KR101985321B1 - Method for producing transgenic plant with increased heavy metal stress tolerance using OsAIR2 gene from Oryza sativa and plant thereof - Google Patents

Abstract

The present invention relates to a method for producing a transgenic plant having increased resistance to heavy metal stress using OsAIR2 gene derived from rice and a plant according to the present invention. More particularly, the transgenic plant overexpressing OsAIR2 gene derived from rice according to the present invention comprises arsenic The plant exhibits stress tolerance and is superior in germination rate and root growth than the control plant grown under the same conditions. Therefore, it can contribute to the development of safe crops with increased heavy metal tolerance and productivity through controlling the expression of genes encoding OsAIR2 protein in plants have.

Description

FIELD OF THE INVENTION [0001] The present invention relates to a method for producing a transgenic plant having increased resistance to heavy metal stress using OsAIR2 gene derived from rice, and to a plant using the OsAIR2 gene,

The present invention relates to a method for producing a transgenic plant having increased resistance to heavy metal stress using rice-derived OsAIR2 gene, and a plant therefor.

Arsenic (As) is naturally present in small amounts in the soil, but accumulation of high levels is a serious threat to human health due to ingestion of food derived from the contaminated crops. In particular, in the case of rice, the most important food in the world, soils and radioactive water arsenic accumulate well, so rice fields are easily contaminated. The accumulation of arsenic in plants directly or indirectly affects plant metabolism including photosynthesis and carbohydrate metabolism, and the root is the first tissue to contact with the compound after exposure to arsenic. Root elongation and proliferation are inhibited by arsenic toxicity, Can lead to yield reduction.

RING (Really Interesting New Gene) E3 ligase is a common sequence (Cys-x2-Cys-x9-Cys-x1-3-His-x2-3-Cys / His-x2- 48-Cys-x2-Cys, where x can be any amino acid), and these structures are bound to two zinc atoms. This structure is generally known to be classified into two basic forms, including RING-H2 and RING-HC, based on the cysteine or histidine position of the RING-finger proteins. In addition, several variants such as RING-D, RING-v, RING-S / T, RING-G and RING-C2 have been found in higher plants including Arabidopsis or rice genomes. The researchers demonstrated that the E3 ubiquitin ligase with the RING-domain plays an important role in adapting to extreme environments, defense mechanisms against pathogens, and plant growth. For example, the function of E3 ubiquitin ligase (i. E., Rma1H1) in red pepper involves the inhibition of the transport of aquaporin to the plasma membrane, and hydrolysis by proteasomes subsequently provides enhanced resistance to drought stress . The Arabidopsis SDIR1 (salt- and drought-induced RING finger1) gene is thought to regulate stress-responsive ABA signaling. Similarly, transgenic rice plants overexpressing the OsSDIR1 gene were found to be resistant to drought.

Korean Patent No. 1740176 discloses a protein and gene for increasing arsenic resistance in plants, and Korean Patent No. 1511190 discloses an OsHIR1 gene and a use thereof for decreasing heavy metal stress tolerance and heavy metal absorption in plants . However, the method for producing transgenic plants having increased resistance to heavy metal stress using the rice-derived OsAIR2 gene of the present invention and the plants therefor have not yet been disclosed.

The present invention is derived by the request as described above, the present inventors have selected the OsAIR2 (Oryza sativa Arsenic-Induced RING E3 ligase 2) gene significantly increased the expression by the arsenic treatment in rice, the OsAIR2 gene-overexpressed The present inventors completed the present invention by confirming that the Arabidopsis thaliana plant is increased in arsenic stress tolerance.

In order to solve the above problems, the present invention provides a method for regulating the expression of the OsAIR2 gene by transforming a plant cell with a recombinant vector comprising a gene encoding rice-derived OsAIR2 ( Oryza sativa Arsenic-Induced RING E3 ligase 2) The present invention provides a method for controlling heavy metal stress tolerance of a plant comprising

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 a gene encoding a rice-derived OsAIR2 ( Oryza sativa Arsenic-Induced RING E3 ligase 2) protein comprising the amino acid sequence of SEQ ID NO: 2; And regenerating the plant from the transformed plant cell. The present invention also provides a method for producing a transgenic plant having regulated heavy metal stress tolerance.

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

The present invention also provides a composition for controlling heavy metal stress tolerance of a plant comprising the gene encoding the OsAIR2 ( Oryza sativa Arsenic-Induced RING E3 ligase 2) protein derived from rice comprising the amino acid sequence of SEQ ID NO: 2 as an active ingredient .

The transgenic plants overexpressing the OsAIR2 gene derived from rice according to the present invention exhibit arsenic stress resistance and are superior in germination and root growth to those of the control plants grown under the same conditions. Therefore, the expression of OsAIR2 protein-encoding gene Can contribute to the development of safe crops with increased heavy metal tolerance and productivity.

FIG. 1 shows the results of 18 OsRFP ( Oryza sativa RING finger proteins) in rice roots treated with arsenic (AsV) stress at various concentrations (50, 100, 150, 200, 250 and 300 μM) for 2 weeks according to an embodiment of the present invention ) Gene expression patterns. The OsActinII gene is an internal control, and the red and green arrows indicate gene up and down regulation.
FIG. 2 shows the amount of OsAIR2 gene expressed in the time course of rice root treated with arsenic (AsV, 1900 μM) stress according to an embodiment of the present invention. The asterisk shows statistical significance using the two-tailed Student t-test compared to the control (0h); * Means P value <0.05, ** means P value <0.01 and *** means P value <0.001.
FIG. 3 is a graph showing the relationship between the translated amino acid sequence of rice-derived OsAIR2 gene and homologous genes thereof (GRMZM2G11930 derived from wheat, Sobic.005G160400 derived from wheat, Pavir.Ha00474 derived from Daphniaceae, Pavir.JI2507, .g, wild lawn-derived Bradi4g16000, and Arabidopsis-derived AT5G22000, 489047 protein). Alignment was performed using ClustalW2 software (http://www.ebi.ac.uk/clustalw/), and asterisks near the red box indicate the conserved (C) or histidine (C) forms of the RING- And Cys and His residues at the metal-ligand position.
FIG. 4 is a graph showing the results of ubiquitin-activating enzyme (E1) and ubiquitin-conjugating enzyme UBC10 (E2) derived from human beings according to an embodiment of the present invention. (A) and the result of ubiquitination of MBP-OsAIR2 protein and E1 and E2 reaction time (B) by confirming the ubiquitination activity of OsAIR2 gene by confirming presence of ubiquitinated band after reaction in reaction buffer. The ubiquitinated bands were detected by immunoblot analysis using anti-His antibodies.
Figure 5 is a graph showing the results of RT-PCR analysis of OsAIR2 The overexpression of OsAIR2 gene was confirmed in overexpressed transgenic plants (35S: OsAIR2-EYEP) and control plants (35S: EYFP ). AtUBC is a loading control.
FIG. 6 is a graph showing the results of OsAIR2 treated with 100, 150 and 200 .mu.M arsenic for 2 weeks, (A) and germination rate (B) of over-expressing transgenic plants (35S: OsAIR2-EYEP) and control plants (35S: EYFP). The asterisk indicates statistical significance using the two-tailed Student t-test; * Means P value <0.05, ** means P value <0.01 and *** means P value <0.001.
FIG. 7 is a graph showing the results of OsAIR2 treated with 100, 150 and 200 .mu.M arsenic for 2 weeks, (A) and root length (B) of over-expressing transgenic plants (35S: OsAIR2-EYEP) and control plants (35S: EYFP). The asterisk indicates statistical significance using the two-tailed Student t-test; * Means P value <0.05, ** means P value <0.01 and *** means P value <0.001.

In order to accomplish the object of the present invention, the present invention provides a recombinant vector comprising a gene encoding OsAIR2 ( Oryza sativa Arsenic-Induced RING E3 ligase 2) protein from rice, transforming plant cells to regulate the expression of OsAIR2 gene Wherein the method comprises the steps of:

The OsAIR2 protein according to the present invention may comprise the amino acid sequence of SEQ ID NO: 2, and the OsAIR2 gene may comprise the nucleotide sequence of SEQ ID NO: 1, but is not limited thereto.

The OsAIR2 protein of the present invention comprises a protein having the 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 physiological activity " means an activity of regulating heavy metal stress tolerance of a plant, and preferably means an activity of regulating an arsenic 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 overexpressing the OsAIR2 gene so as to control the heavy metal stress tolerance of the plant, And may preferably increase the arsenic stress tolerance of the plant, but the present invention 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 method according to one embodiment of the present invention, the heavy metal may be zinc, lead, cadmium, copper, magnesium, manganese, nickel, cobalt, iron, antimony or arsenic, preferably arsenic, Do not.

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 OsAIR2 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 OsAIR2 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 a gene encoding a rice-derived OsAIR2 ( Oryza sativa Arsenic-Induced RING E3 ligase 2) protein comprising the amino acid sequence of SEQ ID NO: 2; And

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

The method according to an embodiment of the present invention is characterized by overexpressing plant cells with a recombinant vector containing OsAIR2 gene derived from rice, thereby increasing tolerance to heavy metal stress of the plant, but not limited thereto.

In the method according to one embodiment of the present invention, the heavy metal may be zinc, lead, cadmium, copper, magnesium, manganese, nickel, cobalt, iron, antimony or arsenic, preferably arsenic, Do not.

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 regulated heavy metal stress tolerance produced 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 heavy metal stress tolerance of a plant comprising the gene encoding the OsAIR2 ( Oryza sativa Arsenic-Induced RING E3 ligase 2) protein derived from rice comprising the amino acid sequence of SEQ ID NO: 2 as an active ingredient .

The composition contains a gene encoding OsAIR2 protein consisting of the amino acid sequence of SEQ ID NO: 2 as an active ingredient. By transforming a plant with the gene or a recombinant vector containing the gene, the plant is able to tolerate heavy metal stress, preferably arsenic It is possible to increase the 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.

EXAMPLES Example 1: OsRFP ( Oryza sativa RING finger protein gene selection

(AsV; Arsenate) treated with various concentrations (50, 100, 150, 200 and 300 μM) of rice (Donganbyeo) plants treated with various concentrations RNA was extracted from the root samples and semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR) analysis was performed to confirm the expression pattern of 18 OsRFP genes. As a result, among the 18 OsRFP genes, Os05g32350 and Os11g36430 In particular, Os11g36430 gene with the highest expression level was selected and named OsAIR2 (Oryza sativa Arsenic-Induced RING E3 ligase 2) (Fig. 1).

In addition, targeting the roots of a rice plant AsV the processing of high concentration (1900μM) Quantitative RT-PCR (quantitative real time- polymerase chain reaction) results confirmed the expression pattern of a time slot OsAIR2 gene through the analysis, the time treated with AsV As a result, the amount of OsAIR2 gene expression was increased and the amount of OsAIR2 gene expression was increased about 15 times after 24 hours (FIG. 2).

Example  2. OsAIR2  Functional analysis of genes as E3 ligase

The OsAIR2 gene is composed of 395 amino acids and the molecular weight of the protein is estimated to be 42.38 kDa. Afterwards, the OsAIR2 amino acid sequence derived from rice and the sequence of Zea mays , Sorghum bicolor , Setaria italica , Panicum (RING-HC type) of the N-terminal region as a result of alignment of the amino acid sequences of homologous genes derived from Arabidopsis lyrata and Arabidopsis thaliana , as well as virgatum , wild grass ( Brachypodium distachyon and Arabidopsis lyrata and Arabidopsis thaliana ) (C-X2-C-X12-C-X1-H-X2-C-X2-C-X11-C-X2-C).

A number of previous studies have shown that the RING domain has E3 ligase activity. For this reason, in order to confirm the E3 ligase activity of the OsAIR2 protein having the RING-HC domain, a fusion protein in which an MBP (maltose-binding protein) was bound to the full-length OsAIR2 gene was expressed in E. coli BL21, The activity of the domain was confirmed by substituting alanine for the 33rd cysteine among the RING domains. Next, the present inventors confirmed that the RING-HC domain of OsAIR2 protein has E3 ubiquitin ligase activity. Recombinant MBP-OsAIR2 and MBP-OsAIR2 ^C33A and non stylized recombinant expressing the empty-MBP each on the E. coli system, each protein purified are derived from human E1 (ubiquitin-activating enzyme) and Arabidopsis-derived E2 (ubiquitin-conjugating enzyme UBC10 ) Was incubated with ubiquitination buffer (50 mM Tris-HCl, pH 7.5, 1 mM MgCl ₂ , 0.05 mM ZnCl ₂ , 1 mM ATP, 0.2 mM DTT, 10 mM phosphocreatine and 0.1 unit of creatine kinase) Lt; / RTI &gt; The ubiquitination activity was measured using an anti-ubiquitin antibody.

As a result, MBP-OsAIR2, while showing a plurality of ubiquitination band, MBP-OsAIR2 ^C33A and Empty-MBP were not in the ubiquitination band (Fig. 4A). In addition, the ubiquitination activity of the MBP-OsAIR2 protein and the E1 and E2 enzymes in the reaction time was examined. As a result, the ubiquitination band gradually increased over time, indicating that the OsAIR2 protein containing the RING-HC domain functions as the E3 ubiquitin ligase (Fig. 4B).

Example  3. OsAIR2 Overexpressed transgenic Arabidopsis arsenic Analysis of response to treatment

It confirmed that the three OsAIR2 gene expression through RT-PCR analysis of OsAIR2 (35S: EYEP lines) were selected through AsV treatment of various concentrations (100, 150, and 200 μM) and the overexpression lines (35S: OsAIR2-EYEP lines # 1, # 2 and # 3) Respectively. As a result, there was no difference in phenotype in the environment without arsenic treatment (in case of no data), but when arsenic was treated, OsAIR2 The germination rate of the overexpressed transformants was significantly increased as compared with the control plants (Fig. 6). Also, OsAIR2 When the root lengths of transgenic plants and control plants were measured and compared with those of 100 μM arsenic, OsAIR2 The overexpressed transgenic plants increased the root length by about 1 cm compared with the control plants, and increased by about 0.3 cm when treated with 200 μM of arsenic (FIG. 7).

<110> KNU-Industry Cooperation Foundation <120> Method for producing transgenic plant with increased heavy metal          stress tolerance using OsAIR2 gene from Oryza sativa and plant          the <130> PN17405 <160> 2 <170> KoPatentin 3.0 <210> 1 <211> 1185 <212> DNA <213> Oryza sativa <400> 1 atggcatctg gaactgatga gaaagccaag atggagggtt tgacatcagc tgcagccttt 60 gttgagggtg ggattcagga tgcatgtgat gatgcatgta gtatctgcct tgaggcgttc 120 tgtgagagtg acccttccac attgactggt tgcaaacacg agttccacct ccaatgcatt 180 cttgaatggt gtcagagaag ttctcagtgt cctatgtgtt ggcagcctat cagtttgaag 240 gatcctacca gtcaagagct gctcgaggca gtggagcgtg aaaggaatgt aaggaccaat 300 caaactcgaa atacaactat atttcatcat cctgctcttg gagattttga ggttcagcat 360 ttacctgttg ttggtaatga tgctgaactt gaagagcgta tattacagca cctagcagca 420 gccgctgcaa tgggaaggtc acaccacctt ggtagaagag aaggacacag gggtcgttcc 480 ggctctcatg gtcgtccaca gttcttagtt ttttcttcgc atccaaacat gccttctgct 540 ggttcagttt cttcatcgtc cgttcaaggg gaagtggata atgaatcaag tcctgtccac 600 acaactggtg aattatcact gcatgctaac acccatgaag aagcaggcaa tcaaagtcct 660 gggatgctta cctacgatgc tgatcaagat gctgttgttt catctggaaa tagtacccct 720 gtatctagcc ctaggttttt caacaggagg cattccactg ggcaatcaac tccagtaaac 780 aacgacagag ctgggccttc agatcttcag tctttctcag actctctgaa gtctcgctta 840 aatgctgtct ctatgaagta caaggaatct attacaaaaa gtactcgagg atggaaggag 900 agactttttt cgcgtcattc atctgtggca gatcttggtt ctgaagtaag aagagaagtt 960 aatgctggaa ttgcatccgt atcaaggatg atggagcgtc tggaaactag aggtagtaat 1020 ggtagaacaa gtgatggccc agcaatatcc acttctgaag ttattcccag tacagaatca 1080 agcaatgaga gagttacaga aaacaatcca actactgcag cgacaagtag tggcaacact 1140 tctgcgtctt ctgccccttg tgttacaaca accggttcaa attag 1185 <210> 2 <211> 394 <212> PRT <213> Oryza sativa <400> 2 Met Ala Ser Gly Thr Asp Glu Lys Ala Lys Met Glu Gly Leu Thr Ser   1 5 10 15 Ala Ala Ala Phe Val Glu Gly Gly Ile Gln Asp Ala Cys Asp Asp Ala              20 25 30 Cys Ser Ile Cys Leu Glu Ala Phe Cys Glu Ser Asp Pro Ser Thr Leu          35 40 45 Thr Gly Cys Lys His Glu Phe His Leu Gln Cys Ile Leu Glu Trp Cys      50 55 60 Gln Arg Ser Ser Gln Cys Pro Met Cys Trp Gln Pro Ile Ser Leu Lys  65 70 75 80 Asp Pro Thr Ser Glu Glu Leu Leu Glu Ala Val Glu Arg Glu Arg Asn                  85 90 95 Val Arg Thr Asn Gln Thr Arg Asn Thr Thr Ile Phe His His Pro Ala             100 105 110 Leu Gly Asp Phe Glu Val Gln His Leu Pro Val Val Gly Asn Asp Ala         115 120 125 Glu Leu Glu Glu Arg Ile Leu Gln His Leu Ala Ala Ala Ala Ala Met     130 135 140 Gly Arg Ser His His Leu Gly Arg Arg Glu Gly His Arg Gly Arg Ser 145 150 155 160 Gly Ser His Gly Arg Pro Gln Phe Leu Val Phe Ser Ser His Pro Asn                 165 170 175 Met Pro Ser Ala Gly Ser Val Ser Ser Ser Ser Val Gln Gly Glu Val             180 185 190 Asp Asn Glu Ser Ser Pro Val His Thr Thr Gly Glu Leu Ser Leu His         195 200 205 Ala Asn Thr His Glu Glu Ala Gly Asn Gln Ser Pro Gly Met Leu Thr     210 215 220 Tyr Asp Ala Asp Gln Asp Ala Val Val Ser Ser Gly Asn Ser Thr Pro 225 230 235 240 Val Ser Ser Pro Arg Phe Phe Asn Arg Arg His Ser Thr Gly Gln Ser                 245 250 255 Thr Pro Val Asn Asn Asp Arg Ala Gly Pro Ser Asp Leu Gln Ser Phe             260 265 270 Ser Asp Ser Leu Lys Ser Arg Leu Asn Ala Val Ser Met Lys Tyr Lys         275 280 285 Glu Ser Ile Thr Lys Ser Thr Arg Gly Trp Lys Glu Arg Leu Phe Ser     290 295 300 Arg His Ser Ser Val Ala Asp Leu Gly Ser Glu Val Arg Arg Glu Val 305 310 315 320 Asn Ala Gly Ile Ala Ser Val Ser Arg Met Met Glu Arg Leu Glu Thr                 325 330 335 Arg Gly Ser Asn Gly Arg Thr Ser Asp Gly Pro Ala Ile Ser Thr Ser             340 345 350 Glu Val Ile Pro Ser Thr Glu Ser Ser Asn Glu Arg Val Thr Glu Asn         355 360 365 Asn Pro Thr Thr Ala Ala Thr Ser Ser Gly Asn Thr Ser Ala Ser Ser     370 375 380 Ala Pro Cys Val Thr Thr Thr Gly Ser Asn 385 390

Claims (9)

A plant comprising the step of overexpressing the OsAIR2 gene by transforming a plant cell with a recombinant vector comprising a gene encoding a rice-derived OsAIR2 ( Oryza sativa Arsenic-Induced RING E3 ligase 2) protein consisting of the amino acid sequence of SEQ ID NO: 2 Of arsenic stress tolerance. delete delete delete Overexpressing the OsAIR2 gene by transforming a plant cell with a recombinant vector comprising a gene encoding a rice-derived OsAIR2 ( Oryza sativa Arsenic-Induced RING E3 ligase 2) protein comprising the amino acid sequence of SEQ ID NO: 2; And
And regenerating the plant from the transformed plant cell. &Lt; RTI ID = 0.0 &gt; 21. &lt; / RTI &gt; delete 6. An improved transgenic plant having increased arsenic stress tolerance produced by the method of claim 5. A transformed seed of a plant according to claim 7. A composition for increasing the arsenic stress tolerance of a plant containing as an active ingredient a gene encoding a rice-derived OsAIR2 ( Oryza sativa Arsenic-Induced RING E3 ligase 2) protein consisting of the amino acid sequence of SEQ ID NO: 2.

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