KR101485831B1 - Gene Implicated in Cold Stress Tolerance and Transformed Plants with the Same - Google Patents

Abstract

The present invention relates to a composition for reinforcing the low temperature resistance of a plant. A nucleotide sequence of the present invention is involved in the resistance of a plant against low temperature stress, and a transgenic plant overexpressed therefrom has excellent resistance against low temperature stress. Accordingly, the transgenic plant of the present invention has excellent resistance against several kinds of stress such as abnormal low temperature and cold-weather damage so as to be usefully used as a novel functional plant having excellent adaptivity to an abnormal climate in a growing area.

Description

Gene Implicated in Cold Stress Tolerance and Transformed Plants with the Same < RTI ID = 0.0 >

The present invention relates to Chinese cabbage rapa ssp . The low-temperature resistance separated from pekinensis) associated BrCSR (Brassica rapa cold stress resistance gene and transgenic plants.

Abiotic stresses such as high concentration of salt accumulation, drying, and low temperature are important factors causing changes in growth and growth pattern, physiological and biochemical changes, and yields in most crops. Among them, low temperature stress causes degradation of enzyme activity, unbalance of energy metabolism, and changes in transcriptome, proteome, and metabolome, and therefore researches for detecting low temperature resistant genes in various crops are actively conducted.

In rice, for example, GPAT (arabidopsis glycerol-3P-acyltransferase), OsCDPK7 (rice calcium-dependent protein kinase), OsDREB1 (rice DRE-binding protein 1), HvCBF4 (barley C-repeat binding factor), OsCIPK03 Like genes such as B-like protein-interaction protein kinase 03 and OsTPP1 (trehalose-6-phosphate phosphatase) were found. In the case of corn, NPK1 (nicotiana protein kinase) and Arabidopsis MYB15 (MYB-like transcription factor) Were identified as representative low-temperature resistance genes. In the case of GPAT gene, it was reported that GPAT gene was introduced into rice and 28% of unsaturated fatty acid was increased. At 17 ℃, the photosynthetic efficiency was increased up to 20%, which minimized the growth of rice in low temperature environment. Inhibition of the MYB15 gene in Arabidopsis has been reported to increase cold resistance by increasing the expression of CBF (C-repeat binding factor) genes associated with low temperature resistance.

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.

Currently, food consumption is continuously increasing due to the increase in population, but cultivable cultivation areas are gradually decreasing due to global warming. Korea is in a temperate climate, but crop damage is frequent due to abnormal low temperature due to global climate change. The degradation of agricultural productivity due to the extreme low temperature 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 low temperature resistance of plants. As a result, the present inventors have completed the present invention by confirming that the nucleotide sequence of SEQ ID NO: 7 is involved in the above-described trait of the plant and that the gene is transformed into a plant, thereby improving the low temperature resistance of the plant.

Accordingly, an object of the present invention is to provide a composition for promoting low temperature resistance of a plant, and a plant cell or plant having enhanced low temperature resistance transformed with the composition.

It is another object of the present invention to provide a method for promoting low temperature resistance 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 promoting low-temperature resistance 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 low temperature resistance of plants. As a result, it was confirmed that the nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 8 was involved in the above-mentioned trait of the plant, and that when the gene was transformed into a plant, the low temperature resistance of the plant could 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 invention, the nucleotide sequence of SEQ ID NO: 7 was isolated from Chinese cabbage of the dielectric, and the name of the gene is BrCSR (Brassica rapa Cold Stress Resistance). The gene contains a core 16 protein domain (NOP16).

According to the present invention, the present inventors have found that when a cold stress is applied to a plant, the expression of BrCSR gene is increased and the resistance to cold stress is increased when the expression of the gene 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. BLAST 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 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.

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, as well as the promoter of the cytosolic triphosphate isomerase (TPI) of Arabidopsis thaliana, the light tolerance promoter of ssRUBISCO, the cytosolic triphosphate isomerase 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 still another aspect of the present invention, there is provided a plant cell transformed with the composition of the present invention and having enhanced low temperature resistance.

According to another aspect of the present invention, there is provided a 'plant' enhanced in low temperature resistance 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 enhancing the low temperature resistance of a plant by increasing the intracellular expression level of a polypeptide having an amino acid sequence represented by SEQ ID NO: 8.

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 promoting low temperature resistance of a plant.

(b) The nucleotide sequence of the present invention is involved in tolerance to cold stress of a plant, and the transgenic plant overexpressing it is excellent in tolerance to various stresses including abnormal cold or cold weather, It can be used as an excellent new functional crop.

1 is a photograph of a fragment of a gene encoding BrCSR amplified from a Chinese cabbage by RT-PCR using BrCSR-F and BrCSR-R primer pairs.
Fig. 2 shows the results of RT-PCR using BrCSR-F and BrCSR-R primer pairs. The gene encoding BrCSR amplified from Chinese cabbage was inserted into pGEM-T easy vector, It is a photograph.
Figure 3 is the sequence of the gene encoding the BrCSR of discrete cabbage by RT-PCR using the GenBank BlastX program and CLC free workbench 3.2.3 program included pyulra (Populus trichocarpa ), Arabidopsis thaliana , grapes ( Vitis vinifera ), and castor ( Ricinus communis sequence and homology.
4 shows the nucleotide sequence and amino acid sequence of the gene encoding BrCSR of Chinese cabbage separated by RT-PCR.
FIG. 5 shows the result of analyzing the gene encoding BrCSR of Chinese cabbage separated by RT-PCR using the NCBI Conserved Domain program.
FIG. 6 is a diagram showing an over-expression vector for transformation (pSL101) overexpressing BrCSR gene of Chinese cabbage separated by RT-PCR.
FIG. 7 shows the results of PCR analysis of the transformation of tobacco transformants produced using the BrCSR overexpression vector.
FIG. 8 shows the results of quantitative real-time RT PCR analysis of the expression level of the BrCSR gene in tobacco transformants transfected with the PCR assay.
FIG. 9 shows the result of analysis of phenotype after cold stress treatment of tobacco transformants showing low temperature resistance.

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 BrCSR gene identification process and the low temperature resistance assay of the present invention will be described.

Example 1. Investigation of low-temperature resistant genes derived from Chinese cabbage

The information of " Brassica rapa EST and Microarray Database" was used to identify the new low temperature resistant 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 / (4 ° C) for 48 hours, total RNA was extracted at each stage of growth, 127,144 Chinese cabbage EST analyzes were performed using a cDNA library, and KBGP-24K oligo chip to provide information that compares and analyzes changes in expression of each gene. Using the " Brassica rapa EST and Microarray Database", the present inventors classify the genes whose expression is strongly increased in the genes responsive to low-temperature stress treatment, and select low-resistance genes among the genes whose function is unknown Respectively.

For identification of low temperature resistant genes in Chinese cabbage, cDNA sequence (AT1G02870) having homology in the 585bp ORF (Open Reading Frame) and Arabidopsis thaliana of the KBGP-24K oligo chip and Brassica rapa subsp. Based on the nucleotide sequence of pekinensis clone KBrB030F10 (AC189302), a pair of primers were prepared, and the sequence thereof is shown in Table 1 below.

[Table 1]

Example 2. Isolation of RNA 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. Isolation of low-temperature resistant genes

To isolate the low-temperature resistant gene, cDNA was synthesized using oligo dT primer and reverse transcriptase using the obtained RNA as a template, and then RT-PCR was performed using the primers 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 of 94 ° C for 30 seconds, 60 ° C for 30 seconds, 72 ° C for 1 minute, and finally a final extension at 72 ° C for 10 minutes After that, the whole reaction was terminated by maintaining the temperature at 4 ° C.

A fragment of the amplified low-temperature resistant gene 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 II (200 μl / ml) was added to the precipitated microbial cells, followed by vortexing for a few seconds, followed by Solution II (200 μl / ml) and carefully mixed 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 low-temperature resistant gene 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 whole sequence of low-temperature resistance 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 585 bp product amplified with BrCSR-F (SEQ ID NO: 1) and BrCSR-R (SEQ ID NO: 2) primer pairs were analyzed using GenBank BlastX program and CLC free workbench 3.2.3 program. Populus trichocarpa ), Arabidopsis ( Arabidopsis thaliana , grapes ( Vitis vinifera ), castor ( Ricinus communis sequence (FIG. 3 and FIG. 4). That is, RT-PCR was performed using the pair of BrCSR-F and BrCSR-R primers for the cabbage RNA, and it was confirmed that the entire gene of the low-temperature resistance gene of the Chinese cabbage was amplified (FIG. In addition, it was confirmed that the nucleotide 16 domain (NOP16) was present in the nucleotide sequence of the isolated gene using the NCBI Conserved Domain program, and it was confirmed that the E-value was 1.14e ^-35 and the accuracy was high (FIG. 5) . 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 a full-length low temperature resistance gene BrCSR isolated from Chinese cabbage .

Example  4. BrCSR  Generation of vectors for gene overexpression

In order to over-express the isolated BrCSR gene, a BrCSR gene of 585 bp was inserted into pH2GW7 (VIB-Ghent University, Belgium) to construct pSL101, a transformation vector (FIG. 6).

Example  5. PCR  Selection of transgenic plants through assay

Transformation of tobacco transgenic plants produced using the pSL101 vector was confirmed. First, a PCR assay was performed on the hygromycin resistance gene (hpt), and all three individuals produced were confirmed 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 the PCR assay are shown in Table 2 below.

[Table 2]

Example  6. Quantitative real - time RT PCR  Analysis of transformants BrCSR  Analysis of gene expression level

The expression level of the BrCSR gene in the body was analyzed by quantitative real-time RT-PCR at the time of low-temperature stress in three individuals identified as the transformant of Example 5. As a result, all three individuals showed about two times higher expression levels than non-transformants (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. BrCSR expression level of each transformant was calculated relative to the expression level of actin gene. The nucleotide sequences of the used primers CSReal-F (SEQ ID NO: 5) and CSReal-R (SEQ ID NO: 6) are shown in Table 3 below.

[Table 3]

Example  7. Low temperature resistance test of tobacco transformants at low temperature stimulation

Hypromycin resistance genes were used to identify the transgenic plants. Three genotypes of transgenic plants were identified by quantitative real-time RT-PCR analysis. Low temperature resistance). The low temperature stimuli can be applied to any non-biological stimuli (moisture, temperature, light, etc.) or biological stimuli (pathogen infection, pest damage, etc.) during the cold treatment of the non-transformant and transformant, ) Was found to be phenotypically distinct in a limited condition (luminous intensity of 250 μmol · m ^-2 · sec ^-1 , photoperiod treatment of 16 hours / treatment of 8 hours, treatment temperature of 4 ° C., humidity of 30 to 50% (Growth stopping, leaf color change, whether or not they were dressed).

As a result, on the fifth day of low temperature treatment, the control non-transgenic tobacco lost the elasticity of leaves and stems and fell down without force, and the leaf color became gradually softened and no longer grows. On the other hand, all three transformants showed the same appearance as before the low temperature treatment and showed no difference in the expression shape (FIG. 9).

<110> UNIVERSITY-INDUSTRY COOPERATION GROUP OF KYUNG HEE UNIVERSITY <120> Gene Implicated in Cold Stress Tolerance and Transformed Plants          with the Same <130> SDP50498 <150> KR 10-2013-0119090 <151> 2013-10-07 <160> 8 <170> Kopatentin 2.0 <210> 1 <211> 19 <212> DNA <213> Artificial Sequence <220> <223> PCR primer <400> 1 atggcgaggt caaggagga 19 <210> 2 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> PCR primer <400> 2 tcagaatgga actaaaacag g 21 <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> 18 <212> DNA <213> Artificial Sequence <220> <223> PCR primer <400> 5 atggcgaggt caaggagg 18 <210> 6 <211> 19 <212> DNA <213> Artificial Sequence <220> <223> PCR primer <400> 6 tcagaatgga actaaaaca 19 <210> 7 <211> 585 <212> DNA <213> Brassica rapa ssp. pekinensis <400> 7 atggcgaggt caaggaggaa gtacaagaac tcaagagcca aagtacgcgt cgcactcccc 60 aagaagaacc caaacatctt caaaccatct ttcaacttcc ctcccaagct ccgcgccctc 120 atggccgacg acgtccccga gtgggacgat caagccagcg tcatccagaa ctacaaatcc 180 ttcggcgtca tctccaaccc taacttgctc ggcatccgat ctcgcaccga ccatatgatc 240 caggacgact ccctcaacgt tcccccacct cctcagccgc cgaccgatga tcctgtggct 300 aaggagttcg agcctatcga ttccggtagc gaactcgagg aagacgatct taagacagca 360 ttagggaagc aaagaaaaga tggaaagagt gctcctttac agccacttac tactatgcaa 420 cgtacgcata tcagacgttt ggttgagaaa catggtgatg acattgagtc catgtaccgg 480 gacaggaagc taaactcgat gcagcattcg gttgcaacac tacagaagct gtgcacaaga 540 tatcacatgt acaaggataa gaaccctgtt ttagttccat tctga 585 <210> 8 <211> 194 <212> PRT <213> Brassica rapa ssp. pekinensis <400> 8 Met Ala Arg Ser Arg Arg Lys Tyr Lys Asn Ser Arg Ala Lys Val Arg   1 5 10 15 Val Ala Leu Pro Lys Lys Asn Pro Asn Ile Phe Lys Pro Ser Phe Asn              20 25 30 Phe Pro Pro Lys Leu Arg Ala Leu Met Ala Asp Asp Val Pro Glu Trp          35 40 45 Asp Asp Gln Ala Ser Val Ile Gln Asn Tyr Lys Ser Phe Gly Val Ile      50 55 60 Ser Asn Pro Asn Leu Leu Gly Ile Arg Ser Ser Thr Asp His Met Ile  65 70 75 80 Gln Asp Ser Ser Leu Asn Val Pro Pro Pro Gln Pro Pro Thr Asp                  85 90 95 Asp Pro Val Ala Lys Glu Phe Glu Pro Ile Asp Ser Gly Ser Glu Leu             100 105 110 Glu Asp Asp Leu Lys Thr Ala Leu Gly Lys Gln Arg Lys Asp Gly         115 120 125 Lys Ser Ala Pro Leu Gln Pro Leu Thr Thr Met Gln Arg Thr His Ile     130 135 140 Arg Arg Leu Val Glu Lys His Gly Asp Asp Ile Glu Ser Met Tyr Arg 145 150 155 160 Asp Arg Lys Leu Asn Ser Met Gln His Ser Val Ala Thr Leu Gln Lys                 165 170 175 Leu Cys Thr Arg Tyr His Met Tyr Lys Asp Lys Asn Pro Val Leu Val             180 185 190 Pro Phe        

Claims (7)

A composition for promoting low temperature resistance of a plant comprising a nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 8. The composition for promoting low temperature resistance of a plant 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 plant cell with enhanced low temperature resistance transformed with the composition of claim 1 or 2. A plant having enhanced low temperature resistance 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 enhancing the low temperature resistance of a plant by increasing the intracellular expression level of a polypeptide having an amino acid sequence represented by SEQ ID NO: 8.

Images