KR101973551B1 - Method for producing transgenic plant with increased environmental stress resistance using BrRH22 gene from Brassica rapa and plant thereof - Google Patents

Abstract

The present invention relates to a method for producing a transgenic plant having increased resistance to environmental stress using a BrRH22 gene derived from Brassica rapa and a plant produced thereby. More specifically, a transgenic plant overexpressing the BrRH22 gene derived from Brassica rapa according to the present invention exhibits salt and drought stress tolerance. Therefore, the BrRH22 gene is expected to be useful for the development of the transgenic plant resistant to environmental stress, and as the plant resistant to environmental stress can be obtained by using the same, it is expected that the present invention can greatly contribute to an increase in yield of profit crops and the like.

Description

FIELD OF THE INVENTION [0001] The present invention relates to a method for producing transgenic plants having increased resistance to environmental stress using BrRH22 gene derived from Chinese cabbage, and a method for producing transgenic plants using BrRH22 gene from Brassica rapa and plant thereof,

The present invention relates to a method for producing a transgenic plant having increased resistance to environmental stress using a BrRH22 gene derived from Chinese cabbage and a plant.

Plants are exposed to a variety of environmental stresses such as low temperature, high temperature, drought, and high salinity, which greatly reduces crop production. Photosynthetic activity in chloroplasts is an important cellular response to plant growth and survival under normal or stress conditions. In particular, photosynthesis acts as a stress sensor, and thus the expression of genes that affect photosynthesis in chloroplasts must be well controlled for plant growth and survival under stress conditions. Gene expression in chloroplasts is regulated primarily at post-transcriptional stages such as RNA processing, intron splicing, RNA editing, RNA degradation and translation. There are about 150 genes in the chloroplast genome, but proteins encoded in more than 3,000 nuclei are transferred to the chloroplast and are involved in the regulation of gene expression in the chloroplast. Thus, the interaction between nuclei and chloroplasts is important for gene regulation between two intracellular organelles.

RH (RNA helicase) is a type of RNA-binding protein that regulates post-transcriptional gene expression by participating in post-transcriptional RNA metabolism and RNA structure formation such as intron splicing, mRNA transport and collapse. The DEAD-box RH (RNA helicase) family is the largest of the six RH Superfamily II (SF II) and is well studied.

There are many reports demonstrating the function of DEAD-box RH present in the nucleus or cytoplasm, but reports on the function of DEAD-box RH present in cytoplasm, such as chloroplasts or mitochondria, are extremely limited to date. The RH3 and HVD1 genes present in the chloroplasts respond to salt and drought stresses, and the RH3 , PLT1 and ABO6 genes, which are DEAD-box RHs that are transferred to chloroplasts or mitochondria, have been reported to play an important role in the regulation of ABA or auxin signaling. These series of studies clearly show that RH plays an important role in plant response to diverse environmental factors in dicotyledonous plants such as Arabidopsis. In particular, chloroplasts are important cell organelles that cause photosynthesis directly linked to the growth of plants. It is important to study the function of DEAD-box RH related to RNA metabolism in chloroplasts.

On the other hand, Korean Patent No. 1052565 discloses a novel RNA helicase LT28 gene and its use, and Korean Laid-Open Patent Application No. 2014-0050218 discloses a method for producing a transgenic plant having enhanced environmental stress tolerance using a BrRZFP1 gene derived from Chinese cabbage Methods and plants therefor. However, the methods for producing transgenic plants with increased resistance to environmental stress using the BrRH22 gene derived from Chinese cabbage of the present invention and the plants therefor have not yet been disclosed.

The present invention has been made in view of the above-mentioned needs, and it has been confirmed that the transgenic plants overexpressing the BrRH22 gene derived from Chinese cabbage belonging to the DEAD-box RH (RNA helicase) exhibit resistance under salt and drought stress conditions , Confirming that the BrRH22 gene has a function of regulating the environmental stress tolerance as described above, thereby completing the present invention.

In order to solve the above-described problems, the present invention provides a method for regulating expression of a BrRH22 gene by transforming a plant cell with a recombinant vector comprising a gene encoding Brassica rapa RNA helicase 22 (BrRC22) derived from Chinese cabbage Lt; RTI ID = 0.0 > environmental stress tolerance. ≪ / RTI >

The present invention also provides a method for producing a recombinant vector comprising the steps of: transforming a plant cell with a recombinant vector comprising the amino acid sequence of SEQ ID NO: 2 and comprising a gene encoding Brassica rapa RNA helicase 22 protein derived from Chinese cabbage; And

And regenerating the plant from the transformed plant cell. The present invention also provides a method for producing a transgenic plant having regulated environmental stress tolerance.

In addition, the present invention provides a transgenic plant having the stress tolerance regulated by the above method and a transformed seed thereof.

The present invention also provides a composition for controlling the environmental stress tolerance of a plant comprising the amino acid sequence of SEQ ID NO: 2, as an active ingredient, a gene coding for a Brassica rapa RNA helicase 22 protein derived from Chinese cabbage.

The transgenic plants overexpressing BrRH22 gene from Chinese cabbage according to the present invention were found to be resistant to salt and drought stress. Therefore, the BrRH22 gene is expected to be useful for the development of a transgenic plant resistant to environmental stress, and it is possible to obtain a plant resistant to environmental stress by using the BrRH22 gene, which can contribute to increase in yield of economic crops It is expected to be.

FIG. 1 shows the result of confirming the domain structure (A) and the intracellular location (B) of Brassica napus BrRH22 protein according to an embodiment of the present invention. The red signal means the automatic fluorescence of the chloroplast, and the scale bar is 1 탆.
FIG. 2 shows the results of measurement of seed germination rate of BrRH22 overexpressing Arabidopsis plants under salt (A) and drought stress (B) conditions according to an embodiment of the present invention. WT is a wild-type Arabidopsis plant and OX1, OX2 and OX3 represent overexpressing Arabidopsis plants.
FIG. 3 is a photograph (A) showing seedling growth of BrømmH22 overexpressing Arabidopsis plants under salt stress conditions according to an embodiment of the present invention and a graph (B) showing fresh weight and root length )to be. A, the scale bar is 1 cm and B * indicates that the increase in root weight and root length of BrJH22 overexpressing Arabidopsis plants (OX1, OX2 and OX3) is statistically significant compared to the wild type (WT) under salt stress conditions .
FIG. 4 is a photograph (A) showing seedling growth of BrømmH22 overexpressed Arabidopsis plant and a graph (B) showing fresh weight and root length under drought stress conditions according to an embodiment of the present invention. A, the scale bar is 1 cm and B * indicates that the increase in root weight and root length of BrRH22 overexpressing Arabidopsis plants (OX1, OX2 and OX3) is statistically significant compared to wild type (WT) under drought stress conditions Means that the p value is less than 0.05.
5 is a photograph showing the survival rate of BrRH22 overexpressed Arabidopsis plants recovered from salt stress according to one embodiment of the present invention.
6 is a photograph showing the survival rate of BrRH22 overexpressed Arabidopsis plants recovered from drought stress according to an embodiment of the present invention.

In order to accomplish the object of the present invention, the present invention includes a step of transforming a plant cell with a recombinant vector comprising a gene encoding Brassica rapa RNA helicase 22 protein derived from Chinese cabbage to regulate the expression of the BrRH22 gene The present invention provides a method for controlling the environmental stress tolerance of a plant.

The BrRH22 protein according to the present invention may comprise the amino acid sequence of SEQ ID NO: 2, and the BrRH22 gene may comprise the nucleotide sequence of SEQ ID NO: 1, but is not limited thereto.

The BrRH22 protein of the present invention comprises a protein having an amino acid sequence represented by SEQ ID NO: 2 and a functional equivalent of the protein. Is at least 70% or more, preferably 80% or more, more preferably 90% or more, still more preferably 90% or more, more preferably 90% or more, Quot; refers to a protein having a homology of at least 95% with a physiological activity substantially equivalent to that of the protein represented by SEQ ID NO: 2. &Quot; Substantially homogenous bioactivity " means an activity that regulates environmental stress tolerance of a plant.

In a method according to an embodiment of the present invention, a plant cell is transformed with a recombinant vector comprising the gene of SEQ ID NO: 1 by overexpression of the BrRH22 gene, so as to control the environmental stress tolerance of the plant, , But is not limited thereto.

By "gene overexpression" is meant that the gene is expressed at a level that is expressed in a wild-type plant. As a method of introducing the gene into a plant, there is a method of transforming a plant using an expression vector containing the gene under the control of a promoter.

In the present invention, " environmental stress " refers to an external factor that lowers the growth or productivity of a plant, and is roughly divided into biotic stress and abiotic stress. Biological stresses are typically pathogens. Abiotic stresses include high salt, drought (dry), low temperature, high temperature and oxidative stress. The term " environmental stress tolerance " refers to a trait that suppresses or delays plant growth or productivity deterioration due to environmental stress.

In the method according to an embodiment of the present invention, the environmental stress may be salt or drought stress, but is not limited thereto.

As used herein, the term " salt tolerance " means that plants that have undergone salt stress at 50 mM to 300 mM NaCl, more preferably 75 mM to 150 mM NaCl, exhibit physiological activity (such as germination or seedling growth) it means.

The term " drought tolerance " in the present invention means a plant having drought stress in 100 to 450 mM of mannitol, preferably 100 mM to 300 mM of mannitol, exhibits a higher survival rate than normal plants or prevents damage to drought- Quot; means exhibiting improved activity.

The term " recombinant " refers to a cell in which a cell replicates a heterologous nucleic acid, expresses the nucleic acid, or expresses a protein encoded by a peptide, heterologous peptide or heterologous nucleic acid. The recombinant cell can express a gene or a gene fragment that is not found in the natural form of the cell in one of the sense or antisense form. In addition, the recombinant cell can express a gene found in a cell in its natural state, but the gene has been modified and reintroduced intracellularly by an artificial means.

In the present invention, the BrRH22 gene sequence can be inserted into a recombinant expression vector. The term " recombinant expression vector " means a bacterial plasmid, a phage, a yeast plasmid, a plant cell virus, a mammalian cell virus, or other vector. In principle, any plasmid and vector can be used if it can replicate and stabilize within the host.

Expression vectors comprising the BrRH22 gene sequences of the invention and appropriate transcription / translation control signals can be constructed by methods known to those skilled in the art. Such methods include in vitro recombinant DNA technology, DNA synthesis techniques, and in vivo recombination techniques. The DNA sequence can be effectively linked to appropriate promoters in the expression vector to drive mRNA synthesis. The expression vector may also 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 a Ti-plasmid vector capable of transferring a so-called T-region to a plant cell when present in a suitable host, such as Agrobacterium tumefaciens. Other types of Ti-plasmid vectors (see EP 0 116 718 B1) are currently used to transfer hybrid DNA sequences to plant cells or protoplasts in which new plants capable of properly inserting hybrid DNA into the plant's genome can be produced have. A particularly preferred form of the Ti-plasmid vector is a so-called binary vector as claimed in EP 0 120 516 B1 and U.S. Patent No. 4,940,838. Other suitable vectors that can be used to introduce the DNA according to the invention into the plant host include viral vectors such as those that can be derived from double-stranded plant viruses (e. G., CaMV) and single- For example, from non -complete plant virus vectors. The use of such vectors may be particularly advantageous when it is difficult to transform the plant host properly.

The expression vector will preferably comprise one or more selectable markers. The marker is typically a nucleic acid sequence having a property that can be selected by a chemical method, and includes all genes capable of distinguishing a transformed cell from a non-transformed cell. Examples include herbicide resistance genes such as glyphosate or phosphinothricin, antibiotics such as kanamycin, G418, Bleomycin, hygromycin, chloramphenicol, Resistant gene, aadA gene, and the like, 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, Clp promoter, but is not limited thereto. The term " promoter " refers to the region of DNA upstream from the structural gene and refers to a DNA molecule to which an RNA polymerase binds to initiate transcription. A " plant promoter " is a promoter capable of initiating transcription in plant cells. A " constitutive promoter " is a promoter that is active under most environmental conditions and developmental conditions or cell differentiation. Constructive promoters may be preferred in the present invention because the choice of transformants can be made by various tissues at various stages. Thus, constitutive promoters do not limit selectivity.

In the recombinant vector of the present invention, conventional terminators can be used. Examples thereof include nopaline synthase (NOS), rice α-amylase RAmy1 A terminator, phaseoline terminator, Agrobacterium tumefaciens (Agrobacterium tumefaciens ) Terminator of the Octopine gene, and the rrnB1 / B2 terminator of E. coli, but the present invention is 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 a terminator is highly desirable in the context of the present invention.

&Quot; Plant cell " used for transformation of a plant may be any plant cell. The plant cell may be any of a cultured cell, a cultured tissue, a culture or whole plant, preferably a cultured cell, a cultured tissue or culture medium, and more preferably a cultured cell.

The term " plant tissue " refers to a variety of cells used in differentiated or undifferentiated plant tissues, such as but not limited to roots, stems, leaves, pollen, seeds, Protoplasts, shoots and callus tissue. The plant tissue may be in planta or may be in an organ culture, tissue culture or cell culture.

The method of delivering the vector of the present invention into a host cell can be carried out by injecting a vector into a host cell by microinjection, calcium phosphate precipitation, electroporation, liposome-mediated transfection, DEAE-dextran treatment, can do.

The present invention also provides a method for producing a recombinant vector comprising the steps of: transforming a plant cell with a recombinant vector comprising the amino acid sequence of SEQ ID NO: 2 and comprising a gene encoding Brassica rapa RNA helicase 22 protein derived from Chinese cabbage; And

And regenerating the plant from the transformed plant cell. The present invention also provides a method for producing a transgenic plant having regulated environmental stress tolerance.

The method according to an embodiment of the present invention is characterized by overexpressing a plant cell with a recombinant vector containing BrRH22 gene derived from Chinese cabbage to increase tolerance to environmental stress of the plant, but is not limited thereto.

In the method according to an embodiment of the present invention, the environmental stress may be salt or drought stress, but is not limited thereto.

The method of the invention comprises the step of transforming a plant cell with a recombinant vector according to the present invention, the transformant is, for example, Agrobacterium tyumeo Pacific Enschede may be mediated by (Agrobacterium tumefiaciens). In addition, the method of the present invention comprises regenerating a transgenic plant from the transformed plant cell. Any of the methods known in the art can be used for regeneration of transgenic plants from transgenic plant cells. The transformed plant cells must be regenerated into whole plants. Techniques for the regeneration of mature plants from callus or protoplast cultures are well known in the art for a number of different species.

In addition, the present invention provides a transgenic plant having the environmental stress tolerance regulated by the above method and a transformed seed thereof.

Wherein the plant is selected from the group consisting of rice, wheat, barley, corn, soybean, potato, wheat, red bean, oats and millet; Vegetable crops selected from the group consisting of Arabidopsis, cabbage, radish, red pepper, strawberry, tomato, watermelon, cucumber, cabbage, melon, squash, onions, onions and carrots; Ginseng, tobacco, cotton, sesame, sugar cane, beet, perilla, peanut and rapeseed; Apple trees, pears, jujubes, peaches, sheep grapes, grapes, citrus fruits, persimmons, plums, apricots and banana; Roses, gladiolus, gerberas, carnations, chrysanthemums, lilies and tulips; And feed crops selected from the group consisting of Ryegrass, Red Clover, Orchardgrass, Alpha Alpha, Tall Fescue, and Fereniallaigrus. Preferably, the plant is selected from the group consisting of Arabidopsis, potato, eggplant, cigarette, pepper, tomato, burdock, ciliaceae, lettuce, bellflower, spinach, modern sweet potato, celery, carrot, parsley, parsley, cabbage, cabbage, It may be a dicotyledonous plant such as melon, cucumber, squash, poultry, strawberry, soybean, mung bean, kidney bean or pea or a terminal plant such as rice, barley, wheat, rye, corn, sugar cane, oats and onion.

The present invention also provides a composition for controlling the environmental stress tolerance of a plant comprising the amino acid sequence of SEQ ID NO: 2, as an active ingredient, a gene coding for a Brassica rapa RNA helicase 22 protein derived from Chinese cabbage.

The composition contains a gene coding for the BrRH22 protein consisting of the amino acid sequence of SEQ ID NO: 2 as an active ingredient. By transforming the gene or a recombinant vector comprising the gene into a plant, it is possible to prevent the environmental stress tolerance of the plant, Or drought stress tolerance.

Hereinafter, the present invention will be described in more detail with reference to Examples. It is to be understood by those skilled in the art that these embodiments are merely illustrative of the present invention and that the scope of the present invention is not limited thereto.

Materials and methods

One. BrRH22 Gene separation and intracellular localization

BrRH22 GFP (green fluorescent protein) was conjugated to the BrRH22 gene and the BrRH22-GFP junction protein was expressed in the tobacco leaves using the CaMV 35S promoter, and confocal microscopy (Carl Zeiss Inc., USA) was used to observe the GFP fluorescence signal.

2. Vector production and Arabidopsis transformation

BrRH22 To construct an overexpressed transformant, the coding region of the BrRH22 cDNA was amplified by PCR (polymerase chain reaction) and inserted into the BamHI / SalI site of the pCambia1303 vector. Transgenic Arabidopsis thaliana transformed Agrobacterium tumefaciens GV3101 ( Agrobacterium tumefaciens GV3101) (Bechtold and Pelletier, 1998, Mol. Biol. 82, 259-266). Selection of the transformed plants was carried out by seeding on MS medium containing hygromycin (50 / / mL) and carbenicillin (250 / / mL) as selection markers after seed harvest and then transformed plants were selected, After additionally screening the transformed lines, the T2 and T3 lines were used for phenotyping.

3. Seed Germination and Transplantation of Transgenic Plants under Various Stress Conditions Seedlings  Analysis of growth

The seeds of A. thaliana wild-type and BrRH22- overexpressing transgenic plants used in the present invention were mixed at a ratio of 2: 1: 1 of vermiculite, peat moss and perlite And cultivated at 23 ° C under long-day conditions (16 hrs condition / 8 hrs dark cycle) at 100 μEm ^-2 s ^-1 . In addition, the Arabidopsis thaliana was grown on MS medium. The germination assay was repeated three times each for 30 to 40 seeds. Based on the MS medium supplemented with 1.5% sucrose, 150-200 mM NaCl and 300-450 mM Of mannitol, respectively, and then grown under normal growth conditions. The seeds were regarded as germinated when the young roots emerged through the seed coat, and the root growth was observed by standing the plate vertically. Fresh weight and root length of seedlings were measured 3 weeks after germination.

4. RNA extraction and RT- PCR

Total RNA was extracted from frozen Arabidopsis samples using the Plant RNeasy extraction kit (Qiagen; www.qiagen.com/) and real-time quantitation was performed using the QuantiTect SYBR Green RT-PCR kit (Qiagen, Germany) Gene 2000 real-time thermal cycling system (Corbett Research www.corbettlifescience.com/). RT-PCR was performed with the same amount of total RNA using a primer set specific for the actin gene. To accurately measure the expression level of the BrRH22 gene after treating 150 mM NaCl and 300 mM mannitol, The stress - treated Arabidopsis thalamus and the stressed Arabidopsis thaliana were studied in the time zone.

Example  One. BrRH22  Identification of cytoplasmic loci in genes

BrRH22, one of the DEAD-box RHs present in the Chinese cabbage genome, contains cTP with 51 amino acids at the N-terminus (Figure 1A). To determine whether BrRH22, which is expected to be transferred to the chloroplast, was present in the chloroplast, the BrRH22-GFP junction protein was expressed in the tobacco leaves and the GFP fluorescence signal was observed. As shown in FIG. 1B, The BrRH22-GFP junction protein was expressed.

Example  2. In salt and drought stress conditions, BrRH22  Promoting seed germination and seedling growth of transgenic Arabidopsis plants overexpressing the gene

The BrRH22 transgenic Arabidopsis plants of the present invention were examined for the function of the BrRH22 gene under salt and drought stress conditions. To investigate the function of BrRH22 gene, BrRH22 transgenic Arabidopsis plants were prepared and the expression pattern of BrRH22 was confirmed by RT-PCR analysis. As a result, when germinating Arabidopsis under normal conditions, germination rate of wild type and transgenic Arabidopsis plants was not significantly different (data not shown), but when germinated under salt and drought stress conditions, the transgenic Arabidopsis plants It was confirmed that the germination period was faster and the germination rate was higher than that of the wild type seed (FIG. 2).

In addition, the seedling growth of transgenic Arabidopsis plants treated with salt and drought stress showed a statistically significant difference in fresh weight and root length compared to the wild type (Figs. 3 and 4) , Salt and drought stress treatment, the survival rate of the transgenic Arabidopsis plants was superior to that of the wild type (Figs. 5 and 6).

<110> INDUSTRY FOUNDATION OF CHONNAM NATIONAL UNIVERSITY <120> Method for producing transgenic plant with increased          environmental stress resistance using BrRH22 gene from Brassica          rapa and plant thereof <130> PN17416 <160> 2 <170> KoPatentin 3.0 <210> 1 <211> 1719 <212> DNA <213> Brassica rapa <400> 1 atgattctct cacgctctgt ctctgttctt cattcctgta gaatttcctc ggcgccgaaa 60 ctcatatccc acaagcttaa ggtatccttc cctctcgctt atggttcttc cgcgttgatc 120 tctctcaatc gatctgaagt gaaatgggtt cgtgtgttcg cgtctgctac tgagactgag 180 actgaagtag agagaaaagg aaacgacacg tttttcgcgg accaagccgt ttcttggaag 240 tctcttgggt tgtcggataa ggtctcagtg gctcttcgag attctgggtt cggaagaccc 300 tctcttactc aggcggtttg cattccatcg atattgtccg ggaaagatgt ggttgttgca 360 gcagaaacag gcagtggcaa aacacatgga tatcttgctc caattatcca ccagttaacc 420 aacaccaaca cccctcttga ttccgaggga gaagaaaggc gggtgccact taagaccatt 480 tctcttgttc tctgccctaa cgtcatgcta tgtgaacaag ttgttcggat ggttaatggt 540 cttcttggag aggacggtca tcctcttctc agagttgaag ctgtttgtgg ctcacagggt 600 tggcctgata aacagcctga tattattgtt tcgacccctg ctgctctttt gaataacatt 660 gagcctaaaa gaaacaggcg tgtggagttt cttcgatctg tcaaatatgt ggtgtttgat 720 gaagctgata tgcttctctg cggaagcttc cagaatcaga ttatccgtct cataaacatg 780 ctgcgttttg atgagaaaca agtgtctcgt ttggctactt ccaaatttgg aaaaccattg 840 gagattgatg cagaatcttc agcacaattt gatttagaga atgaggatga tgcagatttt 900 gaagaagacg ggtctatctc tgatgaagag gaggaagagt atcttgaaga taccacccaa 960 atccctaccg ttgaaaatgg agctggtgct gatattagaa agggttggag aagagtgaga 1020 aagatctata cccgtagcaa gcagtatatt ttcattgcag ccacacttcc ggttaacggg 1080 aagaaaactg caggaggact tttgaaacat atgttccaag atgctgtctg ggttagcgga 1140 aactttcttc accgaaacag tcctagacta aagcagaaat gggttgaagt cacagtcgac 1200 acacaagttg atgctctcat agaggctgta aacaacagta gcacagacag gacgatggtg 1260 tttgcaaata cagttgaggc ggttgaagca gtagccgaca tattggagaa agctagtatc 1320 cagtgctatc gttatcataa gaaccataca cttgaagagc gtgctaatat attagctgat 1380 ttcagagaaa acggtggtgt ctttgtctgt actgacgcag ctgcacgtgg agttgatgtt 1440 cctaacgtct cgcacgttat tcaggcggat ttttctagtt ctgcggtgga ttttcttcac 1500 aggataggtc gaacagctag agctgggcaa tacggaacgg tgacgagtct atacactgag 1560 gctaaccgtg atttagtgga agcaatccgt gaagcagtga agacgggtca gccagtggaa 1620 actgcattta gcaggaaaag aggtttcagg aacaagctaa agaagagagc tttcttgaaa 1680 gccggagaag caaaggagtc tcaagctgtg cgagcttaa 1719 <210> 2 <211> 572 <212> PRT <213> Brassica rapa <400> 2 Met Ile Leu Ser Arg Ser Val Val Ser Le His Ser Cys Arg Ile Ser   1 5 10 15 Ser Ala Pro Lys Leu Ile Ser His Lys Leu Lys Val Ser Phe Pro Leu              20 25 30 Ala Tyr Gly Ser Ser Ala Leu Ile Ser Leu Asn Arg Ser Glu Val Lys          35 40 45 Trp Val Arg P Val Ala Ser Ala Thr Glu Thr Glu Thr Glu Val Glu      50 55 60 Arg Lys Gly Asn Asp Thr Phe Phe Ala Asp Gln Ala Val Ser Trp Lys  65 70 75 80 Ser Leu Gly Leu Ser Asp Lys Val Ser Val Ala Leu Arg Asp Ser Gly                  85 90 95 Phe Gly Arg Pro Ser Leu Thr Gln Ala Val Cys Ile Pro Ser Ile Leu             100 105 110 Ser Gly Lys Asp Val Val Val Ala Glu Thr Gly Ser Gly Lys Thr         115 120 125 His Gly Tyr Leu Ala Pro Ile Ile His Gln Leu Thr Asn Thr Asn Thr     130 135 140 Pro Leu Asp Ser Glu Gly Glu Glu Arg Arg Val Pro Leu Lys Thr Ile 145 150 155 160 Ser Leu Val Leu Cys Pro Asn Val Met Leu Cys Glu Gln Val Val Arg                 165 170 175 Met Val Asn Gly Leu Leu Gly Glu Asp Gly His Pro Leu Leu Arg Val             180 185 190 Glu Ala Val Cys Gly Ser Gln Gly Trp Pro Asp Lys Gln Pro Asp Ile         195 200 205 Ile Val Ser Thr Pro Ala Ala Leu Leu Asn Asn Ile Glu Pro Lys Arg     210 215 220 Asn Arg Arg Val Glu Phe Leu Arg Ser Val Lys Tyr Val Val Phe Asp 225 230 235 240 Glu Ala Asp Met Leu Leu Cys Gly Ser Phe Gln Asn Gln Ile Ile Arg                 245 250 255 Leu Ile Asn Met Leu Arg Phe Asp Glu Lys Gln Val Ser Arg Leu Ala             260 265 270 Thr Ser Lys Phe Gly Lys Pro Leu Glu Ile Asp Ala Glu Ser Ser Ala         275 280 285 Gln Phe Asp Leu Glu Asn Glu Asp Asp Ala Asp Phe Glu Glu Asp Gly     290 295 300 Ser Ile Ser Asp Glu Glu Glu Glu Glu Tyr Leu Glu Asp Thr Thr Gln 305 310 315 320 Ile Pro Thr Val Glu Asn Gly Ala Gly Ala Asp Ile Arg Lys Gly Trp                 325 330 335 Arg Arg Val Lys Ile Tyr Thr Arg Ser Lys Gln Tyr Ile Phe Ile             340 345 350 Ala Ala Thr Leu Pro Val Asn Gly Lys Lys Thr Ala Gly Gly Leu Leu         355 360 365 Lys His Met Phe Gln Asp Ala Val Trp Val Ser Gly Asn Phe Leu His     370 375 380 Arg Asn Ser Pro Arg Leu Lys Gln Lys Trp Val Glu Val Thr Val Asp 385 390 395 400 Thr Gln Val Asp Ala Leu Ile Glu Ala Val Asn As Ser Ser Thr Asp                 405 410 415 Arg Thr Met Val Phe Ala Asn Thr Val Glu Ala Val Glu Ala Val Ala             420 425 430 Asp Ile Leu Glu Lys Ala Ser Ile Gln Cys Tyr Arg Tyr His Lys Asn         435 440 445 His Thr Leu Glu Glu Arg Ala Asn Ile Leu Ala Asp Phe Arg Glu Asn     450 455 460 Gly Gly Val Phe Val Cys Thr Asp Ala Ala Ala Arg Gly Val Asp Val 465 470 475 480 Pro Asn Val Ser His Val Ile Gln Ala Asp Phe Ser Ser Ser Ala Val                 485 490 495 Asp Phe Leu His Arg Ile Gly Arg Thr Ala Arg Ala Gly Gln Tyr Gly             500 505 510 Thr Val Thr Ser Leu Tyr Thr Glu Ala Asn Arg Asp Leu Val Glu Ala         515 520 525 Ile Arg Glu Ala Val Lys Thr Gly Gln Pro Val Glu Thr Ala Phe Ser     530 535 540 Arg Lys Arg Gly Phe Arg Asn Lys Leu Lys Lys Arg Ala Phe Leu Lys 545 550 555 560 Ala Gly Glu Ala Lys Glu Ser Gln Ala Val Arg Ala                 565 570

Claims (9)

A plant drought or a plant drought comprising the step of over-expressing a BrRH22 gene by transforming a plant cell with a recombinant vector comprising a gene encoding Brassica rapa RNA helicase 22 protein derived from Chinese cabbage, which comprises the amino acid sequence of SEQ ID NO: 2 Methods for increasing salt stress tolerance. delete delete delete Overexpressing the BrRH22 gene by transforming the plant cell with a recombinant vector comprising the amino acid sequence of SEQ ID NO: 2 and comprising a gene encoding Brassica rapa RNA helicase 22 protein derived from Chinese cabbage; And
And regenerating the plant from the transformed plant cell. &Lt; RTI ID = 0.0 &gt; 21. &lt; / RTI &gt; delete A transgenic plant having increased drought or salt stress tolerance produced by the method of claim 5. A transformed seed of a plant according to claim 7. A composition for increasing drought or salt stress tolerance of a plant comprising an amino acid sequence of SEQ ID NO: 2 as an active ingredient, the gene encoding Brassica rapa RNA helicase 22 protein derived from Chinese cabbage.

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