KR20150120135A - Shattering-controled transgenic plant and method for producing thereof - Google Patents

Abstract

The present invention relates to a gene controlling shattering properties, a recombinant vector controlling expression of the gene, a composition for controlling shattering properties in monocot plants, a transformant transformed by the composition, and production method thereof. An LOC_Os05g38120 gene of the present invention controls shattering properties of monocot plants, thereby increasing yield of major crops including rice, and providing convenience in cultivation.

Description

TECHNICAL FIELD The present invention relates to a transgenic plant having regulatability, and a method for producing the transgenic plant.

The present invention relates to a transgenic plant having regulated inagitability and a method for producing the same.

Rice, which is a major food resource in Korea, has been continuously studied as a major crop with corn, barley, and wheat all over the world. However, in order to improve suitable varieties and to be more productive in accordance with environmental changes such as climate change It is necessary to continue research and development. To this end, efforts are being made to increase the productivity and to make rice more suitable for the cultivation environment through the development of new varieties by analyzing useful gene functions in addition to the existing breeding methods.

Rice is a major crop and has been selectively cultivated until it becomes the cultivated rice that we used in the wild type rice used by ancient humans. The most prominent trait in the selective edification process is the overgrowth of rice. The wild type rice naturally has a highly infertile trait to propagate the seeds farther away, while the yield of seeds is greatly decreased when the overtaking ability of the cultivated rice is high. As a result, most of the cultivated rice is reduced in the quality of inaggregation. However, when the degree of overgrowth is greatly reduced, there is a problem that the seeds are not easily shaken during the threshing process. There have been some studies that have found related genes in QTL (quantitative locus) analysis by crossing the wild type and similar rice paddies with cultivated rice in order to study the genes related to the inability to overgrowth.

Therefore, research is needed to increase the productivity of rice plants through gene discovery that can regulate inagulation.

Korean Patent Publication No. 10-2013-0132963 Korean Patent No. 10-0795372

The present invention provides a gene that regulates inaglipticity.

The present invention also provides a recombinant vector that regulates the expression of a gene that regulates inaglipticity.

The present invention also provides a transformed cell transformed with a recombinant vector that regulates the expression of a gene that regulates inagulation.

The present invention also provides a composition for promoting or inhibiting in flushing of a terminal plant.

The present invention also provides a transgenic monocotyledonous plant having regulated inability to inoculate.

In addition, the present invention provides a method for producing transgenic monocotyledonous plants having controlled inability to inoculate.

In order to accomplish the above object, the present inventors have made intensive efforts to discover a gene that regulates the inability to infer the plant, and as a result, they have found that the LOC_Os05g38120 gene of rice regulates the inability to inhale and produce a transgenic plant that overexpresses or suppresses the gene Thereby completing the present invention.

The present invention provides a gene (hereinafter referred to as 'the gene of the present invention') that regulates inability to overgrowth in a terminal plant. In the present invention, the gene that regulates inability to inactivate is LOC_Os05g38120 (Os05g0455200) gene of rice. By using this gene, it is possible to increase the yield and cultivation convenience of monocotyledonous plants, especially rice.

The genome DNA size of LOC_Os05g38120 is 4572 bp and the coding sequence size is 1743 bp. It consists of 4 exons and 3 introns and is translated into 581 amino acids to make protein.

In the present invention, the CDS of LOC_Os05g38120 is shown as SEQ ID NO: 1, the protein sequence is shown as SEQ ID NO: 2, and the genome DNA of LOC_Os05g38120 is shown as SEQ ID NO:

The present invention also provides a recombinant vector that regulates the expression of a gene that regulates inaglipticity. The recombinant vector of the present invention can enhance or suppress the infertility according to its structure.

The conventional recombinant vector may be any vector as long as it can introduce the gene of the present invention or introduce RNA i, antisense oligonucleotide or the like to the gene of the present invention, preferably pCAMBIA series, pGA series, pGWB series, For example, a Ti-plasmid such as pGA3383, pCAMBIA3301, pCAMBIA3300, pGA3426, pGA3780, pGWB12, pGWB14, and vectors derived therefrom. Among them, the pGA3426 vector has a maize ubiquitin gene promoter, which is a binary vector capable of overexpressing the gene of interest by cloning the desired gene into this vector and transforming into the plant.

Preferably, the recombinant vector for promoting inability to overexpress the present invention is a recombinant vector comprising the LOC_Os05g38120 gene represented by SEQ ID NO: 1 and a promoter operably linked thereto. The recombinant vector for enhancing inability to inactivate the expression of the gene of interest in the 3 'direction of the present invention.

The exogenous promoter may be anything that is expressed only by the whole body or a specific tissue of the plant. Preferably, the promoter is CaMV (cauliflower mosaic virus), which is used as a promoter for a dicotyledonous plant, Promoter of the 35S RNA gene of the rice plant, and a promoter of the rice plant actin, a promoter of the rice ubiquitin gene and the rice cytochrome C gene (OsOc1) (RbcS: ribulose bisphosphate carboxylase / oxygenase small subunit) promoter, a RolD promoter for inducing the expression of plant roots derived from Agrobacterium, a potato-specific expression-inducing patatin promoter derived from potato, a tomato-derived fruit Mature specific expression-induced PDS (phytoene synthase) promoter. Preferably, the promoter of the present invention is a maize ubiquitin promoter, more preferably a promoter represented by SEQ ID NO: 3.

Preferably, the recombinant vector for inhibition of in vitro fertilization of the present invention comprises a recombinant vector comprising an RNA i capable of inhibiting the expression of the LOC_Os05g38120 gene, or a recombinant vector comprising an antisense oligonucleotide, It is a vector.

As used herein, the term "RNAi" refers to short double-stranded RNA capable of inducing RNAi (RNA interference) phenomenon through cleavage of a specific mRNA. A sense RNA strand having a sequence homologous to the mRNA of the target gene and an antisense RNA strand having a sequence complementary thereto. Since RNA i can inhibit expression of a target gene, it is provided as an efficient gene knockdown method or as a method of gene therapy. RNA i is not limited to a pair of double-stranded RNA portions that are paired with each other, but a pair is formed by a mismatch (corresponding base is not complementary), a bulge (no base corresponding to one chain) And a portion that is not achieved may be included. The recombinant vector for inhibiting the inability to infertility of the present invention can inhibit the expression of the LOC_Os05g38120 gene and thereby inhibit the inability to inappropriately grow. The RNA i sequence of the present invention is the nucleotide sequence shown in SEQ ID NO: 5 or SEQ ID NO: 6.

"LOC_Os05g38120 gene of the present invention" and "functionally equivalent fragment" refer to a fragment or a fragment of a nucleic acid comprising the nucleotide sequence shown in SEQ ID NO: 1, which exhibits substantially equivalent effect to the gene of the present invention. These nucleic acid fragments are 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95% 96%, 97%, 98%, 99% or more of sequence homology. Such a nucleic acid fragment can be easily prepared by molecular biological methods well known in the art.

The recombinant vector may include a sequence and a structure capable of introducing the gene of the present invention into a conventional vector or inhibiting the gene of the present invention. The production of a recombinant vector by introducing the gene of the present invention, Those skilled in the art can easily carry out the present invention in accordance with known methods.

The present invention also provides transformed cells transformed with the recombinant vector.

Such recombinant vectors may be introduced into host cells cultured using well known techniques such as infection, transfection, transfection, electroporation and transformation. Representative examples of a host include, but are not limited to, bacterial cells such as E. coli, Streptomyces and Salmonella typhimurium cells and plant cells.

Any plant cell can be used as "plant cell" used for transformation of a plant. The plant cell may be any type of cultured cell, cultured tissue, cultured organ or whole plant, preferably cultured cell, cultured tissue or culture organ, and more preferably cultured cell. "Plant tissue" refers to a tissue of a differentiated or undifferentiated plant, such as, but not limited to, stem cells, leaves, cancer tissues, and various types of cells used in culture, such as single cells, protoplasts, Callus tissue.

The present invention provides a composition for controlling florability of a terminal plant. The composition for controlling off-gassing of a terminal plant of the present invention may be a composition for promoting in flushing or a composition for inhibiting flushing depending on the kind of the recombinant vector or the type of transformed cells.

Preferably, the composition for enhancing the in flushability of a monocotyledonous plant comprises a recombination vector for promoting inability to inoculate comprising the gene of SEQ ID NO: 1 or a recombinant vector for promoting inability to inoculate comprising the gene of SEQ ID NO: 1 RTI ID = 0.0 > transformed < / RTI > cells.

Preferably, the composition for inhibiting flollability of the monocotyledonous plant comprises a recombinant vector for inhibiting the malleability, comprising RNA i capable of inhibiting the expression of the LOC_Os05g38120 gene, or a recombinant vector for inhibition of malacogenesis comprising the antisense oligonucleotide, Lt; RTI ID = 0.0 > transfected < / RTI > cells. And more preferably a recombinant vector containing the RNA i sequence of SEQ ID NO: 5 or SEQ ID NO: 6 for inhibition of inaggression or a transformed cell transformed with these recombinant vectors.

The composition of the present invention for controlling florability of a terminal plant may enhance or inhibit the infertility, so that the yield of the terminal plant and the convenience of cultivation can be improved.

In addition, the present invention provides a transgenic plant transformed with the vector, the transfected cell or the composition for the control of asymmetry. The gene introduced into the monocotyledonous plant may enhance the inability to infertility, and the infertility can be suppressed by RNA i or antisense oligonucleotide for the gene. This monocotyledonous plant is preferably rice (Oryza sativa L.).

The transgenic monocotyledonous plants of the present invention exhibited improved ingrowthability of 60%, 65%, 70%, 75%, 80%, 85%, 90% and 91%, respectively, as compared with the variants of the nucleotide sequences represented by SEQ ID NO: The transgenic monocotyledonous plants prepared by using the nucleotide sequences having the nucleotide sequences of SEQ ID NO: 1, 2, 3, 4, 5, 92, 93, 94, 95, 96, 97, 98, 99 or more, And may include substantially equivalent levels of transgenic plants.

The transgenic monocotyledonous plants of the present invention having suppressed inaggregation activity are characterized in that the expression of the nucleotide sequence of SEQ ID NO: 1 is 10%, 20%, 30%, 40%, 50%, 60%, 65% , 70%, 75%, 80%, 85%, 90%, or more of the transgenic plant.

Transformation of a plant of the present invention can be carried out by any method known in the art for 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. For example, 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. (Klein et al., 1987, Nature 327, 70), infiltration of plants or mature pollen of various plant elements (DNA or RNA-coated) And infection by (non-integrative) viruses in Agrobacterium tumefaciens mediated gene transfer (EP 0 301 316). Such mutants can be easily produced by molecular biological methods well known in the art.

In addition, the present invention provides a method for producing a terminal plant having improved ingrowth, comprising the step of transfecting a terminal plant with a recombinant vector containing the LOC_Os05g38120 gene and expressing the LOC_Os05g38120 gene which regulates inability to inactivate the plant in the plant.

Preferably, a step of preparing a recombinant vector comprising the nucleotide sequence shown in SEQ ID NO: 1; Introducing the recombinant vector into a terminal plant; And selecting a terminal transgenic plant having the transfection vector introduced therein to improve the inferiority of the transfacial plant to the transgenic plant.

Also, the present invention provides a method for producing an infertility-inhibited terminal plant comprising the step of expressing in a plant a recombinant vector comprising an RNA i or an antisense oligonucleotide capable of inhibiting the expression of the LOC_Os05g38120 gene do.

Preferably, a step of preparing a recombinant vector comprising the RNA i represented by SEQ ID NO: 5 and SEQ ID NO: 6 capable of inhibiting the expression of the LOC_Os05g38120 gene; Introducing the recombinant vector into a terminal plant; And selecting a terminal transgenic plant having introduced the recombinant vector and inhibiting inversion of the terminal plant.

By using the gene of the present invention, it is possible to increase the yield and control the ease of threshing according to the cultivated cultivated area and the cultivation area by controlling the infertility of the terminal plant.

FIG. 1 is a diagram showing the LOC_Os05g38120 gene and the T-DNA insertion position and genotyping results of the 3A-07285 strain as a heatmap.
FIG. 2 shows changes in the transcriptional uptake and inability of LOC_Os05g38120 gene according to T-DNA tagging in the 3A-07285 strain.
Fig. 3 shows the proliferative phenotype of strain 3A-07285, which shows high hygroscopicity of hetero and homo plants compared to the wild type.
FIG. 4 shows the development of the abscission zone and changes in lignin accumulation in the wild type and the 3A-07285 strain.
FIG. 5 is a graph showing the results of confirming an increase in the inability to overexpress upon overexpression of the LOC_Os05g38120 gene.
FIG. 6 is a schematic diagram showing the production of an RNA i vector for the LOC_Os05g38120 gene using the 3'UTR portion and the 5'UTR portion of the LOC_Os05g38120 gene.
FIG. 7 is a graph showing the inhibition of in fl ashing and the inhibition of developmental defects in the transgenic plants in which expression of LOC_Os05g38120 gene was inhibited by RNA i vector in Kasalath, which is a variant of Indica which is highly inferable.

Hereinafter, the present invention will be described in more detail with reference to examples. However, the following embodiments are only examples for helping understanding of the invention, and thus the scope of the present invention is not limited thereto.

< Example  1> Inability to  Increasing system selection

To identify the genes controlling the inability to overtake, phenotypes were observed using a population of active mutants among 100,000 T-DNA mutant populations. In the wild-type Dongjin rice, it was not possible to observe the phenotype of inerting ability, but it was able to select the line with increased inability in the active mutant group 3A-07285. The in flushability was measured using a digital force gauge and the force was measured when the grain of the rice fell from the panicle to confirm the flakiness. It was confirmed that in the heterozygous and homozygous mutants, the degree of infertility increases with increasing gene expression. The homozygous mutation was a digital force gauge to measure the inability to inoculate the seeds before harvesting. The results of the measurements of wild type and heterozygosity were as shown in Table 1.

Table 1. Changes in the floristic behavior of strain 3A-07285

As shown in Table 1, the selected 3A-07285 strains were significantly increased in heterozygosity compared to the wild type strains of the backgroud of Dongjin rice.

< Example  2 > The T- DNA co - segregation  Verification and confirmation of gene activity

The 3A-07285 strain produced by T-DNA insertion was trapped by the pGA2715 vector. Genotyping was performed on the inserted position in this system.

The selected lineage rice seeds were cultured in MSO medium at 28 ° C for 10 days. 100 mg of the leaves of the cultivated individuals were sampled and pulverized by rapid cooling using liquid nitrogen. Genomic DNA was extracted from the crushed leaves using CTAB buffer (2% CTAB, 1.42 M NaCl, 20 mM EDTA (pH 8.0), 100 mM Tris-Cl (pH 8.0)).

The 3A-07285 strain is inserted 4 Kb away from the stop codon of the LOC_Os05g38120 gene and has T-DNA inserted in the LB direction with an engineer site capable of activating the gene. Primers on the genomic DNA sequence and primer sequences for confirming the T-DNA analysis are shown in FIG. 1 and Table 2, respectively.

The expression of LOC_Os05g38120 was observed using primers (SEQ ID NO: 7 and SEQ ID NO: 8) prepared in order to observe whether the expression of LOC_Os05g38120 was increased by T-DNA.

Table 2. 3A-07285 Primers for Genomic Analysis

PCR reactions were performed after 5 minutes at 95 ° C; 30 seconds at 95 ° C, 30 minutes at 55 ° C, 1 minute at 72 ° C for 35 minutes, and finally 7 minutes at 72 ° C. PCR products were confirmed by electrophoresis using agarose gel. The genotype was identified and selected.

In order to confirm whether the expression of the LOC_Os05g38120 gene was actually activated, expression was confirmed using the primers shown in Table 3. As a control, rice ubiquitin 5 primer was used. The primer information is shown in Table 3.

Table 3. LOC_Os05g38120 transcript confirmation primer

Next, two lines of seeds were grown in MSO medium for 10 days, and then 100 mg of leaves of the individuals were harvested and rapidly frozen using liquid nitrogen. The frozen leaf tissue was pulverized and total RNA was isolated using RNA iso buffer (Takara, http://www.takara-bio.com). 2 ug of total RNA was synthesized by Moloney murine leukemia virus reverse transcriptase (Promega, Madison, WI, USA). The synthesized cDNA was observed by PCR and quantitative real time PCR was performed using the synthesized cDNA. The results are shown in Fig.

As shown in FIG. 2, it was confirmed that the real time PCR result in the 3A-07286 strain was consistent with the phenotype with increased inability to inappropriately grow.

It can also be confirmed by the phenotype shown in FIG. 3 that the phenotype of increased ineligibility increases with increasing expression of LOC_Os05g38120 in heterozygous and homozygous individuals compared with the wild type, and it is confirmed that homozygous plants have the highest inversion rate. That is, the above results confirmed that the 3A-07285 strain was co-segregated with the LOC_Os05g38120 gene.

< Example  3> LOC _ Os05g38120  Gene overexpression Mutant Defrag  Confirm change

In the line 3A-07285, which is an active mutant of LOC_Os05g38120 of Example 2, changes in the trait of this line were observed cytologically.

To observe the changes in the abscission zone of the spinal cord, the spinal cord was sampled to determine the change in the FAA fixation solution (formalin (35 (1)) (see Plant Cell Physiol. 2005 Jan; %): Acetic acid: ethanol (70%) = 1: 1: 18). The immobilized sample was paraffin-substituted and the block was cut and terminated to within 10 μm using a LEICA RM2265 section instrument and placed on a slide glass. The rehydrated samples were stained with 0.05% toluidine blue solution and fluroglucinol-HCl capable of staining lignin, and observed with an optical microscope at 10-20 magnification. The results are shown in Fig.

As shown in FIG. 4, when the cells were stained with toluidine blue staining, it was confirmed that the abscission zone was further developed in the active mutant of LOC_Os05g38120, strain 3A-07285. It was also confirmed that the lignin staining showed a remarkable decrease in lignin accumulation around the peel and pedicel of the 3A-07285 homo-mutant.

< Example  4> LOC _ Os05g38120  Genes for gene overexpression Cloning

In order to clone the LOC_Os05g38120 gene, PCR was performed using SEQ ID NO: 14 and SEQ ID NO: 15 shown in Table 4 as a template of KOME cDNA. 20 ng of cDNA template and 5 pmol of each primer were used. PCR reactions were performed after 5 minutes at 95 ° C; 35 seconds at 95 ° C for 30 seconds, 55 ° C for 30 seconds, and 72 ° C for 3 minutes, and finally at 72 ° C for 7 minutes. The PCR product was electrophoresed using an agarose gel, and its size was checked for its size, separated, and cloned into a T-easy vector (a vector for sub-cloning) to decode the base sequence. The cloned DNA was digested with HindIII and SpeI and ligated with the same restriction enzyme digested pGA3426 vector (over expression vector) and ligation at 14 DEG C for 12 hours. The ligation product was mixed with 50 占 퐇 of Top10 E. coli competent cells, transferred to a 1.5 ml tube, and left on ice for 15 minutes. Subsequently, 1 ml of LB liquid medium was further added to the tube after being left to stand in a 37 ° C oven for 1 minute, and then left in a 37 ° C shaking incubator for 1 hour. Then, the cells were plated on ampicillin-resistant LB solid medium and the resulting colonies were awaited for 12 hours and then mini-prepared after cell culture in 1 mL of LB liquid medium. The DNA obtained as miniprep was cleaved with restriction enzymes HindIII and SpeI, and electrophoresis was performed to separate the agarose gel and identify bands. The cloning DNA thus obtained was subjected to base sequence analysis again to select DNA that did not cause an error. The primers are shown in Table 4.

Table 4. Primers for sequencing

< Example  5> LOC _ Os05g38120  Production of Transgenic Transgenic Cells

The vector prepared in Example 4 was extracted. 50 아 of Agrobacterium tumefaciens LB4404 and 10 의 of plant expression vector were mixed and left on ice for 15 minutes. Subsequently, 1 minute was added to liquid nitrogen, and the same procedure was repeated 3 times in a 37 ° C oven for 5 minutes. Then, 1 ml of YEP liquid medium was added, followed by incubation in a 28 ° C shaking incubator for 5 hours. Next, the cells were plated on a tetracycline-resistant LB solid medium and incubated for 36 hours. Cells were cultured in a YEP liquid medium containing 1 mL of tetracycline antibiotics and mini-preparations were performed. The cells were digested with HindIII and SpeI Respectively. The resulting transformed Agrobacterium was used for transgenic rice transformation experiments.

< Example  6> LOC _ Os05g38120  Generation of overexpressed transgenic plants

In the N6D solid medium, Dongjinbyeon was grown in 28 ℃ growth chamber for 10 days under light condition to produce calli of rice. The resulting callus was mixed with the cells cultured for 72 hours with Agrobacterium transformed with the plant expression vector obtained in Example 4 and left in a 22 占 폚 cancer-treated growth chamber in a medium containing N6D-Acetosyringone.

Hygromycin 30 mg / L, hygromycin 40 mg / L, and hygromycin 40 mg / L were used in the N6D solid medium, and the callus contaminated with Agrobacterium was rinsed 5 times with 3 times distilled water containing Sepharose. Lt; RTI ID = 0.0 &gt; subculture. &Lt; / RTI &gt; Selection was made in the 28 ℃ growth room for 2 weeks each for 4 weeks. The divided callus was transferred to the MSR (hygromycin 40 mg / L) solid medium for regeneration and induced to regenerate in a light condition growth chamber for 4 weeks at 28 ° C. The plants were transferred to MS solid medium, grown in a light condition growth room for 7 days, To grow regenerated plants.

< Example  7> LOC _ Os05g38120  Gene overexpression in transgenic plants Infertility  Increase confirmation

FIG. 5 shows the results of confirming the inability to invert the LOC_Os05g38120 gene transgenic plants prepared in Example 6 above. RT-PCR and measurement of in flushability with a digital force gauge were carried out in the same manner as in Examples 1 and 2 above.

As shown in FIG. 5, LOC_Os05g38120 expression was observed in different LOC_Os05g38120 gene overexpressing plants, OX1 to OX5, and the amount of transcript was increased compared to the control plant. In addition, it was confirmed that the inability to overexpress LOC_Os05g38120 gene could increase the overexpression by overexpression of LOC_Os05g38120 gene.

< Example  8> LOC _ Os05g38120  For gene expression inhibition RNAi  Inclusion vector Cloning

In order to reduce the expression of LOC_Os05g38120 gene, PCR was performed on 208 bp of 5'UTR and 286 bp of 3'UTR using the cDNA observed to increase expression in Fig. 2 as a template. GGGGTACC (Kpn1 site) was added to the reverse primer among the primers used for PCR. The PCR product using PFU was subjected to EtOH ppt and enzyme digestion was performed using Kpn1 restriction enzyme. pGA3720 (Amp resistance gene / pGA3591 (RV)) vector was cut out with SmaI to gel elution Amp linker, and pGA3426 vector used as a binary vector backbone was cut into Kpn1 and triple ligation was performed on three products. The ligation-performed DNA was transformed into E. coli Top10, and colonies having both resistance to tetracycline and ampicillin were selected. The selected colonies were cultured in a liquid medium to perform miniprep, digested with Kpn1, and finally sequenced to screen for DNA without error. The primers used in the RNA i construction for reducing the expression of the LOC_Os05g38120 gene are shown in Table 5, and the prepared RNA I has the nucleotide sequence of SEQ ID NO: 5 or SEQ ID NO: 6.

< Example  9> LOC _ Os05g38120  Production of transgenic cells inhibiting gene expression

The vector prepared in Example 5 was extracted. 50 아 of Agrobacterium tumefaciens LB4404 and 10 의 of plant expression vector were mixed and left on ice for 15 minutes. Subsequently, 1 minute was added to liquid nitrogen, and the same procedure was repeated 3 times in a 37 ° C oven for 5 minutes. Then, 1 ml of YEP liquid medium was added, followed by incubation in a 28 ° C shaking incubator for 5 hours. Then, the cells were plated on tetracycline-resistant LB solid medium and incubated for 36 hours. The resulting colonies were cultured in YEP liquid medium supplemented with 1 mL of tetracycline antibiotic. After mini-preparation, Kpn1 was used for enzyme cleavage and the size was confirmed. The resulting transformed Agrobacterium was used for transgenic rice transformation experiments.

< Example  10> LOC _ Os05g38120  Production of Transgenic Plants Suppressing Gene Expression

Kasalath seeds of indica cultivars were grown in N6D solid medium for 28 days at 28 ℃ for 10 days to produce calli of rice. The resulting callus was mixed with the cells cultured for 72 hours with Agrobacterium transformed with the plant expression vector obtained in Example 8 and left in a growth chamber containing 22DC-treated N6D-Acetosyringone.

Hygromycin 30 mg / L, hygromycin 40 mg / L, and hygromycin 40 mg / L were used in the N6D solid medium, and the callus contaminated with Agrobacterium was rinsed 5 times with 3 times distilled water containing Sepharose. Lt; RTI ID = 0.0 &gt; subculture. &Lt; / RTI &gt; Selection was made in the 28 ℃ growth room for 2 weeks each for 4 weeks. The divided callus was transferred to the MSR (hygromycin 40 mg / L) solid medium for regeneration and induced to regenerate in a light condition growth chamber for 4 weeks at 28 ° C. The plants were transferred to MS solid medium, grown in a light condition growth room for 7 days, To grow regenerated plants.

< Example  11> LOC _ Os05g38120  Gene Expression Suppression in Transgenic Plants Fleeing Sustained inhibition confirmation

FIG. 7 shows the results of the inactivation of the LOC_Os05g38120 gene expression-suppressing transgenic plant prepared in Example 10. FIG. RT-PCR and measurement of inability to a digital force gauge were carried out in the same manner as in Examples 1 and 2, and cytological observation was performed in the same manner as in Example 3.

As shown in FIG. 7, LOC_Os05g38120 expression was observed in different LOC_Os05g38120 gene expression suppressing plants 1 to 10, and it was confirmed that the amount of transcript was decreased compared to the control plant. In addition, it was confirmed that the inability to degrade LOC_Os05g38120 gene expression was also decreased in the plants. In addition, the transgenic plants 9 and 10 were confirmed to show that the development of the abscission zone was inferior to that of wild type. From these results, it was confirmed that inhibition of LOC_Os05g38120 gene expression could inhibit the inability to infertility.

<110> University-Industry Cooperation Group of Kyung Hee University <120> Shattering-controlled transgenic plant and method for producing          the <130> P14-009-KHU <160> 21 <170> Kopatentin 2.0 <210> 1 <211> 1743 <212> DNA <213> Oryza sativa <400> 1 atgtcgtccg ccgcgggggg aggaggaggg tacgggggcg gaggcggcga gcatcagcat 60 cagcagcagc agcaccacct gctgcttggg caggcggcgg ggcagctgta ccacgtgccg 120 cagcacagcc ggcgggagaa gctgcggttc ccgcccgacc acccggcgga gtcgccgccg 180 ccgccgccgc ccgggtcgtg gccgctgccc ccggcgttct actcctacgc gtcgtcctcg 240 tcatcgtact cgccgcacag cccgacgctg gcgcacgcgc agttggtggc gcatgggatg 300 ccgccggggg ccgcgacgag cggaggggcc cagatcccga gccagaactt cgcgctgtcg 360 ctgtcgtcgg cgtcctcgaa ccctccgccc acgccgagga ggcagtttgg cggcggcggc 420 ggcggcggcg gggccgccgg gccgtacggg cccttcacgg gctacgccgc cgtgctcggg 480 cggtccaggt tcttgggccc cgcgcagaag ctgctcgagg agatctgcga cgtcggcggc 540 cgccccgcgc agcttgacag gggctccgac gagggtttgc tcgacgtaga cgccatggac 600 gccgcgggaa gcgtcgacca cgagatggac ggcagcgatc gcgccgtcgc ggacgcggtc 660 acggtctccg gcgccgagca gcagtggagg aagactaggc tcatctcgct tatggaagac 720 gtttgcaagc gatacaggca atactaccag caactccaag ctgtagtatc gtcctttgag 780 actgttgcgg gtctgagcaa tgctgctcct tttgcttcca tggctcttag gacaatgtcg 840 aagcatttca agtatttgaa gggcattata ctgaaccagc tgcgcaatac gggcaagggt 900 gctacaaaag atggtctcgg caaggaagac acaacaaact ttgggcttat gggcggcggc 960 gctggcctac taaggggaaa caatgtgaat tcgtttagcc aacctcacaa catatggcgc 1020 ccgcaaagag ggctaccaga gcgtgctgtt tcggttcttc gtgcatggct attcgaacac 1080 ttcctgcacc cgtatccaac tgacagcgat aagcagatgc ttgctaaaca gacaggatta 1140 actaggaacc aggtatcgaa ctggtttatc aatgcaaggg ttaggctctg gaagccaatg 1200 gttgaagaaa ttcacaacct cgagatgagg cagctgcaga agaatccgtc tcttgacaag 1260 aatcagctct ccatgcagca cacccaacat tcgtcggaca gcagcgggaa gccctgtgat 1320 ccatcaaact cgctgcaagg gcaaagtagc agcatgacca ggaatcacag catctccgcc 1380 tcccggcaca tcgaggacgg cctctcccag atgccccatg acatctccgg tcaggtgagc 1440 ttcgcataca acgggctcgc cgcgcatcac agcatcgcga tggcgcacca ccaccaacct 1500 gacctcattg gcaccggtgg tgccgcgaat gctggcggtg tctcactcac ccttggcctt 1560 caccagaaca ataaccgagc ttacattgct gagccccttc cggccgcgct tccgctcaat 1620 cttgcccatc gtttcggact ggaggacgtt agtgatgcct acgtgatgag ctcatttggt 1680 ggtcaggacc ggcatttcac caaggagatc ggtggccatt tgctccatga ctttgttggt 1740 tga 1743 <210> 2 <211> 580 <212> PRT <213> Oryza sativa <400> 2 Met Ser Ser Ala Gly Gly Gly Gly Gly Gly Tyr Gly Gly Gly Gly Gly   1 5 10 15 Glu His Gln His Gln Gln Gln Gln His His Leu Leu Leu Gly Gln Ala              20 25 30 Ala Gly Gln Leu Tyr His Val Pro Gln His Ser Arg Arg Glu Lys Leu          35 40 45 Arg Phe Pro Pro Asp His Pro Ala Glu Ser Pro Pro Pro Pro Pro      50 55 60 Gly Ser Trp Pro Leu Pro Pro Ala Phe Tyr Ser Tyr Ala Ser Ser Ser  65 70 75 80 Ser Ser Tyr Ser Pro His Ser Pro Thr Leu Ala His Ala Gln Leu Val                  85 90 95 Ala His Gly Met Pro Gly Ala Ala Thr Ser Gly Gly Ala Gln Ile             100 105 110 Pro Ser Gln Asn Phe Ala Leu Ser Leu Ser Ser Ala Ser Ser Asn Pro         115 120 125 Pro Pro Thr Pro Arg Arg Gln Phe Gly Gly Gly Gly Gly Gly Gly Gly     130 135 140 Ala Ala Gly Pro Tyr Gly Pro Phe Thr Gly Tyr Ala Ala Val Leu Gly 145 150 155 160 Arg Ser Phe Leu Gly Pro Ala Gln Lys Leu Leu Glu Glu Ile Cys                 165 170 175 Asp Val Gly Gly Arg Pro Ala Gln Leu Asp Arg Gly Ser Asp Glu Gly             180 185 190 Leu Leu Asp Val Asp Ala Met Asp Ala Ala Gly Ser Val Asp His Glu         195 200 205 Met Asp Gly Ser Asp Arg Ala Val Ala Asp Ala Val Thr Val Ser Gly     210 215 220 Ala Glu Gln Gln Trp Arg Lys Thr Arg Leu Ile Ser Leu Met Glu Asp 225 230 235 240 Val Cys Lys Arg Tyr Arg Gln Tyr Tyr Gln Gln Leu Gln Ala Val Val                 245 250 255 Ser Ser Phe Glu Thr Val Ala Gly Leu Ser Asn Ala Ala Pro Phe Ala             260 265 270 Ser Met Ala Leu Arg Thr Met Ser Lys His Phe Lys Tyr Leu Lys Gly         275 280 285 Ile Ile Leu Asn Gln Leu Arg Asn Thr Gly Lys Gly Ala Thr Lys Asp     290 295 300 Gly Leu Gly Lys Glu Asp Thr Thr Asn Phe Gly Leu Met Gly Gly Gly 305 310 315 320 Ala Gly Leu Leu Arg Gly Asn Asn Val Asn Ser Phe Ser Gln Pro His                 325 330 335 Asn Ile Trp Arg Pro Gln Arg Gly Leu Pro Glu Arg Ala Val Ser Val             340 345 350 Leu Arg Ala Trp Leu Phe Glu His Phe Leu His Pro Tyr Pro Thr Asp         355 360 365 Ser Asp Lys Gln Met Leu Ala Lys Gln Thr Gly Leu Thr Arg Asn Gln     370 375 380 Val Ser Asn Trp Phe Ile Asn Ala Arg Val Arg Leu Trp Lys Pro Met 385 390 395 400 Val Glu Glu Ile His Asn Leu Glu Met Arg Gln Leu Gln Lys Asn Pro                 405 410 415 Ser Leu Asp Lys Asn Gln Leu Ser Met Gln His Thr Gln His Ser Ser             420 425 430 Asp Ser Ser Gly Lys Pro Cys Asp Pro Ser Asn Ser Leu Gln Gly Gln         435 440 445 Ser Ser Ser Met Thr Arg Asn His Ser Ser Ser Ala Ser Arg His Ile     450 455 460 Glu Asp Gly Leu Ser Gln Met Pro His Asp Ile Ser Gly Gln Val Ser 465 470 475 480 Phe Ala Tyr Asn Gly Leu Ala Ala His His Ser Ile Ala Met Ala His                 485 490 495 His His Gln Pro Asp Leu Ile Gly Thr Gly Gly Ala Ala Asn Ala Gly             500 505 510 Gly Val Ser Leu Thr Leu Gly Leu His Gln Asn Asn Asn Arg Ala Tyr         515 520 525 Ile Ala Glu Pro Leu Pro Ala Ala Leu Pro Leu Asn Leu Ala His Arg     530 535 540 Phe Gly Leu Glu Asp Val Ser Asp Ala Tyr Val Met Ser Ser Phe Gly 545 550 555 560 Gly Gln Asp Arg His Phe Thr Lys Glu Ile Gly Gly His Leu Leu His                 565 570 575 Asp Phe Val Gly             580 <210> 3 <211> 3341 <212> DNA <213> Oryza sativa <400> 3 tcatctcgca cgtgcttatc gatggcccta catacaccat agcctaacct cttcaaaatt 60 gacaacacca taacaatgta ttgagtgcta cttactttgt aacatactgt aagagacagc 120 ttgtgaattc cttcatggat tatacacatg agaacgtttg aaatagttgt ttggatgaaa 180 cacaaacata tagtaattaa aaaactaaaa aagggaaaaa aaaagctttg ttcggaaact 240 ccaatgaaat atctcaaaag gaaatttgaa gagtcattta tattccatat gactacctat 300 aattatctac gagatttaaa ttcttatgga cacaaattct tcggataaat ttacttgaac 360 ttcggtgaaa gaaccctaaa cacatgtatg catccagaca atgaaaagca agcgtgccat 420 gagattttct atttgggccg tgtttagatt aaaatttttt tcttcaaact tccaactttt 480 tcgttacatc aaatgttagg acatatgcat ggagttttaa atgtagacga aaaaaatcaa 540 ttgcatagtt tgcatgtaaa tcgcgagacg aatcttttga tcctaattac gccatgattt 600 gataatgtga tgctacagta aacatttgct aatgacagat taattagact taataaattc 660 gtctcgtctt ttacatgcgg aatctgtaat ttgttttgtt attagtctac gtttgatact 720 tcaaatatgt gttcatatat ttaaaaaaaa tctagcacac gaactaaaca caaccttgga 780 tggcctccta agactgtctc caacaagtga cccataagag cacccaaacc caaaatgggt 840 ctccgatagt actatttcag cctccaacag agtacctata cagaagaccc attttgcgtg 900 ctataagagg cataacctaa atctgagtat cctctctcct gaagacctat ttgcagtaag 960 tgttctcttt taggtcttat tgttggagaa gaccaaaaat atgtattgaa gtctttgact 1020 gtagcgctat gtaaacatga aatgtgtctt gtattttggg tttcattgtt ggagatagcc 1080 taagttgctt atccggtttt actgcctgcc atataagaac cccatcatcc atatccgaag 1140 aggacatatt tttattgcaa aactttaaaa ttataataat aatgtaacta caatataaca 1200 taactagtac ataactagta gtacatgtaa cgtttgttca agcaattata ctactaccat 1260 agtacgattg cgtataaagt ttttcaattc aaaaatgttg ctgccgtttt ctagatggtg 1320 aaaaacaaac ttcacccatc tccgacgaac tacaaaaccc tcagctcgat ctccgcacag 1380 tcccgggcca cgcccacacc accctgatcc gctcacccct tccaccacag ggtccaggag 1440 gacctcgccc ccctcctcct cctcgtcgca cgaccagacc agcagcacca actccctttc 1500 tgcctttcgc tttcactgct cagtgctgct cctacacacc atttccccgt ccgtccttcg 1560 ccccccgcgt ctcttttccg cacccatctg ctgcagctgc gcacctttaa tttgcagaaa 1620 gaaagaaaga aagaaaggaa agcaaggccc ccccgagcgc aacagctagc ccaacacacc 1680 tcctccatca ccaccgagaa ggcaacgcat atcaaaagcg cgggcgcaaa gcaaagataa 1740 catcagatca ggtcggcgcc ccccgctccc ggctgccgca aagcccaccc gatcgatcga 1800 tcgacgcctc gttctcctcc tcctccgagg ctactctctg cagggcgctc gcttacgtct 1860 gcctctgcgt acgtgcaccg cccagcgcgg gcgccatgtc gtccgccgcg gggggaggag 1920 gagggtacgg gggcggaggc ggcgagcatc agcatcagca gcaccacctg ctgcttgggc 1980 aggcggcggg gcagctgtac cacgtgccgc agcacagccg gcgggagaag ctgcggttcc 2040 cgcccgacca cccggcggag tcgccgccgc cgccgccgcc cgggtcgtgg ccgctgcccc 2100 cggcgttcta ctcctacgcg tcgtcctcgt catcgtactc gccgcacagc ccgacgctgg 2160 cgcacgcgca gttggtggcg catgggatgc cgccgggggc cgcgacgagc ggaggggccc 2220 agatcccgag ccagaacttc gcgctgtcgc tgtcgtcggc gtcctcgaac cctccgccca 2280 cgccgaggag gcagtttggc ggcggcggcg gcggcggcgg ggccgccggg ccgtacgggc 2340 ccttcacggg ctacgccgcc gtgctcgggc ggtccaggtt cttgggcccc gcgcagaagc 2400 tgctcgagga gatctgcgac gtcggcggcc gccccgcgca gcttgacagg ggctccgacg 2460 agggtttgct cgacgtagac gccatggacg ccgcgggaag cgtcgaccac gagatggacg 2520 gcagcgatcg cgccgtcgcg gacgcggtca cggtctccgg cgccgagcag cagtggagga 2580 agactaggct catctcgctt atggaagacg tgagtaatcg caagctatat ttgttgtttc 2640 atatcgctgc ttctgttttt ccaaagaaaa tactatacta gactttactt ttgctaccct 2700 gcattaatta acttcatgaa taagaatcta ataacgaaca cctcatggtc acatcatgga 2760 tgcacttact gttgttgcac agattatttc ttgtgcttgt gatttgatga gggttgatct 2820 ggattagtga cacatctctt ctcttttgga tatatttttg ttctttaata tgtctcaaat 2880 caatcaacag ttcaaggctc ttttaagctc ccttctaaaa ctagcaggag gtgatccgca 2940 gtttatctat aatcaaaagg tagcatacaa ttctgtgaaa atgcacgata gaagctaaat 3000 tcctcaggat gctcttaatt acatcataaa caaaaattca gtttgctagt acttgataag 3060 attccagcag tttgtcctct ttcacgagat aagattcgtt gagtttgtca gcagaaagat 3120 tctgattata tagttactag tacatactga aaatatgttg agcaccagtt tttttatttg 3180 caatggtgtc aaatatgatc ctcgttcctt aattaggaag cttgcgttgg tgtagtgtcc 3240 acatgatcgt aaagtaggtc tgcacacctt gatgtattat taaacaatta attgggttac 3300 atatcgtgat gtcctcttaa cagttacaaa aaaccttgta g 3341 <210> 4 <211> 5961 <212> DNA <213> Oryza sativa <400> 4 tcatctcgca cgtgcttatc gatggcccta catacaccat agcctaacct cttcaaaatt 60 gacaacacca caacaatgta ttgagtgcta cttactttgt aacatactgt aagagacagc 120 ttgtgagttc cttcatggat tatacacatg agaacgtttg aaatagttgt ttggatgaaa 180 cacaaacata tagtaattaa aaaactaaaa aagggaaaaa aaagctttgt tcggaaactc 240 caatgaaata tctcaaaagg aaatttgaag agtcacttat attccatatg actacctata 300 attatctacg agatttaaat tcttatggac acaaattctt cggataaatt tacttgaact 360 ttggtgaaag aaccctaaac acatgtatgc atccagacaa tggaaagcaa gcgtgccatg 420 agattttcta tttgggccgt gtttagatta aaattttttt cttcaaactt ccaacttttt 480 cgttacatca aatgttagga catatgcatg gagttttaaa tgtggacgaa aaaaatcaat 540 tgcatagttt gcatgtaaat cgcgagacga atcctttgag cctaattacg ccatgatttg 600 acaatgtgat gctacagtaa acatttgcta atgacagatt aattagactt aataaattcg 660 tctcgtcttt tacatgcgga atctgtaatt tgttttgtta ttagtctatg tttgatactt 720 caaatatgtg ttcgtatatt taaaaaaaat ctagcacacg aactaaacac aaccttggat 780 ggccttctaa gactgtctcc aacaagtgac ccataagagc acccaaaccc aaaatgggtc 840 tccgatagta ctatttcagc ctccaacaga gtacctatac aaaagaccca ttttgcgtgc 900 tataagaggc ataacctaaa tctgagtatc ctctctcctg aagacctatt tgcagtaagt 960 gttctctttt aggtcttatt gttggagaag accaaaaata tgtattgaac tctttgactg 1020 tagcgctatg caaacgtgaa atggttcttg tattatgggt ttcattgttg gagatagcct 1080 aagttgctta tccggtttta ttgcctgcca tataagaacc ccatcatcca catccgaaga 1140 ggacatattt ttgttgcaaa actttaaaat tataataata atgtaactac aatataacat 1200 aactagtaca tgtaacgttt gttcaagcaa ttatactact accatagtac gattgcgtat 1260 aaagtttttc aattcaaaaa tgttgctgcc gttttctaga tggtgaaaaa caaacttcac 1320 ccatctccga cgaactacaa aaccctcagc tcgatctccg cacagtcccg ggccacgccc 1380 acaccaccct gatccgctca ccccttccac cacagggtcc aggaggacct cgcccccctc 1440 ctcctcctcg tcgcacgacc agaccagcag caccaactcc ctttctgcct ttcgctttca 1500 ctgctcagtg ctgctcctac acaccatttc cccgtccgtc cttcgccccc cgcgtctctt 1560 ttccgcaccc atctgctgca gctgcgcacc tttaatttgc agaaagaaag aaagaaagaa 1620 agaaaggaaa gcaaggcccc cccgagcgca acagctagcc caacacacct cctccatcac 1680 caccgagaag gcaacgcata tcaaaagcgc gggcgcaaag caaagataac atcagatcag 1740 gtcggcgccc cccgctcccg gctgccgcaa agcccacccg atcgatcgat cgacgcctcg 1800 ttctcctcct cctccgaggc tactctctgc agggcgctcg cttacgtctg cctctgcgta 1860 cgtgcaccgc ccagcgcggg cgccatgtcg tccgccgcgg ggggaggagg agggtacggg 1920 ggcggaggcg gcgagcatca gcatcagcag cagcagcacc acctgctgct tgggcaggcg 1980 gcggggcagc tgtaccacgt gccgcagcac agccggcggg agaagctgcg gttcccgccc 2040 gaccacccgg cggagtcgcc gccgccgccg ccgcccgggt cgtggccgct gcccccggcg 2100 ttctactcct acgcgtcgtc ctcgtcatcg tactcgccgc acagcccgac gctggcgcac 2160 gcgcagttgg tggcgcatgg gatgccgccg ggggccgcga cgagcggagg ggcccagatc 2220 ccgagccaga acttcgcgct gtcgctgtcg tcggcgtcct cgaaccctcc gcccacgccg 2280 aggaggcagt ttggcggcgg cggcggcggc ggcggggccg ccgggccgta cgggcccttc 2340 acgggctacg ccgccgtgct cgggcggtcc aggttcttgg gccccgcgca gaagctgctc 2400 gaggagatct gcgacgtcgg cggccgcccc gcgcagcttg acaggggctc cgacgagggt 2460 ttgctcgacg tagacgccat ggacgccgcg ggaagcgtcg accacgagat ggacggcagc 2520 gatcgcgccg tcgcggacgc ggtcacggtc tccggcgccg agcagcagtg gaggaagact 2580 aggctcatct cgcttatgga agacgtgagt aatcgcaagc tatatttgtt gtttcatatc 2640 gctgcttctg tttttccaaa gaaaatacta tactagactt tacttttgct accctgcatt 2700 aattaacttc atgaataaga atctaataac gaacacctca tggtcacatc atggatgcac 2760 ttactgttgt tgcacagatt atttcttgtg cttgtgattt gatgagggtt gatctggatt 2820 agtgacacat ctcttctctt ttggatatat ttttgttctt taatatgtct caaatcaatc 2880 aacagttcaa ggctctttta agctcccttc taaaactagc aggaggtgat ccgcagttta 2940 tctataatca aaaggtagca tacaattctg tgaaaatgca cgatagaagc taaattcctc 3000 aggatgctct taattacatc ataaagaaaa attcagtttg ctagtacttg ataagattcc 3060 agcagtttgt cctctttcac gagataagat tcgttgagtt tgtcagcaga aagattctga 3120 ttatatagtt actagtacat actgaaaata tgttgagcac cagttttttt tatttgcaat 3180 ggtgtcaaat atgatcctcg ttccttaatt aggaagcttg cgttggtgta gtgtccacat 3240 gatcgtaaag taggtctgca caccttgatg tattattaaa caattaattg ggttacatat 3300 cgtgatgtcc tcttaacagt tacaaaaaac cttgtaggtt tgcaagcgat acaggcaata 3360 ctaccagcaa ctccaagctg tagtatcgtc ctttgagact gttgcgggtc tgagcaatgc 3420 tgctcctttt gcttccatgg ctcttaggac aatgtcgaag catttcaagt atttgaaggg 3480 cattatactg aaccagctgc gcaatacggg caagggtgct acaaaagatg gtctcggcaa 3540 ggaagacaca acaaactttg ggcttatggg cggcggcgct ggcctactaa ggggaaacaa 3600 tgtgaattcg tttagccaac ctcacaacat atggcgcccg caaagagggc taccagagcg 3660 tgctgtttcg gttcttcgtg catggctatt cgaacacttc ctgcacccgt aagtatatga 3720 tataaatcat cgcaacagaa tttttctttt taggtatgct aatctggaag cacataactg 3780 aatatgcgta ttgtgcacta attggaaaaa taaaaggaaa aaggacttta tggtatatat 3840 actgcatgtt gcatttttac accgacaagg gtgtacatgg attcctatta gtggagaatt 3900 atccatagac cctacaaatg ggtaatatga aggatattgc aaccaaaccg ttggtgaaaa 3960 ttgttatttt gctttattga aatcttggat tttttttcct cctttagatg cattaaaatg 4020 attgtggttt tagtgcatgg tttacacaag tcattgttgt gcacttacag gggagtcttg 4080 gtctccatca catataacct ttattgttgt ttcaatttta gatgtgattg ccgttgaaca 4140 atcattagtt gcatgttatg tttattggat acctgtatgg gtcccaattg atatttggtt 4200 tgagatgata ctacaacaac ttgaaagtgc attttatttt ccaccaattt agggcaacca 4260 ttgtgtgtag tttctatcca cacataccat atcttcattc ttcttggcaa cttctatttt 4320 ggtacagttt ttcctacagt aaattcctcc ttttgtggcc ctgacgaacc attaattcat 4380 gtttgtacta catcagatag aaaatatata tagtagtaca cagtttcaat ttatacacgt 4440 ggtcttgaaa acttcaatgt ttactaatgc ctattggatg gatagtttac tcgaaaagtg 4500 agtaccatca catactaaca cttgataatt gtgtgtacct tcattttcat tgtggttatt 4560 gccttggcat aatggaggag aatttttgcc ataggacatc caactgactt gctgacttgc 4620 acatggcaac aacctttttg ctgtaattta attaatttgt tccatgcaat aattgttgaa 4680 aatattgttt tttaatctag aattctttcc tatcctacta cacgtaatgt gcaatgaggg 4740 ttggcttgtt gtttatgttt agtttcattt gttcttccat attttgtatg tggtgcttca 4800 tccttgtgca attgtgtatg taccggcatt ctaatgttgg tgcttgattt caggtatcca 4860 actgacagcg ataagcagat gcttgctaaa cagacaggat taactaggaa ccaggtaaac 4920 aaccatttca agtttttctt ttcccaaatt tatatcgatg taactagcta gtccgctaca 4980 gttgtccact agcaatatct ttgggtccat aatgttacta gcgttgtaaa acattttgat 5040 atctcaattc ttgagcaggt atcgaactgg tttatcaatg caagggttag gctctggaag 5100 ccaatggttg aagaaattca caacctcgag atgaggcagc tgcagaagaa tccgtctctt 5160 gacaagaatc agctctccat gcagcacacc caacattcgt cggacagcag cgggaagccc 5220 tgtgatccat caaactcgct gcaagggcaa agtagcagca tgaccaggaa tcacagcatc 5280 tccgcctccc ggcacatcga ggacggcctc tcccagatgc cccatgacat ctccggtcag 5340 gtgagcttcg catacaacgg gctcgccgcg catcacagca tcgcgatggc gcaccaccac 5400 caacctgacc tcattggcac cggtggtgcc gcgaatgctg gcggtgtctc actcaccctt 5460 ggccttcacc agaacaataa ccgagcttac attgctgagc cccttccggc cgcgcttccg 5520 ctcaatcttg cccatcgttt cggactggag gacgttagtg atgcctacgt gatgagctca 5580 tttggtggtc aggaccggca tttcaccaag gagatcggtg gccatttgct ccatgacttt 5640 gttggttgaa gagcagatat gattgtttca ccaaggagat cggtggtgat tattgtagga 5700 tgcagatgta tgatctacct atattgtagt tggaagtagg aggtgaagaa aagaggggca 5760 ttggcacgcc tgtcgtagat ccacgaatgc ttgtcgatat ttgacattgt ggggctatag 5820 agagcatatg actatacctt tgaagacctg tttgtagtgt tgttccctat ttgttctgat 5880 ccgcggttaa gccgtgttaa ccctgtacta ttcatctgac ttgcaaatgt ggttgctccc 5940 actctggcaa ataaagggtg c 5961 <210> 5 <211> 208 <212> DNA <213> Artificial Sequence <220> <223> Loc Os05g38120 5'UTR RNAi <400> 5 caacagctag cccaacacac ctcctccatc accaccgaga aggcaacgca tatcaaaagc 60 gcgggcgcaa agcaaagata acatcagatc aggtcggcgc cccccgctcc cggctgccgc 120 aaagcccacc cgatcgatcg atcgacgcct cgttctcctc ctcctccgag gctactctct 180 gcagggcgct cgcttacgtc tgcctctg 208 <210> 6 <211> 286 <212> DNA <213> Artificial Sequence <220> <223> Loc Os05g38120 3'UTR RNAi <400> 6 ccaaggagat cggtggtgat tattgtagga tgcagatgta tgatctacct atattgtagt 60 tggaagtagg aggtgaagaa aagaggggca ttggcacgcc tgtcgtagat ccacgaatgc 120 ttgtcgatat ttgacattgt ggggctatag agagcatatg actatacctt tgaagacctg 180 tttgtagtgt tgttccctat ttgttctgat ccgcggttaa gccgtgttaa ccctgtacta 240 ttcatctgac ttgcaaatgt ggttgctccc actctggcaa ataaag 286 <210> 7 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> 3A-07285 LS <400> 7 cgtattcgag aaggaagcta 20 <210> 8 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> 3A-07285 R <400> 8 acgaagatag cccattgtta 20 <210> 9 <211> 22 <212> DNA <213> Artificial Sequence <220> <223> L1S <400> 9 ctagagtcga gaattcagta ca 22 <210> 10 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> LOC_Os05g38120 F <400> 10 aggctcatct cgcttatgga 20 <210> 11 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> LOC_Os05g38120 R <400> 11 tcttccttgc cgagaccatc 20 <210> 12 <211> 19 <212> DNA <213> Artificial Sequence <220> <223> ubi F <400> 12 aaccagctga ggcccaaga 19 <210> 13 <211> 24 <212> DNA <213> Artificial Sequence <220> <223> ubi R <400> 13 acgattgatt taaccagtcc atga 24 <210> 14 <211> 23 <212> DNA <213> Artificial Sequence <220> <223> LOC_Os05g38120 FL F <400> 14 aagcttatgt cgtccgccgc ggg 23 <210> 15 <211> 28 <212> DNA <213> Artificial Sequence <220> <223> LOC_Os05g38120 FL R <400> 15 actagttcaa ccaacaaagt catggagc 28 <210> 16 <211> 23 <212> DNA <213> Artificial Sequence <220> <223> Tnos R <400> 16 ccatctcata aataacgtca tgc 23 <210> 17 <211> 24 <212> DNA <213> Artificial Sequence <220> <223> pUbi F3 <400> 17 cagctatatg tggatttttt tagc 24 <210> 18 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> LOC_Os05g38120 5'UTR RNAi F <400> 18 caacagctag cccaacacac 20 <210> 19 <211> 28 <212> DNA <213> Artificial Sequence <220> <223> LOC_Os05g38120 5'UTR RNAi R <400> 19 ggggtaccca gaggcagacg taagcgag 28 <210> 20 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> LOC_Os05g38120 3'UTR RNAi F <400> 20 ccaaggagat cggtggtgat t 21 <210> 21 <211> 30 <212> DNA <213> Artificial Sequence <220> <223> LOC_Os05g38120 3'UTR RNAi R <400> 21 ggggtaccct ttatttgcca gagtgggagc 30

Claims (14)

A recombinant vector comprising a gene represented by SEQ ID NO: 1 and a promoter operably linked to the gene. A transformed cell transformed with a recombinant vector comprising a gene represented by SEQ ID NO: 1 and a promoter operably linked to the gene, 3. The composition of claim 1 or 2, wherein the promoter is a promoter represented by SEQ ID NO: 3. A method for producing a terminal plant having improved flushing ability comprising the step of transforming a terminal plant with the composition for improving full flushing according to claim 1 or 2. 5. The method of claim 4, further comprising the step of:
Preparing a recombinant vector comprising a gene represented by SEQ ID NO: 1 and a promoter operably linked to the gene;
Introducing the recombinant vector into a terminal plant; And
Selecting a transgenic plant overexpressing the gene of SEQ ID NO: 1 upon introduction of the recombinant vector; [6] The method according to claim 5, wherein the terminal plant is rice (Oriza sativa). 10. A transgenic terminal plant transformed with the composition for promoting inagulation promotion according to claim 1 or 2. 1. A composition for inhibiting the in fl ascent of a terminal plant comprising a recombinant vector comprising an RNA i or an antisense oligonucleotide which inhibits expression of a gene represented by SEQ ID NO: 1. 9. The composition according to claim 8, wherein the recombinant vector comprises a recombinant vector comprising RNA i which inhibits gene expression represented by SEQ ID NO: 1. The composition according to claim 9, wherein the RNA i is a sequence represented by SEQ ID NO: 5 or 6. 10. A method for producing a terminal plant having suppressed inagulation, comprising the step of transforming a terminal plant with the composition for inhibiting the growth of a plant according to any one of claims 8 to 10. 12. The method according to claim 11, which comprises the following steps:
Preparing a recombinant vector comprising the RNA i represented by SEQ ID NO: 5 or SEQ ID NO: 6 which inhibits the expression of the gene represented by SEQ ID NO: 1;
Introducing the recombinant vector into a terminal plant; And
Selecting a transgenic plant having a gene expression inhibition represented by SEQ. ID. No. 1 according to the introduction of the recombinant vector; [12] The method according to claim 11, wherein the terminal plant is rice (Oriza sativa). 10. A transgenic monocotyledonous plant transformed with the composition for the prevention of aspiration-suppressing disease according to any one of claims 8 to 10.

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