KR20230036814A - Genes Related to drought stress and Salt Stress Resistances and Transformed Plants with the Same - Google Patents

Abstract

The present invention relates to genes related to salt stress resistance of plants and uses thereof, and by regulating the expression of a BSP1 protein involved in plant growth, it is possible to allow the plants to significantly grow without dying under dry and high salt stress conditions. Therefore, as the transformed plants in which the BSP1 protein coding gene has been removed show excellent growth ability even under the dry or salt stress conditions, the effect of increasing yield can be derived in an environment where crops are difficult to grow.

Description

Genes Related to drought stress and Salt Stress Resistances and Transformed Plants with the Same}

The present invention relates to genes related to salt stress resistance of plants and their uses, and more particularly, to compositions and traits capable of enhancing resistance to salt stress by inducing deletion of a gene encoding BSP1 to suppress protein expression. It is to provide a conversion plant and a method for producing the same.

When plants are exposed to high salt stress, osmotic pressure imbalance and intracellular ion concentration imbalance occur due to the accumulation of intracellular Na+ ions, resulting in poor development and photosynthetic disorders. Plants, as sessile organisms, build complex defense systems in response to these external stresses. Representatively, there are an ion influx control into the cytoplasm (influx system), a control to pump ions out of the cell (efflux system), and a system that stores ions in the vacuole.

The SOS signal transduction system is the most representative of the ion efflux system of plants. Proteins involved in the SOS signaling system include SOS1, SOS2, and SOS3, SOS1 is a membrane protein and has Na + / H + antiporter activity, SOS2 is a serine / threonine protein kinase, SOS3 is EF-hand-type-calcium binding known to be a protein. When Arabidopsis thaliana is placed under high salt stress conditions, the concentration of Ca ²⁺ in the cells increases, and the increased Ca ²⁺ binds to the SOS3 protein in the cell membrane, and then binds to the SOS2 protein present in the cytoplasm. After forming a complex, the complex phosphorylates and activates SOS1, a Na + / H + antiporter, so that Na ⁺ efflux occurs.

Ubiquitin is a small 8-kDa protein present in all eukaryotes that attaches to a target protein through a series of processes of E1 (Ub-activating enzyme), E2 (Ub-conjugating enzyme), and E3 (Ub-ligase). is known to be Depending on the number of ubiquitin attached, it is monoubiquitination or polyubiquitination, and in the case of polyubiquitination, it is degraded by the 26S proteasome complex. Through this process, plants can selectively degrade useless or harmful proteins to respond to various stresses. Arabidopsis thaliana is known that about 5% of all genes are related to ubiquitination, and among them, there are two E1 and at least 37 E2, and in the case of E3 Although more than 1400 are known to exist, only a few enzymes have been reported to be involved in salt stress.

Salinity stress has a direct effect on crop productivity in the widest range among environmental stresses, and has been pointed out as a major stress that reduces crop production. Therefore, it is most urgent to secure stable food resources by identifying new genes through understanding and consideration of these stress signal transduction mechanisms and developing salt stress-resistant plants through their regulation.

The present inventors, while researching to develop plants with excellent growth under dry stress or salt stress conditions, confirmed that the transgenic plants in which the BSP1 protein coding gene was removed showed excellent growth even under dry stress and salt stress conditions, thereby achieving the present invention. completed.

Having been made in view of the above problems, an object of the present invention is to provide a composition capable of enhancing resistance to drying stress and salt stress.

In addition, another object of the present invention is to provide a transgenic plant with improved dryness and salt stress resistance.

Another object of the present invention is to provide a method for producing transgenic plants with improved resistance to dryness and salt stress.

In order to achieve the above object, the present invention provides a composition for improving drying and salt stress resistance of plants, comprising a nucleic acid molecule that inhibits the expression of a BSP1 (Binder of SOS1 Protein 1) protein-encoding gene.

The BSP1 protein may be represented by the amino acid sequence of SEQ ID NO: 1.

The BSP1 protein coding gene may be represented by SEQ ID NO: 2.

The nucleic acid molecule may be any one or more selected from the group consisting of T-DNA, siRNA, shRNA, miRNA, ribozyme, antisense nucleic acid sequence and transposon.

The plant is rice, wheat, barley, corn, soybean, potato, wheat, red bean, oat, sorghum, Arabidopsis, cabbage, radish, pepper, strawberry, tomato, watermelon, cucumber, cabbage, melon, pumpkin, green onion, onion, Carrot, ginseng, tobacco, cotton, sesame, sugarcane, sugar beet, perilla, peanut, rapeseed, apple tree, pear tree, jujube tree, peach, lamb, grape, tangerine, persimmon, plum, apricot, banana, rose, gladiolus, It may be any one selected from the group consisting of gerbera, carnation, chrysanthemum, lily, tulip, ryegrass, red clover, orchard grass, alfalfa, tall fescue and perennial ryegrass.

In order to achieve the above other object, the present invention is a dry and salt stress in which the expression of the BSP1 (Binder of SOS1 Protein 1) protein coding gene is suppressed or the BSP1 (Binder of SOS1 Protein 1) protein coding gene is knocked out. Transgenic plants with enhanced resistance are provided.

The BSP1 protein coding gene may be represented by SEQ ID NO: 2.

The plant is rice, wheat, barley, corn, soybean, potato, wheat, red bean, oat, sorghum, Arabidopsis, cabbage, radish, pepper, strawberry, tomato, watermelon, cucumber, cabbage, melon, pumpkin, green onion, onion, Carrot, ginseng, tobacco, cotton, sesame, sugarcane, sugar beet, perilla, peanut, rapeseed, apple tree, pear tree, jujube tree, peach, lamb, grape, tangerine, persimmon, plum, apricot, banana, rose, gladiolus, It may be any one selected from the group consisting of gerbera, carnation, chrysanthemum, lily, tulip, ryegrass, red clover, orchard grass, alfalfa, tall fescue and perennial ryegrass.

The present invention, in order to achieve the above object, (a) suppressing the expression of the BSP1 protein-encoding gene in the plant or knocking out the BSP1 protein-encoding gene; A method for producing a converted plant is provided.

The inhibition of expression of the BSP1 protein coding gene or the knock-out of the BSP1 protein coding gene is selected from the group consisting of T-DNA, siRNA, shRNA, miRNA, ribozyme, antisense nucleic acid sequence and transposon for the BSP1 protein coding gene Any one or more of them may be used.

The BSP1 protein coding gene may be represented by SEQ ID NO: 2.

According to the present invention, in order to increase the resistance of plants to dryness and salt stress, as a transgenic plant inhibiting the expression of BSP1 protein and a method for producing the same, by controlling the expression of the BSP1 protein involved in plant growth, drying and high salt stress It can be made to grow vigorously without dying under the conditions. Therefore, as the transgenic plants in which the BSP1 protein coding gene has been removed show excellent growth ability even in dry or salt stress conditions, the effect of increasing yield can be derived in an environment where crops are difficult to grow.

Figure 1a shows T-DNA insertion into BSP1 (At3g52030) mutant alleles.
Figure 1b is a photograph taken to compare the degree of cotyledon greening of wild-type plants (Col-0), bsp1-1 mutant, bsp1-2 mutant and hos15-2 mutant under 0 or 125 mM NaCL conditions.
1C is a graph showing the cotyledon greening rate (% Cotyledon greening) of wild-type plants (Col-0), bsp1-1 mutant, bsp1-2 mutant, and hos15-2 mutant.
1d shows seedlings grown from wild-type plants (Col-0), bsp1-1 mutants, bsp1-2 mutants, and sos2-2 mutants under 0 or 125 mM NaCL conditions, and then transplanted into soil, and 300 mM NaCl aqueous solution was added to 4 These are photos taken on days 1, 8, and 12, and grown for 14 days.
Figure 2 confirms the resistance of wild-type plants (Col-0), bsp1-1 mutant, bsp1-2 mutant, hos15-2 mutant and ost1-3 mutant under dry stress conditions, before drought stress , This is the result of measuring the survival rate of each group after 14 days of no watering (14-days drought) and 48-hrs of rewatering (48-hrs of rewatering).
Figure 3a is a result of Western blotting (western blotting) of proteins separated by immunoprecipitation using a GFP antibody after expressing BSP1-GFP or ASK1-HA with an HA antibody.
Figure 3b is the result of analyzing tobacco variants expressing SGT1a, SOS1, RAR1 or BSP1 by Split Luciferase assay.

The present outbreak is described in more detail below.

One aspect of the present invention relates to a composition for improving drying and salt stress resistance of a plant, comprising a nucleic acid molecule that inhibits the expression of BSP1 (Binder of SOS1 Protein 1) protein.

The BSP1 (Binder of SOS1 Protein 1) protein of the present invention can be represented by the amino acid sequence of SEQ ID NO: 2, and among the entire sequence, the F-BOX domain of amino acid residues 20 to 66 and amino acid residues 107-144, 213-246 , 249-287, 290-331, 334-374 of the WD-40 repeat sequence. Although not limited thereto, in an experimental example of the present invention, resistance to drying and salt stress of the BSP1 coding gene was analyzed. In order to fully elucidate the functions of the above genes, mutants for the bsp1-1 and bsp1-2 genes were prepared to confirm that they have resistance to dryness and salt stress, as well as that the F-BOX domain of the BSP1 protein is cullin 1-based By confirming that the BSP1 protein interacts with SOS1 and interacts as a substrate for E3 ligase, it was confirmed that inhibition of BSP1 functions as a negative regulator of dry and salt stress signaling in plants.

In the present invention, the gene encoding the BSP1 protein refers to a gene including a sequence encoding the BSP1 protein, and includes both genomic DNA or cDNA encoding the BSP1 protein. Preferably, it may be the Arabidopsis-derived At3g52030 gene (GenBank accession no: At3g52030), and more preferably, it may be the one represented by the nucleotide sequence represented by SEQ ID NO: 2. In addition, homologues of the above nucleotide sequences are included within the scope of the present invention. Specifically, the gene is a nucleotide sequence having 70% or more, more preferably 80% or more, still more preferably 90% or more, and most preferably 95% or more sequence homology with the nucleotide sequence of SEQ ID NO: 2, respectively. can include The "percentage of sequence homology" for polynucleotides is determined by comparing two optimally aligned sequences with a comparison region, wherein a portion of the polynucleotide sequence in the comparison region is a reference sequence (addition or deletion) for the optimal alignment of the two sequences. may include additions or deletions (i.e., gaps) compared to (not including).

In the present invention, the term "salt stress resistance" is a property that a plant endures under a high salt concentration condition, and means the resistance of a plant at a high salt concentration. Salt stress tolerance is the maximum salt concentration that crops can tolerate without losing productivity, and higher salt concentrations have a negative effect on crop productivity. The salt concentration generally refers to soil salinity or salinity of irrigation water.

In the present invention, the term "dry stress resistance" is a property that plants endure under dry conditions, and means resistance of plants in drought. When a prolonged drought occurs, varieties with strong stress resistance are required, and the dry stress resistance of crops means that the decline in plant growth or productivity is suppressed or delayed under dry stress conditions.

In the present invention, the BSP1 protein is not limited to the protein consisting of the amino acid sequence represented by SEQ ID NO: 1 described in the attached sequence listing, and may be functionally equivalent to the protein having the amino acid sequence of SEQ ID NO: 1. The term “functional equivalent” refers to one having at least 60%, specifically 70%, more specifically 80% or more sequence homology with the amino acid sequence represented by SEQ ID NO: 1 as a result of addition, substitution or deletion of amino acids. It means a protein exhibiting substantially the same activity as BSP1 of the present invention.

The functional equivalent includes, for example, an amino acid sequence variant in which some of the amino acids of the amino acid sequence of SEQ ID NO: 1 are substituted, deleted, or added. Substitutions of amino acids are preferably conservative substitutions. Examples of conservative substitutions of naturally occurring amino acids are; Aliphatic amino acids (Gly, Ala, Pro), hydrophobic amino acids (Ile, Leu, Val), aromatic amino acids (Phe, Tyr, Trp), acidic amino acids (Asp, Glu), basic amino acids (His, Lys, Arg, Gln, Asn ) and sulfur-containing amino acids (Cys, Met). Deletion of amino acids is preferably located in a region not directly involved in the activity of BSP1 of the present invention. In addition, the scope of the functional equivalent includes protein derivatives in which a part of the chemical structure of the protein is modified while maintaining the basic skeleton of BSP1 and its physiological activity. For example, fusion proteins made by fusion with other proteins such as GFP (Green Fluorescent Protein) while maintaining structural changes and physiological activity to change the stability, storage stability, volatility or solubility of the protein of the present invention are included. .

The nucleic acid molecule that inhibits the expression of BSP1 of the present invention may be one or more selected from the group consisting of T-DNA, siRNA, shRNA, miRNA, ribozyme, peptide nucleic acids (PNA), antisense nucleic acid sequences, and transposons. And, it may be an RNAi vector containing them, but is not limited thereto. The RNAi vector may include any vector commonly used in the art, and may be, for example, a pANDA-β RNAi vector (Miki and Shimamoto, 2004).

In the present specification, the term "siRNA" refers to a short double-stranded RNA capable of inducing RNAi (RNA interference) through cleavage of a specific mRNA. It consists of a sense RNA strand having a sequence homologous to the mRNA of the target gene and an antisense RNA strand having a sequence complementary thereto. Since siRNA can suppress the expression of a target gene, it is provided as an efficient gene knockdown method or as a method of gene therapy.

The siRNA is not limited to complete pairing of double-stranded RNA parts that pair with each other, but is paired by mismatch (corresponding bases are not complementary), bulge (there is no base corresponding to one chain), etc. Unfinished parts may be included. The total length is 10 to 100 bases, preferably 15 to 80 bases, and most preferably 20 to 70 bases. The siRNA end structure can be either a blunt end or a cohesive end structure, as long as the expression of the target gene can be suppressed by the RNAi effect. As for the sticky end structure, both a structure with 3 ends protruding and a structure with 5 ends protruding are possible. The number of protruding bases is not limited. For example, the number of bases may be 1 to 8 bases, preferably 2 to 6 bases. In addition, siRNA is a small molecule RNA (e.g., natural RNA molecules such as tRNA, rRNA, viral RNA or artificial RNA) attached to the protruding part of one end to the extent that the expression suppression effect of the target gene can be maintained. molecule) may be included. The siRNA terminal structure does not have to have a truncated structure on both sides, and may have a stem loop structure in which one end portion of the double-stranded RNA is connected by a linker RNA. The length of the linker is not particularly limited as long as it does not interfere with forming a pair of stem portions.

In the present specification, the term "short hairpin RNA (shRNA)" refers to a single strand composed of 50-70 nucleotides, and forms a stem-loop structure in vivo. Long RNAs of 19-29 nucleotides complementary to both sides of the loop of 5-10 nucleotides are base-paired to form a double-stranded stem.

As used herein, the term "miRNA (microRNA)" regulates gene expression, and is 20-50 nucleotides in length, preferably 20-45 nucleotides, more preferably 20-40 nucleotides, and even more preferably 20 nucleotides in length. - means a single-stranded RNA molecule composed of 30 nucleotides, most preferably 21-23 nucleotides. miRNA is an oligonucleotide that is not expressed intracellularly and has a short stem-loop structure. miRNAs have full or partial homology with one or two or more messenger RNAs (mRNAs), and suppress target gene expression through complementary binding with the mRNAs.

As used herein, the term "ribozyme" is a type of RNA and refers to an RNA molecule having a function such as an enzyme that recognizes a specific RNA base sequence and cuts it itself. A ribozyme consists of a region that binds with specificity to a complementary nucleotide sequence of a target messenger RNA strand and a region that cleaves the target RNA.

As used herein, the term "peptide nucleic acid (PNA)" refers to a molecule that has properties of both nucleic acid and protein, and is complementary to DNA or RNA. PNA is a DNA-like nucleobase linked by peptide bonds and was first reported in 1999 (Nielsen PE, Egholm M, Berg RH, Buchardt O, "Sequenceselective recognition of DNA by strand displacement with a thymine-substituted polyamide ", Science 1991, Vol. 254: pp1497-1500]). PNA is not found in nature and is artificially synthesized through chemical methods.

PNA hybridizes with natural nucleic acids of complementary nucleotide sequence to form double strands. PNA/DNA duplexes are more stable than DNA/DNA duplexes, and PNA/RNA duplexes are more stable than DNA/RNA duplexes when they are the same length. As the peptide backbone, N-(2-aminoethyl)glycine repeatedly linked by amide bonds is most commonly used. In this case, the backbone of the peptide nucleic acid has an electrical is neutral with The spatial size and distance between the four nucleotide bases present in PNA are almost the same as those of natural nucleic acids. PNA is not only chemically more stable than natural nucleic acids, but also biologically stable because it is not degraded by nucleases or proteases.

As used herein, the term "antisense nucleic acid sequence" refers to DNA or RNA or derivatives thereof containing a nucleotide sequence complementary to a specific mRNA sequence, and binds to a complementary sequence in mRNA to inhibit translation of mRNA into protein. If there is a feature to do, it is not particularly limited thereto. The antisense nucleic acid sequence of the present invention refers to a DNA or RNA sequence that is complementary to mRNA of a target gene and capable of binding to mRNA of a target gene, and is capable of performing translation, cytoplasmic translocation (trans cation), and maturation of the mRNA of the target gene. ) or other essential activities for overall biological function. The length of the antisense nucleic acid sequence is 6 to 100 bases, preferably 10 to 40 bases, and is also referred to as an antisense oligonucleotide.

The antisense nucleic acid sequence can be modified at one or more bases, sugars or backbone positions to enhance efficacy (De Mesmaeker et al., Curr Opin Struct Biol., 5(3):343-55, 1995 ). The oligonucleotide backbone can be modified with phosphorothioates, phosphotriesters, methyl phosphonates, short-chain alkyls, cycloalkyls, short-chain heteroatomic, heterocyclic intersugar linkages, and the like. In addition, antisense nucleic acids may contain one or more substituted sugar moieties. Antisense oligonucleotides can include modified bases. Modified bases include hypoxanthine, 6-methyladenine, 5-methyl pyrimidine (especially 5-methylcytosine), 5-hydroxymethylcytosine (HMC), glycosyl HMC, gentobiosyl HMC, 2-aminoadenine, 2 -Thiouracil, 2-thiothymine, 5-bromouracil, 5-hydroxymethyluracil, 8-azaguanine, 7-deazaguanine, N6 (6-aminohexyl)adenine, 2,6-diaminopurine, etc. there is

As used herein, the term "T-DNA" refers to a DNA segment transferred to the nucleus of a host plant cell as transfer DNA in a tumor-inducing (Ti) plasmid of Agrobacterium species. T-DNA has a repeating sequence of 25 bp at both ends, and transfer starts at the left border and ends at the right border. Bacterial T-DNA is about 20,000 bp in length and is used to cause insertional mutagenesis by disrupting the target gene by their insertion, and the inserted T-DNA sequence not only mutates but also marks the target gene. do.

In the present invention, the dry and salt stress resistance enhancing effect is characterized by enhancing the resistance to dry and salt stress conditions in the plant by suppressing the action of the BSP1 protein or its coding gene involved in the growth process of the target plant.

According to the present invention, when the expression of the gene encoding BSP1 (Binder of SOS1 Protein 1) was suppressed, it was confirmed that the growth and germination and survival rate of plants increased under high salinity conditions or drought conditions.

Specifically, the present invention prepared a transgenic Arabidopsis thaliana in which the BSP1 protein-coding gene was knocked out using wild-type Arabidopsis thaliana and T-DNA, and analyzed the phenotypes for each of them under dry stress conditions and salt stress conditions. As a result, it was found that the growth and survival of the transgenic Arabidopsis thaliana in which the BSP1 protein-encoding gene was knocked out was superior to that of wild-type Arabidopsis thaliana under high salinity and dry conditions. These results mean that the transgenic plants in which the BSP1 protein-encoding gene is knocked out have excellent resistance to external environmental stress, and in particular, growth and growth can proceed normally under salt stress and dry stress.

In addition, the present inventors have identified for the first time that the BSP1 protein coding gene has a function of negatively regulating the SOS1 gene related to salt stress in plants and is degraded as a substrate of Cullin-1 based E3 ligase. That is, it can be seen that it has a function of regulating the system for leaching ions out of the cell.

The plant is not particularly limited, rice, barley, wheat, corn, soybean, potato, rice crops, such as oats, sorghum; vegetable crops including Chinese cabbage, radish, pepper, strawberry, tomato, watermelon, cucumber, cabbage, melon, pumpkin, green onion, onion and carrot; Specialty crops including ginseng, tobacco, cotton, sesame, sugar cane, sugar beet, perilla, peanut and rapeseed; fruit trees including apple trees, pear trees, jujube trees, peaches, kiwi trees, grapes, tangerines, persimmons, plums, apricots and bananas; flowers including roses, gladiolus, gerberas, carnations, chrysanthemums, lilies and tulips; And it may be selected from the group consisting of feed crops including ryegrass, red clover, orchard grass, alpha alpha, tall fescue and perennial ryegrass, but is not limited thereto, and if the plant has improved resistance to drying and salt stress Can be used without limitation. Preferably, the plant is Arabidopsis, soybean, tobacco, eggplant, red pepper, potato, tomato, Chinese cabbage, radish, cabbage, lettuce, peach, pear, strawberry, watermelon, melon, cucumber, carrot, celery and rapeseed dicotyledons It may be a plant, most preferably Arabidopsis thaliana.

Another aspect of the present invention is that the expression of the BSP1 (Binder of SOS1 Protein 1) protein coding gene is suppressed or the BSP1 (Binder of SOS1 Protein 1) protein coding gene is knocked out (knock-out) dry and salt stress resistance is improved It relates to transgenic plants. Preferably, it is a transgenic plant in which the BSP1 protein coding gene having the nucleotide sequence represented by SEQ ID NO: 2 is knocked out, and resistance to dryness and salt stress is enhanced. In a specific example of the present invention, it was confirmed that BSP1 mutant plants were prepared and resistance significantly increased compared to wild-type plants under dry and salt stress.

The plant of the present invention is a plant with improved resistance to dryness and salt stress, and is transformed with the composition of the present invention. In the present invention, the "plant body" is interpreted as a concept including not only mature plants, but also plant cells, plant tissues, and plant seeds capable of growing into mature plants.

The plant is not particularly limited, rice, barley, wheat, corn, soybean, potato, rice crops, such as oats, sorghum; vegetable crops including Chinese cabbage, radish, pepper, strawberry, tomato, watermelon, cucumber, cabbage, melon, pumpkin, green onion, onion and carrot; Specialty crops including ginseng, tobacco, cotton, sesame, sugar cane, sugar beet, perilla, peanut and rapeseed; fruit trees including apple trees, pear trees, jujube trees, peaches, kiwi trees, grapes, tangerines, persimmons, plums, apricots and bananas; flowers including roses, gladiolus, gerberas, carnations, chrysanthemums, lilies and tulips; And it may be selected from the group consisting of feed crops including ryegrass, red clover, orchard grass, alpha alpha, tall fescue and perennial ryegrass, but is not limited thereto, and if the plant has improved resistance to drying and salt stress Can be used without limitation. Preferably, the plant is Arabidopsis, soybean, tobacco, eggplant, red pepper, potato, tomato, Chinese cabbage, radish, cabbage, lettuce, peach, pear, strawberry, watermelon, melon, cucumber, carrot, celery and rapeseed dicotyledons It may be a plant, most preferably Arabidopsis thaliana.

In addition, the present invention may provide seeds of the transgenic plants. Preferably, the seed may be a seed of a dicotyledonous plant, most preferably a seed of Arabidopsis.

Another aspect of the present invention is, (a) inhibiting the expression of the BSP1 protein-encoding gene in the plant or knocking out the BSP1 protein-encoding gene; Transformation with enhanced resistance to dryness and salt stress comprising It provides a method for producing plants.

The method of suppressing the expression of the BSP1 protein-encoding gene or knocking out the BSP1 protein-encoding gene can be performed using any method known in the art, for example, the target nucleic acid sequence is a spacer (non-spacer). -coding DNA) and cloned as an inverted repeat (inverted repeat) can be achieved by the introduction and expression of the gene construct into the plant, more detailed information Grierson et al. (1998) WO 98/53083; See Waterhouse et al. (1999) WO 99/53050. Also, any of the known "gene silencing" methods can be used to achieve the same effect. Examples of such methods for reducing endogenous gene expression include short interfering RNA (siRNA), which is RNA-mediated gene silencing of gene expression, nucleic acid sequences in the sense direction of a nucleic acid sequence of a target gene, or antisense ( Introduction of antisense directional nucleic acid sequences into plants, ribozymes, insertional mutagenesis (eg, T-DNA insertion or transposon insertion), miRNA (microRNA), etc., but are not limited thereto. . In one embodiment of the present invention, a T-DNA insertion method was used.

The methods described above are examples of various methods for the reduction or substantial elimination of the expression of a specific gene in a plant of an endogenous gene, and one skilled in the art can use suitable methods, for example, for silencing to reduce the expression of an endogenous gene in a whole plant or part thereof. Modifications to the above-mentioned methods can be easily achieved using promoters, and such methods are known in the art.

In addition, the transgenic plant of the present invention can be produced through the introduction of a target gene into a plant, which can be achieved using a vector containing the target gene, and the method of delivering the vector of the present invention into a plant host cell is known in the industry. For example, gene gun-mediated transformation method (bombardment), Agrobacterium-mediated transformation method, microinjection method, calcium phosphate precipitation method, electroporation method, liposome-mediated transfection method, DEAE-dextran treatment method, PEG treatment methods, etc., but are not limited thereto.

The vector is also referred to as a recombinant vector, and the recombinant vector is a gene cloned as an inverted repeat of a BSP1 protein-coding gene nucleic acid sequence capable of suppressing or reducing the expression of a BSP1 protein-coding gene, a short interfering RNA (siRNA) gene , a nucleic acid sequence in the sense direction or an antisense direction of the nucleic acid sequence of the target gene, a ribozyme coding gene, a mutagenesis gene (eg, T-DNA or transposon) or a miRNA (microRNA) gene may be an inserted vector, but is not limited thereto. Methods for inhibiting gene expression using a recombinant vector containing a gene sequence capable of inhibiting or reducing the expression of a target gene are known in the art, examples of which are as described above.

In the present invention, "recombinant" refers to a cell that replicates a heterologous nucleic acid, expresses the nucleic acid, or expresses a peptide, a protein encoded by a heterologous peptide or a heterologous nucleic acid. Recombinant cells can express genes or gene segments not found in the cell's native form, either in sense or antisense form. A recombinant cell may also express a gene found in the cell in its natural state, but the gene has been reintroduced into the cell by artificial means as a modified one.

In the present invention, a gene sequence capable of suppressing or reducing the BSP1 protein-encoding gene can be inserted into a recombinant vector. The term "recombinant vector" refers to a bacterial plasmid, phage, yeast plasmid, plant cell virus, mammalian cell virus, or other vector. In general, any plasmid and vector may be used provided that it is capable of replicating and stabilizing in the host. An important characteristic of the expression vector is that it has an origin of replication, a promoter, a marker gene and a translation control element.

An expression vector comprising a gene sequence capable of inhibiting or reducing the expression of the gene encoding the BSP1 protein of the present invention and appropriate transcriptional/translational control signals can be constructed by methods well known to those skilled in the art. The methods include in vitro recombinant DNA technology, DNA synthesis technology and in vivo recombinant technology, and the like. The DNA sequence can be effectively linked to a suitable promoter in an expression vector to direct mRNA synthesis. In addition, the expression vector may include a ribosome binding site and a transcription terminator as a translation initiation site.

A preferred example of the recombinant vector of the present invention is the Ti-plasmid vector, which is capable of transferring a part of itself, the so-called T-region, into plant cells when present in a suitable host such as Agrobacterium tumefaciens. Another type of Ti-plasmid vector (see EP 0 116 718 B1) is currently used to transfer hybrid DNA sequences into plant cells, or protoplasts, from which new plants can be produced that properly integrate the hybrid DNA into the plant's genome. there is. A preferred form of the Ti-plasmid vector is a binary vector. Other suitable vectors that can be used to introduce DNA according to the present invention into plant hosts include viral vectors, such as those that can be derived from double-stranded plant viruses (eg, CaMV) and single-stranded viruses, gemini viruses, and the like. For example, it may be selected from incomplete plant viral vectors. The use of such vectors can be particularly advantageous when properly transforming a plant host is difficult.

Expression vectors will preferably include one or more selectable markers. The marker is a nucleic acid sequence having a characteristic that can be selected by a conventional chemical method, and includes all genes capable of distinguishing transformed cells from non-transformed cells. Examples include herbicide resistance genes such as glyphosate or phosphinothricin, antibiotics such as kanamycin, G418, bleomycin, hygromycin, and chloramphenicol. Resistance genes, aadA genes, etc., but are not limited thereto.

In the recombinant vector of the present invention, the promoter may be CaMV 35S, actin, ubiquitin, pEMU, MAS, histone promoter, or Clp promoter, but is not limited thereto. The term "promoter" refers to a region of DNA upstream from a structural gene and refers to a DNA molecule to which RNA polymerase binds to initiate transcription. A "plant promoter" is a promoter capable of initiating transcription in a plant cell. A "constitutive promoter" is a promoter that is active under most environmental conditions and states of development or cell differentiation. Constitutive promoters may be preferred in the present invention because selection of transformants may be made by various tissues at various stages. Thus, constitutive promoters do not limit selection possibilities.

In the recombinant vector of the present invention, common terminators can be used, such as nopaline synthase (NOS), rice α-amylase RAmy1 A terminator, phaseoline terminator, Agrobacterium tumefaciens ), the terminator of the Octopine gene, and the rrnB1/B2 terminator of Escherichia coli, but are not limited thereto. Regarding the need for terminators, it is generally known that such regions increase the certainty and efficiency of transcription in plant cells. Therefore, the use of terminators is highly preferred in the context of the present invention.

In the case of transforming plant cells with the recombinant vector of the present invention, the transformation may be mediated by, for example, Agrobacterium tumefiaciens.

Methods for delivering the recombinant vector of the present invention into host cells include gene gun-mediated transformation method (bombardment), Agrobacterium-mediated transformation method, microinjection method, calcium phosphate precipitation method, electroporation method, liposome-mediated transfection method. The vector can be injected into the host cell by the method, DEAE-dextran treatment method, PEG treatment method, or the like.

It may include regenerating a transgenic plant from the transformed plant cell. Any method known in the art may be used as a method for regenerating a transgenic plant from the transgenic plant cell.

The transformed plant cells must be regenerated into whole plants. Techniques for regenerating mature plants from callus or protoplast culture can be used without particular limitation as long as they are well known in the art for a number of different species.

As described above, suppression of BSP1 protein-coding gene expression or knockout of BSP1 protein-coding gene in plants identified in the present invention can effectively enhance the growth and growth of plants or crops even under dry and salt stress conditions, and conventional plants It can be confirmed that the negative regulator function of SOS1, which plays an important role in relation to the system for leaching ions out of the cell, is also effectively performed.

Hereinafter, the present invention will be described in more detail with reference to preferred embodiments. However, these examples are intended to explain the present invention in more detail, and it will be apparent to those skilled in the art that the scope of the present invention is not limited thereto.

Examples 1 and 2. Securing mutant plants

T-DNA introduced mutant plants of Arabidopsis thaliana BSP1 (At3g52030) were obtained. Seeds bsp1-1 mutant (SALK_064279) (Example 1) and bsp1-2 mutant (GK-221-F01.01) (Example 2) of the allele with T-DNA inserted into the BSP1 gene were prepared in Arabidopsis. It was obtained from the Center for Biological Resources (ABRC) (FIG. 1A).

The loss-of-function of the BSP1 gene was confirmed by genotyping PCR for the two mutants. As a result, it was confirmed that the T-DNA was introduced into the BSP1 gene and knocked out plants, and it was confirmed that the bsp1-1 mutant and the bsp1-2 mutant had T-DNA located in the first and ninth exons, respectively (Fig. 1a ).

Comparative Example 1. Securing hos15-2 plants

The T-DNA insertion transformation mutant hos15-2 (GK_785B10) was purchased from the Arabidopsis Bioresource Center (ABRC). Then, a homozygous line was selected through genotyping and a line was constructed.

Comparative Example 2. securing sos2-2 plants

The T-DNA insertion transformation mutant sos2-2 (CS6528) was purchased from the Arabidopsis Biological Resource Center (ABRC). Then, a homozygous line was selected through genotyping and a line was constructed.

Comparative Example 3. Obtaining ost1-3 plants

The T-DNA insertion transformation mutant ost1-3 (SALK_008068) was purchased from the Arabidopsis Bioresource Center (ABRC). Then, a homozygous line was selected through genotyping and a line was constructed.

Experimental Example 1. Isolation and identification of high salt resistant bsp1 mutants

48 bsp1-1 mutants of Example 1 and 48 bsp1-2 mutants of Example 2 were obtained, and the seeds were surface sterilized, and then MS medium (1/2 Murashige and Skoog (MS) salt, 1.5% sugar and 0.6% agar, pH 5.7), and seedlings grown for 3 weeks were transferred to soil and samples were collected 14 days after planting (MS).

High salt (125 mM NaCl) was seeded in MS medium (1/2 Murashige and Skoog (MS) salt, 1.5% sugar and 0.6% agar, pH 5.7) containing 125 mM NaCl, respectively, at 22 ° C for 16 hours, They were grown in a plant growth chamber with an 8-hour dark cycle. The seedlings grown for 3 weeks were transplanted into the soil and grown by giving a 300 mM NaCl solution on the 4th, 8th and 12th days, and samples were taken on the 14th day to analyze the phenotype.

The degree of greening was recorded by distinguishing normal greening (including dark green cotyledons) and damaged greening (including white, yellow or light green cotyledons) from 48 seeds in each group before transplanting into soil, and normal seeds cultured in MS medium. The greening rate was calculated as a percentage of the cotyledon of the plant and presented in the graph (% Cotyledon greening). Data are presented as mean ± standard error of three technical replicates.

Figure 1b is a photograph measured to compare the degree of cotyledon greening of a wild-type plant (Col-0), a bsp1-1 mutant, a bsp1-2 mutant, and a hos15-2 mutant under 0 or 125 mM NaCL conditions, and Figure 1c is a wild-type It is a graph showing the cotyledon greening rate (% Cotyledon greening) of plants (Col-0), bsp1-1 mutant, bsp1-2 mutant and hos15-2 mutant. hos15-2 was used as a high salt sensitive control.

1b and 1c, all bsp1 allelic mutants (bsp1-1 mutant, bsp1-2 mutant) in the wild-type plant (Col-0) were better than the wild-type plant (Col-0) and the hos15-2 mutant under high salt conditions. It can be confirmed that the germination rate and growth rate are significantly and remarkably high.

In addition, it can be confirmed that the greening rate of cotyledons in the bsp1-1 mutant and the bsp1-2 mutant is significantly higher than that of the wild-type plant (Col-0) and the hos15-2 mutant under high salt conditions.

Through the above results, it was confirmed that the resistance of the plant to salt stress can be increased through the deletion of the gene encoding BSP1.

1d shows seedlings grown from wild-type plants (Col-0), bsp1-1 mutants, bsp1-2 mutants, and sos2-2 mutants under 0 or 125 mM NaCL conditions, and then transplanted into soil, and 300 mM NaCl aqueous solution was added to 4 These are photos taken on days 1, 8, and 12, and grown for 14 days. sos2-2 was used as a high salt sensitive control.

As shown in FIG. 1D, after 14 days in high salt conditions, wild-type plants (Col-0) and sos2-2 mutants turned yellow and almost died, but bsp1-1 mutants and bsp1-2 mutants survived and were still green. you can see what stands out.

Through these results, it can be confirmed that the BSP1 coding gene has resistance to salt stress.

Experimental Example 2. Confirmation of drying stress resistance of bsp1 mutants

48 bsp1-1 mutants of Example 1 and 48 bsp1-2 mutants of Example 2 were obtained, and the seeds were surface sterilized, and then MS medium (1/2 Murashige and Skoog (MS) salt, 1.5% sugar and 0.6% agar, pH 5.7), seedlings grown for 3 weeks were transplanted to soil, grown for 1 week, water was not supplied for 14 days, and then water was supplied again, and the phenotype was monitored after 48 hours. They were grown in the same soil and conditions per pot so that all experienced the same drying stress.

Figure 2 confirms the resistance of wild-type plants (Col-0), bsp1-1 mutant, bsp1-2 mutant, hos15-2 mutant and ost1-3 mutant under dry stress conditions, before drought stress , This is the result of measuring the survival rate of each group after 14 days of no watering (14-days drought) and 48-hrs of rewatering (48-hrs of rewatering). hos15-2 and ost1-3 were used as controls for desiccation stress.

As shown in Figure 2, as shown in Figure 1d, the wild-type plant (Col-0) and the ost1-3 mutant showed a survival rate of less than 20% under dry stress conditions, but the bsp1-1 mutant and the bsp1-2 mutant It can be confirmed that they survive and have an excellent survival rate of 78% or more.

Through these results, it can be confirmed that the BSP1 coding gene has resistance to drying stress.

Experimental Example 3. Confirmation of the relationship between BSP1 and Cullin-1 based E3 ligase

Most proteins with the F-BOX domain serve as substrates for Cullin-1 based E3 ligase, and at this time, the F-BOX domain is known to play a role in proteasome-dependent degradation of proteins by binding to ASK1, an adapter protein. .

Since the BSP1 protein of the present invention contains the F-DOX domain in its sequence, in order to confirm whether it acts as a substrate for the Cullin-1 based E3 ligase, the interaction between the F-BOX domain and ASK1 was tested by Co-IP (Co-immunoprecipitation) analyzed.

transgenic tobacco

To analyze the correlation between BSP1 and ASK1, PCR products amplified from total cDNA were cloned into DONR207 plasmid (Invitrogen, USA). The resulting entry clone was used as a gateway virus vector, pMDC83 (Curtis et al., (2003) Plant physiology 133: 462-469), pH7RWG2.0 (Karimi et al., (2002) Trends in plant science 7: 193-195), pGW-HA (Nakagawa et al., (2007) J Biosci Bioeng. 104:34-41), pDEST-GWVYNE or pDEST-GWVYCE and fused with a GFP tag. To transiently express the GFP-fused BSP1 domain deletion mutant constructs in wild tobacco, each entry clone was prepared using Gateway technology and LR reactions were performed with the pMDC83 vector. Similar to the above BSP1, ASK1 was also cloned into an entry clone, cloned into a gateway binary vector, pH7RWG2.0, pDEST-GWVYNE or pEG203, and fused with an RFP, YFPN or HA tag. All constructs prepared from the PCR reaction were verified through DNA sequencing, and all constructs were introduced into the wild type by the floral dip method. At this time, in order to confirm the interaction between BSP1 and ASK1, a mutant was prepared by combining the protein expressions of BSP1-GFP and ASK1-HA (hemagglutinin), and the expressed protein was '+' and the non-expressed protein was '-' marked as

Coimmunoprecipitation (Co-IP)

Tobacco prepared through the above process ( Nicotiana benthamiana ) 1 ml of extraction buffer (50 mM Tris-HCl, pH 7.5, 150 mM NaCl, 1 mM EDTA, 1% NP-40, 10% glycerol, 0.3 Membrane-enriched total protein was prepared using % sodium deoxycholate, 2 mM sodium orthovanadate (Na3V04), 2 mM NaF, 50 µM MG132, 1 mM phenyl-methyl sulfonyl-fluoride (PMSF) and 1x protease inhibitor cocktail). extracted. After vigorously vortexing for 1 minute, placed on ice for 5 minutes, then centrifuged at 15,000 rpm and 4°C for 10 minutes. The supernatant was reacted with 20 μl of agarose beads coupled with an anti-GFP antibody (Invitrogen, USA) at 4° C. for 4 hours while rotating 360 degrees. After the reaction, the agarose beads were collected and washed 6 times with washing buffer (50 mM Tris-HCl, pH 7.5, 150 mM NaCl, 1 mM EDTA, 1% NP-40 and 1 mM DDT). The immunoreaction mixture was divided in half and analyzed by Western blot. The presence of ASK1-HA in the immunoreaction mixture was confirmed using an anti-HA monoclonal antibody (Roche, Switzerland), and the presence of BSP1-GFP was detected using an anti-GFP antibody. The expression level of each protein was measured by the Enhanced Chemiluminescence (ECL) method.

Figure 3a is a result of Western blotting (western blotting) of proteins separated by immunoprecipitation using a GFP antibody after expressing BSP1-GFP or ASK1-HA with an HA antibody.

As shown in Figure 3a, it can be confirmed that the BSP1 protein and the ASK1 protein interact with each other in the plant. It was confirmed that the BSP1 protein is degraded as a substrate for Cullin-1 based E3 ligase in plants.

Experimental Example 4. Confirmation of correlation between BSP1 protein and SOS1

Split Luciferase assay

SGT1a or SOS1 cDNA was cloned into pCambia1300-nLUC and RAR1 or BSP1 cDNA into pCambia1300-cLUC, respectively. SOS1-nLUC and BSP1-cLUC were transformed into Agrobacterium tumefacien strain GV 3101 and infiltrated into tobacco leaves. After 3 days, 1 mM luciferin was sprayed onto the abaxial surface of the tobacco leaves, and after incubation in the dark for 5 minutes, LUC fluorescence signals were analyzed with a low-light cooled CCD imaging device (Andor iXon; Andor, Northern Ireland).

Figure 3b is the result of analyzing tobacco mutants expressing SGT1a, SOS1, RAR1 or BSP1 by Split Luciferase assay. CLuc-RAR1 and SGT1a-NLuc were used as positive controls.

As shown in Figure 3b, it can be confirmed that BSP1 and SOS1 interact with each other in the plant. SOS1 and BSP1 were fused to the N-terminus of NLuc and the C-terminus of CLuc, and then a split luciferase assay was performed. 35S::CLuc-BSP1 and 35S::SOS1-NLuc were introduced into wild tobacco through transformed Agrobacterium.

SOS1 is related to the salt stress response, and it can be seen that the BSP1 protein of the present invention functions as a negative regulator of SOS1 in plants.

<110> Konkuk University Glocal Industry-Academic Collaboration Foundation <120> Genes Related to drought stress and Salt Stress Resistances and Transformed Plants with the Same <130> HPC10062 <160> 2 <170> KoPatentIn 3.0 <210> 1 <211> 454 <212> PRT <213> artificial sequence <220> <223> BSP1 (Binder of SOS1 Protein 1, AT3G52030) <400> 1 Met Gly Arg Thr Glu Thr Gly Asp Glu Ser Ser Ala Arg Lys Met Arg 1 5 10 15 Arg Lys Val Pro Thr Ser Ile Glu Ser Leu Asp Ala Asp Ile Leu Cys 20 25 30 Ile Ile Phe Ser Phe Leu Asp Leu Phe Asp Leu Val His Cys Thr Val 35 40 45 Val Cys Asn Ser Trp Asn Ala Val Ile Lys Arg Leu Lys Leu Leu Gln 50 55 60 Ala Ser Cys Arg Lys Met His His Leu Gly Ser Asp Ser Pro Ser Ser 65 70 75 80 Ser Thr Ser Leu Asp Arg Pro Ala Glu Ile Asp Val Glu Asp Phe Ala 85 90 95 Met Lys His His Lys Met Ala Leu Leu Arg Gly Arg Ile Glu Ile Glu 100 105 110 Arg Trp Glu Ala His Ser His Arg Val Ser Gln Cys Arg Met Lys Lys 115 120 125 Gly Leu Leu Leu Thr Gly Val Gly Asp Lys Val Met Arg Leu Trp Ser 130 135 140 Leu Lys Ser Tyr Lys Cys Met Glu Glu Tyr Ser Leu Pro Asp Ala Ser 145 150 155 160 Ser Leu Ile Asp Phe Asp Phe Asp Glu Ser Lys Lys Leu Glu Val Val 165 170 175 Ser Leu Ala Trp Glu Tyr Phe Asp Ser Val Val Val Val Val Val Glu 180 185 190 Ile Val Gly Leu Val Gly Thr Arg Ile Ser Ile Trp Arg Arg Asn Gly 195 200 205 Gln Arg Ser Ile Phe Pro Ser Arg Ala Gly Thr Phe Pro Lys Gly Leu 210 215 220 Cys Met Arg Tyr Ile Asp Pro Glu Ala Val Val Gly Cys Glu Asp Gly 225 230 235 240 Thr Ala Arg Val Phe Asp Met Tyr Ser Lys Thr Cys Ser Gln Ile Ile 245 250 255 Arg Thr Gln Gly Gly Pro Ile Thr Cys Leu Ser Leu Ser Asp Asn Gln 260 265 270 Leu Phe Leu Ser Gly Ser Ser Leu Gly Arg Val Thr Val Ser Asp Pro 275 280 285 Leu Met Asp Gln Pro Val Ala Thr Leu Lys Ser Thr Ile Thr Ala Gly 290 295 300 Gly Ile Gln Thr Ile Cys Phe Asn Gln Gly Thr Asn Leu Ala Phe Ile 305 310 315 320 Gly Thr Thr Gly Gly Tyr Val Ser Cys Trp Asp Leu Arg Lys Met Asp 325 330 335 Arg Leu Trp Glu Lys Arg Val Ser Pro Asn Val Val Tyr Ser Ile Gln 340 345 350 Gln Leu Arg Asn Asp Thr Ser Val Met Val Ala Gly Gly Ile Asp Gly 355 360 365 Val Leu Arg Met Ile Asp Gln Lys Ser Gly Arg Val Leu Ser Arg Val 370 375 380 Ile Met Asp Asp Lys Phe Ser Thr Thr Ser Thr Arg Asn Asn Gln Val 385 390 395 400 Val Ile Glu Lys Arg Arg Gly Lys Arg Val Ser Gln Asp Met Glu Ile 405 410 415 Asp Lys Ile Glu Arg Lys Val Arg Pro Gln Ile Ser Cys Ile Ala Met 420 425 430 Gly Met Lys Lys Met Val Thr Ala His Asn Gly Lys Cys Ile Ser Val 435 440 445 Trp Lys Phe Asn Leu Ser 450 <210> 2 <211> 2356 <212> DNA <213> artificial sequence <220> <223> cDNA; BSP1 (Binder of SOS1 Protein 1, AT3G52030) <400> 2 atggggagaa cagagaccgg cgacgaatcg tcggcgagaa agatgcgacg taaagtaccc 60 acgtcgatcg aatctctaga cgccgatata ctgtgcatca tcttctcttt tcttgatttg 120 ttcgacctcg ttcattgtac cgtcgtctgc aattcctggt actcccttca atccaaatcc 180 ctagcttctt ttttctcatg gttgcgaatc ccgtgttgac cttattcgaa cttttttctt 240 attgaggaat gctgttatca aaaggctgaa gctgctacaa gcttcttgcc ggaagatgca 300 ccatctcggt tccgactctc ccagtagctc cacgtcactt gatcgtccag ctgagataga 360 tgtggaggat tttgcgatga agcatcacaa gatggcattg ttaagggggtc gaatcgaaat 420 tgagcgatgg gaagctcatt ctcaccggtg aggcaatttt tccattggga ttgtttttca 480 atgttccaga gattttgttt actcttgcca tgtttctaat gttctctatg tcagggtttc 540 tcagtgtcga atgaagaaag gtttgcttct tactggtgtc ggtgataagg ttaggatttt 600 ttttagtata caattcgatt aggtattgta tgtgttatct aagtagcact ggctaagtta 660 atctgactac tttgatcagg ttatgcgcct ctggtcatta aaaagctaca aatgcatgga 720 agaatactcc cttcctgatg cttcttcctt gattgacttc gattttgatg agagtaaggt 780 aaaccttctc taaaggtcat ggatattact taaactggga atgagctaaa aatggattta 840 atgatcaatg atgatacaga aacctcagtc aagtcactga catgaatttc tgaaaccacc 900 agtatagcag tataatctgt ttccatgtca cagtttgcct cacctgttgt agctgaatat 960 ttccttttgg ctaaatcttg ctattttcta tagaaattgg aagttgtatc tctagcatgg 1020 gagtattttg attctgttgt tgttgttgtt gttgagattg ttggcttggt tggtacccgc 1080 attagcatat ggaggagaaa tggacaaaga agcatttttc catctcgtgc aggcaccttt 1140 cctaaaggcc tttgtatgcg gtatgtcaga agatactctc ctgatatcaa gttcctgaga 1200 acctatctgt ttctatagaa agatcaactg atattccatg ttactgacaa aagtcggtgt 1260 gatacttcaa catggctacc tagcccctct gcccctgtaa ggttgttggc ttctctatcc 1320 acactgataa gttgtatgtt ttctttttca accagttaca ttgacccaga agctgtagtt 1380 ggatgcgagg atggaacagc tcgtgtattt gatatgtaca gtaagacatg ttcacagatc 1440 attaggtaag ctatgtatta tgtgtctgtt gtgttatgga acaacacagt gaaactggaa 1500 aagaacggtt tttcattatc aaaattgttt acaatttctt ttgagaaagc taaatcacaa 1560 ttttcctatc agattggtta ataactatgt atgtaattgc aggactcagg gtgggccaat 1620 aacatgctta tctctaagcg ataatcaact atttcttagt gggtcgtcac tcgggagagt 1680 aacagtatct gaccctttga tggatcaacc agtggccact cttaaatcca cgataactgc 1740 aggaggtaaa ctcaaactca atgtcagatt gcatctccag tgacatttct taaagcaacc 1800 tccattctct acaggtatac agacaatctg ttttaaccaa ggcactaatc tcgccttcat 1860 tggaacaact ggtggatatg tttcatgctg ggacctcagg tatgttaaaa tagtcgtcta 1920 aggttttttt caaagatgca agttttaatt cactatctta agacgttata accaactatt 1980 atctaacagg aaaatggata gattgtggga aaaacgagtg agtcctaatg tcgtgtactc 2040 aatacaacag ctaaggaatg acacatcggt tatggtagct ggtggaatag acggtgtgct 2100 gcgtatgatt gatcagaaga gtggaagggt tttatctaga gttataatgg acgataaatt 2160 ctccacaaca tcaacaagga acaaccaagt ggtgatagag aaaagaagag gaaaaagagt 2220 ttctcaagac atggagatg ataaaatcga aaggaaagtt cggcctcaaa tcagttgcat 2280 agcaatgggg atgaagaaga tggtaacggc tcacaatggt aaatgcataa gtgtatggaa 2340 gttcaatctc tcgtaa 2356

Claims (11)

A composition for improving drying and salt stress resistance of a plant, comprising a nucleic acid molecule that inhibits the expression of a Binder of SOS1 Protein 1 (BSP1) protein-encoding gene. According to claim 1,
The BSP1 protein is a composition for improving drying and salt stress resistance of plants, characterized in that represented by the amino acid sequence of SEQ ID NO: 1. According to claim 1,
The BSP1 protein coding gene is a composition for improving drying and salt stress resistance of plants, characterized in that represented by SEQ ID NO: 2. According to claim 1,
The nucleic acid molecule is at least one selected from the group consisting of T-DNA, siRNA, shRNA, miRNA, ribozyme, peptide nucleic acid (PNA), antisense oligonucleotide, and RNAi vector Drying of plants, characterized in that and a composition for enhancing salt stress resistance. According to claim 1,
The plant is rice, wheat, barley, corn, soybean, potato, wheat, red bean, oat, sorghum, Arabidopsis, cabbage, radish, pepper, strawberry, tomato, watermelon, cucumber, cabbage, melon, pumpkin, green onion, onion, Carrot, ginseng, tobacco, cotton, sesame, sugarcane, sugar beet, perilla, peanut, rapeseed, apple tree, pear tree, jujube tree, peach, lamb, grape, tangerine, persimmon, plum, apricot, banana, rose, gladiolus, Gerbera, carnation, chrysanthemum, lily, tulip, ryegrass, red clover, orchard grass, alfalfa, tall fescue and perennial ryegrass A composition for improving dryness and salt stress resistance of plants, characterized in that any one selected from the group consisting of . A transgenic plant in which the expression of the BSP1 (Binder of SOS1 Protein 1) protein coding gene is suppressed or the BSP1 (Binder of SOS1 Protein 1) protein coding gene is knocked out, with improved resistance to drying and salt stress. According to claim 6,
The BSP1 protein coding gene is represented by SEQ ID NO: 2, characterized in that a transgenic plant with improved dryness and salt stress resistance. According to claim 6,
The plant is rice, wheat, barley, corn, soybean, potato, wheat, red bean, oat, sorghum, Arabidopsis, cabbage, radish, pepper, strawberry, tomato, watermelon, cucumber, cabbage, melon, pumpkin, green onion, onion, Carrot, ginseng, tobacco, cotton, sesame, sugarcane, sugar beet, perilla, peanut, rapeseed, apple tree, pear tree, jujube tree, peach, lamb, grape, tangerine, persimmon, plum, apricot, banana, rose, gladiolus, Transformation with improved dry and salt stress resistance, characterized in that any one selected from the group consisting of gerbera, carnation, chrysanthemum, lily, tulip, ryegrass, red clover, orchard grass, alfalfa, tall fescue and perennial ryegrass plant. (a) suppressing the expression of the BSP1 protein-encoding gene or knocking out the BSP1 protein-encoding gene in the plant; a method for producing a transgenic plant with enhanced resistance to dryness and salt stress. According to claim 10,
The inhibition of expression of the BSP1 protein coding gene or the knock-out of the BSP1 protein coding gene is selected from the group consisting of T-DNA, siRNA, shRNA, miRNA, ribozyme, antisense nucleic acid sequence and transposon for the BSP1 protein coding gene Method for producing a transgenic plant with enhanced resistance to drying and salt stress, characterized in that using any one or more of the. According to claim 10,
The BSP1 protein coding gene is a method for producing a transgenic plant with enhanced resistance to drying and salt stress, characterized in that represented by SEQ ID NO: 2.

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