KR102026766B1 - A novel genes for enhancing drought resistance and their uses - Google Patents

Abstract

The present invention relates to a drought resistant OsPYL / RCAR7 gene and its use. Specifically, the plant transformed with the OsPYL / RCAR7 gene isolated from rice is confirmed to have an effect on improving the drying resistance. It can be used to produce plants having good drying resistance without any influence.

Description

A novel genes for enhancing drought resistance and their uses}

The present invention relates to OsPYL / RCAR7, a drought resistant rice gene, and the use thereof.

Higher plants are immovable and are subject to various environmental factors such as drought, high salt, heavy metals, cold, heat shock and ozone during their lifetime. These abiotic stresses are a limiting factor in the growth and development of crops. Among these stresses, water shortage is the most serious environmental factor considered to be the major cause of crop production decline. Consumption of water has continued to increase worldwide, and the availability of clean water can be an important issue for higher plants as well as humans. Plants operate a variety of defense strategies, such as regulating signaling network pathways or inducing stress-responsive genes to increase resistance to short-term or long-term water shortages. Cellular or genetic defense mechanisms against water stress are well known (Shinozaki and Yamaguchi-shinozaki, 2007). However, there is still a lack of knowledge of the biological functions of stress-related genes involved in stress tolerance or sensitivity in higher plants. Therefore, functional studies of stress response genes are important to increase crop productivity.

On the other hand, since plants are frequently exposed to various environmental stresses such as drought, salt, cold, heat and pests, they have developed physiological, biochemical and molecular defense mechanisms to cope with the harmful effects of stress. abscisic acid (ABA) signal transduction regulation allows plants to overcome this stress. Reportedly, plants that pretreated and stressed abscis acid are more resistant to stress than those that do not, while mutant plants that do not produce or respond to absic acid are weak in stress. Therefore, by using proteins involved in the reaction of abscitic acid, it is expected that the plant can be improved in resistance to environmental stress. On the other hand, although stress resistance is increased by abscis acid, the action of abscis acid may inhibit plant growth.

Therefore, the method of improving the drying resistance of plants has been the subject of major challenges, and researches on the use of proteins involved in the reaction of abscic acid have been conducted (Korean Patent Publication No. 10-2010-0040789), but the situation is still insufficient. to be.

Accordingly, the present inventors searched for a drought resistant gene derived from a plant, and found a drought resistant rice gene OsPYL / RCAR7 [ Oryza sativa PYR1 (PYRABACTINE RESISTANCE 1) -LIKE / REGULATORY COMPONENTS OF ABA RECEPTOR7]. The gene completes the present invention by revealing that the gene acts as a receptor for abscitic acid only under high stress, and thus shows resistance.

Korean Patent Publication No. 10-2010-0040789

It is an object of the present invention to enhance drought resistance comprising a gene encoding OsPYL / RCAR7 [Oyza sativa PYR1 (PYRABACTINE RESISTANCE 1) -LIKE / REGULATORY COMPONENTS OF ABA RECEPTOR7] protein derived from rice, consisting of the amino acid sequence represented by SEQ ID NO: 2. It is to provide a recombinant vector for.

In addition, another object of the present invention to provide a transformed plant transformed with the recombinant vector.

In addition, another object of the present invention is to provide a method for producing the transformed plant.

In addition, another object of the present invention is to provide a method of increasing drought resistance by overexpressing a gene encoding a rice-derived OsPYL / RCAR7 protein consisting of the amino acid sequence represented by SEQ ID NO: 2 in a plant.

In order to achieve the above object, the present invention provides a recombinant vector for improving drought resistance comprising a gene encoding an OsPYL / RCAR7 protein derived from rice consisting of the amino acid sequence represented by SEQ ID NO: 2.

The present invention also provides a transformed plant transformed with the recombinant vector.

The present invention also provides a method for producing the transformed plant.

In addition, the present invention provides a method for increasing drought resistance by overexpressing a gene encoding a rice-derived OsPYL / RCAR7 protein consisting of the amino acid sequence represented by SEQ ID NO: 2 in a plant.

Oryza sativa PYR1 (PYRABACTINE RESISTANCE 1) -LIKE / REGULATORY COMPONENTS OF ABA RECEPTOR7] derived from rice plants transformed with a recombinant vector for enhancing drought resistance, which contains a gene encoding a protein, does not change its growth state. Drought resistance can be enhanced, making environmentally resistant crops useful for development.

1 shows the results of verifying the effects of various ABA receptors on ABA signaling.
Figure 2 is a result of verifying the interaction with OsPP2C according to the ABA concentration in order to confirm whether the OsPYL / RCAR7 protein has a function as an ABA receptor: A: yeast two hybridization, B: inhibiting OsPP2C activity of OsPYL / RCAR7
Figure 3 shows the results of analyzing the phenotype of the transgenic rice overexpressing OsPYL / RCAR7. A: Growth photo, B: Quantitative analysis of major agricultural traits
4 is a diagram showing a vector structure for expressing OsPYL / RCAR7 gene in rice.
5 is a diagram confirming the drought stress resistance in the transgenic rice overexpressing the OsPYL / RCAR7 gene. A: Growth photograph, B: Survival test after drought treatment, C: Water loss test

Hereinafter, the present invention will be described in detail.

The invention is drought tolerance comprising a gene encoding OsPYL / RCAR7 [ Oryza sativa PYR1 (PYRABACTINE RESISTANCE 1) -LIKE / REGULATORY COMPONENTS OF ABA RECEPTOR7] protein derived from rice, consisting of the amino acid sequence represented by SEQ ID NO: 2 Provide a recombinant vector for promotion.

The term "drought tolerance" in the present invention is a property that plants can tolerate drought damage, that is, tolerate drought, also referred to as dry resistance or drought resistance.

The gene is preferably composed of the nucleotide sequence represented by SEQ ID NO: 1, but is not limited thereto. In addition to the gene encoding the gene Absecon seusan (Abscisic acid, ABA) receptor OsPYL / RCAR7 [Oryza sativa PYR1 ( PYRABACTINE RESISTANCE 1) -LIKE / REGULATORY COMPONENTS OF ABA RECEPTOR7] protein, unlike other receptor Absecon seusan high concentration of It is characterized by its function as a receptor only in absic acid. Thus, normal growth in normal non-stressed environment, and contributes to promoting plant resistance only in a high stress environment.

In addition, variants of the above nucleotide sequences may be included within the scope of the present invention. Specifically, the gene has a base sequence having a sequence homology of at least 70%, more preferably at least 80%, even more preferably at least 90%, most preferably at least 95% with the nucleotide sequence of SEQ ID NO: 1, respectively. It may include. The "% sequence homology" for a polynucleotide is identified by comparing two optimally arranged sequences with a comparison region, wherein part of the polynucleotide sequence in the comparison region is the reference sequence (addition or deletion) for the optimal alignment of the two sequences. It may include the addition or deletion (ie, gap) compared to).

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, a heterologous peptide, or a heterologous nucleic acid. Recombinant cells can express genes or gene fragments that are not found in their natural form in either the sense or antisense form. Recombinant cells can also express genes found in natural cells, but the genes are modified and reintroduced into cells by artificial means.

The term “vector” is used to refer to a DNA fragment (s), a nucleic acid molecule, that is delivered into a cell. Vectors can replicate DNA and be reproduced independently in host cells. The term "expression vector" is often used interchangeably with "recombinant vector." The term “recombinant vector” refers to a recombinant DNA molecule comprising a coding sequence of interest and a suitable nucleic acid sequence necessary to express a coding sequence operably linked in a particular host organism. Promoters, enhancers, termination signals and polyadenylation signals available in eukaryotic cells are known.

Vectors of the invention can typically be constructed as vectors for cloning or expression. In addition, the vector of the present invention can be constructed using prokaryotic or eukaryotic cells as hosts. For example, when the recombinant vector of the present invention is an expression vector and the prokaryotic cell is a host, a strong promoter (for example, a pLλ promoter, a trp promoter, a lac promoter, a T7 promoter, a tac promoter, etc.) capable of promoting transcription may be used. It is common to include ribosomal binding sites and transcription / detox termination sequences for initiation of translation.

On the other hand, vectors that can be used in the present invention are plasmids (eg, pSC101, ColE1, pBR322, pUC8 / 9, pHC79, pGEX series, pET series and pUC19, etc.) often used in the art, phage (e.g., λgt4λB, λ) -Can be produced by manipulating viruses, λΔz1 and M13, etc.) or viruses (eg SV40, etc.).

On the other hand, when the recombinant vector of the present invention is an expression vector and the eukaryotic cell is a host, a promoter derived from the genome of the mammalian cell (e.g., a metallothionine promoter) or a promoter derived from a mammalian virus (e.g., Adenovirus late promoter, vaccinia virus 7.5K promoter, SV40 promoter, cytomegalovirus promoter and tk promoter of HSV) can be used and generally have a polyadenylation sequence as a transcription termination sequence.

Vectors of the present invention may include antibiotic resistance genes commonly used in the art as optional markers, for example ampicillin, gentamicin, carbenicillin, chloramphenicol, streptomycin, kanamycin, geneticin, neomycin And resistance genes for tetracycline and bastard herbicides.

The present invention also provides a transgenic plant transformed with a recombinant vector for improving drought resistance, including a gene encoding an OsPYL / RCAR7 protein derived from rice, which consists of the amino acid sequence represented by SEQ ID NO: 2.

Preferably, the plant has increased drought resistance by OsPYL / RCAR7 gene expression, but is not limited thereto.

The plant is preferably a monocotyledonous plant, most preferably rice ( Oryza sativa L. ), but is not limited thereto.

 Plant transformation refers to any method of transferring DNA to a plant. Such transformation methods do not necessarily have a period of regeneration and / or tissue culture. Transformation of plant species is now common for plant species, including both dicotyledonous plants as well as monocotyledonous plants. In principle, any transformation method can be used to introduce hybrid DNA according to the invention into suitable progenitor cells. Method is calcium / polyethylene glycol method for protoplasts (Krens, FA et al., 1982, Nature 296, 72-74; Negrutiu I. et al., June 1987, Plant Mol. Biol. 8, 363-373), protoplasts Electroporation (Shillito RD et al., 1985 Bio / Technol. 3, 1099-1102), microscopic injection into plant elements (Crossway A. et al., 1986, Mol. Gen. Genet. 202, 179-185 ), (DNA or RNA-coated) particle bombardment of various plant elements (Klein TM et al., 1987, Nature 327, 70), Agrobacterium tumulopasis by plant infiltration or transformation of mature pollen or vesicles And infection with (incomplete) virus (EP 0 301 316) in en mediated gene transfer. Preferred methods according to the invention include Agrobacterium mediated DNA delivery. Especially preferred is the use of the so-called binary vector technology as described in EP A 120 516 and US Pat. No. 4,940,838.

In addition, the present invention

(1) preparing a recombinant vector comprising a gene encoding the OsPYL / RCAR7 protein of claim 1;

(2) transforming the vector into a plant using Agrobacterium; And

(3) It provides a method for producing a transformed plant comprising the step of selecting the transformant.

In addition, the present invention provides a method of increasing drought resistance by overexpressing a gene encoding a rice-derived OsPYL / RCAR7 protein consisting of an amino acid sequence represented by SEQ ID NO: 2 in a plant.

Example 1 OsPYL / RCAR7 Protein Selection of Rice

<1-1> ABA Signaling Activity Assay of Rice ABA Receptor

CDNA was synthesized after total RNA isolation from Dongjin rice. PCR was performed using these cDNA as a template and primers prepared for OsPYL / RCARs. PCR products were purified using Qiagen's Gel extraction kit. The purified PCR products were cloned into Invotrogen's pENTR-D TOPO vector and they were cloned into overexpression vector pGEM3XHA. OsPYL / RCAR gene rice protoplast expression vector and Rab16a promoter-luciferase-conjugated ABA signaling reporter vector were simultaneously transformed into protoplasts and examined for responsiveness to ABA.

Specifically, rice protoplast isolation and transformation were performed as follows, and after transfection, luciferase assay was performed to analyze the reactivity with ABA upon overexpression of OsPYL / RCAR protein.

First, in order to isolate the rice protoplasts and transform the Rab16a promoter-luciferase binding vector, the peeled rice seed was sterilized with 70% ethanol and 40% sodium hypochlorite, followed by solid medium (1 / 2MS, 0.8 % agar, pH5.7) was planted in a plant culture vessel, and cultured for 7 days at 28 ℃ culture chamber. After cutting the leaf sheath of rice into 1 ~ 2 mm size, enzyme solution (1.5% cellulose R-10, 0.75% macerozyme R-10, 0.6M mannitol, 10mM MES, 0.1% BSA, 3.4) mM CaCl 2, 5 mM β-mecaptoethanol, pH5.7) was treated for 4 hours to remove cell walls.

After removing the leaf sheath of the rice using a 0.45㎛ sieve, the suspension was prepared using a W5 solution (W5 solution; 0.1% glucose, 0.9% NaCl, 2 mM MES, 0.08% KCl, 125 mM CaCl2, pH5.7). After washing, the protoplasts were precipitated by centrifugation. The precipitated protoplasts were released using MaMg solution (MaMg solution; 600mM mannitol, 15mM MgCl2, 5mM MES, pH5.6) and finally 300μl of the protoplasts with the constructed ABA receptor overexpression vector DNA (4μl) Rab16 promoter-luciferase binding vector DNA (4 μL), mixed with UBQ10 promoter DNA (1 μL, Renillia luciferase) as an internal control and transformed with 330 μL PEG It was.

To assay luciferase activity, the transformed rice protoplasts were suspended and lysed using 100 μL passive lysis buffer (Promega, USA). 10 μl of the dissolved protoplasts was examined for luciferase activity using the Promega Dual luciferase assay kit.

As a result, as shown in Figure 1, OsPYL / RCAR genes showed significantly different reactivity to ABA.

As a result, OsPYL / RCAR7 showed the lowest ABA reactivity compared to the other receptors, which showed a similar level of reactivity to the control that was not treated with absic acid (FIG. 1).

<1-2> Requirement Test of Apsis Acid of OsPYL / RCAR7 Gene

OsPYL / RCAR7 showed the same activity as the control even when 5 μM of acetic acid was treated in the protoplast transient expression system. Therefore, in order to confirm whether it can actually function as an acetic acid receptor, yeast two hybridization, BiFC, etc. The function was confirmed.

For yeast two hybridization, cDNAs of OsPP2C08, 30, 51 were cloned into pGAD7 carrier and OsPYL / RCAR1, OsPYL / RCAR5, OsPYL / RCAR6, and OsPYL / RCAR7 were cloned into pGBKT7. Each combination of OsPYL / RCAR and OsPP2C was transformed into yeast strain AH109. Transformed yeast cells were confirmed whether the interaction of yeast cell growth according to the addition of ABA in the synthetic medium in which leucine, tryptophan, histidine was removed.

 As a result of the yeast two hybridization experiments, unlike OsPYL / RCAR interacting at 0.1 or 1uM, OsPYL / RCAR7 only interacted with all of the examined OsPP2Cs and 10uM absic acid (FIG. 2A). In addition, in order to confirm that OsPYL / RCAR7 exhibits intrinsic properties of inhibiting the activity of OsPP2C, the expression method of OsPYL / RCAR7, OsPP2C, and Rab16a promoter-luciferase in the rice protoplasts was performed simultaneously in the same experimental method as in FIG. Reactivity to was observed. Although significantly lower than OsPYL / RCAR5, it was found that there was an inhibitory ability against OsPP2C8 and OsPP2C30 when 5uM acetic acid was treated (FIG. 2B). Summarizing the above results, it can be seen that OsPYL / RCAR7 is an acetic acid receptor having high concentration of acetic acid demand and low activity.

Example 2 Preparation of OsPYL / RCAR7 Gene Expression Vector and Transformant

<2-1> Construction of OsPYL / RCAR7 Gene Expression Vector

The plant transformation vector pGA2897, which is regulated by the ubiquitin promoter and selects a transformant with Basa, was used to constantly express the OsPYL / RCAR7 gene. Primer specific for OsPYL / RCAR7 gene to introduce OsPYL / RCAR7 gene into pGA2897 vector (Forward: 5'-CACCATGGCCACCAAGCGCGCGTAC-3 ', SEQ ID NO: 3' Reverse: 5'-CTATCTGCTTGTCAG CTGAAAGATATCCTG-3 ', SEQ ID NO: 4) Was produced. PCR was carried out using cDNA synthesized from Dongjin rice as a template and using the prepared primers. PCR products were purified using Qiagen's Gel extraction kit, and purified PCR products were cloned into Invotrogen's pENTR-D TOPO vector. In order to introduce OsPYL / RCAR7 gene cloned into pENTR-D TOPO vector into pGA2897, a plant transformation vector, the reaction was performed using LR clonase of Invitrogen. Finally, to confirm that the OsPYL / RCAR7 gene is introduced into pGA2897, the introduction of the OsPYL / RCAR7 gene through sequencing using a primer specific for the ubiquitin promoter possessed by the pGA2897 vector (5'-GCATATGCAGCAGCTATATGTGG-3 ', SEQ ID NO: 5). It was confirmed.

<2-2> Transformation of OsPYL / RCAR7 Gene Expression Transformant

The OsPYL / RCAR7 gene expression vector prepared in <Example 2-1> was transformed into Agrobacterium and then transformed into rice to prepare a transformed rice.

Specifically, the OsPYL / RCAR7 / pGA2897 vector, which is produced so that the OsPYL / RCAR7 gene can be expressed at all times, was used as A. tumefaciens. LBA4404) to freeze and thaw 2-3 times to transform, then transformed by heat shock method at 37 ℃ for 2 days in YEP medium (Yeast 10g, NaCl 5g, peptone 10g, Agar 15g / 1L) Incubation confirmed colonies. Colonies confirmed to be transformed by colony PCR were prepared using AB medium (AB buffer (K ₂ HPO ₄ 60 g, NaH ₂ PO ₄ 20 g / 1 L), AB Salts (NH ₄ Cl 60 g, MgSO ₄ ㆍ 7H) for transformation into rice. ₂ O 6g, and cultured at 3g KCl, CaCl ₂ and _{2H2O 0.265g, FeSO 4 .7H 2 O} 50mg / 1L, Glucose 5g / 1L).

Dongjin rice seed used for rice transformation used brown rice peeled. Soybean seeds were soaked in 70% ethanol for 1 minute and then sterilized for 40 minutes in 50% solution. At this time, it is more preferable to use 10uL Tween 20 dropped per 30mL of lacx.

After disinfecting the disinfectant solution, rice seeds were washed 5 times with sterile water. When the disinfectant solution was washed well, water was removed from the rice seed using filter paper and the rice seed was soaked in 2N6 medium. At this time, rice seed was put in the medium so that the contact with the medium was good, and the seed was faced upward. 2N6 medium contains 4g / L and 30g / L sucrose, 0.5g / L proline, 0.5g / L glutamine, 0.3g / L casamino acids, 0.01g / L myo Solid medium (pH5.8) with added -inositol, 2 mg / L 2,4-D, 4 g / L phytagel. The culture temperature was 28 ℃ and treated with cancer.

Incubation was carried out for 5 days at 2N6 to remove endosperm when callus was induced. The draining oil is carefully removed using tweezers so that the callus is not damaged, and the cotyledons and roots should be left intact to prevent the callus from being damaged.

Agrobacterium was prepared by inserting the gene of interest, incubated in AB solid medium for 3 days, and then collected in AAM liquid medium to make a suspension. The concentration of the suspension was measured by using an absorbance machine. First, OD ₅₉₅ = 0.1, and then diluted 10-fold with AAM to OD ₅₉₅ = 0.01.

The suspension was inoculated with Agrobacterium by immersion for 2 minutes to remove the endosperm, and the solution was gently shaken for 2 minutes. At the end of the inoculation, only the suspension was poured out of the callus and the filtrate was removed using filter paper. This is a rapid process and care is taken not to allow the callus to dry too long on filter paper. Callus was co-cultured by transferring to 2N6-AS medium. 2N6-AS medium was adjusted to pH5.2 by adding 10g / L glucose and 100uM acetosyringone to 2N6 medium. On the medium, the filter paper was laid down one by one in advance so that the medium component was sufficiently infiltrated, and then the inoculated callus was placed on the filter paper. The filter paper is intended to prevent excessive growth of Agrobacterium during the co-culture and the callus receding. Co-culture was performed by cancer culture at 25 ℃.

At the end of the co-culture, Agrobacterium, which was responsible for inserting useful genes into the callus, was no longer needed and was removed by washing. The washing was performed with a solution containing 500 mg of antibiotic carbenicillin in 1 L of sterile distilled water. After washing the callus about 3 times, all of the filtrate was removed using a filter paper. In the case of using the Agrobacterium LBA4404 strain, it may be more preferable not to perform the above washing operation because no excessive growth of Agrobacterium was achieved during the coculture period. Callus was transferred to 2N6-CH medium in which antibiotics cefotaxime 200 mg / L and Hygromycin 40 mg / L were added to 2N6 medium. At this time, the cotyledons and roots were removed using tweezers, and the callus was divided into 2 ~ 3 pieces and the culture area was moved to maximize the area of contact with the medium. The culture temperature was incubated for 14 days in the cancer incubator at 28 ℃ to induce transformed callus.

After 14 days, when the transformed callus was split and grown in 2N6-CH medium to which the antibiotic hygromycin was added, it was passaged to MSR medium, which is a regeneration medium capable of inducing chute and root. The distinction of transformed callus was judged by the presence of newly differentiated callus mass, and the browned and dead callus was discarded. Dairy plants grown in MSR regeneration medium were transferred to aquatic soils and grown in greenhouses.

<Example 3> Drought Resistance Analysis of Rice Transfected with OsPYL / RCAR7 Gene

<3-1> Main Agricultural Characterization of OsPYL / RCAR7 Overexpressed Transgenic Rice

Apsis acid receptor overexpressing plants have characteristics such as growth inhibition and yield decrease, and thus have many disadvantages in their utility. Therefore, in order to confirm whether the OsPYL / RCAR7 overexpressing transformants show the same effect, the main agricultural trait characteristics of the overexpressing transformants were assayed. In four strains of OsPYL / RCAR7 overexpressing transformants prepared in Example 2, the ear length and soy size were almost the same as those of the control, and the average value of C14-1 and 3 in the number of ear and total seed weight was reduced compared to the control. However, there was no statistically significant difference, and the two different strains showed the same numbers. Therefore, OsPYL / RCAR7 was determined to show no growth inhibition unlike the general acetic acid receptor overexpressing plants (FIG. 3).

<3-2> OsPYL Of RCAR7  Overexpression Transgenic rice  Drought Resistance Characteristic Test

 In order to determine whether OsPYL / RCAR7 has very low abscitic acid receptor activity and the drought resistance effect of the overexpressed transgenic rice under the same growth conditions as that of the control, as well as other abscitic acid receptors, Resistance was assayed.

As a result, OsPYL / RCAR7 overexpressed transgenic rice showed higher survival rate compared to the control when drought treatment in the pot, and water loss assay (water loss assay) confirmed that the water loss is delayed compared to the control, thereby improving drought resistance It was confirmed that (Fig. 4).

<110> Republic of Korea <120> A novel genes for enhancing drought resistance and their uses <130> P17R12C1171 <160> 5 <170> KoPatentIn 3.0 <210> 1 <211> 621 <212> DNA <213> Artificial Sequence <220> <223> OsPYL / RCAR7 <400> 1 atgaacggcg ctggtggtgc gggaggagca gcggcaggga agttgccaat ggtgagccac 60 cgacgggtgc agtgcagact agcggacaag cggtgtgagc tccgggagga agagatggag 120 tatatccggc agttccaccg ccacgagccc agcagcaacc agtgcacctc gttcgtcgcc 180 aagcatatca aggcgcccct ccaaaccgtt tggtcactag tgaggaggtt tgatcagcca 240 caacttttca aaccttttgt gagaaagtgc gtaatgcgag aaaacattat tgtgaccgga 300 tgtgttaggg aggtcaatgt tcaaagcggg cttccagcca caaggagcac tgagaggtta 360 gagttgcttg atgataacga acatatcctc aaagtcaagt ttattggggg cgatcatatg 420 ttgaagaatt actcatccat cctaaccatc cactctgagg tcatcgatgg ccaacttgga 480 acattggtgg tcgaatcatt tgtagtggat attccagatg ggaacaccaa agacgacata 540 tgctatttca tcgagaacgt tctcaggtgc aaccttatga cccttgctga tgtgtcagag 600 gagcgccttg ccaatccttg a 621 <210> 2 <211> 206 <212> PRT <213> Artificial Sequence <220> <223> OsPYL / RCAR7 <400> 2 Met Asn Gly Ala Gly Gly Ala Gly Gly Ala Ala Ala Gly Lys Leu Pro   1 5 10 15 Met Val Ser His Arg Arg Val Gln Cys Arg Leu Ala Asp Lys Arg Cys              20 25 30 Glu Leu Arg Glu Glu Glu Met Glu Tyr Ile Arg Gln Phe His Arg His          35 40 45 Glu Pro Ser Ser Asn Gln Cys Thr Ser Phe Val Ala Lys His Ile Lys      50 55 60 Ala Pro Leu Gln Thr Val Trp Ser Leu Val Arg Arg Phe Asp Gln Pro  65 70 75 80 Gln Leu Phe Lys Pro Phe Val Arg Lys Cys Val Met Arg Glu Asn Ile                  85 90 95 Ile Val Thr Gly Cys Val Arg Glu Val Asn Val Gln Ser Gly Leu Pro             100 105 110 Ala Thr Arg Ser Thr Glu Arg Leu Glu Leu Leu Asp Asp Asn Glu His         115 120 125 Ile Leu Lys Val Lys Phe Ile Gly Gly Asp His Met Leu Lys Asn Tyr     130 135 140 Ser Ser Ile Leu Thr Ile His Ser Glu Val Ile Asp Gly Gln Leu Gly 145 150 155 160 Thr Leu Val Val Glu Ser Phe Val Val Asp Ile Pro Asp Gly Asn Thr                 165 170 175 Lys Asp Asp Ile Cys Tyr Phe Ile Glu Asn Val Leu Arg Cys Asn Leu             180 185 190 Met Thr Leu Ala Asp Val Ser Glu Glu Arg Leu Ala Asn Pro         195 200 205 <210> 3 <211> 25 <212> DNA <213> Artificial Sequence <220> <223> OsPYL / RCAR7 forward primer <400> 3 caccatggcc accaagcgcg cgtac 25 <210> 4 <211> 30 <212> DNA <213> Artificial Sequence <220> <223> OsPYL / RCAR7 reverse primer <400> 4 ctatctgctt gtcagctgaa agatatcctg 30 <210> 5 <211> 23 <212> DNA <213> Artificial Sequence <220> <223> ubiquitin promoter primer <400> 5 gcatatgcag cagctatatg tgg 23

Claims (8)

A recombinant vector for enhancing resistance to dryness, comprising a gene encoding an OsPYL / RCAR7 protein derived from rice, consisting of the amino acid sequence represented by SEQ ID NO: 2.
The method of claim 1, wherein the gene is a recombinant vector for drying resistance, characterized in that consisting of the nucleotide sequence represented by SEQ ID NO: 1.
A transformed plant transformed with the recombinant vector of claim 1.
The transgenic plant of claim 3, wherein the plant has increased resistance to dryness by expression of OsPYL / RCAR7 gene.
The transgenic plant of claim 3, wherein the plant is a monocotyledonous plant.
The transformed plant according to claim 5, wherein the monocotyledonous plant is rice ( Oryza sativa L. ).
(1) preparing a recombinant vector comprising a gene encoding the OsPYL / RCAR7 protein of claim 1;
(2) transforming the vector into a plant using Agrobacterium; And
(3) A method for producing a transformed plant comprising the step of selecting the transformant.
A method of increasing dry resistance by overexpressing a gene encoding a rice-derived OsPYL / RCAR7 protein consisting of an amino acid sequence represented by SEQ ID NO: 2 in a plant.

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