KR101567543B1 - Method for improving salt-resistance of plant by overexpression of cold regulated 15A protein and the transgenic plant using thereof - Google Patents

Abstract

The present invention provides a method for increasing the salt tolerance of plants through the regulation of the expression of the low temperature resistant protein 15A (COR15A), and using the same to provide transgenic plants having increased salt tolerance. According to the present invention, it is possible to provide a transgenic plant having resistance to growth inhibition by high salt stress and cell damage by providing a method for producing a transformed plant having a salt tolerance through promotion of low temperature resistant protein expression.

Description

FIELD OF THE INVENTION [0001] The present invention relates to a method for enhancing the salt tolerance of a plant by promoting the expression of a low-temperature resistant protein, and a method for improving the salt tolerance of the plant by overexpression of cold regulated 15A protein and the transgenic plant using the same.

The present invention provides a method for increasing the salt tolerance of plants through the regulation of the expression of the low temperature resistant protein 15A (COR15A), and using the same to provide transgenic plants having increased salt tolerance.

Genetic modification technology is used to inject genes such as herbicide tolerance, pest resistance, high sugar content, high nutrient content, growth rate control, toxic substance resistance, environmental resistance, and useful microorganisms, It can contribute to the dramatic improvement of productivity through development of crops that are resistant to the environment and pests and diseases. It can contribute to the increase of foodstuffs by producing crops with high nutrients and functional components (anti-cancer effects, etc.) I have shown the possibility.

Efforts to improve crop yields through the development of new plant species can be divided into two trials. One is to reduce crop yield loss by breeding or processing crop varieties with increased resistance to abiotic stress conditions such as drought, cold weather or salt, or biological stress conditions resulting from pest or disease-causing pathogens will be. However, even if such an attempt is worthwhile, it does not provide fundamentally improved crop yields in the absence of stress conditions.

Another second attempt is breeding or processing new varieties of crops with increased primary yield. Traditional breeding programs have achieved substantive gains in improved yields in early variant crops. In general, the pattern experienced is accompanied by an actual incremental gain in yield initially followed by an incremental further enhancement which is less or more difficult to obtain.

A more recently developed approach based on molecular biology techniques has been developed to achieve a substantial improvement in crop yield by altering the timing, location, or level of expression of a plant gene that plays an important role in plant growth and / Provides talent. Substantial progress has been made over the past 20 years in identifying plant genes responsible for plant growth and / or development. Because of the complexity of how plant growth regulation is ultimately related to yield characteristics, it is still unclear which of these genes can be a clear trail to improve crop yield.

Recent genomic-level gene analysis methods such as DNA microarray analysis have been used to identify stress-adaptive genes in Arabidopsis. The COR15A gene has been reported to be induced by ABA hormone in addition to cold weather as a cold inducing gene in chloroplasts in previous studies. However, no direct relationship between COR15A and high salt stress has been reported.

In Korean Patent Registration No. 571131, 'a gene coding for a transcription factor of a plant', a gene derived from a monocotyledonous plant such as rice, which encodes a transcription factor specific for a stress tolerance gene, is identified and a new environmental stress tolerant plant . However, the patent literature does not disclose any salt tolerant transgenic plants that promoted the expression of COR15A protein isolated from Arabidopsis thaliana.

Korean Patent Registration No. 571131

Under these technical backgrounds, the present inventors have made intensive efforts to develop a method for improving the salt tolerance of plants by promoting the expression of the low temperature resistant protein 15A (COR15A) and a transgenic plant using the same.

Finally, it is an object of the present invention to provide a method for over-expressing the COR15A gene in a transformed plant to improve the salt tolerance of the plant.

It is another object of the present invention to provide a transgenic plant produced by the above method.

In order to solve the above technical problem,

According to one aspect of the present invention, there is provided a method for producing a recombinant vector comprising: (a) operably binding a COR15A (cold regulated 15A) gene isolated from Arabidopsis thaliana to an over-expression promoter and inserting it into a vector; (b) transforming the plant using the vector into which the COR15A gene is inserted; And (c) over-expressing the COR15A gene in the transgenic plant. A method of enhancing plant resistance to high salt stress may be provided.

In one embodiment, the COR15A gene may have the DNA sequence shown in SEQ ID NO: 1.

In one embodiment, the over-expression promoter may be a flower cabbage mosaic virus promoter (CaMV 35S promoter).

In one embodiment, the high salt stress may be a salt concentration of at least 50 mM.

According to another aspect of the present invention, a transgenic plant produced by the above method can be provided.

According to the present invention, a transgenic plant having resistance to growth damage and cell damage by high salt stress can be provided by providing a method for producing a transformed plant having a salt tolerance through promotion of low temperature resistant protein expression.

FIG. 1 shows the results of comparing the expression level of COR15A in response to high salt stress and the expression level of COR15A in Arabidopsis overexpressing COR15A and wild type Arabidopsis .
FIG. 2 shows the results of analysis of chlorophyll amount and ion leaking ratio, which is a measure of cytotoxicity of COR15A overexpressing Arabidopsis in response to high salt stress.
Fig. 3 shows phenotypic analysis results of COR15A overexpressing Arabidopsis and wild type Arabidopsis in response to high salt stress.
Figure 4 shows the growth inhibition resistance of COR15A overexpressing Arabidopsis and wild-type Arabidopsis in response to high salt stress.

Hereinafter, the present invention will be described in more detail.

In the present invention, the term "COR15A gene" is a kind of low-temperature resistant gene isolated from Arabidopsis thaliana and is associated with cold tolerance of organelles such as chloroplasts and protoplasts of Arabidopsis thaliana. In particular, CBF1 binds to the CRT / DRE sequence in response to low temperature, leading to the regulation (increase in expression) of the COR gene.

In the present invention, the term " specific " or " specific " means the ability to suppress only the target gene without affecting other genes in the cell.

In the present invention, the term " salt tolerance " or " salt resistance "means a property in which a plant can tolerate a high salt environment. When a plant is exposed to high salt, an imbalance in osmotic pressure and an imbalance in ion concentration occur within the cell. In addition, as the secondary stress, oxidative stress, cell membrane breakdown, nutritional disorder, metabolic disorder, photosynthetic disorder and the like are caused, and the salt tolerance can be confirmed by the resistance to the above various obstacles.

According to one aspect of the present invention, there is provided a method for producing a recombinant vector comprising: (a) operably binding a COR15A (cold regulated 15A) gene isolated from Arabidopsis thaliana to an over-expression promoter and inserting it into a vector; (b) transforming the plant using the vector into which the COR15A gene is inserted; And (c) over-expressing the COR15A gene in the transgenic plant. A method of enhancing plant resistance to high salt stress may be provided.

In one embodiment, the over-expression promoter operably linked to the COR15A gene may be increased in expression under high salt stress of greater than 50 mM. The ensuring of salt tolerance of plants can be achieved by various biochemical pathways or signal transduction pathways.

In one embodiment, the COR15A gene may have the DNA sequence shown in SEQ ID NO: 1. The COR15A gene is preferably substantially identical to the DNA sequence shown in SEQ ID NO: 1, but is not limited thereto.

In the present invention, the term "substantial identity" of the term DNA sequence means that the polynucleotide has a sequence identity of 60% or more, usually 70% or more, more usually , More preferably 80% or more, and most preferably 90% or more sequence identity. Those of ordinary skill in the art will appreciate that these values may be suitably adjusted to measure the corresponding identity of the proteins encoded by the two nucleotide sequences, taking into account codon degeneration, amino acid similarity, read frame localization, and the like.

An example of an algorithm suitable for determining percent sequence identity and sequence similarity is the BLAST algorithm. Software for performing BLAST analysis is available on the National Center for Biotechnology Information website.

In one embodiment, the gene expression is preferably under the control of a promoter. Promoters suitable for operatively linking to the COR15A gene using the methods disclosed herein may be from the same species or from different species of plants to be transformed. In addition, the promoter may originate from the same species or may originate from a different species with respect to the gene of the present invention to be used. A promoter for use in the methods of the invention may also include a chimeric promoter that may comprise one or more combinations of promoters having a common expression profile as described below.

Methods for identifying and characterizing promoter regions in plant genomic DNA are well known to those of ordinary skill in the art. Examples of such promoter sequences include a promoter (AAP1) for an amino acid permiease gene, a promoter for an oleate 12-hydroxylase: desaturase gene (e.g., a promoter designated as LFAH12 from Rescuelella pendarelis) , 2S2 albumin gene promoter (e.g., 2S2 promoter from Arabidopsis thaliana), fatty acid elongase gene promoter (FAEl) (e.g., FAE1 promoter from Arabidopsis thaliana), and lipocortiladone gene promoter (LEC2) E.g., the LEC2 promoter from Arabidopsis thaliana).

In one embodiment, the over-expression promoter may be a flower cabbage mosaic virus promoter (CaMV 35S promoter). The promoter for over-expression may be selected depending on the kind of the crop or the plant tissue to be expressed. As the promoter, a normal expression promoter, a tissue-specific promoter, a development-stage-specific promoter, a conditionally-expressing promoter, have.

In the present invention, the term " vector "or" expression vector "means a DNA strand that is typically double stranded that can insert it into an external DNA. The vector or replicon may be, for example, a plasmid or a viral origin. The vector contains a "replicon" polynucleotide sequence that promotes autonomous replication of the vector within the host cell. In the present invention, the term " replicon "also includes polynucleotide sequences that target or otherwise facilitate recombination of vector sequences into the host chromosome.

Also, transformation of the viral vector DNA into the host cell may result in the conversion of the viral DNA into the viral RNA vector molecule, even though the external DNA may initially be inserted into, for example, a DNA viral vector. External DNA is defined as heterologous DNA, defined, for example, as heterologous DNA, a DNA that is not found naturally in the host cell, which replicates vector molecules and encodes a selectable or selectable marker or transgene do. The vector is used to deliver foreign or heterologous DNA into a suitable host cell. Within the host cell, the vector can replicate independently or inconsistently with the host chromosomal DNA and can produce several vector copies and the inserted DNA.

In addition, the vector can target insertion into the host chromosome of an exogenous or heterologous DNA. The vector may contain an essential component which permits the transcription of the inserted DNA into the mRNA molecule or induces the replication of the inserted DNA into multiple copies of the RNA. Some expression vectors further comprise a sequence component that is close to the inserted DNA that allows translation of the mRNA into the protein molecule. Thus, many mRNA molecules and polypeptides encoded by the inserted DNA can be synthesized rapidly.

In one embodiment, the high salt stress may be a salt concentration of at least 50 mM. The high salt stress means the same effect as inhibiting the growth of plants due to the salt concentration which is excessively present in the soil, and is expressed as various units in the technical field to which the present invention belongs. Overexpression of the COR15A gene according to the present invention can improve the resistance (i.e., salt tolerance) of transformed plants under high salt stress of 50 mM or more.

According to another aspect of the present invention, a transgenic plant produced by the above method can be provided. In one embodiment, the plant may be Arabidopsis or rapeseed.

In the present invention, the term "transformation" refers to the introduction of DNA into cells. In the present invention, the term "plant" includes whole plants, plant organs (eg, leaves, stems, flowers, roots, etc.), seeds and plant cells (including tissue culture cells) and their offspring. The classes of plants that can be used in the methods of the present invention are broadly broad, such as a family of higher plants generally applicable to transgenic techniques including both monocotyledons and dicotyledons, and certain lower plants such as algae. This includes plants of varying levels of purity, including polyploids, diploids and haploids.

Typically, when the vector is a plasmid, the vector also includes a selectable marker gene, for example, a kanamycin gene encoding a resistance to kanamycin. The most common method of plant transformation is the transformation of the target transgene into a plant transformation vector and then into Agrobacterium tumifaciens containing the helper Ti-plasmid. Agrobacterium cells containing the transgene vector can be cultured with leaf pieces of the plant to be transformed.

Transformation of cultured plant host cells can be accomplished via Agrobacterium tumefaciens as described above. Cultures of host cells that do not have tight cell membrane barriers are transformed using the modified calcium phosphate method.

However, other methods for introducing DNA into cells, such as polybrene, protoplast fusion, electroporation, and direct microinjection into the nucleus, may also be used. Transformed plant cells can be grown on a medium containing an optical hormone such as naphthaleneacetic acid and benzyladenine for induction of cells, for example, with kanamycin and appropriate amounts of callus and shoot induction, Markers can be selected. Thereafter, plant cells are regenerated and the resulting plants can be transferred to soil using techniques well known to those of ordinary skill in the art.

In addition to the methods described above, a number of methods are well known in the art for transferring cloned DNA into a wide variety of plant species, including asiatic plants, spore plants, terminal leaves and dicotyledonous plants.

Representative examples include electroporation-promoted DNA uptake by protoplasts; Treatment of protoplasts with polyethylene glycol; And bombardment of cells with micro-firing containing DNA.

In the present invention, the term "primer" refers to a nucleic acid that is annealed to a complementary target DNA strand by nucleic acid hybridization to form a hybrid between the primer and the target DNA strand, followed by a polymerase, It is an isolated nucleic acid that extends along the target DAN strand by an enzyme. The primer pairs of the present invention represent their use for amplification of a target nucleic acid sequence by, for example, polymer chain reaction (PCR) or other conventional nucleic acid amplification methods.

The probe and the primer are generally at least 11 nucleotides, preferably at least 18 nucleotides, more preferably at least 24 nucleotides, and most preferably at least 30 nucleotides in length. Such probes and primers hybridize specifically to the target sequence under very stringent hybridization conditions.

Methods for making and using probes and primers are well known in the art. PCR-primer pairs can be derived from known sequences, for example, using a computer program such as Primer (Version 0.5, ⓒ 1991, Whitehead Institute for Biomedical Research, Cambridge, Mass.) Developed for this purpose.

Nucleic acid amplification can be used for any of a variety of nucleic acid amplification methods known in the art to which the present invention belongs, including polymerase chain reaction (PCR). Various amplification methods are known to the ordinarily skilled artisan. The PCR amplification method is developed to amplify up to 22 kb of genomic DNA and up to 42 kb of bacteriophage DNA.

Hereinafter, the present invention will be described in more detail by way of examples. However, these embodiments are not intended to limit the scope of the present invention.

Experimental methods 1. Target plants and growth conditions

Arabidopsis thaliana L. Heynh of Colombia ecotype (Col-0) was used for the experiments of the present invention. The plants contained 2% sucrose and 0.25% phyta-gel (pH 5.8) under long-day conditions of 22 ° C (light condition of 80 μEm ^-2 s ^-1 for 16 hours and dark conditions for 8 hours), phytohormones Lt; RTI ID = 0.0 > (MSO). ≪ / RTI > To induce simultaneous germination, the seeds were cultured for 3 days under 4 ° C dark conditions and then transferred to the growth chamber.

Experimental method 2. RNA extraction and reverse transcriptase (RT) -PCR

For quantitative RT-PCR analysis of Arabidopsis , total RNA was extracted from Arabidopsis at 10 days using TRIzol reagent (Invitrogen, Carlsbad, Calif., USA). (Chen et < RTI ID = 0.0 > al., ≪ / RTI > 3) using Primin (5'- ATGGCGATGTCTTTCTC-3 'and 5'-TTTGTGGCATCCTTAG- 3') and Actin2 (5'- ATGGCTGAGGCTGATGATATTCAA- 3 'and 5'- TTAGAAACATTTTCTGTGAACGATT- 2010), and RT-PCR was performed. The obtained PCR product was analyzed by electrophoresis after sequencing.

Experimental method 3. Generation of transgenic plants

To amplify the full length COR15A cDNA, RT-PCR was performed on total RNA obtained from wild-type (col-0) oil plants. The reaction was carried out using PCR primers 5'-AGGATCCATGGCGATGTCTTTCTC-3 'and 5'-AGTCGACTTTGTGGCATCCTTAG-3'. The RT-PCR product was cloned into pGEM-T Easy (Promega, Madison, Wis.). COR15A cDNA was treated with BamHI and SalI, and the resulting 416 bp fragment was inserted into a pCAMBIA 1301 plasmid to construct a 35Spro: COR15A-smGFP construct. Recombinant plasmids were introduced into Agrobacterium tumefaciens GV3101 and used for transformation of Arabidopsis plants through the floral dip method. An isotype T3 line containing a single T-DNA insert was used for analysis and the transgenic plants were maintained as described above.

Experimental method 4. Chlorophyll fluorescence measurement and ion leak test method

For in vivo chlorophyll fluorescence assays, 10-day-old Arabidopsis plants (transgenic plants number 14) grown in MSO plates were supplemented with 200 mM NaCl for 1 to 3 days under 16 h light / 8 h dark conditions at 22 ° C Lt; / RTI > MSO solution. NaCl-treated plants were acclimated for 15 min and chlorophyll fluorescence was detected by Yu et al. (Handy REA, Hansatech Instruments, UK) using the plant efficiency analyzer according to the method described in the literature (2002).

Ionic leaks have been reported by Scarpeci et al. (2008). The conditions of the wild-type plant and the NaCl-treatment are the same as those of the chlorophyll fluorescence measurement method. 10 days old wild-type and COR15A overexpressed Arabidopsis plants were cultured in 1/4 MSO solution (control) or in 1/4 MSO solution supplemented with 200 mM NaCl for 1 to 12 hours under long-day conditions at 22 占 폚. After the incubation, the plants were washed with ddH ₂ O, after transferred into test tubes containing ddH ₂ O 25 ml of conductivity meter (Orion 115Aplus; ThermoElectron) measuring the electrical conductivity of a solution (C1) by using and for 2 hours And cultured at room temperature. The tube was then heated for 15 minutes, cooled at room temperature and again the electrical conductivity (C2) was measured. The REL was calculated according to the formula C1 / C2 X 100% (Cao et al., 2007).

Experimental method 5. Phenotypic analysis of transgenic plants

For NaCl stress analysis at the oily plant stage, 4-day old non-transformed wild-type and T3-transformed Arabidopsis oil plants were transferred to a new 1/2 MSO plate with or without 200 mM NaCl. After an additional 10 days of vertical growth, the weight of the top part, the length of the main muscle and the number of branching points were measured respectively. For salt stress of soil growth, 2 - week - old plants were treated with 300 mM NaCl solution every 2 days until 14th.

Experimental results 1. Induction of COR15A transcript by high salt stress

RNA was extracted from wild type Arabidopsis treated with 200 mM NaCl for 0, 1, 3, and 6 hours, respectively. CDNA was synthesized by reverse transcriptase and PCR was performed using the template strand. The expression amount of COR15A was 0 hour (Fig. 1, A). In addition, under normal conditions, it was confirmed that the present exemplary result of comparing the expression level of the COR15A overexpression of the wild-type Arabidopsis and COR15A built according to the embodiment of the invention, over-expressing Arabidopsis increases the amount of expression of COR15A than the wild type (WT) Arabidopsis.

In other words, it was confirmed that COR15A did not only react to low-temperature stress (cold weather) but also increased the amount of transcript in response to high salt stress.

Experimental results 2. Increased resistance to cell damage by high salt stress of COR15A overexpressing plants

The present inventors planted the wild-type Arabidopsis and COR15A overexpressed Arabidopsis in MSO medium for 10 days in a 200 mM NaCl solution for 10 days in order to determine the degree of plant survival caused by high salt stress induced cell damage. 3 days.

As a result, on day 2, the wild type Arabidopsis was weaker than the COR15A overexpressing Arabidopsis , but the whitening progressed more, and on the third day, the whitening progressed more prominently in the wild type Arabidopsis than the COR15A overexpressing Arabidopsis 2).

In addition, the present inventors numerically quantified the amount of chlorophyll for the objective and quantitative evaluation. The changes in chlorophyll under high salt stress from day 1 to day 3 showed that the wild type Arabidopsis was slightly lower than the COR15A overexpressing Arabidopsis on the second day and the difference was remarkably large on the third day (See FIG. 2B, *; meaning that the significance value is lower than 0.05).

Chlorophyll is a key molecule in photosynthesis, and it corresponds to a photosynthetic pigment that acts as an antenna to absorb light energy. Therefore, the amount of chlorophyll is directly related to the activity of the plant including the chlorophyll. The presence of chlorophyll in the COR15A overexpressing Arabidopsis is less than that of the wild type, and the cell activity is relatively more maintained .

When plant cells are damaged by abiotic stress, the ions inside the cells flow out to the outside of the cell, which can be confirmed by measuring ion flux. When measuring the leakage of the non-ionic early treatment of high salt stress, ion outflow at 1 time ratio it is confirmed in wild-type Arabidopsis overexpressing Arabidopsis COR15A than three hours after it appears, but only narrowly ion leakage ratio high. After 9 hours, the ionic efflux ratio of wild type Arabidopsis was higher than that of COR15A overexpressed Arabidopsis. However, after 12 hours, there was no difference in the ion outflow ratio among the plants (see C in FIG. 2).

Plants that have undergone high salt stress induce ionic fluxes due to degeneration of early cell membranes due to high salt stress, and the chlorophyll breakdown proceeds under constant stress. According to the above experimental results, Overexpressed Arabidopsis has high chlorophyll levels and low ion flux ratios even under high salt stress conditions compared to wild type Arabidopsis . Therefore, it was confirmed that the COR15A gene increased the expression level under the high salt stress condition, thereby increasing the resistance to the high salt stress.

Experimental results 3. Increased resistance to growth inhibition by high salt stress of COR15A overexpressing plants

The present inventors observed the overall phenotype of the plants by transferring wild-type Arabidopsis and COR15A overexpressing Arabidopsis at 4 days to 1/2 MSO medium containing 200 mM NaCl and growing them under long-day conditions in order to examine the resistance to growth inhibition caused by high salt stress .

After 10 days from the transfer of the plant with the 1/2 MO 2 medium containing NaCl, the weight of the top part of the plant (the weight of the upper part of the surface) was measured. As a result, it was confirmed that the COR15A overexpressed Arabidopsis weighed more than the wild type Arabidopsis (See FIG. 3 A), but the length of the primary root showed similar values irrespective of the presence or absence of the high salt stress treatment (see FIG. 3B). The content of chlorophyll was measured for wild-type Arabidopsis is was confirmed that the level of 2/3 to 3/4 of the COR15A overexpressed Arabidopsis (see Fig. 3 C), generation of the inner circle or absence (the number of branch points of the inner circle) , And that the wild-type Arabidopsis was inhibited by a slight difference from the COR15A overexpressing Arabidopsis (see Fig. 3 D).

As described above, it was confirmed that COR15A overexpressed Arabidopsis showed higher resistance to high salt stress than wild type Arabidopsis , based on the weight of the top part, the content of chlorophyll, and the occurrence of side lines corresponding to the representative phenotype of the plant.

Two-week-old plants cultured in soil were treated with 300 mM NaCl solution at 2-day intervals, and wild-type Arabidopsis and COR15A overexpressed Arabidopsis And confirmed the difference in resistance to high salt stress. As a result, the wild type Arabidopsis survived 9.5% while the COR15A overexpressed Arabidopsis survived 90.5% and 95.2%, respectively (see FIG. 4). Although there is a difference in the resistance to high salt stress at the growth stage of each plant, the COR15A gene plays an important role in resistance of plants to high salt stress.

Therefore, according to the present invention, by providing a method for producing a transformed plant having a salt tolerance through promotion of expression of the low temperature resistant protein 15A (COR15A), the transgenic plant having resistance to growth inhibition by high salt stress and cell damage Can be provided.

While the present invention has been particularly shown and described with reference to exemplary embodiments thereof, it is to be understood that the invention is not limited to the disclosed embodiments, but, on the contrary, It will be obvious that it is not. It is therefore intended that the scope of the invention be defined by the claims appended hereto and their equivalents.

                         SEQUENCE LISTING <110> GYEONGSANG NATIONAL UNIVERSITY OFFICE OF ACADEMY AND        INDUSTRY COLLABORATION   <120> Method for improving salt-resistance of plant by overexpression        of cold regulated 15A protein and the transgenic plant using        the <130> NPF-24591 <160> 1 <170> PatentIn version 3.2 <210> 1 <211> 755 <212> DNA <213> Arabidopsis thaliana <400> 1 atcgatctat aaaacgatac tattggagat tagattcttc tcatctcact ttgttcatct 60 aaaaactcct cctttcattt ccaaacaaaa acttcttttt attctcacat cttaaagatc 120 tctctcatgg cgatgtcttt ctcaggagct gttctcactg gtatggcttc ttctttccac 180 agcggagcca agcagagcag cttcggcgct gtcagagtcg gccagaaaac tcagttcgtc 240 gtcgtttctc aacgcaagaa gtcgttgatc tacgccgcta aaggtgacgg caacatcctc 300 gatgacctca acgaggccac aaagaaagct tcagatttcg tgacggataa aacaaaagag 360 gcattagcag atggtgagaa agcgaaagac tacgttgttg aaaaaaacag tgaaaccgca 420 gatacattgg gtaaagaagc tgagaaagct gcggcgtatg tggaggagaa aggaaaagaa 480 gccgcaaaca aggcggcaga gttcgcggag ggtaaagcag gagaggctaa ggatgccaca 540 aagtagggtc ttacctaatc agttaatttc aagcacttaa actcgtagat atattgatcc 600 atatcctctc tcttcatgtt taatagtact tacaataaga tgagtctgtt gtaatttcta 660 ttaatttcac atcacaactg aaataagata tggtatccac agtcaccgtc acattcttta 720 atgttttgca aaatattcaa tagacaaaat taaat 755

Claims (6)

(a) inserting a COR15A (cold regulated 15A) gene isolated from Arabidopsis thaliana into a vector operably linked to an over-expression promoter;
(b) transforming the plant using the vector into which the COR15A gene is inserted; And
(c) overexpressing the COR15A gene in the transformed plant,
The COR15A gene has the nucleotide sequence shown in SEQ ID NO: 1,
A method of enhancing plant resistance to high salt stress with a salt concentration of greater than 50 mM.
delete The method according to claim 1,
Wherein the promoter for overexpression is a flower cabbage mosaic virus promoter (CaMV 35S promoter).
The method according to claim 1,
Wherein the plant is a Arabidopsis thaliana or a rapeseed.
delete delete

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