KR101485825B1 - Gene Implicated in Salt Stress Tolerance and Transformed Plants with the Same - Google Patents

Abstract

The present invention relates to a composition for reinforcing the salt resistance of a plant. A nucleotide sequence of the present invention is involved in the salt stress of a plant, and a transgenic plant overexpressed therefrom has excellent resistance against salt stress on farmland caused by changes in climatic conditions such as global warming, so that the transgenic plant can be usefully used as a novel functional plant which can excellently adapt to high salinity farming conditions.

Description

≪ Desc / Clms Page number 1 > Gene Implicated in Salt Stress Tolerance and Transformed Plants with the Same &

The present invention relates to Chinese cabbage rapa ssp . related BrSSR ( Brassica rapa Salt Stress Resistance) gene and a transgenic plant isolated from wild-type pekinensis .

The problem of human food shortage is caused by an increasing population, and in order to overcome this problem, efforts are being made to increase arable land that can be cultivated through wasteland clearing and reclamation projects. Wasteland and reclaimed land have a lot of abiotic stresses compared with general farmland, especially salinization in soil, which adversely affects crop growth. High concentrations of salts are known to cause ion stress and osmotic stress and lead to secondary stresses such as oxidative stress and malnutrition .

Around 900 million hectares of land are reported to be affected by salt, especially for countries where irrigation is essential for agriculture. Salt accumulation is a very important factor that determines the yield of crops. It is an important factor.

There are many factors that inhibit the growth of crops, but the most potent factor is the increase in Na ⁺ ion concentration in the body. The plants are different, such as compartmentalized mechanism into the Na ⁺ ion absorption control of the mechanism, Na ⁺ keep it said ions in a museum the mechanism for re-recovered, from the root, Na ⁺ out of the ion ejection mechanism, vacuoles to adapt to salt stress environment We are setting up a mechanism.

Recent global climate change is expected to result in a 30% reduction in current arable land area within 20 years and a further 50% reduction by 2050, over the next 20 years.

Therefore, in terms of stable supply of food for the future, the identification of salt-related genes and improvement of salt tolerance of crops through the study of mechanisms are treated as the top priority in plant biotechnology research. Since the first study of the tolerance of salt resistance traits by Lyon in 1941, various resistance genes have been found in various crops and studies on its expression network have been actively carried out.

For example, an increase in salt tolerance of rice with HVA1, a late embryogenesis abundant protein (LEA) protein of barley, and an overexpression of rice rice-dependent protein kinase (OsCDPK7) (DREB1, dehydration-responsive element-binding protein 1 (DREB1)) in the Arabidopsis thaliana, the overexpression of Na ⁺ / H ⁺ (DREB1A, DREB1B) are very effective in improving the low temperature resistance of crops.

On the other hand, Chinese cabbage is one of the four major vegetables of Korea as the main ingredient of kimchi and is included in the lacquer ( Brassica ). Rice (Brassica) is an important crop in foreign countries because it includes crops with excellent industrial economics such as oilseed rape, mustard. Brassica crops are susceptible to damage by salinization in the soil, resulting in reduced yields due to growth disturbances, which can be devastating to farmers and consumers.

The decrease in agricultural productivity due to salt accumulation is serious, and development of good varieties resistant to such environment is very urgent for securing stable food resources.

Accordingly, the present inventors have made extensive efforts to find genes capable of improving the salt tolerance of plants. As a result, it has been confirmed that the nucleotide sequence of SEQ ID NO: 7 is involved in the above-described trait of the plant, and when the gene is transformed into a plant, the salt tolerance of the plant can be enhanced.

Accordingly, an object of the present invention is to provide a composition for enhancing salt tolerance of a plant, and a plant cell or plant having enhanced salt tolerance transformed with the composition.

Another object of the present invention is to provide a method for enhancing the salt tolerance of a plant.

Other objects and advantages of the present invention will become more apparent from the following detailed description of the invention, claims and drawings.

According to one aspect of the present invention, there is provided a composition for enhancing salt tolerance of a plant comprising a nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 8.

The present inventors have made extensive efforts to find genes capable of improving the salt tolerance of plants. As a result, it was confirmed that when the nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 8 is involved in the above-described trait of the plant, and when the gene is transfected and overexpressed in the plant, the salt stress resistance of the plant can be enhanced.

According to a preferred embodiment of the present invention, the nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 8 of the present invention consists of the nucleotide sequence of SEQ ID NO: According to the present invention, the nucleotide sequence of SEQ ID NO: 7 was identified in the genome of Chinese cabbage, and the name of this gene was named BrSSR ( Brassica rapa Salt Stress Resistance). The gene includes a domain of unknown function 581 (DUF16).

According to the present invention, the present inventors have found that when a salt stress is applied to a plant, the expression of the BrSSR gene is increased, and when the expression of the gene is increased, tolerance to such salt stress is increased.

It is clear to a person skilled in the art that the nucleotide sequence used in the present invention is not limited to the nucleotide sequence described in the attached sequence listing.

Variations in nucleotides do not cause changes in the protein. Such nucleic acids include functionally equivalent codons or nucleic acid molecules comprising a codon coding for the same amino acid, or a codon coding for a biologically equivalent amino acid. Considering the mutation having the above-mentioned biological equivalent activity, the nucleic acid molecule used in the present invention is interpreted to include a sequence showing substantial identity with the sequence described in the sequence listing. The above-mentioned substantial identity is determined by aligning the sequence of the present invention with any other sequence as much as possible and analyzing the aligned sequence using an algorithm commonly used in the art. Homology, more preferably 70% homology, even more preferably 80% homology, and most preferably 90% homology. Alignment methods for sequence comparison are well known in the art. Various methods and algorithms for alignment can be found in prior art [Smith and Waterman, Adv. Appl. Math. 2: 482 (1981); Needleman and Wunsch, J. Mol. Bio. 48: 443 (1970); Pearson and Lipman, Methods in Mol. Biol. 24: 307-31 (1988); Higgins and Sharp, Gene 73: 237-44 (1988); Higgins and Sharp, CABIOS 5: 151-3 (1989); Corpet et al., Nuc. Acids Res. 16: 10881-90 (1988); Huang et al., Comp. Appl. BioSci. 8: 155-65 (1992) and Pearson et al., Meth. Mol. Biol. 24: 307-31 (1994). The NCBI Basic Local Alignment Search Tool (BLAST) (Altschul et al., J. Mol. Biol. 215: 403-10 (1990)) is accessible from NCBI (National Center for Biological Information) It can be used in conjunction with sequence analysis programs such as blastx, tblastn and tblastx. BLSAT is available on the Internet (http://www.ncbi.nlm.nih.gov/BLAST/). The sequence homology comparison method using this program can be found on the Internet (http://www.ncbi.nlm.nih.gov/BLAST/blast_help.html).

According to another aspect of the present invention, the present invention provides a nucleic acid comprising (a) a nucleotide sequence disclosed in the present invention; (b) a promoter operably linked to said nucleotide sequence and forming RNA molecules in plant cells; And (c) a recombinant expression vector for plant expression comprising a poly A signal sequence which acts on plant cells to cause polyadenylation of the 3'-terminal of the RNA molecule. The present invention also provides a composition for promoting salt tolerance of a plant.

As used herein, the term " operably linked " refers to a functional linkage between a nucleic acid expression control sequence (e.g., an array of promoter, signal sequence, or transcription factor binding site) and another nucleic acid sequence, Transcription < / RTI > and / or translation of the other nucleic acid sequences.

The vector system of the present invention can be constructed through various methods known in the art, and specific methods for this are disclosed in Sambrook et al., Molecular Cloning, A Laboratory Manual, Cold Spring Harbor Laboratory Press (2001).

The promoter suitable for the present invention may be any of those conventionally used in the art for the gene introduction of a plant, and examples thereof include SP6 promoter, T7 promoter, T3 promoter, PM promoter, corn ubiquitin promoter, (CaMV) 35S promoter, nopaline synthase (nos) promoter, Piguet mosaic virus 35S promoter, water crab basil lipid viral promoter, commeline yellow moth virus promoter, ribulose-1,5-bis-phosphate carboxamide (TPI) promoter, adenine phosphoribosyl transferase (APRT) promoter of Arabidopsis, and the tryptophan synthase promoter of ssRUBISCO, a cysteine triphosphate isomerase (TPI) promoter of Arabidopsis thaliana, do.

In addition, the polyA signal sequence causing the 3'-terminal polyadenylation according to the present invention is derived from the nopaline synthase gene of Agrobacterium tumefaciens (NOS 3 'end) (Bevan et al. Nucleic Acids Research, 11 (2): 369-385 (1983)), derived from the Octopine synthase gene of Agrobacterium tumefaciens, the 3'end of the protease inhibitor I or II gene of tomato or potato, CaMV 35S terminator and an OCS terminator (octopine synthase terminator) sequence.

Alternatively, the vector may further carry a gene encoding a reporter molecule (e.g., luciferase and beta -glucuronidase). In addition, the vector of the present invention can be used as a selection marker for a gene resistant to antibiotics (e.g. neomycin, carbenicillin, kanamycin, spectinomycin, hygromycin etc.) (for example, neomycin phosphotransferase (nptII) Mycophosphotransferase (hpt), and the like).

According to another aspect of the present invention, there is provided a salt tolerant plant cell transformed with the composition of the present invention.

According to another aspect of the present invention, there is provided a salt tolerant enhanced plant transformed with the composition of the present invention.

(Methods of Enzymology, Vol. 153, (1987)) to produce transgenic plant cells and transgenic plants of the present invention. Plasmids can be transformed by inserting exogenous polynucleotides into carriers such as plasmids and viruses, and Agrobacterium bacteria can be used as a mediator (Chilton et al., Cell 11: 263-271, 1977) Direct exogenous polynucleotides can be introduced into plant cells to transform plants (Lorz et al., Mol. Genet. 199: 178-182; 1985). For example, when a vector not containing a T-DNA region is used, electroporation, microparticle bombardment, and polyethylene glycol-mediated uptake may be used.

Generally, a method commonly used for transforming plants is to infect plant cells or seeds with Agrobacterium tumefaciens transformed with an exogenous polynucleotide. One of ordinary skill in the art can cultivate or plant transformed plant cells or seeds under suitable known conditions to develop into plants.

As used herein, the term " plant (or plant) " is understood to mean not only mature plants but also plant cells, plant tissues and plant seeds that develop into mature plants.

According to a preferred embodiment of the present invention, the plant of the present invention is a food crop including rice, wheat, barley, corn, soybean, potato, wheat, red bean, oats and millet; Vegetable crops including Arabidopsis, cabbage, radish, pepper, strawberry, tomato, watermelon, cucumber, cabbage, melon, squash, onions, onions and carrots; Special crops including ginseng, tobacco, cotton, sesame, sugar cane, beet, perilla, peanut and rapeseed; Apple trees, pears, jujubes, peaches, sheep, grapes, citrus, persimmon, plums, apricots and banana; Roses, gladiolus, gerberas, carnations, chrysanthemums, lilies and tulips; And feed crops including ragras, red clover, orchardgrass, alpha-alpha, tall fescue and perennialla grass.

According to another aspect of the present invention, there is provided a method for increasing the intracellular expression level of a polypeptide having an amino acid sequence represented by SEQ ID NO: 8 to improve the salt tolerance of a plant.

The nucleic acid molecule, the recombinant vector for plant expression, and the method for introducing the same into a plant have already been described above, so that description thereof is omitted in order to avoid excessive redundancy.

The features and advantages of the present invention are summarized as follows:

(a) The present invention provides a composition for enhancing salt tolerance of a plant.

(b) The nucleotide sequence of the present invention is involved in tolerance to salt stress of a plant. Transgenic plants overexpressing the plant are highly resistant to salt stress caused by changes in climatic conditions due to global warming and the like, It can be used as a new functional crop with high adaptability to cultivation conditions.

FIG. 1 is a photograph of a fragment of a gene encoding BrSSR amplified from Chinese cabbage by RT-PCR using each pair of BrSSR-F and BrSSR-R primers.
FIG. 2 shows the result of RT-PCR using BrSSR-F and BrSSR-R primer pairs. The gene encoding BrSSR amplified from Chinese cabbage was inserted into pGEM-T easy vector, It is a photograph.
Figure 3 shows a sequence of a gene encoding BrssR of Chinese cabbage isolated by RT-PCR using a GenBank BlastX program and a CLC free workbench 3.2.3 program for wild strawberry ( Fragaria vesca ), Camelina sativa , ( Arabidopsis thaliana ), soybean ( Glycine max ), and cucumber ( Cucumis sativus ) sequences.
Fig. 4 shows the nucleotide sequence and amino acid sequence of the gene encoding BrSSR of Chinese cabbage separated by RT-PCR.
FIG. 5 shows the result of analysis of the gene encoding BrSSR of Chinese cabbage isolated by RT-PCR using the NCBI Conserved Domain program.
FIG. 6 is a diagram showing a transcriptional overexpression vector (pSL94) overexpressing the BrSSR gene of Chinese cabbage isolated by RT-PCR.
FIG. 7 shows the results of PCR analysis of the transformation of tobacco transformants produced using the BrSSR overexpression vector.
FIG. 8 shows the results of quantitative real-time RT PCR analysis of the expression level of the BrSSR gene in tobacco transformants transfected with the PCR assay.
FIG. 9 shows the results of analysis of phenotype after salt stress treatment of tobacco transformants showing salt tolerance.

Hereinafter, the present invention will be described in more detail with reference to Examples. These embodiments are only for illustrating the present invention, and thus the scope of the present invention is not construed as being limited by these embodiments.

The Brassica rapa ssp. Pekinensis inbred line 'CT001' used in the present invention was sampled from the cabbage grown in an aseptic condition and used for RNA isolation. Hereinafter, the BrSSR gene identification process and salt tolerance test of the present invention will be described.

Example 1: Investigation of salt resistance-related genes derived from Chinese cabbage

The information of " Brassica rapa EST and Microarray Database" was used to identify the new salt tolerance genes derived from Chinese cabbage. The above-mentioned database was prepared for 3 weeks by growing the Chinese cabbage ( Brassica rapa ssp. Pekinensis inbred line 'Chiifu') for 3 weeks in the growth phase (luminous intensity 290 μmol · m ^-2 · sec ^-1 , photoperiod treatment 16 h / ℃), growth humidity (40 ~ 70%), total RNA was extracted by salt stress (250mM NaCl) for 48 hours and KBGP-24K oligo chip was used to compare the expression of each gene. Provide the analyzed information. Using the " Brassica rapa EST and Microarray Database", the present inventors classify the genes whose expression is strongly increased in the response genes during the salt stress treatment, and selected the target genes having a complete intestine and unknown function.

To identify the novel resistance genes in Chinese cabbage, the cDNA sequence (AT2G44670) having homology in the 285bp open reading frame (ORF) and Arabidopsis thaliana in the KBGP-24K oligo chip (AT2G44670) and the Brassica rapa subsp. A pair of primers were prepared based on the nucleotide sequence of the pekinensis clones (AC189399, AC189261), and the sequence thereof is shown in Table 1 below.

[Table 1]

Example 2: RNA isolation from Chinese cabbage

The leaf of Chinese cabbage ( Brassica rapa ssp. Pekinensis ) was disrupted by using liquid nitrogen, and 1 ml of trizol (Trizol, Gibco BRL) solution was added and mixed well and left at room temperature for 5 minutes. After adding 200 μl of chloroform, the mixture was centrifuged at 13,000 rpm for 10 minutes at 4 ° C, and the supernatant was transferred to a new tube. Isopropanol (500 μl) was added and centrifuged at 13,000 rpm for 15 minutes to precipitate. Isopropanol was removed and the precipitate was washed with 75% ethanol. Then, it was well dried without ethanol and dissolved in sterilized distilled water treated with DEPC (diethyl pyrocarbonate).

Example  3: Segregation of salt-related genes

To isolate the salt tolerance-related gene, cDNA was synthesized using oligo dT primer and reverse transcriptase using the obtained RNA as a template, and then RT-PCR was performed with the primer prepared in [Table 1] Respectively.

The cDNA synthesis program conditions are as follows.

1 의 of RNA was added to the oligo dT primer and stored at 70 째 C to remove the RNA secondary structure, followed by reverse transcription at 45 째 C for 30 minutes, and then kept at 94 째 C for 5 minutes to inactivate the reverse transcriptase used. Then, 1 μg of the synthesized cDNA was subjected to PCR using the primers shown in Table 1 and tag polymerase (Taq polymerase). The PCR program was run for 40 cycles at 94 ° C for 30 seconds, at 61 ° C for 30 seconds and at 72 ° C for 30 seconds, followed by final extension at 72 ° C for 10 minutes , And the whole reaction was terminated at 4 ° C.

As a result of the experiment, the amplified salt resistance related gene fragment was confirmed by electrophoresis (FIG. 1). The fragments amplified with each of the primers were cloned into a vector pGEM-T easy vector for PCR product to analyze their respective base sequences to construct a recombinant vector.

Experimental Example  1. E. coli using recombinant vector. coli DH10 Transformation of β

E. coli was inoculated to LB DH10β 50㎖ OD ₅₇₀ at 37 ℃ = 0.375-0.400, the culture broth was centrifuged at 3,000 rpm for 10 minutes, and the resulting precipitate was dissolved in 0.1 M CaCl ₂ solution and then stored in ice for 30 minutes. Thereafter, the mixture was centrifuged at 3,000 rpm for 7 minutes, and the precipitate was dissolved in 0.1 M CaCl _2. The recombinant vector of Example 3 was mixed with ice for 30 minutes, and then thermally shocked at 42 ° C for 90 seconds. Thereafter, the cells were left on ice for 10 minutes, and then 700 L of LB medium was added, followed by shaking culture at 37 캜 for 1 hour. After centrifuging the culture at 12000 rpm for several seconds, only 100 μl was left, the supernatant was removed, and the precipitate was dissolved with the remaining supernatant. Thereafter, a precipitate dissolved in a solid LB medium supplemented with ampicillin (50 mg / L), X-gal (200 mg / L in NN dimethylformamide) and IPTG (200 mg / L) was developed, Transformed colonies showing resistance to the solid medium and forming white colonies were selected after incubation.

Experimental Example  2: Recombination DNA  Detection of isolation and gene insertion

The selected transformant colonies were inoculated in 3 ml of LB medium supplemented with antibiotics and cultured at 37 占 폚 for 12 hours, and the culture broth was centrifuged at 4 占 폚 and 12,000 rpm for 10 minutes. Solution I (200 μl / ml) is added to the precipitated microbial cells, followed by vortexing for several seconds. Afterwards, add Solution II (200 μl / ml) and mix carefully to completely break the cell membrane. And allowed to stand at room temperature for 5 minutes. After that, Solution III (200 μl / ml) was added and mixed well. After allowing to stand for 10 minutes on ice, supernatant was obtained by centrifugation at 12,000 rpm for 10 minutes at 4 ° C. Recombinant DNA was precipitated at -20 ° C for 2 hours by adding 1/10 NaOAc and 2.5-fold 100% ethanol to the obtained supernatant, and centrifuged to obtain a precipitate. The precipitate was washed with 70% ethanol and dissolved in sterile distilled water.

To confirm the insertion of the resistance-related genes amplified from Chinese cabbage into the pGEM-T easy vector, restriction enzyme EcoR I was confirmed by treating the isolated recombinant DNA (FIG. 2). The nucleotide sequence was examined using T7 and SP6 primers close to the insertion site.

Experimental Example  3. Identification of the entire sequence of the salt tolerance-related gene

Sequencing of the PCR fragments cloned into the pGEM-T easy vector for sequencing was performed using the GenBank BLAST program. The nucleotide sequence and amino acid sequence of the 285 bp product amplified with BrSSR-F (SEQ ID NO: 1) and BrSSR-R (SEQ ID NO: 2) primers were analyzed using GenBank BlastX program and CLC free workbench 3.2.3 program. ( Fragaria vesca , Camelina sativa , Arabidopsis thaliana , soybean ( Glycine max ), cucumber ( Cucumis sativus sequence (Fig. 3 and Fig. 4). That is, RT-PCR was performed using BrSSR-F and BrSSR-R primers for cabbage RNA, and it was confirmed that the entire genes of the salt tolerance related genes of Chinese cabbage were amplified (FIG. 4). In addition, it was confirmed that the gene isolated by using the NCBI Conserved Domain program had a functionally undisclosed domain 581 (Domain of unknown function 581, DUF 16), and that the E-value was 2.23e ^-31 , . The starting codon and the termination codon were determined based on the nucleotide sequence of the identified SEQ ID NO: 7 to determine the amino acid sequence of SEQ ID NO: 8, which was named as the full length salt tolerant gene BrSSR isolated from Chinese cabbage.

Example  4. BrSSR  Generation of vectors for gene overexpression

In order to over-express the isolated BrSSR gene, a brSRS gene of 285 bp was inserted into pH2GW7 (VIB-Ghent University, Belgium) to construct pSL94 for transformation (Fig. 6).

Example  5. PCR  Selection of transgenic plants through assay

Transformation of tobacco transformants produced using pSL94 vector was confirmed. First, a PCR assay was performed on the hygromycin resistance gene (hpt), and all of the six individuals produced were identified as transformants (FIG. 7). The PCR program was run for 40 cycles of 94 ° C for 30 seconds, 60 ° C for 30 seconds, and 72 ° C for 1 minute, after the initial denaturation time of 94 ° C for 2 minutes, After extension, the whole reaction was terminated by maintaining at 4 ° C. The nucleotide sequences of the primers HPT-F (SEQ ID NO: 3) and HPT-R (SEQ ID NO: 4) used for PCR assays are shown in Table 2.

[Table 2]

Example  6. Quantitative real - time RT PCR  Analysis of Transformants BrSSR  Analysis of gene expression level

The expression level of the BrSSR gene in the body was analyzed by quantitative real-time RT-PCR in the case of salt stress in 6 individuals identified as transformants in Example 5. As a result, the expression level was about 1.9 to 3.8 times higher than that of the non-transformant (CON) (FIG. 8). The PCR program was first denatured at 95 ° C for 10 minutes, followed by cDNA synthesis for 30 minutes at 42 ° C. Thereafter, the reaction was repeated 35 times at 95 캜 for 10 seconds, 59 캜 for 15 seconds, and 72 캜 for 20 seconds. In the expression analysis method, fluorescence intensity data according to the amount of amplification was measured at the end of each cycle, and the measured values were analyzed by a Rotor-Gene 6000 analysis software. The BrSSR expression level of each transformant was calculated relative to the expression level of the actin gene. The nucleotide sequences of the used primers SSReal-F (SEQ ID NO: 5) and SSReal-R (SEQ ID NO: 6) are shown in Table 3.

[Table 3]

Example  7. Salt tolerance test of tobacco transformants when salt stress was issued

Hypromycin resistant genes were used to identify the transgenic plants. Six genotypes of transgenic plants were identified by quantitative real-time RT PCR analysis. Were examined under the condition of 250 mM NaCl salt treatment. Salt stress is caused by the presence of any non-biological stimuli (moisture, temperature, light, etc.) or biological stimuli (pathogen infection, pest damage, etc.) during the course of salt treatment, (Light intensity of 250 μmol · m ^-2 · sec ^-1 , photoperiod treatment for 16 hours, treatment for 8 hours, growth temperature of 22 ° C., humidity of 30 to 50%). (Growth stopping, leaf color change, whether or not they were dressed).

As a result, at the 5th day of the salt treatment, the control group, non-transformant, lost its elasticity of leaves and stems and collapsed. As the leaf color became gradually softened, it began to change to yellow and showed no growth. On the other hand, all of the transgenic 6 individuals remained the same as before the salt treatment, showing no significant difference in the expression form (FIG. 9).

<110> UNIVERSITY-INDUSTRY COOPERATION GROUP OF KYUNG HEE UNIVERSITY <120> Gene Implicated in Salt Stress Tolerance and Transformed Plants          with the Same <130> SDP50497 <150> KR 10-2013-0119094 <151> 2013-10-07 <160> 8 <170> Kopatentin 2.0 <210> 1 <211> 22 <212> DNA <213> Artificial Sequence <220> <223> PCR primer <400> 1 atggcttcgt attactctgc gt 22 <210> 2 <211> 19 <212> DNA <213> Artificial Sequence <220> <223> PCR primer <400> 2 ctacgcgacc acgagtgtt 19 <210> 3 <211> 16 <212> DNA <213> Artificial Sequence <220> <223> PCR primer <400> 3 tttccactgc gagtac 16 <210> 4 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> PCR primer <400> 4 tgtcgagaag tttctgatcg a 21 <210> 5 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> PCR primer <400> 5 atggcttcgt attactctgc g 21 <210> 6 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> PCR primer <400> 6 cacgagtgtt cctgtccgta 20 <210> 7 <211> 285 <212> DNA <213> Brassica rapa ssp. pekinensis <400> 7 atggcttcgt attactctgc gtttttcggc agtgaagagc cacactttct ggaatcatgc 60 tctctttgca gtaaacacct tggtcctaac tccgacatct tcatgtacag aggagacacg 120 gcgttttgta gcaatgagtg tagagaagaa cagattgaat ctgatgaagc caaggagaga 180 cgctggaaac tgtctgctag atctctccgt aaaaaatctt ctgaagctgc aaaagattcg 240 gccgccggaa aaaacgtacg gacaggaaca ctcgtggtcg cgtag 285 <210> 8 <211> 94 <212> PRT <213> Brassica rapa ssp. pekinensis <400> 8 Met Ala Ser Tyr Tyr Ser Ala Phe Phe Gly Ser Glu Glu Pro His Phe   1 5 10 15 Leu Glu Ser Cys Ser Leu Cys Ser Lys His Leu Gly Pro Asn Ser Asp              20 25 30 Ile Phe Met Tyr Arg Gly Asp Thr Ala Phe Cys Ser Asn Glu Cys Arg          35 40 45 Glu Glu Gln Ile Glu Ser Asp Glu Ala Lys Glu Arg Arg Trp Lys Leu      50 55 60 Ser Ala Arg Ser Leu Arg Lys Lys Ser Ser Glu Ala Ala Lys Asp Ser  65 70 75 80 Ala Ala Gly Lys Asn Val Thr Gly Thr Leu Val Val Ala                  85 90

Claims (7)

8. A composition for promoting salt tolerance of a plant comprising a nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 8. The composition according to claim 1, wherein the nucleotide sequence is a nucleotide sequence of SEQ ID NO: 7. (a) the nucleotide sequence of claim 1 or 2; (b) a promoter operably linked to said nucleotide sequence and forming RNA molecules in plant cells; And (c) a recombinant vector for plant expression comprising a poly A signal sequence that acts in plant cells to cause polyadenylation of the 3'-terminal of the RNA molecule. A salt tolerant plant cell transformed with the composition of claim 1 or 2. A salt tolerant plant transformed with the composition of claim 1 or 2. 6. The method according to claim 5, wherein the plant is selected from the group consisting of food crops including rice, wheat, barley, corn, soybean, potato, wheat, red bean, oats and millet; Vegetable crops including Arabidopsis, cabbage, radish, pepper, strawberry, tomato, watermelon, cucumber, cabbage, melon, squash, onions, onions and carrots; Special crops including ginseng, tobacco, cotton, sesame, sugar cane, beet, perilla, peanut and rapeseed; Apple trees, pears, jujubes, peaches, sheep, grapes, citrus, persimmon, plums, apricots and banana; Roses, gladiolus, gerberas, carnations, chrysanthemums, lilies and tulips; And feed crops including rice grains, red clover, orchardgrass, alpha-alpha, tall fescue, and fermented rice grains. A method for increasing the intracellular expression level of a polypeptide having an amino acid sequence represented by SEQ ID NO: 8 to enhance the salt tolerance of a plant.

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