Application of GhPSKR1 gene in regulation and control of plant salt tolerance

Abstract

The invention belongs to the technical field of plant breeding, and particularly relates to application of a GhPSKR1 gene in regulation and control of plant salt tolerance. The nucleotide sequence of the GhPSKR1 gene provided by the invention is as shown in SEQ ID NO. 1, and the amino acid sequence of the protein expressed by the GhPSKR1 gene is as shown in SEQ ID NO. 2. A research result proves that the growth inhibition of a GhPSKR1 gene silent plant under salt stress treatment is more obvious, the salt damage characteristic is more obvious, sodium ion accumulation in leaves and roots is increased, and the GhPSKR1 gene silent plant is more sensitive to salt stress; under salt stress treatment, the root length of a GhPSKR1 gene overexpression transgenic homozygous plant is remarkably elongated compared with that of wild arabidopsis thaliana, and the GhPSKR1 gene has good stress resistance, so that the GhPSKR1 gene plays a positive regulation role in regulating plant salt stress.

Description

Technical Field

[0001] The invention belongs to the technical field of plant breeding, and particularly relates to application of a GhPSKR1 gene in regulating plant salt tolerance. Background Art

[0002] As a type of soil widely distributed around the world, saline-alkali land is mainly concentrated in arid and coastal areas. As an important raw material for the textile industry, cotton is gradually expanding its planting area to saline-alkali land to cope with the decreasing high-quality arable land resources. However, cotton is not a halophyte and its salt tolerance is relatively limited. As soil salinity increases, cotton growth will suffer from severe salt stress, which is manifested in stunted growth and development, reduced yield, and deterioration of fiber quality. Currently, there is a lack of research on the salt tolerance mechanism of cotton, especially at the molecular level.

[0003] The specific phenotypes of salt stress threats to cotton growth are: growth inhibition, manifested as short plants, fewer and smaller leaves; yellowing, which is due to high salt affecting cell water absorption and nutrient transport; salt stress delays flowering, reduces the number of flowers, reduces fruiting rate, and significantly affects cotton yield; especially critically, salt stress also significantly shortens cotton root length, weakens the root system's ability to absorb water and nutrients, and further hinders the overall growth and yield formation of cotton. Therefore, when planting cotton in saline-alkali land, it is necessary to conduct further research to reduce the adverse effects of salt stress on cotton growth and yield and ensure the sustainable development of the cotton industry.

[0004] Phytosulfokine is a new type of plant peptide growth regulator. Mature phytosulfokine is a sulfonated short peptide composed of five acids [Tyr(SO3H)-Ile-Tyr(SO3H)-Thr-Gln], in which the tyrosine is sulfonated. Phytosulfokine regulates a variety of physiological and biochemical reactions in plants, including in vitro differentiation of plant cells, cell growth, regulation of root apical meristem cells and quiescent central cells, development of pollen tube cells, and regulation of plant immunity. Phytosulfokine in plants transmits signals to the cell through the leucine-rich receptor kinase protein family member PSKR on the cell membrane as a receptor. The phytosulfokine receptor PSKR reported in Arabidopsis has dual signal perception functions of kinase activity and guanylate cyclase activity. However, there are few reports on the research of PSKR in plant stress resistance. Among them, the English name of phytosulfokine is phytosulfokine, PSK. The English name of leucine-rich receptorlike kinase is leuine-rich receptorlike kinase protein, LRR-RLK.

[0005] Currently, there is a need to develop a new product application of the GhPSKR gene in cotton stress resistance. Summary of the invention

[0006] In order to develop a new application of the GhPSKR gene in cotton stress resistance, the present invention provides an application of the GhPSKR1 gene in regulating plant salt tolerance. To achieve the above purpose, the present invention adopts the following technical solution.

[0007] The first aspect of the present invention provides an application of a GhPSKR1 gene in regulating plant salt tolerance, wherein the nucleotide sequence of the GhPSKR1 gene is shown in SEQ ID NO.1.

[0008] The present invention discloses for the first time the positive regulatory role of the GhPSKR gene in cotton salt tolerance and provides excellent regulatory targets and theoretical guidance for cotton salt tolerance cultivation. The present invention provides an application of the GhPSKR1 gene in regulating plant salt tolerance and develops a new product application of the GhPSKR gene in cotton stress resistance.

[0009] Preferably, the amino acid sequence of the protein expressed by the GhPSKR1 gene is shown in SEQ ID NO.2.

[0010] Preferably, the GhPSKR1 gene in the plant is overexpressed by genetic engineering methods to improve the salt tolerance of the plant.

[0011] Preferably, the genetic engineering method is: using the GhPSKR1 gene to construct a recombinant expression vector, and introducing the recombinant expression vector into a plant to improve the salt tolerance of the plant.

[0012] Preferably, the method of introducing the recombinant expression vector into the plant includes direct DNA transformation through any one of Ti plasmid, Ri plasmid and plant virus vector; or introducing the recombinant expression vector into the plant through any one of microinjection, electroporation and Agrobacterium-mediated. The present invention cloned the cotton GhPSKR1 gene, constructed a transgenic Arabidopsis plant overexpressing GhPSKR1 using transgenic technology, and constructed a GhPSKR1 silenced plant using VIGS technology, verified the function of GhPSKR1, and clarified that GhPSKR1 can improve the salt tolerance of plants. It is conducive to a deeper study of the response mechanism of plants to salt stress signals, lays a good molecular foundation for effectively improving plant salt tolerance, and has great value in exploring the signal regulation network of plants under salt stress.

[0013] The second aspect of the present invention provides a recombinant expression vector containing the GhPSKR1 gene.

[0014] Preferably, the recombinant expression vector is obtained by inserting the DNA fragment of the GhPSKR1 gene into the Sma I and Kpn I restriction sites of the super1300-GFP vector.

[0015] The plant RNA is reverse transcribed to obtain a template DNA, and PCR amplification is performed based on the template DNA using a primer pair as shown in SEQ ID NO.3 and SEQ ID NO.4, thereby obtaining the DNA fragment of the GhPSKR1 gene.

[0016] The primer in the primer pair shown in SEQ ID NO.3 is the upstream primer F1, and its nucleotide sequence is as follows:

[0017] 5'-gtaccagattacgctcatatgATGGGGGCTCAAGGCTGCTG-3'.

[0018] The primer in the primer pair shown in SEQ ID NO.4 is the downstream primer R1, and its nucleotide sequence is as follows:

[0019] 5'-actggcctccatggccatatgTCAAACACTAGATAACGATGTC-3'.

[0020] Furthermore, a method for constructing a recombinant expression vector containing the GhPSKR1 gene comprises the following steps:

[0021] The sequenced PGADT7-GhPSKR1 plasmid was used as template for amplification.

[0022] The obtained amplified product was double-digested and connected with the plant binary transformation vector super1300-GFP vector double-digested with Sma I and Kpn I, transformed, and the correct positive clone plasmid was extracted and verified, which was the recombinant expression vector 35S::GhPSKR1-GFP.

[0023] Among them, according to the multiple cloning site of the plant binary transformation vector super1300-GFP and the coding region sequence of the GhPSKR1 gene, a forward primer: upstream primer F1 and a reverse primer: downstream primer R1 for amplifying the entire coding region of the GhPSKR1 gene were designed to obtain a 35S::GhPSKR1-GFP recombinant expression vector. The specific construction method is as follows:

[0024] The GhPSKR1 gene is used as a template, and PCR amplification is performed using a forward primer and a reverse primer to obtain a product containing the GhPSKR1 gene; the product containing the GhPSKR1 gene and super1300 are respectively digested using Sma I and Kpn I to obtain a digestion product and a vector frame, respectively, and the digestion product and the vector frame are recovered; the digestion product and the vector frame are connected to obtain a recombinant expression vector 35S::GhPSKR1-GFP, that is, the DNA fragment between the Sma I and Kpn I digestion sites of super1300 is replaced with the GhPSKR1 gene as shown in SEQ ID NO.1, and the other sequences of super1300 are kept unchanged.

[0025] Among them, the plant binary transformation vector super1300-GFP is a recombinant expression vector connected to the GFP gene, which is recorded in the doctoral dissertation "Research on metabolic characteristics of in vitro growth of Cuscuta and Agrobacterium mis genes in its genome, Zhang Yuexia, "China Agricultural University", 2020". With the author's consent, the public can obtain it from the research group of Li Zhaohu of China Agricultural University to repeat the experiments in the present invention. It cannot be used for other purposes. It is referred to as "super1300" in the present invention).

[0026] The structure of the recombinant expression vector 35S::GhPSKR1-GFP is as follows Figure 3 shown.

[0027] The third aspect of the present invention provides a method for improving plant salt tolerance, which overexpresses the GhPSKR1 gene in the plant, or increases the activity of the protein expressed by the GhPSKR1 gene in the plant, or increases the content of the protein expressed by the GhPSKR1 gene in the plant, so as to improve the salt tolerance of the plant.

[0028] In the above method, increasing the expression level of the protein GhPSKR1 gene and / or the activity of the protein GhPSKR1 and / or the content of the protein GhPSKR1 in the target plant is to overexpress the protein GhPSKR1 in the plant.

[0029] Preferably, the overexpression of the GhPSKR1 gene in the plant is achieved by transforming the plant with the GhPSKR1 gene mediated by Agrobacterium to improve the salt tolerance of the plant.

[0030] Preferably, the plant is a monocot or a dicot. More preferably, the plant is a plant of the family Cruciferae and / or Malvaceae. More preferably, the plant is a plant of the genus Gossypium and / or Arabidopsis. Most preferably, the plant is Arabidopsis and / or cotton.

[0031] The fourth aspect of the present invention provides a method for breeding salt-tolerant plants, which overexpresses the GhPSKR1 gene in plants to obtain salt-tolerant transgenic plants.

[0032] Wherein, the plant is M1) or M2) or M3) or M4) or M5).

[0033] M1) Monocotyledonous or dicotyledonous plants.

[0034] M2) Cruciferae.

[0035] M3) Arabidopsis thaliana.

[0036] M4) cotton plants.

[0037] M5) Cotton.

[0038] The fifth aspect of the present invention provides a protein, which is named cotton polypeptide receptor GhPSKR1, referred to as GhPSKR1 protein, derived from cotton, and is any of the following proteins:

[0039] A1) comprises or is a protein having an amino acid sequence of SEQ ID NO.2.

[0040] A2) A fusion protein obtained by connecting a tag to the N-terminus and / or C-terminus of the amino acid sequence shown in SEQ ID NO.2.

[0041] A3) A protein having more than 90% identity and the same function as the protein shown in A1) 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.

[0042] The above proteins can be artificially synthesized, or their encoding genes can be synthesized first and then expressed biologically.

[0043] In the above proteins, the protein 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 or purification of the target protein. The protein tag can be a Flag tag, a His tag, an MBP tag, an HA tag, a myc tag, a GST tag and / or a SUMO tag.

[0044] In the above-mentioned proteins, 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, Perresidue gap cost and Lambda ratio to 11, 1 and 0.85 (default values) respectively, and searching for the identity of a pair of amino acid sequences, the identity value (%) can be obtained.

[0045] In the above proteins, the above 90% or more identity may be at least 91%, 92%, 95%, 96%, 98%, 99% or 100% identity.

[0046] The biomaterials related to the above-mentioned GhPSKR1 protein also fall within the protection scope of the present invention. The present invention also provides new uses of the biomaterials related to the GhPSKR1 protein.

[0047] The present invention provides an application of a biomaterial related to the GhPSKR1 protein in regulating plant stress resistance, wherein the biomaterial is any one of the following:

[0048] C1) Nucleic acid molecule encoding GhPSKR1 protein.

[0049] C2) An expression cassette containing the nucleic acid molecule described in C1).

[0050] C3) A recombinant expression vector containing the nucleic acid molecule described in C1), or a recombinant expression vector containing the expression cassette described in C2).

[0051] 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 expression vector described in C3).

[0052] 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), or a transgenic plant cell line containing the recombinant expression vector described in C3).

[0053] C6) A transgenic plant tissue containing the nucleic acid molecule described in C1), or a transgenic plant tissue containing the expression cassette described in C2), or a transgenic plant tissue containing the recombinant expression vector described in C3).

[0054] 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), or a transgenic plant organ containing the recombinant expression vector described in C3).

[0055] C8) A transgenic plant containing the nucleic acid molecule described in C1), or a transgenic plant containing the expression cassette described in C2), or a transgenic plant containing the recombinant expression vector described in C3).

[0056] C9) Tissue culture produced from regenerable cells of the transgenic plant described in C8).

[0057] C10) Protoplasts produced from the tissue culture described in C9).

[0058] C11) A recombinant expression vector or recombinant microorganism that inhibits the expression of the gene of the above-mentioned GhPSKR1 protein and / or inhibits the activity of the above-mentioned GhPSKR1 protein and / or reduces the content of the above-mentioned GhPSKR1 protein.

[0059] Wherein, the nucleic acid molecule can be DNA, such as cDNA, genomic DNA or recombinant DNA. The nucleic acid molecule can also be RNA, such as mRNA or hnRNA.

[0060] In the above biological material, the nucleic acid molecule in C1) is any one of the following:

[0061] B1) The DNA molecule shown in SEQ ID NO.1.

[0062] B2) The coding sequence is the DNA molecule shown in SEQ ID NO.1.

[0063] B3) A DNA molecule that hybridizes with the DNA molecule defined in B1) or B2) under stringent conditions and encodes the GhPSKR1 protein.

[0064] The above stringent conditions are hybridization and washing the membrane twice in a 2×SSC, 0.1% SDS solution at 68°C, each time for 5 minutes, and hybridization and washing the membrane twice in a 0.5×SSC, 0.1% SDS solution at 68°C, each time for 15 minutes.

[0065] In the above biological materials, the expression cassette described in C2) refers to a DNA capable of expressing the GhPSKR1 protein in a host cell, and the DNA may include not only a promoter for initiating transcription of the GhPSKR1 gene, but also a terminator for terminating transcription of the GhPSKR1 gene.

[0066] Furthermore, the expression cassette may also include an enhancer sequence. Promoters that can be used in the present invention include, but are not limited to: the promoter of the GhPSKR1 gene itself, a constitutive promoter, a tissue, organ and development-specific promoter, and an inducible promoter. Examples of promoters include, but are not limited to, the constitutive promoter 35S of cauliflower mosaic virus, the wound-inducible promoter from tomato, leucine aminopeptidase ("LAP", Chao et al. (1999) Plant Physiol 120:979-992); the chemically inducible promoter from tobacco, pathogenesis-related 1 (PR1) (induced by salicylic acid and BTH (benzothiadiazole-7-thiocarboxylic acid S-methyl ester)); the tomato proteinase inhibitor II promoter (PIN2) or the LAP promoter (both inducible by methyl jasmonate); heat shock promoters (U.S. Pat. No. 5,187,267); tetracycline-inducible promoters (U.S. Pat. No. 5,057,422); seed-specific promoters, such as millet seed-specific promoter pF128 (Chinese Patent CN101063139B), promoters specific for seed storage proteins (e.g., phaseolusin, napin, oleosin and soybean beta-glutamic acid); conglycin promoter (Beach et al. (1985) EMBO J. 4: 3047-3053). They can be used alone or in combination with other plant promoters. All references cited herein are cited in their entirety. Suitable transcription terminators include, but are not limited to, the terminator of the GhPSKR1 gene itself, the Agrobacterium nopaline synthase terminator (NOS terminator), the cauliflower mosaic virus CaMV 35S terminator, the tmL terminator, the pea rbcS E9 terminator, and the nopaline and octopine synthase terminators (see, for example, Odell et al. (1985) Nature 313: 810; Rosenberg et al. (1987) Gene, 56: 125; Guerineau et al. (1991) Mol. Gen. Genet, 262: 141; Proudfoot (1991) Cell, 64: 671, Sanfacon et al. Genes Dev, 5: 141, Mogen et al. (1991) Plant Genet, 262: 141; Cell, 2: 1261; Munroe et al. (1990) Gene, 91: 151; Ballad et al. (1989) Nucleic Acids Res. 17: 7891; Joshi et al. (1987) Nucleic Acid Res., 15: 9627).

[0067] In the above biological materials, the recombinant expression vector in C3) may contain the DNA molecule for encoding protein GhPSKR1 shown in SEQ ID NO.1. Existing plant expression vectors may be used to construct a recombinant expression vector containing the gene of the GhPSKR1 protein or the gene expression cassette of the protein GhPSKR1. The plant expression vector may be a Gateway system vector or a binary expression vector, such as super1300, pGWB411, pGWB412, pGWB405, pBin438, pCAMBIA1302, pCAMBIA2300, pCAMBIA2301, pCAMBIA1301, pCAMBIA1300, pBI121, pCAMBIA1391-Xa or pCAMBIA1391-Xb. When GhPSKR1 is used to construct a recombinant expression vector, any enhancing, constitutive, tissue-specific or inducible promoter can be added before its transcription initiation nucleotide, such as the cauliflower mosaic virus (CAMV) 35S promoter, the ubiquitin gene Ubiqutin promoter (pUbi), etc., which can be used alone or in combination with other plant promoters; in addition, when the gene of the present invention is used to construct a plant expression vector, an enhancer can also be used, including a translation enhancer or a transcription enhancer. These enhancer regions can 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 signal and the start codon are extensive and can be natural or synthetic. The translation initiation region can come from the transcription initiation region or the structural gene.

[0068] In order to facilitate the identification and screening of transgenic plant cells or plants, the plant expression vector used can be processed, such as adding genes that can be expressed in plants and encode enzymes or luminescent compounds that can produce color changes (GUS gene, luciferase gene, etc.), antibiotic resistance markers (gentamicin marker, kanamycin marker, etc.) or chemical resistance marker genes (such as herbicide resistance genes), etc.

[0069] In a specific embodiment of the present invention, C3) the recombinant expression vector is a 35S::GhPSKR1-GFP recombinant expression vector, which is obtained by replacing the DNA fragment between the Sma I and Kpn I restriction sites of super1300 with the GhPSKR1 gene of SEQ ID NO.1, while keeping the other sequences of super1300 unchanged.

[0070] In the above biological materials, the recombinant expression vector C11) is the recombinant expression vector pYL156-GhPSKR1, and the pYL156-GhPSKR1 is a recombinant expression vector obtained by replacing the DNA fragment between the EcoR I and BamH I restriction sites of the pYL156 vector with the DNA molecule shown at positions 1-311 of SEQ ID NO.1, while keeping the other sequences of the pYL156 vector unchanged.

[0071] In the above biological materials, the recombinant microorganism can specifically be yeast, bacteria, algae and fungi; for example, the bacteria can be Agrobacterium GV3101.

[0072] In the above biological materials, the transgenic plant organs can be roots, stems, leaves, flowers, fruits and seeds of transgenic plants.

[0073] Among the above biological materials, the tissue culture can be derived from roots, stems, leaves, flowers, fruits, seeds, pollen, embryos and anthers.

[0074] Among the above biological materials, the transgenic plant cell lines, transgenic plant tissues and transgenic plant organs do not include propagation materials.

[0075] The present invention further provides a product for regulating plant stress resistance, which contains the above-mentioned GhPSKR1 protein or biological materials related thereto.

[0076] The use of the above-mentioned GhPSKR1 protein or biological materials related thereto in any of the following also falls within the protection scope of the present invention:

[0077] D1) Application in breeding transgenic plants with enhanced stress resistance.

[0078] D2) Application in the preparation and cultivation of transgenic plant products with enhanced stress resistance.

[0079] D3) Application in breeding gene silenced plants with reduced stress resistance.

[0080] D4) Application in the preparation and cultivation of gene-silenced plant products with reduced stress resistance.

[0081] D5) Application in plant breeding.

[0082] Among the above applications, the application in plant breeding can specifically be hybridizing plants containing GhPSKR1 protein or biological materials related thereto (such as the gene GhPSKR1 encoding the GhPSKR1 protein) with other plants to perform plant breeding.

[0083] Compared with the prior art, the present invention has the following beneficial effects:

[0084] The present invention provides an application of the GhPSKR1 gene in regulating plant salt tolerance. The research results of the present invention confirm that the growth inhibition of GhPSKR1 gene-silenced plants (especially cotton) under salt stress treatment is more obvious, the salt damage characteristics are more significant, the sodium ion accumulation in the leaves and roots increases, and they are more sensitive to salt stress; and under salt stress treatment, the root length of the GhPSKR1 gene-overexpressing transgenic homozygous plants (especially Arabidopsis) is significantly elongated compared with the wild-type Arabidopsis, and has good stress resistance, which indicates that the GhPSKR1 gene plays a positive regulatory role in regulating plant salt stress. The present invention reveals for the first time the positive regulatory role of GhPSKR in cotton salt tolerance, and provides excellent regulatory targets and theoretical guidance for cotton salt-tolerant cultivation. The present invention provides an application of the GhPSKR1 gene in regulating plant salt tolerance, and develops a new product application of the GhPSKR gene in cotton stress resistance. BRIEF DESCRIPTION OF THE DRAWINGS

[0085] Figure 1 It is an agarose gel electrophoresis diagram of the gene amplification product of the GhPSKR1 gene described in the examples of the present invention.

[0086] Figure 2 Schematic diagram of the structure of the recombinant expression vector 35S::GhPSKR1-GFP described in the examples of the present invention.

[0087] Figure 3 This is a subcellular localization map of the GhPSKR1 protein described in the examples of the present invention.

[0088] Figure 4 This is an agarose gel electrophoresis diagram of the amplified products at positions 2296-2596 of the GhPSKR1 gene described in the examples of the present invention.

[0089] Figure 5 The silencing efficiency and salt stress phenotype of the VIGS-GhPSKR1 silenced plants in the embodiment of the present invention; wherein, Figure 5 Figure (A) is a diagram showing the results of identifying the gene silencing efficiency of the GhPSKR1 gene in cotton described in the examples of the present invention; Figure 5 Figure (B) is a growth phenotype result diagram of the stress resistance verification of the GhPSKR1 gene silenced plants described in the examples of the present invention.

[0090] Figure 6 Schematic diagram of the sodium ion content in the leaves and roots of the GhPSKR1 gene silenced plant described in the embodiment of the present invention; wherein, Figure 6 (A) shows the sodium ion content in leaves of VIGS-GFP and VIGS-GhPSKR1 after salt treatment; Figure 6(B) shows the sodium ion content in the roots of VIGS-GFP and VIGS-GhPSKR1 after salt treatment.

[0091] Figure 7 This is a diagram showing the Western Blot identification results of the homozygous transgenic Arabidopsis thaliana strain overexpressing GhPSKR1 described in the examples of the present invention.

[0092] Figure 8 : is a graph comparing the growth conditions of the homozygous transgenic Arabidopsis thaliana strain overexpressing GhPSKR1 and the wild-type Arabidopsis thaliana under salt stress conditions described in the examples of the present invention; wherein, Figure 8 The plants from left to right in the figure are wild-type Arabidopsis (WT), GhPSKR1 overexpressing transgenic Arabidopsis homozygous line OE1, and GhPSKR1 overexpressing transgenic Arabidopsis homozygous line OE2.

[0093] Fig. 9 It is a schematic diagram of the statistical results of root length of the homozygous transgenic Arabidopsis thaliana strain overexpressing GhPSKR1 and the wild-type Arabidopsis thaliana under salt stress conditions described in the examples of the present invention. DETAILED DESCRIPTION

[0094] The present invention is described in detail below in conjunction with the accompanying drawings and specific examples, but should not be construed as limiting the present invention. Unless otherwise specified, the technical means used in the following examples are conventional means well known to those skilled in the art, and the materials, reagents, etc. used in the following examples, unless otherwise specified, can be obtained from commercial sources.

[0095] Virus-induced gene silencing is an effective reverse genetics technique widely used in the identification of plant genome functions. Virus-induced gene silencing can successfully silence endogenous genes in different parts of the plant, providing a practical means for studying gene functions at different growth stages. In the embodiments of the present invention, a GhPSKR1 gene silenced plant is provided by a virus-induced gene silencing method to demonstrate the function of the gene. Among them, the English name of virus-induced gene silencing is virus-induced gene silencing, VIGS.

[0096] The present invention confirms that the growth inhibition of GhPSKR1-silenced cotton plants is more obvious under salt stress treatment, the salt damage characteristics are more obvious, the sodium ion accumulation in the leaves and roots is increased, and the plants are more sensitive to salt stress; and under salt stress treatment, the root length of GhPSKR1-overexpressing transgenic homozygous Arabidopsis plants is significantly elongated compared with wild-type Arabidopsis, and the plants have good stress resistance, which indicates that the GhPSKR1 gene plays a positive regulatory role in regulating plant salt stress.

[0097] To this end, the embodiment of the first aspect of the present invention proposes an application of the GhPSKR1 gene in regulating plant salt tolerance.

[0098] Example 1: Discovery, cloning, localization of the GhPSKR1 gene and construction of a recombinant vector for overexpression of GhPSKR1

[0099] 1.1 Discovery of the GhPSKR1 gene

[0100] In this example, the salt-related genes in the VIGS cDNA library were screened and searched using the cotton database. A protein was obtained from the cotton variety "Guoxin No. 3", which was named GhPSKR1 protein. The amino acid sequence of the GhPSKR1 protein is shown in SEQ ID NO.2:

[0101]

[0102] The gene encoding the GhPSKR1 protein is named as GhPSKR1 gene, and the nucleotide sequence of its open reading frame is shown in SEQ ID NO.1:

[0103]

[0104] For the screening method, please refer to "Using VIGS technology to study the function of cotton stress resistance genes, Li Fangjun, "China Agricultural University", 2014".

[0105] For details on the cotton variety "Guoxin No. 3", please refer to "Guoxin No. 3 Insect-Resistant Cotton Cultivation Technology, Feng Sulian, "Hebei Agriculture", 2011, 06".

[0106] 1.2 Cloning of GhPSKR1 gene

[0107] In this example, the GhPSKR1 gene was cloned and amplified by reverse transcription PCR, confirmed by electrophoresis, and the obtained clone was further sequenced.

[0108] (1) RNA from cotton leaves was extracted using the Adelaide kit and the first-strand cDNA was synthesized using the M-mLV reverse transcription kit. The obtained first-strand cDNA was used as a template to amplify the full-length GhPSKR1 gene.

[0109] Among them, the Adelaide kit was purchased from Beijing Adelaide Biotechnology Co., Ltd. and the M-mLV reverse transcription kit was purchased from Takara.

[0110] (2) Design two specific primers: upstream primer F1 and downstream primer R1, perform PCR amplification, and obtain PCR products.

[0111] Among them, using Novozymes Max Super-Fidelity DNA Polymerase Kit was used to amplify the full length of GhPSKR1 gene.

[0112] The nucleotide sequence of the upstream primer F1 is shown in SEQ ID NO.3:

[0113] 5'-gtaccagattacgctcatatgATGGGGGCTCAAGGCTGCTG-3'.

[0114] The nucleotide sequence of the downstream primer R1 is shown in SEQ ID NO.4:

[0115] 5'-actggcctccatggccatatgTCAAACACTAGATAACGATGTC-3'.

[0116] The PCR amplification reaction system is shown in Table 1:

[0117] Table 1 PCR reaction system

[0118]

[0119] The PCR amplification reaction program is shown in Table 2:

[0120] Table 2 PCR amplification program

[0121] Temperature / ℃ time Cycle times 95 3 minutes 1 time 95 15 seconds 35 times 52 15 seconds 35 times 72 40 seconds 35 times 72 5 minutes 1 time

[0122] (3) The PCR amplification product was electrophoresed on a 1.5% agarose gel.

[0123] (4) After the electrophoresis, the target band was cut out under ultraviolet light, and recovered and purified using an agarose gel DNA recovery kit, wherein the agarose gel DNA recovery kit was purchased from Kangwei Biotechnology Co., Ltd.

[0124] (5) The recovered fragments were recombined with the pGADT7 vector. The recombination reaction system was as shown in Table 3. The reaction was carried out at 37° C. for 30 min.

[0125] Table 3 Ligation reaction system

[0126]

[0127] (6) Take 5 μL of the ligation product and transform E. coli DH5α by heat shock method, and screen positive clones on LB solid plate containing 50 mg / L ampicillin. The heat shock method refers to "J. Sambrook, et al., translated by Huang Peitang, et al., Molecular Cloning Experiment Guide (3rd edition), Science Press, 2002 edition".

[0128] (7) Five clones were selected and sent to Beijing Qingke Sequencing Company for sequencing to obtain the required full-length gene CDS, that is, the GhPSKR1 gene.

[0129] Agarose gel electrophoresis results Figure 1 shown.

[0130] The sequencing results showed that the sequence of the GhPSKR1 gene was 3039 bp in length and encoded a complete ORF reading frame of 1012 amino acids.

[0131] 1.4 Subcellular localization of GhPSKR1 gene

[0132] Tobacco epidermal cells were used to study the subcellular localization of GhPSKR1 protein.

[0133] Agrobacterium expressing super100-GhPSKR1-GFP plasmid was cultured overnight in 10 ml YEP liquid medium containing 50 mg / ml Kan and 25 mg / ml Gen. Centrifuged at 6000 rpm for 10 min, the supernatant was removed, and the suspension was resuspended to OD = 0.6. The resuspended bacterial solution was injected into tobacco leaves. Two days later, the injected tobacco leaves were observed under a laser confocal microscope, and the excitation light of GFP was 488 nm.

[0134] Figure 3 The figure is a subcellular localization diagram of the GhPSKR1 protein described in the examples of the present invention, which shows that GhPSKR1 is localized on the cell membrane.

[0135] Example 2: Obtaining GhPSKR1 silenced plants using VIGS method and studying their phenotypes under salt stress

[0136] In this example, a VIGS-GhPSKR1 silencing vector was constructed by the VIGS method, and the VIGS-GhPSKR1 silencing vector was further used to obtain GhPSKR1 silenced plants, and their phenotypes under salt stress were studied.

[0137] 2.1 Construction of VIGS-GhPSKR1 silencing vector

[0138] (1) According to the steps in Example 1, total RNA was extracted from leaves of the cotton variety "Guoxin No. 3" and reverse transcribed into cDNA.

[0139] (2) Using the cDNA obtained in step (1) as a template, PCR amplification was performed using the following primer pairs to obtain a PCR amplification product.

[0140] The nucleotide sequence of primer F1 is shown in SEQ ID NO.5:

[0141] 5'-gtgagtaaggttacc gaattc TGTGGTCAGATGGATAGAGA-3'. The underline in the sequence is the restriction site of EcoR I.

[0142] The nucleotide sequence of primer R1 is shown in SEQ ID NO.6:

[0143] 5'-gagacgcgtgagctc ggtacc AAGAATGTTACTTGACTTTA-3'. The underline in the sequence is the restriction site of BamHI.

[0144] (3) Double digest the PCR amplification product obtained in step (2) with restriction endonucleases EcoR I and BamHI, and recover the digestion product.

[0145] (4) Double digest the pYL156 vector with restriction endonucleases EcoR I and BamHI to recover the vector backbone. The pYL156 vector is also called pTRV2:RNA2 vector, which is described in the non-patent document "Gao X, 2013, Functional algenomic analysis of cotton genes with agrobacterium-mediated virus-induced gene silencing."

[0146] (5) The enzyme digestion product of step (3) and the vector backbone of step (4) are connected to obtain the recombinant plasmid pYL156-GhPSKR1. The connection system is as described in the example.

[0147] (6) The recombinant vector pYL156-GhPSKR1 was sequenced and verified.

[0148] The sequencing results showed that the recombinant plasmid pYL156-GhPSKR1 was obtained by replacing the DNA fragment between the EcoR I and BamHI restriction sites of the pYL156 vector with a partial GhPSKR1 gene fragment shown in positions 1-300 of SEQ ID NO.1, while keeping the other sequences of the pYL156 vector unchanged, namely, the VIGS-GhPSKR1 silencing vector.

[0149] Figure 5 Figure (A) shows the results of identifying the gene silencing efficiency of the GhPSKR1 gene in cotton as described in the examples of the present invention.

[0150] Depend on Figure 5 It can be seen that compared with VIGS-GFP plants, the GhPSKR1 gene in VIGS-GhPSKR1 plants was indeed silenced.

[0151] 2.1 Obtaining VIGS-GhPSKR1 silenced plants

[0152] (1) pYL156-GhPSKR1, pYL156-GFP, pTRV-RNA1 and pYL156-GhCLA1 were transformed into Agrobacterium GV3101 by electroporation to obtain recombinant bacteria pYL156-GhPSKR1 / GV3101, recombinant bacteria pYL156-GFP / GV3101, recombinant bacteria pTRV1 / GV3101 and recombinant bacteria pYL156-GhCLA1 / GV3101.

[0153] Among them, pYL156-GhPSKR1 is the VIGS-GhPSKR1 silencing vector constructed in step 2.1 of this example.

[0154] pYL156-GFP, pTRV1 (pTRV-RNA1), and pYL156-GhCLA1 are all described in the non-patent document “Gao X, 2013, Functional genomic analysis of cotton genes with agrobacterium-mediatedvirus-induced gene silencing.”

[0155] (2) The recombinant bacteria were cultured in LB liquid medium containing antibiotics at 28°C on a shaking platform.

[0156] The formula composition of the LB liquid culture medium containing antibiotics is: 50 μg / mL kanamycin, 25 μg / mL gentamicin, 10 mM MES at pH 5.6, 20 μM acetosyringone, and the solvent is water.

[0157] (3) Resuspend the recombinant bacteria with VIGS solution and adjust the concentration of the bacterial solution to OD 600 =1.5.

[0158] The formula of VIGS solution is: 10 mM MES at pH 5.6, 10 mM MgCl2, 200 μM acetosyringone, and the solvent is water)

[0159] (4) The recombinant bacteria pYL156-GhPSKR1 / GV3101, pYL156-GFP / GV3101 and pYL156-GhCLA1 / GV3101 of specific concentrations were mixed with the bacterial solution of the recombinant bacteria pTRV1 / GV3101 at a volume ratio of 1:1 to obtain mixed solution 1, mixed solution 2 and mixed solution 3.

[0160] (5) Using a 1 mL needle-free syringe, the lower surfaces of the cotyledons of different cotton varieties "Xinshi No. 17" were filled with mixed solutions 1, 2, and 3, and cultured for 2 weeks to obtain VIGS-GhPSKR1 silenced plants, VIGS-GFP control plants, and VIGS-GhCLA1 indicator plants, respectively.

[0161] (6) About two weeks after the VIGS-GhCLA1 indicator plants injected with mixture 3 showed albino phenotype, RNA was extracted from the leaves of the plants injected with mixture 1 and mixture 2, and cDNA was reverse transcribed. The gene silencing efficiency was analyzed by fluorescent real-time quantitative PCR. The relative expression level was calculated using 2 -ΔΔCt Method calculation.

[0162] The primer pairs and PCR procedures used are as follows:

[0163] The nucleotide sequence of primer F2 is shown in SEQ ID NO.7:

[0164] 5′-AAGATGTCAACCTTGCCCG-3′.

[0165] The nucleotide sequence of primer R2 is shown in SEQ ID NO.8:

[0166] 5'-GGTGAGAACCAAAGCAGTTAGG-3'.

[0167] PCR program: denaturation at 94°C for 30 s; denaturation at 94°C for 5 s, annealing at 60°C for 35 s, 40 cycles.

[0168] Among them, the cotton Actin9 gene was used as a control, and the primer pairs used to identify the cotton Actin9 gene were as follows:

[0169] The nucleotide sequence of primer F3 is shown in SEQ ID NO.9:

[0170] 5′-GCCTTGGACTATGAGCAGGA-3′.

[0171] The nucleotide sequence of primer R3 is shown in SEQ ID NO.10:

[0172] 5′-AAGAGATGGCTGGAAGAGGA-3′.

[0173] Figure 5 Figure (A) is a diagram showing the results of identifying the gene silencing efficiency of the GhPSKR1 gene in cotton described in an embodiment of the present invention.

[0174] Figure 5 The results in Figure (A) show that the expression level of GhPSKR1 gene in the plants injected with mixed solution 1 was significantly lower than that in the plants injected with mixed solution 2, that is, VIGS-GhPSKR1 silenced plants with silenced GhPSKR1 gene and VIGS-GFP control plants were successfully obtained by the above method.

[0175] Among them, mixed solution 1 contains pYL156-GhPSKR1 / GV3101 and pTRV1 / GV3101 bacterial solutions.

[0176] Mixture 2 contains pYL156-GFP / GV3101 and pTRV1 / GV3101 bacterial solutions.

[0177] 2.3 Phenotypic study of VIGS-GhPSKR1 silenced plants under salt stress

[0178] (1) The VIGS-GhPSKR1 silenced plants and VIGS-GFP control plants obtained in step 2.2 of this example were subjected to 250 mM NaCl stress treatment, and the growth phenotype was observed.

[0179] The stress treatment method is as follows: when cotton grows to the three-leaf stage, VIGS-GhPSKR1 silenced plants and VIGS-GFP control plants are treated with 250 mM NaCl.

[0180] (2) The VIGS-GhPSKR1 silenced plants and VIGS-GFP control plants obtained in step 2. of this example were subjected to 250 mM NaCl stress treatment, and the leaves and roots of the plants were taken to detect the sodium ion content in the leaves and roots of the plants.

[0181] Figure 5 Figure (B) in the figure is a growth phenotype result diagram of the stress resistance verification of the GhPSKR1 gene silenced plants described in the examples of the present invention. The results show that under salt stress, the VIGS-GhPSKR1 silenced plants have more obvious growth inhibition, smaller leaves, and more significant salt damage characteristics such as wilting and curling at the leaf margins compared with the VIGS-GFP control plants, and are more sensitive to salt stress; in the control group without NaCl treatment, there is no difference in growth characteristics between the VIGS-GhPSKR1 silenced plants and the VIGS-GFP control plants.

[0182] Figure 6 The figure is a schematic diagram of the sodium ion content in the leaves and roots of the GhPSKR1 gene silenced plants described in the examples of the present invention. The results show that the sodium ion accumulation in the leaves and roots of the VIGS-GhPSKR1 silenced plants is increased compared with the VIGS-GFP control plants.

[0183] Based on the above results, the use of VIGS gene silencing technology in cotton significantly reduced the expression of the GhPSKR1 gene, and the gene silencing efficiency reached 73.9%; compared with the VIGS-GFP control plants, the growth inhibition of VIGS-GhPSKR1 silenced plants under salt stress was more obvious, the salt damage characteristics were more obvious, the sodium ion accumulation in the leaves and roots increased, and they were more sensitive to salt stress.

[0184] The above experimental results show that the GhPSKR1 gene plays a positive regulatory role in cotton salt stress, etc.

[0185] Example 3: Construction of GhPSKR1 overexpression vector, acquisition of overexpression transgenic plants and study of their phenotype under salt stress

[0186] 3.1 Construction of GhPSKR1 overexpression vector

[0187] (1) Using the GhPSKR1 gene as a template, PCR amplification was performed using a forward primer and a reverse primer to obtain a product containing the GhPSKR1 gene, and the amplification was performed as described in Example 1. The forward primer is the upstream primer F1, and the reverse primer is the downstream primer R1.

[0188] (2) The product containing the GhPSKR1 gene and the super1300-GFP vector were digested with Sma I and Kpn I, respectively, to obtain the digestion product and the vector frame, and then recovered them.

[0189] (3) The enzyme digestion product and the vector framework are connected to obtain a recombinant expression vector 35S::GhPSKR1-GFP recombinant vector, i.e., a GhPSKR1 overexpression vector. The connection reaction is shown in Example 1.

[0190] The recombinant expression vector 35S::GhPSKR1-GFP was obtained by replacing the DNA fragment between the Sma I and Kpn I restriction sites of super1300 with the GhPSKR1 gene of SEQ ID NO.1, while keeping the other sequences of super1300 unchanged. The structural schematic diagram of the recombinant expression vector 35S::GhPSKR1-GFP is shown in the figure Figure 3 shown.

[0191] 3.2 Obtaining GhPSKR1 overexpressing transgenic plants

[0192] (1) The recombinant expression vector 35S::GhPSKR1-GFP obtained in step 3.1 was introduced into the Agrobacterium strain GV3101 to obtain recombinant Agrobacterium.

[0193] (2) The recombinant Agrobacterium obtained in (1) was cultured in a shaking incubator at 28°C in LB liquid medium containing 50 μg / mL kanamycin and 25 μg / mL gentamicin for 24 h.

[0194] (3) Centrifuge at 4000 rpm for 10 min, collect the recombinant Agrobacterium and resuspend it with resuspension solution. Adjust the concentration of the resuspended bacterial solution to OD 600 =0.8. The formula of the resuspension solution is: 50 mM MES at pH 5.6, 5 wt% sucrose, and the solvent is water.

[0195] (4) Add 500 μl / L of silwetL-77 to the bacterial suspension, then use it to wet the Arabidopsis inflorescence that has not yet turned white. Wrap the Arabidopsis in a black plastic bag to maintain humidity, lay it flat, and culture it in the dark at 20°C for 24 h. Then remove the plastic bag, restore light, and culture the plants according to the conventional method until they are strong, and harvest mature T0 generation seeds.

[0196] Among them, silwetL-77 was purchased from: Beijing Coolaibo Technology Co., Ltd.; item number: CS9791.

[0197] (5) Using 1 / 2MS medium containing 50 mg / L hygromycin to culture T0 seeds and select positive plants from them. Among them, positive plants are characterized by healthy dark green true leaves and roots extending into the medium.

[0198] (6) The obtained positive plants were self-pollinated to obtain T1 generation seeds, and then the T1 generation seeds were cultured in 1 / 2MS medium containing 25 mg / L hygromycin and positive plants were selected from them. The screening standard was that the ratio of positive plants was greater than 3:1.

[0199] (7) The positive plants obtained in (6) are self-pollinated to obtain T2 seeds, and the T2 seeds are cultured in 1 / 2MS medium containing 25 mg / L hygromycin and positive plants are selected from them. The screening criterion is that all the plants in the strain are positive.

[0200] (8) Self-pollinating the positive plants obtained in (7) to obtain T3 seeds, cultivating the T3 seeds, and obtaining T3 transgenic GhPSKR1 Arabidopsis plants.

[0201] (9) When the T3 generation GhPSKR1-transfected Arabidopsis plants grew to the fourth week, leaf samples were taken with a hole puncher, the leaf tissue was ground with liquid nitrogen, and SDS plant total protein extract (250 mM Tris-HCl pH 6.8, 4% SDS, 40% glycerol, 0.1% bromophenol blue, 4% β-mercaptoethanol) was added. After mixing, the mixture was denatured at 95 degrees or above for 10 minutes, and the supernatant was taken as the denatured total protein for SDS-PAGE detection.

[0202] The formula composition of the SDS plant total protein extract is: 250mM Tris-HCl with pH 6.8, 4wt% SDS, 40wt% glycerol, 0.1wt% bromophenol blue, and 4wt% β-mercaptoethanol.

[0203] GhPSKR1 transgenic Arabidopsis plants are GhPSKR1 overexpressing transgenic Arabidopsis.

[0204] Figure 7 The Western Blot identification result of the homozygous transgenic Arabidopsis thaliana strain overexpressing GhPSKR1 described in the embodiment of the present invention is shown in Figure 1. The results show that the homozygous transgenic Arabidopsis thaliana strain OE1 and the homozygous transgenic Arabidopsis thaliana strain overexpressing GhPSKR1 OE2 were successfully obtained through this embodiment.

[0205] 3.3 Phenotypic study of hPSKR1 overexpressing transgenic plants under salt stress

[0206] (1) After vernalization at 4°C for 72 h, the seeds of the GhPSKR1 overexpressing transgenic Arabidopsis homozygous line OE1, the GhPSKR1 overexpressing transgenic Arabidopsis homozygous line OE2, and the wild-type Arabidopsis (WT) obtained in step 3.2 were moved into a 20°C greenhouse with 16 h light / 8 h dark and a light intensity of 60 μmol / m 2 / s, and cultured in a culture room with a humidity of 65±5% for 4 days.

[0207] (2) The Arabidopsis seedlings were then transferred to a culture medium containing 0 mM or 100 mM NaCl and continued to grow for 8 days. Their growth phenotypes were observed and the changes in their root length were statistically analyzed.

[0208] Figure 8 : is a graph comparing the growth conditions of the homozygous transgenic Arabidopsis thaliana strain overexpressing GhPSKR1 and the wild-type Arabidopsis thaliana under salt stress conditions described in the examples of the present invention; wherein, Figure 8 The plants from left to right in the figure are wild-type Arabidopsis (WT), GhPSKR1 overexpressing transgenic Arabidopsis homozygous line OE1, and GhPSKR1 overexpressing transgenic Arabidopsis homozygous line OE2.

[0209] Fig. 9 It is a schematic diagram of the statistical results of root length of the homozygous transgenic Arabidopsis thaliana strain overexpressing GhPSKR1 and the wild-type Arabidopsis thaliana under salt stress conditions described in the examples of the present invention.

[0210] The results showed that on the MS medium as the normal control group, there was no significant difference in root length between wild-type Arabidopsis (WT) and GhPSKR1 overexpression transgenic Arabidopsis homozygous lines OE1 and OE2. However, under salt stress treatment, the root length of GhPSKR1 overexpression transgenic Arabidopsis homozygous lines OE1 and OE2 was significantly elongated compared with wild-type Arabidopsis (WT). This shows that GhPSKR1 overexpression transgenic Arabidopsis homozygous lines OE1 and OE2 have good stress resistance. The GhPSKR1 gene plays a positive regulatory role in regulating plant salt stress. The GhPSKR1 gene can be used to improve plant salt tolerance.

[0211] The present invention cloned the cotton GhPSKR1 gene, constructed a transgenic Arabidopsis plant overexpressing GhPSKR1 by using transgenic technology, and constructed a GhPSKR1 silenced plant by using VIGS technology, verified the function of the GhPSKR1 gene, clarified that the GhPSKR1 gene can improve the salt tolerance of plants, facilitated a deeper study of the response mechanism of plants to salt stress signals, laid a good molecular foundation for effectively improving the salt tolerance of plants, and was of great value for exploring the signal regulation network of plants under salt stress.

[0212] The present invention discloses for the first time the positive regulatory role of the GhPSKR gene in cotton salt tolerance and provides excellent regulatory targets and theoretical guidance for cotton salt tolerance cultivation. The present invention provides an application of the GhPSKR1 gene in regulating plant salt tolerance and develops a new product application of the GhPSKR gene in cotton stress resistance.

[0213] It should be noted that when the present invention involves a numerical range, it should be understood that the two endpoints of each numerical range and any numerical value between the two endpoints can be selected. In order to avoid redundancy, the present invention describes a preferred embodiment.

[0214] Although preferred embodiments of the present invention have been described, those skilled in the art may make additional changes and modifications to these embodiments once the basic inventive concepts are known, and all such changes and modifications fall within the scope of the present invention.

Claims

1. Application of the GhPSKR1 gene in regulating plant salt tolerance, characterized in that: The nucleotide sequence of the GhPSKR1 gene is shown in SEQ ID NO.

1.

2. The use according to claim 1, characterized in that: The amino acid sequence of the protein expressed by the GhPSKR1 gene is shown in SEQ ID NO.

2.

3. The use according to claim 1, characterized in that: The GhPSKR1 gene in plants is overexpressed by genetic engineering methods to improve the salt tolerance of plants.

4. The use according to claim 3, characterized in that: The genetic engineering method comprises: constructing a recombinant expression vector using the GhPSKR1 gene, and introducing the recombinant expression vector into plants to improve the salt tolerance of the plants.

5. A recombinant expression vector containing the GhPSKR1 gene according to claim 1.

6. The recombinant expression vector according to claim 5, characterized in that The recombinant expression vector is obtained by inserting the GhPSKR1 gene into the Sma I and Kpn I restriction sites of the super1300-GFP vector.

7. A method for improving plant salt tolerance, characterized in that: The GhPSKR1 gene as claimed in claim 1 is overexpressed in a plant, or the activity of the protein expressed by the GhPSKR1 gene as claimed in claim 2 in a plant is increased, or the content of the protein expressed by the GhPSKR1 gene as claimed in claim 2 in a plant is increased, so as to improve the salt tolerance of the plant.

8. The method according to claim 7, characterized in that The GhPSKR1 gene is overexpressed in plants by transforming the plants into the GhPSKR1 gene through Agrobacterium-mediated method, so as to improve the salt tolerance of the plants.

9. The method according to claim 8, characterized in that The plants include cotton and / or Arabidopsis plants.

10. A method for breeding salt-tolerant plants, characterized in that: Overexpressing the GhPSKR1 gene as claimed in claim 1 in a plant to obtain a salt-tolerant transgenic plant; The plant is cotton.

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