Wang, B., Li, L., Liu, M., Peng, D., Wei, A., Hou, B., Lei, Y., & Li, X. (2022). TaFDL2-1A confers droughtstress tolerance by promoting ABA biosynthesis, ABA responses, and ROSscavenging in transgenic wheat. The Plant journal : for cell and molecularbiology, 112(3), 722–737. https: / / doi.org / 10.1111 / tpj.15975”.

[0046] The pBI121 vector in the following examples is described in the document “Yu, M., Yu, Y., Song, T., Zhang, Y., Wei, F., Cheng, J., Zhang, B., & Zhang, X. (2022). Characterization of the voltage-dependent anion channel (VDAC) gene family in wheat (TriticumaestivumL.) and its potential mechanism in response to drought and salinitystresses. Gene, 809, 146031. https: / / doi.org / 10.1016 / j.gene.2021.146031”.

[0047] The wheat variety JW1 in the following examples is described in the document “Yu, Y., Song, T., Wang, Y., Zhang, M., Li, N., Yu, M., Zhang, S., Zhou, H., Guo, S., Bu, Y., Wang, T., Xiang, J., & Zhang, X. (2023). The wheat WRKY transcription factor TaWRKY1-2Dconfers drought resistance in transgenic Arabidopsis and wheat (Triticumaestivum L.). International journal of biological macromolecules, 226, 1203–1217. https: / / doi.org / 10.1016 / j.ijbiomac.2022.11.234”.

[0048] The plant overexpression vector pCAMBIA3301 in the following examples is described in the document “Du, L., Huang, X., Ding, L., Wang, Z., Tang, D., Chen, B., Ao, L., Liu, Y., Kang, Z., & Mao, H. (2023). TaERF87 and TaAKS1 synergistically regulate TaP5CS1 / TaP5CR1-mediated proline biosynthesis to enhance drought tolerance in wheat. The Newphytologist, 237(1), 232–250. https: / / doi.org / 10.1111 / nph.18549”.

[0049] Embodiment 1, wheat TaSSRP1 Gene cloning, tissue specificity and salt stress expression pattern analysis 1. Take wheat variety JW1 seeds and disinfect them with 70% alcohol, then rinse with sterile water. The seedlings are cultured in a light incubator (70% humidity, 18000 Lux light intensity) at a temperature of 20-25°C and a light / dark cycle of 16h / 8h in 1 / 2 strength Hoagland solution to obtain two-week-old wheat seedlings.

[0050] 2. Two-week-old wheat seedlings were treated under 200 mM NaCl stress, and leaves were collected at 0, 1, 6, 12, and 24 h after treatment; leaves at 0 h were used as gene cloning templates and stress treatment controls.

[0051] 3. Two-week-old wheat seedlings were cultured in a greenhouse under the following culture conditions: temperature of 15-20°C, light / dark cycle of 16h / 8h, cultured until wheat matured and bloomed 7 days later, and wheat root, stem, leaf, ear and grain tissues were collected. All samples were quickly stored at -80°C for subsequent RNA extraction.

[0052] 4. Take the wheat variety JW1 seedling material treated with NaCl for 0h, extract total RNA and reverse transcribe it into cDNA. Use cDNA as a template and use primers 5'-ACGCGACCACACAGGATAAGAT-3' and 5'-TGAGAACGATCACTGGTCGCTT-3' for PCR amplification to obtain the PCT product (TaSSRP1cDNA), and sequence the PCR product.

[0053] The sequencing results showed that the nucleotide sequence of the PCR product was as shown in SEQ ID NO: 1-868, among which, 21-839 was TaSSRP1 The gene coding sequence encodes the TaSSRP1 protein whose amino acid sequence is sequence 2.

[0054] 5. Take the wheat variety JW1 seedling materials treated with NaCl at different times or in different tissues, extract total RNA and reverse transcribe it into cDNA, use cDNA as a template, and use primers 5'-ACCTTCGACAGCGCCGAG-3' and 5'-GGAGATGAAGAGTCGGGGCT-3' to perform real-time fluorescence quantitative PCR (qRT-PCR) in a fluorescence quantitative instrument to detect TaSSRP1 Gene expression level.

[0055] The results are as follows Figure 1 A and Figure 1 As shown in B, the results show that wheat TaSSRP1 The gene was highly expressed in roots and leaves and was significantly upregulated under salt stress, suggesting that TaSSRP1 may play a role in regulating root and leaf development and salt stress response.

[0056] Embodiment 2, TaSSRP1 Construction of Arabidopsis overexpression lines and analysis of their salt tolerance one, TaSSRP1 Construction of overexpression Arabidopsis lines 1. Replace the 21st to 839th position of sequence 1 TaSSRP1The coding region sequence was connected between the BamHI and SaCl restriction sites of the plant overexpression vector pBI121 to obtain the recombinant vector pBI121-TaSSRP1. The schematic diagram of the structure of the recombinant vector pBI121-TaSSRP1 is shown in Figure 1 As shown in C.

[0057] 2. The recombinant vector pBI121-TaSSRP1 was transformed into GV3101 Agrobacterium (Shanghai Weidi Biotechnology Co., Ltd., CAT#: AC1001). After identification, GV3101 Agrobacterium containing the recombinant vector pBI121-TaSSRP1 was obtained and recorded as the recombinant bacterium GV3101 / pBI121-TaSSRP1.

[0058] 3. Use the recombinant bacteria GV3101 / pBI121-TaSSRP1 to transfer the TaSSRP1 The gene was transferred into Arabidopsis thaliana (Columbia type), and T0 seeds were harvested; T0 seeds were screened using MS medium containing kanamycin to obtain T1 seedlings; T1 seedlings were individually transferred to substrate soil and cultured until maturity, and T2 seeds were harvested; T2 seeds were screened using MS medium containing kanamycin, and T2 resistant seedlings with a segregation ratio of resistant (green seedlings) to sensitive (albino seedlings) of 3:1 were selected.

[0059] T0 represents the Arabidopsis seeds harvested in the current period after floral dip infection; T1 represents the seeds obtained in the current period of transformation and the plants grown from them; T2 represents the seeds produced by self-pollination of T1 and the plants grown from them.

[0060] 4. Extract T2 generation TaSSRP1 Total RNA from Arabidopsis thaliana overexpressing lines OE1, OE2, OE3 and wild-type Arabidopsis was reverse transcribed into cDNA. PCR amplification was performed using primers 5'-ATGCTGCTGCTTAATCCGGC-3' and 5'-GGAGATGAAGAGTCGGGGCT-3' to amplify the cDNA sequence of Arabidopsis thaliana. Actin2 The gene was an internal reference gene, and the primer sequences of the internal reference gene were as follows: 5′-CTAAGCTCTCAAGATCAAAGGC-3′ and 5′-AACATTGCAAAGAGTTTCAAGG-3′.

[0061] The results are as follows Figure 1 As shown in D, the results show that the internal reference gene TaSSRP1 The target fragment was detected in both the overexpressing Arabidopsis lines and wild-type Arabidopsis. TaSSRP1 It was detected in overexpressing Arabidopsis lines but not in wild-type Arabidopsis.

[0062] The positive T2 generation Arabidopsis seedlings were transplanted into the matrix soil and cultured until maturity to obtain T3 generation single copy transgenic Arabidopsis seeds. Finally, three T3 generation Arabidopsis TaSSRP1 Overexpressing Arabidopsis lines OE1, OE2, and OE3 were used for the following salt tolerance analysis.

[0063] two, TaSSRP1 Analysis of salt tolerance in overexpressing Arabidopsis Test materials: Wild-type Arabidopsis thaliana (WT), T3 generation TaSSRP1 There were 40 strains of each of the overexpressing Arabidopsis lines OE1, OE2, and OE3.

[0064] Experimental method: After the seeds were sterilized, they were sown on MS solid medium and cultured at 22°C, 16 hours of light / 8 hours of darkness to obtain three-week-old Arabidopsis seedlings. Three-week-old Arabidopsis seedlings were subjected to salt stress treatment. The salt stress treatment method was as follows: Arabidopsis seedlings were cultured in pots filled with nutrient soil (substrate: vermiculite = 3:1) at 22°C, 16 hours of light / 8 hours of darkness. During the culture process, 300mM NaCl aqueous solution was used for irrigation in the pots. 200mL of 300mM NaCl aqueous solution was irrigated each time, twice a day, for 13 consecutive days. After 13 days of salt stress treatment, photos were taken, and then the total chlorophyll content (including chlorophyll a and chlorophyll b) in the leaves of each Arabidopsis strain was determined using the Solebold Chlorophyll Content Kit (Cat. No.: BC0995). At the same time, normal water supply was used as a control.

[0065] The results show that after 13 days of salt stress, the leaves of WT were severely wilted, while TaSSRP1 Overexpressing Arabidopsis lines all showed relatively mild symptoms ( Figure 1 E). Under normal water supply conditions, WT and TaSSRP1 There was no difference in chlorophyll content between the overexpression lines; under salt stress conditions, TaSSRP1 The total chlorophyll content in the overexpressing Arabidopsis lines was significantly higher than that in the WT ( Figure 1 F). The above results show that overexpression of wheat TaSSRP1 Genes can enhance salt tolerance in Arabidopsis thaliana.

[0066] Embodiment 3, TaSSRP1 Construction of overexpression wheat lines and analysis of their salt tolerance one, TaSSRP1 Construction of overexpression wheat lines 1. Replace the 21st to 839th position of sequence 1 TaSSRP1 The coding region sequence was connected between the BamHI and SpeI restriction sites of the plant overexpression vector pCAMBIA3301 to obtain the recombinant vector pCAMBIA3301-TaSSRP1.

[0067] 2. The recombinant vector pCAMBIA3301-TaSSRP1 was transformed into EHA105 Agrobacterium (Shanghai Weidi Biotechnology Co., Ltd., CAT#: AC1010). After identification, EHA105 Agrobacterium containing the recombinant vector pCAMBIA3301-TaSSRP1 was obtained and recorded as the recombinant bacteria GV3101 / pCAMBIA3301-TaSSRP1.

[0068] 3. The recombinant vector pCAMBIA3301-TaSSRP1 was transformed into the immature embryo callus of the wheat variety Fielder by Agrobacterium infection. After screening, pre-differentiation, differentiation, PCR identification and generation, three T3 transgenic wheat lines were finally obtained, named OE1, OE2 and OE3.

[0069] 4. At the three-leaf stage, wild-type wheat Fielder and T3 transgenic wheat lines OE1, OE2 and OE3 were taken, total RNA was extracted and reverse transcribed into cDNA, and then real-time fluorescence quantitative PCR (qRT-PCR) was performed on a fluorescence quantitative instrument using primers 5'-ACCTTCGACAGCGCCGAG-3' and 5'-GGAGATGAAGAGTCGGGGCT-3' to detect TaSSRP1 Gene expression level.

[0070] The results are as follows Figure 2 As shown in A, the results showed that compared with wild-type wheat, the T3 generation TaSSRP1 Transgenic wheat lines OE1, OE2 and OE3 TaSSRP1 The expression levels of TaSSRP1 Transgenic wheat lines OE1, OE2 and OE3 were used for the salt tolerance analysis described below.

[0071] two, TaSSRP1 Analysis of salt tolerance in overexpressing wheat Test materials: Wild-type Arabidopsis thaliana (WT), T3 generation TaSSRP1 There were 40 overexpressing wheat lines OE1, OE2, and OE3.

[0072] Experimental method: After the seeds were sterilized, they were sown on MS solid culture medium and cultured for 2 days at 23°C, 16 hours of light / 8 hours of darkness. Then they were moved into pots containing 250 grams of nutrient soil: vermiculite = 1:1. They were continued to be cultured at 23°C, 16 hours of light / 8 hours of darkness, and humidity>50% until the three-leaf stage and then subjected to salt stress treatment. The salt stress treatment method was as follows: the wheat plants were moved into pots containing nutrient soil (matrix: vermiculite = 3:1) and cultured at 23°C, 16 hours of light / 8 hours of darkness. During the culture process, 300 mM NaCl aqueous solution was used for irrigation in the pots. Each irrigation was 200 mL of 300 mM NaCl aqueous solution, twice a day, for 20 consecutive days. After 20 days of salt stress treatment, the phenotype was observed and photographed. The MDA content and proline content in the leaves were determined using the Solebold MDA content and proline content determination kit. At the same time, normal water supply was used as a control.

[0073] The results are as follows Figure 2 As shown in B, the results show that under salt stress conditions, TaSSRP1 The growth of the overexpressing wheat lines was better than that of the WT, but the WT wilted severely. TaSSRP1 There was no difference in MDA and proline content between the overexpression wheat lines; under salt stress conditions, TaSSRP1 The MDA content in the overexpression wheat lines was significantly lower than that in the WT ( Figure 2 C), while for the proline content index, TaSSRP1 The overexpression wheat lines were significantly higher than WT ( Figure 2 D). The above results indicate that overexpression in wheat TaSSRP1 Gene could enhance salt tolerance in wheat.

[0074] Example 4: Effects of TaSSRP1 on the activity of stress and antioxidant enzyme gene promoters 1. In order to understand the molecular mechanism of TaSSRP1 regulating salt tolerance, the PlantCARE database was used to analyze wheat stress response ( TaP5CS1 , TaDREB1 , TaERF3 ) and antioxidant enzymes ( TaPOD , TaCAT and TSA ( Fe The promoters of the genes (2500 bp upstream of ATG) were analyzed and the distribution of DRE elements (specific elements bound by TaSSRP1 transcription factor) in the promoters of these genes was mapped. Figure 3 As shown in A, it is found that except TaPOD All genes except TaSSRP1 Combined DRE elements.

[0075] 2. In order to study whether TaSSRP1 can activate the activity of TaP5CS1, TaDREB1, TaERF3, TaPOD, TaCAT and TaSOD (Fe) promoters, the promoter sequence fragments of these genes were cloned into the pGreen II 0800 vector containing the Renilla luciferase (REN) gene driven by the CaMV35S promoter by homologous recombination to obtain the pGreen II0800 recombinant vector (pGreen II 0800-TaP5CS1, pGreen II 0800-TaDREB1, pGreen II 0800-TaERF3, pGreen II 0800-TaPOD, pGreen II 0800-TaCAT or pGreen II0800-TaSOD (Fe)). In the same way, the TaSSRP1 CDS sequence was cloned into the pGreenII 62-SK vector to obtain the pGreenII62-SK-TaSSRP1 recombinant vector ( Figure 3 B). pGreen II 0800-TaP5CS1, pGreen II0800-TaDREB1, pGreen II 0800-TaERF3, pGreen II 0800-TaPOD, pGreen II 0800-TaCAT, pGreen II 0800-TaSOD (Fe), and pGreenII 62-SK-TaSSRP1 plasmids were transformed into Agrobacterium (strain: GV3101 containing pSoup-19), and then Agrobacterium containing pGreen II 0800 recombinant vectors were mixed with Agrobacterium containing pGreenII 62-SK-TaSSRP1 plasmid at a volume ratio of 1:1, centrifuged, resuspended in infiltration buffer (150 mm Acetosyringone, 10 mm MgCl2, 10 mm MES, pH 5.6), and finally injected into Nicotiana benthamiana leaves for transient expression. The dual luciferase detection system was used to detect the fluorescence ratio of LUC and REN using a microplate reader 2 days after transfection, and at least three biological replicates were tested for each combination.

[0076] The results are as follows Figure 3 As shown in B, the results show that: TaPOD Gene, TaSSRP1 The above results indicate that TaSSRP1 may be able to activate the expression of stress and antioxidant enzyme genes to regulate wheat salt tolerance.

[0077] Example 5: Subcellular localization, transcriptional activation activity and interaction protein analysis of wheat TaSSRP1 1. Subcellular localization of wheat TaSSRP1 Will TaSSRP1 The coding region sequence without the stop codon was connected to the Pcambia1302-GFP vector to obtain the recombinant vector Pcambia1302-GFP-TaSSRP1 。 The recombinant vector Pcambia1302-GFP-TaSSRP1 was transformed into the GV3101 Agrobacterium strain to obtain the recombinant bacteria GV3101 / Pcambia1302-GFP-TaSSRP1. The recombinant bacteria GV3101 / Pcambia1302-GFP-TaSSRP1 was injected into Nicotiana benthamiana leaves for transient expression, and the recombinant bacteria GV3101 / Pcambia1302-GFP was used as a control. The GFP signal was observed 48 hours later using an Olympus laser confocal microscope.

[0078] The results are as follows Figure 4 As shown in A, the results showed that compared with the control, the fluorescence signal of TaSSRP1 was mainly concentrated in the cell nucleus, indicating that TaSSRP1 was mainly localized in the cell nucleus.

[0079] 2. Wheat TaSSRP1 transcriptional activation activity The coding sequence of the full-length TaSSRP1 protein (1-272 aa), the coding sequence of the N-terminal truncated fragment of the TaSSRP1 protein (1-127 aa at the N-terminus), and the coding sequence of the C-terminal truncated fragment of the TaSSRP1 protein (128-272 aa at the C-terminus) were ligated into the pGBKT7 bait vector to obtain the recombinant vectors pGBKT7- TaSSRP1 (1-272 aa), pGBKT7- TaSSRP1 (N-terminal 1-127 aa) and pGBKT7- TaSSRP1 (C-terminal 128-272 aa), and then transformed into AH109 yeast strain. To evaluate the transcriptional activation activity, pick a single clone and incubate in SD / -Trp liquid medium at 30°C and 180rpm overnight. Spot 1.2μL of yeast liquid on SD / -Trp and SD / -Trp / -His / -Ade solid medium, invert and culture for 1-3 days to observe the growth of yeast colonies. If the yeast can grow on the above two media, it means that the protein has transcriptional activation activity.

[0080] The results are as follows Figure 4 As shown in B, the results showed that all recombinant vector-transformed AH109 strains could grow in SD / -Trp medium, but only the N-terminal AH109 strain could grow in SD / -Trp / -His / -Ade medium. The above results indicate that the full-length TaSSRP1 protein has no transcriptional activation activity, and the transcriptional activation region is at the N-terminus.

[0081] 3. Yeast two-hybrid screening and verification of wheat TaSSRP1 The plasmid containing pGBKT7-TaSSRP1 was transformed into AH109 yeast cells for further yeast two-hybrid screening. Single clones were picked and placed in a 2 mL centrifuge tube (containing 1 mL SD / -Trp liquid medium) and shaken at 200 rpm overnight. Then, 100 μL was pipetted into 40 mL SD / -Trp liquid medium and cultured for 15 h to a concentration of OD 600nm About 0.8. Centrifuge at 5000rpm for 5min at room temperature, remove the supernatant, and then resuspend the bacterial block with SD / -Trp liquid culture medium (4mL). Add 4mL of resuspended strain, 1mL of library strain, and 45mL of 2×YPDA to a 2 L sterile conical flask in sequence. Culture slowly at 30℃ and 50rpm for 22h. Then observe whether yeast conjugates appear under an optical microscope. If conjugates appear, proceed to the next step, otherwise continue to culture with shaking. Centrifuge at 4500rpm for 4min, remove the supernatant, and then resuspend the precipitate with 50mL of 0.5×YPDA. Repeat the above centrifugation and supernatant removal operations, and resuspend the bacteria with 10mL of 0.5×YPDA. Spread the resuspended bacterial solution on SD / -Trp / -His / -Leu / -Ade solid culture medium and culture at 30℃ for 4d. Single clones were picked and placed in SD / -Trp / -His / -Leu / -Ade liquid medium and cultured at 30°C for 2 days. Yeast plasmids were extracted using the Tiangen yeast plasmid extraction kit. After the collected plasmids were transformed into DH5α Escherichia coli, single clones were picked and sent for sequencing after shaking. The sequencing results showed that the genes screened from the cDNA library had complete CDS sequences. The recombinant plasmids containing pGBKT7-TaSSRP1 and pGADT7-prey were co-transformed into the AH109 yeast strain and cultured on SD / -Trp / -Leu and SD / -Trp / -Leu / -His / -Ade / +X-α-Gal solid media for rotation verification.

[0082] The results are as follows Figure 4 As shown in C, the results showed that the AH109 yeast strain co-transfected with pGADT7-TaNF-YC6 and pGBKT7-TaSSRP1 could grow on SD / -Trp / -Leu and SD / -Trp / -Leu / -His / -Ade / + X-α-Gal medium, indicating that TaSSRP1 and TaNF-YC6 candidate interacting proteins interacted with each other in the yeast system.

[0083] The interaction between TaSSRP1 and TaNF-YC6 proteins was further verified in plant cells. The TaSSRP1 and TaNF-YC6 genes were connected to the pCAMBIA1302-nYFP and pCAMBIA1302-cYFP vectors, respectively, to obtain the recombinant vectors pCAMBIA1302-nYFP-TaSSRP1 and pCAMBIA1302-cYFP-TaNF-YC6, respectively. The recombinant vectors pCAMBIA1302-nYFP-TaSSRP1 and pCAMBIA1302-cYFP-TaNF-YC6 were co-transformed into Agrobacterium GV3101 and transiently co-expressed in tobacco. After 48 hours, the YFP fluorescence signal was observed using a laser confocal microscope. If the YFP fluorescence signal exists, it means that TaSSRP1 and TaNF-YC6 proteins interact in tobacco.

[0084] The results are as follows Figure 4 As shown in D, the results showed that the interaction between TaSSRP1 and TaNF-YC6 was further verified by BiFC. YFP fluorescence was observed when TaSSRP1 and TaNF-YC6 were co-transfected into tobacco, while no YFP fluorescence was observed in the control combination, indicating that there was an interaction between TaSSRP1 and TaNF-YC6 in tobacco cells.

[0085] 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.

Claims

1. Use of the protein in any of the following A1)-A4): A1) Regulate plant stress tolerance; A2) Cultivate plants resistant to reverse gene; A3) Preparation of products for regulating plant stress tolerance; A4) Preparation of products for breeding plants resistant to reverse gene mutation; The protein is any one of the following B1)-B4): B1) The amino acid sequence is the protein shown in SEQ ID NO:2; B2) A fusion protein having the same function obtained by connecting a tag to the N-terminus and / or C-terminus of the amino acid sequence shown in SEQ ID NO: 2; B3) A protein having the same function obtained by replacing and / or deleting and / or adding one or more amino acid residues in the amino acid sequence shown in SEQ ID NO: 2; B4) A protein that has 80% or more identity with the amino acid sequence shown in SEQ ID NO: 2 and has the same function.

2. Use of a biological material related to the protein of claim 1 in any of the following A1)-A4): A1) Regulate plant stress tolerance; A2) Cultivate plants resistant to reverse gene; A3) Preparation of products for regulating plant stress tolerance; A4) Preparation of products for breeding plants resistant to reverse gene mutation; The biological material is any one of the following E1) to E5): E1) a nucleic acid molecule encoding the protein according to claim 1; E2) an expression cassette containing the nucleic acid molecule described in E1); E3) a recombinant vector containing the nucleic acid molecule described in E1), or a recombinant vector containing the expression cassette described in E2); E4) a recombinant microorganism containing the nucleic acid molecule described in E1), or a recombinant microorganism containing the expression cassette described in E2), or a recombinant microorganism containing the recombinant vector described in E3); E5) A recombinant host cell containing the nucleic acid molecule of E1), or a recombinant host cell containing the expression cassette of E2), or a recombinant host cell containing the recombinant vector of E3).

3. The use according to claim 2, characterized in that: E1) The nucleic acid molecule is any of the following: F1) the DNA molecule shown in SEQ ID NO: 21-839; F2) A DNA molecule that has 75% or more identity with the nucleotide sequence defined in F1) and encodes the protein.

4. The use according to any one of claims 1 to 3, characterized in that: The stress resistance is salt resistance.

5. The use according to any one of claims 1 to 4, characterized in that: The plant is a dicotyledonous plant or a monocotyledonous plant.

6. A method for cultivating plants resistant to gene reversal, comprising the following steps: increasing the content and / or activity of the protein of claim 1 in a target plant to obtain a transgenic plant; the transgenic plant has higher stress tolerance than the target plant.

7. The method according to claim 6, characterized in that The stress resistance is salt resistance.

8. The method according to claim 7, characterized in that: The method for increasing the content and / or activity of the protein described in claim 1 in the target plant is to overexpress the protein described in claim 1 in the target plant.

9. The method according to any one of claims 6 to 8, characterized in that: The overexpression method is to introduce the gene encoding the protein in claim 1 into the target plant.

10. The method according to any one of claims 6 to 9, characterized in that: The plant is a dicotyledonous plant or a monocotyledonous plant.

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