KR102448536B1 - ABA receptor mutants and use thereof - Google Patents

Abstract

The present invention relates to an ABA receptor mutant protein, a gene encoding the same, a recombinant vector comprising the gene, a plant transformed with the recombinant vector, and a plant with improved resistance to drying or environmental stress, and a method for enhancing drying or environmental stress resistance of the plant. it's about
The use of the ABA receptor mutant of the present invention increases the ABA-dependent and/or non-dependent response, so it is possible to regulate the ABA signaling activity involved in drying and environmental stress resistance. By using the vector containing the ABA receptor mutant gene of the present invention, a transgenic plant having improved resistance to drying or environmental stress can be prepared.

Description

ABA receptor mutants and use thereof

The present invention relates to an ABA receptor mutant protein, a gene encoding the same, a recombinant vector comprising the gene, a plant transformed with the recombinant vector, and a plant with improved resistance to drying or environmental stress, and a method for enhancing drying or environmental stress resistance of the plant. it's about

Abscisic acid (ABA) is a plant hormone that modulates signal transduction associated with abiotic stress responses. The ABA signaling pathway has been exploited to improve plant stress responses and related harvesting traits through a number of approaches. Direct use of ABA in plants improves the plant's water use efficiency. Therefore, there is growing interest in ABA agonists as ABA agonist molecules may be beneficial in improving crop yields. Supplementary methods of activating the ABA pathway include increasing the sensitivity of plants to ABA through genetic methods. For example, conditional antisense of the farnesyl transferase beta subunit gene, which increases the ABA sensitivity of plants, improves yields under moderate drought in both Canola and Arabidopsis.

Therefore, the present inventors study the ABA receptor mutation that activates ABA signal independent of ABA in order to enhance the drying or environmental stress resistance of plants, and the present invention by identifying the sequence and combination of ABA receptor mutations with excellent ABA signaling activity completed.

Republic of Korea Patent Registration No. 10-1686429

The problem to be solved by the present invention is ABA in which leucine (L) at position 93 in the amino acid sequence of SEQ ID NO: 1 is substituted with phenylalanin (F), tyrosine (Y) or tryptophan (W) To provide a receptor mutant protein.

Another object of the present invention is to provide an ABA receptor mutant gene encoding the ABA receptor mutant protein.

Another object of the present invention is to provide a recombinant vector comprising the ABA receptor mutant gene.

Another object of the present invention is to provide a transgenic plant with enhanced resistance to dry or environmental stress transformed with the recombinant vector.

Another object of the present invention is to provide a method for enhancing the resistance to drying or environmental stress of a plant, comprising the step of transforming the plant with a recombinant vector to overexpress the ABA receptor mutant protein.

In one aspect of the present invention in order to solve the above problems, leucine (L) at position 93 in the amino acid sequence of SEQ ID NO: 1 is phenylalanine (phenylalanin, F), tyrosine (Y) or tryptophan (tryptophan) , W) substituted ABA receptor mutant protein.

In the present invention, "abcisic acid (ABA)" refers to eye dormancy, seed dormancy and/or maturation, leaf and fruit detachment, and response to a wide variety of biological stresses (eg, low temperature, high temperature, salinity and drought). , a multifunctional plant hormone implicated in various plant-protective functions. ABA also regulates stomatal blocking by a mechanism independent of CO ₂ concentration.

In the present invention, "ABA receptor" is a protein having the amino acid sequence of SEQ ID NO: 1 as OsPYL/RCAR5 of rice. It binds to PP2C via ABA of the ABA receptor protein and mediates ABA-dependent signaling. Specifically, the amino acid sequence of SEQ ID NO: 1 may be encoded by the nucleotide sequence of SEQ ID NO: 2.

The "mutant protein" of the present invention is a protein in which some amino acids of OsPYL/RCAR5 of rice are substituted. Specifically, in the amino acid sequence of the OsPYL / RCAR5 protein of SEQ ID NO: 1, leucine (L) at position 93 is substituted with an amino acid having a phenyl ring residue.

The ABA receptor mutation is characterized in that it interacts with PP2C51 in an ABA-independent manner to induce ABA-independent signaling.

In an embodiment of the present invention, random mutations were made using the OsPYL/RCAR5 gene, which has a high demand for ABA among Abscisic acid (ABA) receptors in rice, and ABA-independent and PP2C51 and PP2C51 using a yeast-to-hybrid system Sixteen interacting OsPYL/RCAR5 mutants were selected. As a result of confirming the nucleotide sequence of the 16 OsPYL/RCAR5 mutants, mutations occurred at several amino acid sites. Therefore, in order to identify the causative amino acid of the ABA-independent interaction, only one amino acid mutation was induced, and the interaction with PP2C51 was confirmed using the yeast-to-hybrid system. was confirmed. Specifically, it was confirmed that the amino acid mutation sequence L93F, E100V, N102I, N102S, and N102Y single amino acid mutations of OsPYL / RCAR5 affecting the interaction with PP2C51 independent of ABA was 93 L, 100 E, 102 N. . In addition, we analyzed whether the mutant OsPYL/RCAR5 protein of these three amino acids actually affects ABA signaling in plants. L93F, E100V, and N102I mutations were overexpressed in rice protoplasts to confirm the expression of the ABA response reporter. L93F and N102I increased the ABA signaling activity compared to the OsPYL/RCAR5 control even in the absence of ABA, but the E100V mutant showed similar signaling activity to the OsPYL/RCAR5 control without the mutation.

In order to select a mutant with an optimized ABA-independent activity, the 93rd amino acid leucine (L), selected as the main causative amino acid for ABA-independent signaling, is substituted with an amino acid having different biochemical properties, and ABA signal transduction activity As a result of the analysis, when the 93rd leucine (Leucine, L) is substituted with an amino acid having a phenyl ring structure residue (phenylalanine (phenylalanin, F), tyrosine (tyrosine, Y) and tryptophan, W)), ABA signal transduction It was confirmed that the activity was increased, and the activity of L93W was found to be the highest.

Therefore, in the present invention, the “amino acid having a phenyl ring residue” is phenylalanine (F), tyrosine (Y), or tryptophan (W), and specifically tryptophan (W).

In addition, in an embodiment of the present invention, a mutant was prepared in which asparagine (N) at position 102, selected as an amino acid for regulating ABA-independent activity, was substituted with an amino acid of a residue having different biochemical properties, and thus the ABA signal As a result of analyzing the transfer activity, in the absence of ABA treatment, activity was increased in all mutations except for the mutation in which the N102 amino acid was substituted with alanine (Alanine, A), and in particular, tyrosine (Y) or glutamic acid (Glutamic acid, E) ) showed the greatest activity in the mutant substituted with

Therefore, the ABA receptor mutant protein of the present invention may be one in which asparagine (N) at position 102 in the amino acid sequence of SEQ ID NO: 1 is substituted with tyrosine (Tyrosine, Y) or glutamic acid (E).

In an embodiment of the present invention, a double mutant was prepared by combining N102 and L93 amino acid mutations to construct OsPYL/RCAR5 with an optimized ABA-independent signaling response. Specifically, a double mutant was prepared by combining N102I or N102Y with L93F or L93W, and ABA-independent and dependent ABA signaling activity was compared and analyzed with a single amino acid mutation.

As a result of analyzing the ABA signaling activity in the absence of ABA using the rice protoplast transient expression system, the double mutant L93W and N102Y showed higher activity than the single mutant. On the other hand, the double mutants of L93F and N102I showed similar levels of activity as those of the N102Y single mutant.

That is, it was confirmed that the two mutations caused additional activity enhancement. In particular, in OsPYL/RCAR5 [L93W, N102Y], the activity was much higher than the individual L93W and N102Y mutations, so the two mutations interact synergistically to enhance ABA signaling activity. confirmed that it did.

In another aspect of the present invention, there is provided an ABA receptor mutant gene encoding the ABA receptor mutant protein.

The ABA receptor mutant gene of the present invention encodes an ABA receptor mutant protein in which leucine (L) at position 93 in the amino acid sequence of SEQ ID NO: 1 is substituted with an amino acid having a phenyl ring residue.

In addition, in the amino acid sequence of SEQ ID NO: 1, leucine (L) at position 93 is substituted with an amino acid having a phenyling residue, and at the same time asparagine at position 102 (Asparagine, N) is tyrosine (Tyrosine, Y) or It may be one encoding an ABA receptor mutant protein substituted with glutamic acid (E). Specifically, at the same time asparagine at the 102nd position (Asparagine, N) may be substituted with tyrosine (Tyrosine, Y) to encode an ABA receptor mutant protein.

In the above-described example, it was confirmed that ABA-independent ABA signaling activity was enhanced during overexpression of ABA receptor mutant proteins (L93W and N102Y single mutations and L93W and N102Y double mutations) in the rice protoplast transient expression system.

Therefore, by overexpressing the ABA receptor mutant of the present invention, it is possible to increase the ABA signaling activity in a plant independent of ABA, and through activation of the ABA signal, it is possible to enhance or improve the drying or environmental stress resistance of the plant.

In another aspect of the present invention, there is provided a recombinant vector comprising the ABA receptor mutant gene.

The recombinant vector of the present invention contains an ABA receptor mutant gene.

Description of the ABA receptor mutant gene of the present invention is as described above.

In the present invention, "recombinant" refers to a cell in which the cell replicates a heterologous nucleic acid, expresses the nucleic acid, or expresses a peptide, a heterologous peptide, or a protein encoded by the heterologous nucleic acid. Recombinant cells can express genes or gene segments not found in the native form of the cell, either in sense or antisense form. In addition, the recombinant cell can express a gene found in a cell in a natural state, but the gene is modified and re-introduced into the cell by artificial means.

As used herein, "vector" means a DNA preparation having a DNA sequence operably linked to a suitable regulatory sequence capable of expressing the DNA in a suitable host. A vector can be a plasmid, a phage particle, or simply a potential genomic insert. Upon transformation into an appropriate host, the vector can replicate and function independently of the host genome, or in some cases can be integrated into the genome itself. In particular, "vector for plant transformation" refers to a recombinant DNA molecule comprising a desired coding sequence and an appropriate nucleic acid sequence essential for expressing a coding sequence operably linked in a specific plant host organism. Promoters, enhancers, termination signals and polyadenylation signals available in plant cells are known to those skilled in the art. The vector of the present invention is most preferably a vector for plant transformation, but is not limited thereto.

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 a prokaryotic cell or a eukaryotic cell as a host. For example, when the recombinant vector of the present invention is an expression vector and a prokaryotic cell is used as a host, a strong promoter capable of proceeding transcription (eg, pLλ promoter, trp promoter, lac promoter, T7 promoter, tac promoter, etc.) , a ribosome binding site for initiation of translation and a transcription/translation termination sequence.

In another aspect of the present invention, there is provided a transgenic plant having improved resistance to dry or environmental stress transformed with the recombinant vector.

Description of the recombinant vector in the present invention is as described above.

The plants are rice ( Oryza sativa L.), wheat ( Triticum aestivum L.), barley ( Hordeum vulgare var. hexastichon), sorghum ( Sorghum bicolor M.), corn ( Zea mays L.), millet ( Panicum miliaceum L.). ), soybean ( Glycine max L.) or potato ( Solanum tuberosum ) may be, for example, rice ( Oryza sativa L.), but may be, but is not limited thereto.

The transgenic plant according to the present invention can be obtained through a sexual reproduction method or asexual reproduction method, which is a conventional method in the art. More specifically, the plant of the present invention can be obtained through sexual reproduction, which is a process of producing seeds through the pollination process of flowers and propagating from the seeds. In addition, it can be obtained through the asexual reproduction method, which is a process of induction, rooting, and soil acclimatization of callus according to a conventional method after transforming a plant with the recombinant expression vector according to the present invention. That is, the explants of plants transformed with the recombinant expression vector of the present invention are plated in a suitable medium known in the art, and then cultured under appropriate conditions to induce the formation of callus, and when shoots are formed, transferred to a hormone-free medium incubate After about 2 weeks, the shoots are transferred to a rooting medium to induce roots. The transformed plant according to the present invention can be obtained by acclimatizing the roots by transplanting them into soil after induced. In the present invention, the transgenic plant may include a whole plant as well as a tissue, cell or seed obtainable therefrom.

In the above-described embodiment of the present invention, it was confirmed that ABA-independent ABA signaling activity was enhanced during overexpression of ABA receptor mutant proteins (L93W and N102Y single mutations and L93W and N102Y double mutations) in the rice protoplast transient expression system.

It is possible to improve or enhance resistance to drying or environmental stress through ABA-independent ABA signaling in plants.

Therefore, the transgenic plant overexpressing the ABA receptor mutant gene has excellent ABA signaling activity involved in drying stress resistance, and thus drying or environmental stress resistance can be enhanced or improved.

As used herein, "dry stress resistance" means "drought resistance" or "dry tolerance", including any of its variations, in which a plant recovers from a period of drought stress (i.e., little or no water for a period of several days). refers to ability. Typically, the drought stress will be at least 5 days, eg depending on the plant species, eg 18 to 20 or more days (eg at least 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20 days).

As used herein, “environmental stress” refers to a plant, plant cell, or non-living (“abiotic”) physical or chemical agent having a detrimental effect on the metabolism, growth, development, reproduction or survival (collectively, “growth”) of the plant. exposure, etc. Stress can be, for example, an environmental factor such as water (eg, flood, drought, or dehydration), anaerobic conditions (eg, lower levels of oxygen or higher levels of CO ₂ ), abnormal osmotic conditions, salinity, or Temperature (eg, heating/heating, cooling, freezing, or freezing), nutrient deficiencies or exposure to contaminants, or by hormones, second messengers or other molecules may impose on plants. Anaerobic stress results from, for example, a decrease in the level of oxygen sufficient to produce a stress response (hypoxia or anoxia). Flood stress can be due to prolonged or transient immersion of plants, plant parts, tissues or isolated cells in liquid medium, such as during the rainy season, feng shui season, or excessive plant stimulation, and the like. Cooling stress or heating stress may occur due to a decrease or increase in temperature, respectively, from the optimum range of growth temperature for a particular plant species. Such optimal growth temperature ranges are readily determined or known to those skilled in the art. Dehydration stress can be caused by water loss, reduced turgor pressure, or reduced water content of a cell, tissue, organ or whole plant. Drought stress may be caused or associated with a deprivation of water or a reduced supply of water to a cell, tissue, organ or organism. Salt-induced stress (salt-stress) may be associated with or induced by perturbations in the osmotic potential of the cell's intracellular or extracellular environment.

The term "environmental stress resistance" as used herein refers to an increased resistance or tolerance of a plant to environmental stress as compared to a plant under normal conditions and the ability to perform in a relatively superior manner when under environmental stress conditions.

In another aspect of the present invention, there is provided a method for improving resistance to drying or environmental stress of a plant, comprising the step of transforming the plant with a recombinant vector to overexpress the ABA receptor mutant protein.

A method for overexpressing the protein is to transform a plant with a linked recombinant vector containing an ABA receptor mutant gene.

The description of the ABA receptor mutant gene, plant, and transformation in the present invention is the same as described above.

It was confirmed that ABA-independent and dependent ABA signaling activity was increased in plant protoplasts using the ABA receptor mutant of the present invention. Therefore, it will be possible to develop environmental disaster-resistant crops using these mutant proteins, thereby enabling the supply of agricultural products with improved productivity.

1 is a result of selecting 16 ABA-independent interaction-promoting OsPYL/RCAR5 mutations using a yeast two hybrid.
Figure 2 is the result of confirming the ABA-independent interaction enhancement of a single amino acid mutation finally using Yeast two Hybrid.
3 is a result of analyzing the expression of the ABA response reporter by overexpressing L93F, E100V, and N102I mutations in rice protoplasts to determine whether the mutant OsPYL/RCAR5 protein actually affects ABA signaling in plants.
Figure 4 shows the results of analyzing the ABA signaling activity (ABA untreated/treated) in each mutant by substituting the 102nd leucine amino acid to select the ABA optimal response mutant.
5 is a comparison result of ABA signaling activity enhancement of single amino acid mutations and double amino acid mutations.

Hereinafter, the configuration and effects of the present invention will be described in more detail through examples. These examples are only for illustrating the present invention, and the scope of the present invention is not limited by these examples.

<Example 1> Mutation selection of ABA receptors (OsPYL/RCAR5) in rice

A library consisting of approximately 2.5×10 ⁷ colonies by making random mutations using PCR technique targeting the OsPYL/RCAR5 gene (SEQ ID NO: 2), which has a high demand for ABA among Abscisic acid (ABA) receptors in rice. (library) was created. The gene of SEQ ID NO: 2 encodes the amino acid sequence of SEQ ID NO: 1 (Table 1).

division order
OsPYL/RCAR5 amino acid sequence
(SEQ ID NO: 1) MVGLVGGGGWRVGDDAAGGGGGGAVAAGAAAAAEAEHMRRLHSHAPGEHQCSSALVKHIK
APVHLVWSLVRSFDQPQRYKPFVSRCVVRGGDLEIGSVREVNVKTGLPATTSTERLELLD
DDEHILSVKFVGGDHRLRNYSSIVTVHPESIDGRPGTLVIESFVVDVPDGNTKDETCYFV
EAVIKCNLTSLAEVSERLAVQSPTSPLEQ* OsPYL/RCAR5
CDS gene sequence
(SEQ ID NO:2) ATGGTGGGGCTTGTGGGAGGAGGAGGTTGGAGGGTCGGGGATGATGCGGCGGGTGGGGGT
GGGGGAGGAGCGGTGGCGGCGGGGGCTGCGGCGGCGGCGGAGGCGGAGCACATGCGGAGG
CTCCACAGCCACGCCCCCGGCGAGCACCAGTGCAGCTCCGCGCTCGTCAAGCACATCAAG
GCTCCTGTTCACCTCGTGTGGTCGCTGGTGCGGAGCTTCGACCAGCCGCAGAGGGTACAAG
CCGTTCGTCAGCCGCTGCGTCGTGCGCGGCGGCGACCTCGAGATCGGCAGCTGTGCGCGAG
GTCAACGTCAAGACCGGCCTCCCGGCGACCACCAGCACGGAGAGGCTCGAGCTGCTCGAC
GACGACGAGCACATCCTCAGCGTCAAGTTCGTCGGCGGCGACCACCGCCTCAGGAACTAC
TCATCCATCGTAACTGTCCATCCGGAGAGCATCGATGGAAGACCAGGGACGCTTGTGATT
GAATCATTTGTGGTGGACGTGCCTGATGGAAATACAAAGGACGAGACATGCTACTTTGTC
GAGGCCGTGATCAAGTGCAACTTAACATCTCTCGCCGAGGTATCAGAGCGGCTAGCAGTT
CAGTCACCCACCTCGCCACTTGAACAGTAG

A mutant that interacts with OsPP2C51 in an ABA-independent manner was selected by performing a Yeast two Hybrid. About 8×10 ⁶ colonies were screened in the selection medium without addition of ABA, and 144 colonies were first selected, and the same mutation was removed through sequencing of the primary selection colonies to reproduce the final ABA-independent interaction. 16 selection colonies were secured (Table 2, FIG. 1).

<Example 2> ABA-independent interaction-inducing single amino acid mutation selection

As a result of confirming amino acid mutations through sequencing of 16 mutants selected in Example 1, it was found that two or more amino acids were mutated, resulting in an ABA-independent interaction.

A single amino acid mutation was prepared for all mutation sites by analyzing the nucleotide sequence of the mutation, and an amino acid mutation sequence affecting the ABA-independent interaction was identified by performing a yeast two hybrid analysis.

As a result of analyzing the interaction of OsPYL/RCAR5 with OsPP2C51 under ABA-treated or untreated conditions of L93F, E100V, N102I, N102S, and N102Y single amino acid mutations, the amino acid mutation sequence that affects the interaction with PP2C51 independent of ABA is 93rd It was confirmed that L, 100th E, 102nd N (Fig. 2).

<Example 3> ABA-independent interaction-inducing single amino acid mutation signaling characteristics assay

The single amino acid mutation (L93F) selected in Example 2 was overexpressed using the rice protoplast transient expression system, and the effect on ABA signaling in planta was tested.

In the non-ABA treatment group, control OsPYL/RCAR5 and three types of OsPYL/RCAR5 mutant proteins (E100V, L93F, N102I amino acid mutations) were compared and analyzed whether they actually affect ABA signaling in plants. L93F, E100V, and N102I mutations were overexpressed in rice protoplasts to confirm the expression of the ABA response reporter. L93F and N102I increased the ABA signaling activity compared to the OsPYL/RCAR5 control even without ABA treatment, but the E100V mutant showed similar signaling activity to the OsPYL/RCAR5 control without the mutation ( FIG. 3 ).

<Example 4> Selection of ABA receptor single amino acid mutations optimized for ABA-independent and dependent signaling responses

In Example 3, when L93F and N102I mutations were overexpressed in rice protoplasts, it was confirmed that ABA signaling activity was excellent in ABA-untreated groups. prepared and analyzed for ABA signaling activity in these mutants to select optimal mutants.

1. The 93rd leucine (L) mutation (L93)

In Examples 2 and 3, the 93th amino acid leucine, selected as the main causative amino acid for ABA-independent signaling, was substituted with amino acids of residues having different biochemical properties, and thus ABA signaling activity was analyzed.

Leucine (Leucine, L), which is the 93rd amino acid of SEQ ID NO: 1, is arginine (Arginine, R), glutamic acid (E), glutamine (Glutamine, Q), serine (Serine, S), proline (Proline, P) , Alanine (A), phenylalanine (Phenylalanine, F), methionine (M), tryptophan (Tryptophan, W), or tyrosine (Tyrosine, Y) to create a mutant and use the rice protoplast transient expression system Thus, the ABA signaling activity according to each amino acid mutation was analyzed.

Among the mutations for L93 amino acids, ABA signaling activity was significantly increased in L93F, L93Y, and L93W in the absence of ABA treatment, and the activity of L93W was the highest among them. In the case of ABA treatment, a slight decrease in activity in L93Y was observed, and no significant change was observed in other mutations (FIG. 4).

That is, when the 93th leucine of SEQ ID NO: 1 is substituted with amino acids (phenylalanin, F), tyrosine (Y) and tryptophan (W) having a residue of a phenyl ring structure, ABA signaling activity is increase was confirmed.

2. 102nd Asparagine (N) Mutation (N102)

In Examples 2 and 3, asparagine (Asparagine, N) at position 102, selected as the main causative amino acid for ABA-independent signaling, was substituted with amino acids of residues having different biochemical properties, and ABA signaling according to this Activity was assayed.

Specifically, asparagine (Asparagine, N), which is the 102nd amino acid of SEQ ID NO: 1, is arginine (Arginine, R), glutamic acid (E), serine (Serine, S), threonine (Threonine, T), alanine (Alanine, A), Isoleucine (I), and tyrosine (Tyrosine, Y) substituted mutants were prepared, and ABA signaling activity according to each amino acid mutation was analyzed using the rice protoplast transient expression system.

In the absence of ABA treatment, activity was increased in all mutations except for the mutation in which the N102 amino acid was substituted with alanine (Alanine, A). In particular, tyrosine (Y) and glutamic acid (E) showed the greatest activity. It was. There was no significant difference in activity during ABA treatment, but ABA signaling activity was slightly increased in arginine (R), threonine (Threonine, T), isoleucine (Isoleucine, I), and tyrosine (Y) mutants, and glutamic acid (Glutamic acid, E) and serine (Serine, S) mutants no change, alanine (Alanine, A) mutant activity was significantly reduced (Fig. 5).

<Example 5> ABA-independent and ABA-dependent signaling response optimization ABA receptor double mutant selection

A double mutant was prepared by combining the N102 and L93 amino acid mutations to optimize the ABA-independent signaling response. Specifically, a double mutant was constructed by combining N102I or N102Y with L93F or L93W and confirmed for ABA-independent and dependent ABA signaling activity. As a result of analyzing the ABA signaling activity in the absence of ABA using the rice protoplast transient expression system, the double mutant L93W and N102Y showed higher activity than the single mutant. On the other hand, the double mutants of L93F and N102I showed similar levels of activity as those of the N102Y single mutant.

That is, it was found that the double mutation of L93 and N102 caused additional activity enhancement. In particular, in OsPYL/RCAR5 [L93W, N102Y], the activity was much higher than that of individual L93W and N102Y mutations. It was confirmed that there was a synergistic interaction.

<110> REPUBLIC OF KOREA (MANAGEMENT : RURAL DEVELOPMENT ADMINISTRATION) <120> ABA receptor mutants and use thereof <130> DP20200058 <160> 2 <170> KoPatentIn 3.0 <210> 1 <211> 209 <212> PRT <213> Artificial Sequence <220> <223> OsPYL/RCAR5 <400> 1 Met Val Gly Leu Val Gly Gly Gly Gly Trp Arg Val Gly Asp Asp Ala 1 5 10 15 Ala Gly Gly Gly Gly Gly Gly Ala Val Ala Ala Gly Ala Ala Ala Ala 20 25 30 Ala Glu Ala Glu His Met Arg Arg Leu His Ser His Ala Pro Gly Glu 35 40 45 His Gln Cys Ser Ser Ala Leu Val Lys His Ile Lys Ala Pro Val His 50 55 60 Leu Val Trp Ser Leu Val Arg Ser Phe Asp Gln Pro Gln Arg Tyr Lys 65 70 75 80 Pro Phe Val Ser Arg Cys Val Val Arg Gly Gly Asp Leu Glu Ile Gly 85 90 95 Ser Val Arg Glu Val Asn Val Lys Thr Gly Leu Pro Ala Thr Thr Ser 100 105 110 Thr Glu Arg Leu Glu Leu Leu Asp Asp Asp Glu His Ile Leu Ser Val 115 120 125 Lys Phe Val Gly Gly Asp His Arg Leu Arg Asn Tyr Ser Ser Ile Val 130 135 140 Thr Val His Pro Glu Ser Ile Asp Gly Arg Pro Gly Thr Leu Val Ile 145 150 155 160 Glu Ser Phe Val Val Asp Val Pro Asp Gly Asn Thr Lys Asp Glu Thr 165 170 175 Cys Tyr Phe Val Glu Ala Val Ile Lys Cys Asn Leu Thr Ser Leu Ala 180 185 190 Glu Val Ser Glu Arg Leu Ala Val Gln Ser Pro Thr Ser Pro Leu Glu 195 200 205 Gln <210> 2 <211> 630 <212> DNA <213> Artificial Sequence <220> <223> OsPYL/RCAR5 CDS <400> 2 atggtggggc ttgtgggagg aggaggttgg agggtcgggg atgatgcggc gggtgggggt 60 gggggaggag cggtggcggc gggggctgcg gcggcggcgg aggcggagca catgcggagg 120 ctccacagcc acgcccccgg cgagcaccag tgcagctccg cgctcgtcaa gcacatcaag 180 gctcctgttc acctcgtgtg gtcgctggtg cggagcttcg accagccgca gaggtacaag 240 ccgttcgtca gccgctgcgt cgtgcgcggc ggcgacctcg agatcggcag cgtgcgcgag 300 gtcaacgtca agaccggcct cccggcgacc accagcacgg agaggctcga gctgctcgac 360 gacgacgagc acatcctcag cgtcaagttc gtcggcggcg accaccgcct caggaactac 420 tcatccatcg taactgtcca tccggagagc atcgatggaa gaccagggac gcttgtgatt 480 gaatcatttg tggtggacgt gcctgatgga aatacaaagg acgagacatg ctactttgtc 540 gaggccgtga tcaagtgcaa cttaacatct ctcgccgagg tatcagagcg gctagcagtt 600 cagtcaccca cctcgccact tgaacagtag 630

Claims (7)

An ABA receptor mutant protein in which leucine (Leucine, L) at position 93 in the amino acid sequence of SEQ ID NO: 1 is substituted with phenylalanine (phenylalanin, F), tyrosine (tyrosine, Y) or tryptophan (tryptophan, W). The method of claim 1,
The mutant protein is characterized in that it induces ABA-independent signaling. The method of claim 1,
At the same time, in the amino acid sequence of SEQ ID NO: 1, asparagine at position 102 (Asparagine, N) is substituted with tyrosine (Tyrosine, Y) or glutamic acid (Glutamic acid, E), a mutant protein. An ABA receptor mutant gene encoding the mutant protein of any one of claims 1 to 3. A recombinant vector comprising the ABA receptor mutant gene of claim 4. [Claim 6] Dry or transformed rice with enhanced resistance to environmental stress, transformed with the recombinant vector of claim 5. A method for enhancing resistance to drying or environmental stress in rice, comprising the step of transforming rice with the recombinant vector of claim 5 to overexpress the ABA receptor mutant protein.

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