KR101719789B1 - OsASGR2 gene for controlling growth of plants and uses thereof - Google Patents

Abstract

The present invention relates to an OsASGR2 gene that regulates plant growth and a method for regulating plant growth using the same. Since the transgenic plants overexpressing the OsASGR2 gene according to the embodiment of the present invention have confirmed that the growth of the plant length or the length of the liver is decreased, the OsASGR2 gene can be used to cultivate crop varieties having infertility .

Description

The OsASGR2 gene controlling plant growth and its use {

The present invention relates to OsASGR2 (Oryza sativa ABA Suppressed GRAS2) gene which regulates the growth of a plant and a method for regulating plant growth using the same.

Plant growth refers to a process in which the seeds of a plant germinate to form roots, stems, and leaves, and then gradually increase in size and weight while performing photosynthesis or nitrogen assimilation. Since not only plants but also all organisms are composed of many cells, the growth of living organisms means that the number of cells is increased by cell division.

Plant growth and development is regulated by plant hormones. Plant hormones that affect the growth of organs such as shoots, roots, roots, and fruit are called plant growth hormones, and the growth of plants is under the control of growth hormone. Examples of such plant growth hormones include oxine, gibberellin, cytokinin, ethylene, abscisic acid, and brassinoside. Among these, abscisic acid and gibberellin are hormones which act antagonistically in various physiological processes such as seed germination and length growth of plants.

On the other hand, rice can be seen to fall down due to heavy rains and strong winds during ripening when the ears are heavy after heading, and lodging resistance is said to be strong against the collapse of these crops. The thickness of the stem and the stem of the crop plays the greatest role in the growth. Especially, varieties with weak stem, large stem, and large ears have very weak characteristics in the dressing.

Studies have been carried out to develop varieties with strong resistance to overgrowth. Varieties with strong resistance to drought are required to have a strong stem and a small height. As a part of this research, Korean Patent No. 0781073 discloses a rice-derived gene RAP7 (Rice After Pollination clone 7) which is used to improve the rice seedling resistance, but the OsASGR2 gene of the present invention is a plant There is no evidence for the presence of growth regulatory functions.

Accordingly, the inventors of the present invention completed the present invention by confirming that rice-derived OsASGR2 gene, whose expression is inhibited by abscisic acid treatment, is involved in the regulation of plant growth, while studying a method for regulating plant growth.

Korean Patent Publication No. 0781073

It is an object of the present invention to provide an OsASGR2 (Oryza sativa ABA Suppressed GRAS2) gene that regulates plant growth.

Another object of the present invention is to provide an OsASGR2 protein which regulates plant growth.

Yet another object of the present invention is to provide a recombinant vector comprising the OsASGR2 gene.

Yet another object of the present invention is to provide a plant transformed with said recombinant vector.

It is a further object of the present invention to provide a transformed seed obtained from said transformed plant.

Yet another object of the present invention is to provide a method for controlling the growth of a plant by overexpressing a gene encoding OsASGR2 protein in the plant.

One embodiment of the present invention provides an OsASGR2 (Oryza sativa ABA Suppressed GRAS2) gene comprising the nucleotide sequence of SEQ ID NO: 1, which regulates plant growth.

The OsASGR2 gene may be isolated from rice ( Oryza sativa L. ).

The OsASGR2 gene was identified as GRAS (Gibberellic Acid Insensitive, Repressor of Gibberellic Acid Insensitive), which is involved in the signal transduction of GA (gibberellin), a hormone known to regulate length growth, among genes affected by ABA (abscisic acid) Scarecrow) gene.

In one embodiment of the present invention, the OsASGR2 gene was confirmed to be a transcription factor that is inhibited by the treatment of ABA (abscisic acid), which is a stress hormone, and that the gene is regulated by ABA and GA (gibberellin).

The gene consisting of the nucleotide sequence of SEQ ID NO: 1 consists of 1,854 nucleotides.

The OsASGR2 gene includes a gene consisting of the nucleotide sequence of SEQ ID NO: 1 and its equivalent. The equivalents include homologous genes having homology.

The OsASGR2 gene consisting of the nucleotide sequence of SEQ ID NO: 1 or its homologous gene is encoded as a protein.

The "homologous gene" is a gene having a sequence homology of preferably 80% or more, more preferably 90% or more, still more preferably 95% or more, most preferably 99% or more, with the nucleotide sequence of SEQ ID NO: 1 Refers to a gene that exhibits substantially the same function as the OsASGR2 gene of the present invention. Sequence homology can be analyzed by methods known in the art.

The "substantially homogeneous function" means that it is involved in the regulation of plant growth. Such functional equivalents include, for example, amino acid sequence variants in which some of the amino acids of the amino acid sequence are substituted, deleted or added. Substitution of amino acids is preferably conservative substitution. Examples of conservative substitutions of amino acids present in nature are as follows: (Gly, Ala, Pro), hydrophobic amino acids (Ile, Leu, Val), aromatic amino acids (Phe, Tyr, Trp), acidic amino acids (Asp, Glu), basic amino acids (His, Lys, Arg, Gln, Asn ) And sulfur-containing amino acids (Cys, Met). The deletion of the amino acid is preferably located at a site that is not directly involved in the activity of OsASGR2 of the present invention. The functional equivalents also include protein derivatives in which the basic skeleton of OsASGR2 and some chemical structures of the protein are modified while maintaining its physiological activity. These include, for example, fusion proteins made by fusion with other proteins such as GFP, while retaining the structural modification and physiological activity to change the stability, storage stability, volatility or solubility of the protein of the present invention.

The OsASGR2 gene can be amplified by polymerase chain reaction (PCR) using primers set forth in SEQ ID NO: 3 and SEQ ID NO: 4.

As used herein, the term "growth" means an increase in plant length. Among the plants, the term rice, in particular, means the length of the rice, the length of the rice field, and the length of the rice field. "Soybean" refers to the length from the ground to the head of the ears of Isaac. The term "grassland" means the length from the ground to the longest leaf of the longest stem. "Head" means the length .

The regulation of the growth according to the present invention means controlling the length of the plant, and preferably means decreasing the growth of the plant length or the length of the liver.

Another embodiment of the present invention provides an OsASGR2 protein that regulates the growth of a plant comprising the amino acid sequence of SEQ ID NO: 2. The present invention includes homologous proteins having homology with the above proteins.

The OsASGR2 protein consisting of the amino acid sequence of SEQ ID NO: 2 consists of 617 amino acids.

The "homologous protein" is a protein having preferably 80% or more, more preferably 90% or more, more preferably 95% or more, and most preferably 99% or more homologous to the amino acid sequence of SEQ ID NO: 2 Refers to a protein that exhibits substantially the same function as the OsASGR2 protein of the present invention.

Another embodiment of the present invention provides a recombinant vector comprising the OsASGR2 gene.

The recombinant vector is preferably a recombinant vector for overexpressing the OsASGR2 gene.

As used herein, 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.

As used herein, the term "vector" is used to refer to a DNA fragment (s), nucleic acid molecule, which is delivered into a cell. The vector replicates the DNA and can be independently regenerated in the host cell. The term "expression vector" is often used interchangeably with a "recombinant vector ". The term "recombinant vector" means a recombinant DNA molecule comprising a desired coding sequence and a suitable nucleic acid sequence necessary for expressing a coding sequence operably linked in a particular host organism. Promoters, enhancers, termination signals and polyadenylation signals available in eukaryotic cells are known.

The vector of the present invention can typically be constructed as a vector for cloning or expression. In addition, the vector of the present invention can be constructed by using prokaryotic cells or eukaryotic cells as hosts. 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 (for example, pL? Promoter, trp promoter, lac promoter, T7 promoter, tac promoter, etc.) , Ribosome binding sites for initiation of detoxification, and transcription / translation termination sequences.

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, glufosinate ammonium or phosphinothricin, kanamycin, G418, Bleomycin, hygromycin, ), Chloramphenicol (chloramphenicol), but are not limited thereto.

In the recombinant vector of the present invention, the promoter may be CaMV 35S, actin, ubiquitin, pEMU, MAS, or histone 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. Since the selection of the transformant can be carried out by various tissues at various stages, the expression promoter is always preferable in the present invention. Thus, the constant expression promoter does not limit the selectivity.

In one embodiment of the present invention, the recombinant vector pCAMBIA1300-OsASGR2 prepared by inserting the OsASGR2 gene into the pCAMBIA1300 vector is exemplified, but the present invention is not limited to this specific vector.

Another embodiment of the present invention provides a transgenic plant transformed with said recombinant vector.

The plant may be any one selected from the group consisting of rice, barley, wheat, sorghum, corn, chrysanthemum, and petunia. It is most preferable that the plant is rice. However, .

Transformation of a plant means any method of transferring DNA to a plant. Such transformation methods do not necessarily have a regeneration and / or tissue culture period. Transformation of plant species is now common for plant species, including both terminal plants as well as dicotyledonous plants. In principle, any transformation method can be used to introduce the hybrid DNA according to the present invention into suitable progenitor cells. The method is based on the 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) (Shillito RD et al., 1985 Bio / Technol. 3, 1099-1102), microinjection into plant elements (Crossway A. et al., 1986, Mol. Gen. Genet. 202,179-185 (Klein et al., 1987, Nature 327, 70), the infiltration of plants or the transformation of mature pollen or vesicles into Agrobacterium tumefaciens Infection by viruses (non-integrative) in virus-mediated gene transfer (EP 0 301 316), and the like. A preferred method according to the present invention comprises Agrobacterium mediated DNA delivery. Particularly preferred is the use of so-called binary vector techniques as described in EP A 120 516 and U.S. Pat. No. 4,940,838.

The plant cell into which the vector of the present invention is introduced is not particularly limited to a specific form as long as the cell can be regenerated as a plant. These cells include, for example, cultured cell suspension, protoplasts, leaf sections and callus. In addition, the present invention provides a plant with reduced length growth which is produced according to the above method and regenerated through tissue culture.

Another embodiment of the present invention provides a transformed seed obtained from said transgenic plant.

In one embodiment of the present invention, seeds of transgenic rice T1 genera in which overexpression of OsASGR2 gene was confirmed were cultured in hygromycin-containing medium to select transformed seeds.

Another embodiment of the present invention is a method for regulating the growth of a plant by overexpressing a gene comprising a nucleotide sequence of SEQ ID NO: 1 encoding OsASGR2 (Oryza sativa ABA Suppressed GRAS2) protein comprising the amino acid sequence of SEQ ID NO: 2 to provide.

The method of overexpressing the gene preferably includes, but is not limited to, transforming the plant with a recombinant vector operably linking the gene to a promoter.

According to the above method, the growth of the plant can be regulated by overexpressing the gene comprising the nucleotide sequence of SEQ ID NO: 1, thereby increasing the intracellular level of the OsASGR2 protein comprising the amino acid sequence of SEQ ID NO: 2.

The intracellular level refers to the amount present in a cell, which can be controlled by various methods known to those skilled in the art. For example, intracellular levels may be regulated through, but not limited to, regulation at the transcription stage or post-transcription stage. The control at the transcription stage may be carried out by a method for promoting the expression of a gene known to a person skilled in the art, for example, by preparing a recombinant expression vector comprising the OsASGR2 gene of SEQ ID NO: 1 or a homologous gene thereof in the promoter, Or a method of inserting an expression control sequence for promoting the expression of the gene in the vicinity of the OsASGR2 gene of SEQ ID NO: 1 or a homologous gene thereof. Modulation in post-transcriptional steps may be accomplished by methods known in the art for promoting protein expression, for example, by promoting the stability of mRNA transcribed from the OsASGR2 gene of SEQ ID NO: 1 or its functional equivalent, A method of promoting stability or a method of promoting the activity of a protein or protein.

The plant may be any one selected from the group consisting of rice, barley, wheat, sorghum, corn, chrysanthemum, and petunia. It is most preferable that the plant is rice. However, .

The regulation of the growth may be, but not limited to, reducing plant growth or length of the liver.

In one embodiment of the present invention, a recombinant vector containing the OsASGR2 gene was introduced into Agrobacterium and transformed into rice using the introduced Agrobacterium. OsASGR2 gene overexpressing transgenic plants were obtained and the phenotype was observed. As a result, the plant height was reduced by about 10-15%, the liver was reduced by about 20-25%, and the internode elongation was suppressed . Therefore, it was confirmed that the OsASGR2 gene overexpressing transformant showed a decrease in the growth of the plant or liver.

Since transgenic plants overexpressing the OsASGR2 gene (Oryza sativa ABA Suppressed GRAS2) according to the embodiment of the present invention have been confirmed to have decreased length growth of the plant or the liver, the OsASGR2 gene can be used for the transformation of plants resistant to strong rain and wind Can be cultivated.

Fig. 1 shows microarray results comparing the expression patterns of GRAS family genes (left) and the shape of rice roots treated with ABA (abscisic acid) for 3 days.
FIG. 2 is a graph showing the results of OsASGR2 gene expression analysis by RT-PCR;
Con, control roots; And
ABA, roots treated with 3 μM ABA (abscisic acid) for 72 hours.
FIG. 3 is a graph showing changes in the expression pattern of OsASGR2 gene according to time (0, 5, 30, 60, 180, 360 minutes) after treatment with ABA (abscisic acid)
4 is a graph showing the expression pattern of OsASGR2 gene according to the seed germination time.
5 is a graph showing the degree of OsASGR2 gene expression in a GA (gibberellin) signal transduction mutant compared with the stem of wild type rice;
T65, GA receptor;
gid, GA non-responsive mutant; And
slr, GA slender.
6 is a schematic diagram of a recombinant vector overexpressing the OsASGR2 gene according to an embodiment of the present invention.
FIG. 7 shows the expression of transgene in OsASGR2 overexpressed transgenic rice by RT-PCR;
DJ, wild type Dongjin;
T63-12, OsASGR2 overexpressed transgenic rice;
T64-4, OsASGR2 overexpressed transgenic rice;
T64-6, OsASGR2 overexpressed transgenic rice; And
Hig, hygromycin phosphotransferase gene.
FIG. 8 is a photograph showing a comparison of the phenotypic phenotypes of plants two months after emergence from packaging:
DJ, wild type Dong Jin-jin; And
OsASGR2ox, OsASGR2 overexpressed transgenic rice.
FIG. 9 is a photograph showing a cross-sectional phenotype of a 2-month old plant after packaging in a package;
DJ, wild type Dong Jin-jin; And
OsASGR2ox, OsASGR2 overexpressed transgenic rice.
10 is a graph comparing the lengths of (A) plant height, (B) culumn length and (C) internode length of a plant 2 months old from the packaging;
DJ, wild type rice (Dongjin);
OsASGR2 ox (T63-4, T63-12, T64-4, T64-6), OsASGR2 overexpressing transgenic rice; And
N1 to N6, 1st to 6th intervals.

Hereinafter, the present invention will be described in more detail in the following Examples. It should be noted, however, that the following examples are illustrative only and do not limit or limit the scope of the present invention. It will be understood by those of ordinary skill in the art that various changes in form and details may be made therein without departing from the spirit and scope of the invention as defined by the appended claims.

< Example  1> OsASGR2  Gene selection and expression analysis

<1-1> OsASGR2  Selection of genes

ABA (abscisic acid), a stress hormone, significantly suppresses root growth and root development in rice seedlings. Gene expression profiling was analyzed by microarray method in the roots of Dongjin rice treated for 3 days under the condition of ABA. Among the genes affected by ABA, GRAS (Gibberellic Acid Insensitive, Repressor of Gibberellic Acid Insensitive, and Scarecrow), which is involved in the signal transduction of GA (gibberellin), a hormone known to regulate length growth, (Oryza sativa ABA Suppressed GRAS2) gene (LOC_Os06g03710) which is remarkably suppressed by ABA was selected (FIG. 1).

Next, in order to confirm whether the selected OsASGR2 gene was related to root growth, 3 μM of abscisic acid was treated for 3 days in the roots of rice, and the expression level of OsASGR2 gene in the root of rice treated with the abscisic acid Were analyzed by RT-PCR. At this time, as a control group, roots of wild type Dongjinbyeon, which did not have abscisic acid treatment, were used. For the RT-PCR analysis, a single cDNA chain was synthesized using dT ₂₀ and reverse transcriptase (Superscript III, Invitrogen) as a template for total RNA isolated from the root of rice. PCR was performed using cDNA as a template using a nucleotide sequence-specific primer of OsASGR2 (Table 1) and Inclone Taq polymerase (clone IN5001-5000), wherein the PCR cycling parameter was 95 ° C-30 Sec, 58 [deg.] C for 30 seconds, and 72 [deg.] C for 40 seconds. After the PCR reaction, the amplified PCR products were developed on a 1% agarose gel using an electrophoresis apparatus and the band of the reaction product developed under UV was confirmed. As a result, it was confirmed that OsASGR2 gene expression was decreased in the roots of ABA-treated rice (Fig. 2).

In addition, the expression of OsASGR2 gene in leaves and roots of rice was remarkably reduced to the minimum level within 3 hours after ABA treatment through the public expression database of rice gene (RiceXPro2, http://ricexpro.dna.affrc.go.jp/) (Fig. 3).

SEQ ID NO: primer  designation primer  direction The base sequence (5 '- &gt; 3') 3 OsASGR2-1 Forward GGCCAAGAAGGAGGAGTACAG 4 OsASGR2-2 Reverse CGTGATGGCGTTGAGTATCC

<1-2> OsASGR2  Analysis of gene expression pattern

The expression pattern of the selected OsASGR2 gene was analyzed using the public database (Rice Oligonucleotide Array Database (http://www.ricearray.org/).

The expression of OsASGR2 gene was analyzed by seed germination time (0, 1, 3, 12, 24 hours). As a result, it was confirmed that the expression of OsASGR2 gene was increased during seed germination (FIG. 4).

It was also found that the expression level of the OsASGR2 gene was increased in the GA receptor (T65) and the GA signaling mutants ( gid1 , gid2 , slr1 ) (Fig. 5). This suggests that OsASGR2 gene expression is regulated by GA as well as ABA.

These results suggest that the OsASGR2 gene is regulated by ABA and GA in the root of rice and may regulate length growth in relation to the action of GA. Since CDS of OsASGR2 gene does not contain intron, gene was cloned from rice genomic DNA by PCR method and the whole nucleotide sequence was determined.

Specifically, genomic DNA was extracted from the leaves of Dongjin rice paddy using Inclone Genomic DNA prep kit (Inclone 1003-0200), and a pair of the following primers specific for the sequence of OsASGR2 containing restriction enzyme BamHI recognition sequence (Table 2) And Taq polymerase (PrimeSTAR HS DNA Polymerase, Takara). PCR was performed using the extracted genomic DNA as a template. At this time, the PCR was performed by repeating a series of steps consisting of heating the PCR amplification conditions at 94 ° C for 5 minutes and then heating at 94 ° C for 40 seconds, 60 ° C for 40 seconds, and 72 ° C for 40 seconds in total for 40 cycles Then, it was heated at 72 캜 for 7 minutes. Amplified PCR products were separated by pure PCR using a PCR kit (QIAGEN), labeled with Biodye Terminator version 3.1 (ABI Prism), and sequenced using an automatic sequencer (ABI3730, Applied Biosystems) Respectively. The nucleotide sequence of the OsASGR2 gene (1,854 bp) thus determined was set forth in SEQ ID NO: 1, and the amino acid sequence of the OsASGR2 protein encoded by the nucleotide sequence was set forth in SEQ ID NO: 2.

SEQ ID NO: primer  designation primer  direction The base sequence (5 '- &gt; 3') 5 OsASGR2-3 Forward GGACTCTAGAGGATCCATGTTGGCGGGTTGCTCG 6 OsASGR2-4 Reverse CGGTACCCGGGGATCCTTAGCTTTGCTGAGAATG

< Example  2> OsASGR2  Genetically overexpressed transgenic rice production

<2-1> OsASGR2  Generation of gene-overexpressing recombinant vector

OsASGR2 gene was ligated to the downstream of the 35S promoter of Cauliflower mosaic virus to produce a plant expression vector that is normally expressed.

Specifically, the OsASGR2 gene PCR product amplified in Example 1-1 was digested with restriction enzyme BamHI, inserted into the BamHI site downstream of the 35S promoter of the pCAMBIA1300 vector having hygromycin resistance, and the OsASGR2 gene overexpressing vector Respectively. The thus constructed OsASGR2 overexpressing vector pCAMBIA1300-OsASGR2 is shown in Fig.

<2-2> OsASGR2  Generation of overexpressed transgenic rice plants

The recombinant vector prepared in Example 2-1 was transformed into Agrobacterium. Agrobacterium (LBA4404 / OsASGR2) was inoculated into Dongjin pit callus and transformants were selected in a medium containing hygromycin (30 / / ℓ) and then grown in a greenhouse.

<2-3> In transgenic rice OsASGR2  Overexpression of genes

RT-PCR was performed using the RNA extracted from the leaves of transgenic rice selected in Example 2-2 as a template to determine whether the OsASGR2 gene was overexpressed.

Specifically, the leaves of the original cultivars or transgenic rice grown for 5 months in the package were ground with liquid nitrogen, and total RNA was isolated using RNA mini extraction kit (Inclone). 500 ng of the separated RNA was used as a template and a pair of primers specific for OsGRAS2 (OsASGR2-5 and OsASGR2-6) or hygromycin phosphotransferase (hgh) specific for HGH-1 and HGH -2) and Maxime RT PCR kit (Intron Bio 25131) (Table 3). RT-PCR conditions were first heated at 45 ° C for 30 minutes to synthesize first cDNA, and then heated at 94 ° C for 3 minutes under PCR amplification conditions, followed by 30 seconds at 94 ° C, 30 seconds at 62 ° C, And a heating step was repeated 25 times in total, followed by further heating at 72 ° C for 5 minutes. The amplified PCR product was developed on a 1% agarose gel using an electrophoresis apparatus and the band of the reaction product developed under UV was confirmed.

SEQ ID NO: primer  designation primer  direction The base sequence (5 '- &gt; 3') 7 OsASGR2-5 Forward GCGTCCTCGCCATGCACCG 8 OsASGR2-6 Reverse CACGGTGTACAGCGGCTGG 9 HGH-1 Forward CGTCTGCTGCTCCATACAAG 10 HGH-2 Reverse GTGCTTGACATTGGGGAGTT

As a result, it was confirmed that OsASGR2 gene was expressed at a high level in transgenic rice (Fig. 7).

< Example  3> OsASGR2  Phenotypic analysis of overexpressed transgenic rice

Seeds of transgenic rice T1 genera in which overexpression of OsASGR2 gene was confirmed in Example 2-3 were selected in a medium containing hygromycin (30 μg / l). After that, the selected seeds were transplanted and grown in the GMO package, and the length of the plant height and the length of the culumn length were compared two months after the outbreak.

As a result, the OsASGR2 transgenic rice was distinctively smaller than the wild type Dongjin rice (Fig. 8), the internode length was short (Fig. 9), the plant height was reduced by about 10-15% , And liver by about 20-25% (Fig. 10). That is, when OsASGR2 gene was overexpressed by comparing the lengths of liver and internodes (FIGS. 10B and 10C), it was confirmed that there was an effect of suppressing interdental extension and reducing the height.

                         SEQUENCE LISTING <110> REPUBLIC OF KOREA (MANAGEMENT: RURAL DEVELOPMENT        ADMINISTRATION)   <120> OsASGR2 gene for controlling growth of plants and uses thereof <130> P15R12D1427 <160> 10 <170> PatentIn version 3.2 <210> 1 <211> 1854 <212> DNA <213> Oryza sativa <400> 1 atgttggcgg gttgctcgtt ctcgtcgtcg aggcatcaga tgagcaccgc gcagcgtttc 60 gacatcctcc cctgcggctt ctccaagcgc ggcagccgcg gcgacggcgc cgccccgcgg 120 gtcgccggcg acgccaggag cggcgccacc acctgctcct tccggacgca ccccgcgccg 180 ccggtcaccc agtccgtgtc ctggggcgcc aagccggagc ccggcggcaa tggcaatggc 240 gcccaccgcg ccgttaagcg ggcgcatgac gaggacgcgg tcgaggagta tggccccatt 300 gttcgcgcca agcggacgcg gatgggcggc gacggcgatg aggtatggtt ccatcaatcc 360 attgcaggga cgatgcaagc gacggcggcg ggagaaggag aggaggcgga ggaggagaag 420 gtcttcttgg tgccgagcgc ggcggcgttc ccgcacggca tggccgccgc ggggccatcg 480 ctggccgcgg ccaagaagga ggagtacagc aagtcgccgt ccgactcgtc gtcctcgtcg 540 ggcacggacg gcggctcgtc ggcgatgatg ccgccgccgc agccgcccga gttcgacgcg 600 aggaacggcg tgccggcgcc ggggcaggcg gagcgggagg cgctggagct ggtgcgcgcg 660 ctcaccgcgt gcgccgactc cctctccgcc ggcaaccacg aggccgccaa ctactacctg 720 gcccggctcg gcgagatggc ctcgccggcg gggcccacgc cgatgcaccg cgtggccgcc 780 tacttcaccg aggcgctcgc gctccgcgtc gtgcgcatgt ggccgcacat gttcgacatc 840 ggcccgccgc gggagctcac cgacgacgcc ttcggcggcg gcgacgacga cgccatggcg 900 ctgcggatac tcaacgccat cacgcccatc ccgaggttcc tgcacttcac gctcaacgag 960 cgcctcctcc gcgagttcga ggggcacgag cgcgtccacg tcatcgactt cgacatcaag 1020 caggggctcc aatggccggg cttgctccag agcctggccg cgcgggcggt gcctccggcg 1080 cacgtgcgga tcaccggagt cggcgagtcg aggcaggagc tgcaggagac gggagcgcgg 1140 ctggcgcgcg tcgccgccgc gctcggcctg gcgttcgagt tccacgccgt ggtcgaccgg 1200 ctcgaggacg tccgcctgtg gatgctccac gtcaagcgcg gcgagtgcgt ggccgtgaac 1260 tgcgtcctcg ccatgcaccg cctgctccgc gacgacgccg cgctgaccga cttcctgggg 1320 ctagcgcgca gcacgggcgc caccatcctc ctcctcggcg agcacgaggg cggcggcctc 1380 aactcgggga ggtgggaggc gcggttcgcg cgcgcgctgc ggtactacgc cgcggcgttc 1440 gacgcggtgg acgcggcggg gctgccggag gcgagccccg cgagggccaa ggcggaggag 1500 atgttcgcgc gggagatccg caacgcggtg gcgttcgagg gccccgagcg gttcgagcgc 1560 cacgagagct tcgccgggtg gcggcggcgc atggaggacg gcggcgggtt caagaacgcc 1620 ggcatcggcg agcgcgaggc gatgcagggg cgcatgatcg cgaggatgtt cgggccggac 1680 aagtacaccg tgcaggcgca cggcggcggc ggcagcggcg gcggcgaggc gctcacgctc 1740 cggtggctgg accagccgct gtacaccgtg acggcgtgga cgccggcggg cgacggcgcg 1800 ggaggcagca ccgtgtcggc gtccacaaca gcatcacatt ctcagcaaag ctaa 1854 <210> 2 <211> 617 <212> PRT <213> Oryza sativa <400> 2 Met Leu Ala Gly Cys Ser Phe Ser Ser Ser His Gln Met Ser Thr 1 5 10 15 Ala Gln Arg Phe Asp Ile Leu Pro Cys Gly Phe Ser Lys Arg Gly Ser             20 25 30 Arg Gly Asp Gly Ala Ala Pro Arg Val Ala Gly Asp Ala Arg Ser Gly         35 40 45 Ala Thr Thr Cys Ser Phe Arg Thr His Pro Ala Pro Pro Val Thr Gln     50 55 60 Ser Val Ser Trp Gly Ala Lys Pro Glu Pro Gly Gly Asn Gly Asn Gly 65 70 75 80 Ala His Arg Ala Val Lys Arg Ala His Asp Glu Asp Ala Val Glu Glu                 85 90 95 Tyr Gly Pro Ile Val Arg Ala Lys Arg Thr Arg Met Gly Gly Asp Gly             100 105 110 Asp Glu Val Trp Phe His Gln Ser Ile Ala Gly Thr Met Gln Ala Thr         115 120 125 Ala Ala Gly Glu Gly Glu Glu Ala Glu Glu Glu Lys Val Phe Leu Val     130 135 140 Pro Ser Ala Ala Phe Pro His Gly Met Ala Ala Ala Gly Pro Ser 145 150 155 160 Leu Ala Ala Ala Lys Lys Glu Glu Tyr Ser Lys Ser Pro Ser Asp Ser                 165 170 175 Ser Ser Ser Gly Thr Asp Gly Gly Ser Ser Ala Met Met Pro Pro             180 185 190 Pro Gln Pro Pro Glu Phe Asp Ala Arg Asn Gly Val Pro Ala Pro Gly         195 200 205 Gln Ala Glu Arg Glu Ala Leu Glu Leu Val Arg Ala Leu Thr Ala Cys     210 215 220 Ala Asp Ser Leu Ser Ala Gly Asn His Glu Ala Ala Asn Tyr Tyr Leu 225 230 235 240 Ala Arg Leu Gly Glu Met Ala Ser Pro Ala Gly Pro Thr Pro Met His                 245 250 255 Arg Val Ala Ala Tyr Phe Thr Glu Ala Leu Ala Leu Arg Val Val Arg             260 265 270 Met Trp Pro His Met Phe Asp Ile Gly Pro Pro Arg Glu Leu Thr Asp         275 280 285 Asp Ala Phe Gly Gly Asp Asp Asp Ala Met Ala Leu Arg Ile Leu     290 295 300 Asn Ale Ile Thr Pro Ile Pro Arg Phe Leu His Phe Thr Leu Asn Glu 305 310 315 320 Arg Leu Leu Arg Glu Phe Glu Gly His Glu Arg Val Val His Ile Asp                 325 330 335 Phe Asp Ile Lys Gln Gly Leu Gln Trp Pro Gly Leu Leu Gln Ser Leu             340 345 350 Ala Ala Arg Ala Val Pro Ala His Val Arg Ile Thr Gly Val Gly         355 360 365 Glu Ser Arg Glu Glu Leu Gln Glu Thr Gly Ala Arg Leu Ala Arg Val     370 375 380 Ala Ala Ala Leu Gly Leu Ala Phe Glu Phe His Ala Val Val Asp Arg 385 390 395 400 Leu Glu Asp Val Arg Leu Trp Met Leu His Val Lys Arg Gly Glu Cys                 405 410 415 Val Ala Val Asn Cys Val Leu Ala Met His Arg Leu Leu Arg Asp Asp             420 425 430 Ala Ala Leu Thr Asp Phe Leu Gly Leu Ala Arg Ser Thr Gly Ala Thr         435 440 445 Ile Leu Leu Leu Gly Gly His Glu Gly Gly Gly Leu Asn Ser Gly Arg     450 455 460 Trp Glu Ala Arg Phe Ala Arg Ala Leu Arg Tyr Tyr Ala Ala Ala Phe 465 470 475 480 Asp Ala Val Asp Ala Ala Gly Leu Pro Glu Ala Ser Pro Ala Arg Ala                 485 490 495 Lys Ala Glu Glu Met Phe Ala Arg Glu Ile Arg Asn Ala Val Ala Phe             500 505 510 Glu Gly Pro Glu Arg Phe Glu Arg His Glu Ser Phe Ala Gly Trp Arg         515 520 525 Arg Arg Met Glu Asp Gly Gly Gly Phe Lys Asn Ala Gly Ile Gly Glu     530 535 540 Arg Glu Ala Met Gln Gly Arg Met Ile Ala Arg Met Phe Gly Pro Asp 545 550 555 560 Lys Tyr Thr Val Gln Ala His Gly Gly Gly Gly Ser Gly Gly Gly Gly Glu                 565 570 575 Ala Leu Thr Leu Arg Trp Leu Asp Gln Pro Leu Tyr Thr Val Thr Ala             580 585 590 Trp Thr Pro Ala Gly Asp Gly Ala Gly Gly Ser Thr Val Ser Ala Ser         595 600 605 Thr Thr Ala Ser His Ser Gln Gln Ser     610 615 <210> 3 <211> 21 <212> DNA <213> Artificial sequence <220> <223> OsASGR2-1 <400> 3 ggccaagaag gaggagtaca g 21 <210> 4 <211> 20 <212> DNA <213> Artificial sequence <220> <223> OsASGR2-2 <400> 4 cgtgatggcg ttgagtatcc 20 <210> 5 <211> 34 <212> DNA <213> Artificial sequence <220> <223> OsASGR2-3 <400> 5 ggactctaga ggatccatgt tggcgggttg ctcg 34 <210> 6 <211> 34 <212> DNA <213> Artificial sequence <220> <223> OsASGR2-4 <400> 6 cggtacccgg ggatccttag ctttgctgag aatg 34 <210> 7 <211> 19 <212> DNA <213> Artificial sequence <220> <223> OsASGR2-5 <400> 7 gcgtcctcgc catgcaccg 19 <210> 8 <211> 19 <212> DNA <213> Artificial sequence <220> <223> OsASGR2-6 <400> 8 cacggtgtac agcggctgg 19 <210> 9 <211> 20 <212> DNA <213> Artificial sequence <220> <223> HGH-1 <400> 9 cgtctgctgc tccatacaag 20 <210> 10 <211> 20 <212> DNA <213> Artificial sequence <220> <223> HGH-2 <400> 10 gtgcttgaca ttggggagtt 20

Claims (10)

delete delete A recombinant vector comprising the OsASGR2 gene represented by SEQ. ID. NO: 1, which reduces the plant length or the growth of the length of the liver. A transgenic plant having reduced length growth of a plant or liver transformed with the recombinant vector of claim 3. 5. The transgenic plant according to claim 4, wherein the plant is any one selected from the group consisting of rice, barley, wheat, sorghum, corn, chrysanthemum and petunia. A transformed seed obtained from the transgenic plant of claim 4. A method for regulating plant growth by overexpressing a gene comprising a nucleotide sequence of SEQ ID NO: 1 encoding OsASGR2 (Oryza sativa ABA Suppressed GRAS2) protein comprising the amino acid sequence of SEQ ID NO: 2. 8. The method according to claim 7, wherein the gene overexpression comprises transforming the plant with a recombinant vector in which the gene is operably linked to a promoter. 8. The method according to claim 7, wherein the plant is any one selected from the group consisting of rice, barley, wheat, sorghum, corn, chrysanthemum, and petunia. 8. The method according to claim 7, wherein the regulation of the growth reduces the plant growth of the plant or the length of the liver.

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