KR20140050122A - Loc_os02g05840 gene, recombinant vector comprising the same, transformed plants thereby and method for preparation thereof for a yield-enhancing of oryza sativa - Google Patents

Abstract

The present invention relates to LOC_Os02g05840 gene for enhancing yield potential of rice, a recombinant vector comprising the same, transformed plants thereby and a preparation method thereof. A LOC_Os02g05840 gene of the present invention regulates a number of ears, stem development, and flowering time which are traits involved in quantity increase thereby increasing yield potential of main grains including rice and bio-mass and accordingly increasing productivity and being used for developing novel feed grains.

Description

LOC_Os02g05840 gene for enhancing the yield of rice, a recombinant vector containing the gene, a transgenic plant by the same, and a method for producing the same, and a method for producing the LOC_Os02g05840 gene, a recombinant vector comprising the same, Oryza sativa}

The present invention relates to a LOC_Os02g05840 gene for enhancing the yield of rice, a recombinant vector containing the gene, a transgenic plant and a method for producing the same.

Rice (Oryza sativa), together with barley and corn, is one of the most important food crops in the world. The demand for rice is increasing due to the increase in population, lack of agricultural land due to industrialization, and climate change, but the development of new varieties for increasing the production is insufficient. In order to solve this problem, new breeds have been developed through the introduction of useful genes out of existing breeding methods. In fact, the development of molecular biology has transformed many plants including rice into new transgenic plants through the isolation and manipulation of useful genes. The discovery of these useful genes and the development of new varieties using them can be useful for the solution of food problems and the whole industry.

Rice is one of the most important crops in the world for the purpose of seed production. In recent years, the entire gene sequence has been revealed, and a lot of efforts have been made to acquire a large number of useful genes and to preoccupy them. As a technique for improving the size and growth of seeds in the past, there has been known a method of controlling plant cell growth by modifying the concentration or the catalytic activity of a cell cycle control protein in a plant cell (US Patent No. 6,087,175) A method of regulating the expression level of a target gene in a plant cell by introducing an expression vector in which a target gene is bound to a plant cell to promote a nucleotide encoding a synthetic zinc finger protein (US Patent No. 7,151,201) . Also, there is a technology (Korean Patent Registration No. 10-0794395) in which the gene of cytochrome P450 protein derived from Arabidopsis thaliana is transformed into plant cells to increase the size of plant seeds.

The shape and size of the organisms such as the stem and leaf of the plant are important factors that increase the biomass and affect the agricultural productivity. Research on stemming mechanism of major grains such as rice, maize, wheat, and millet is in the active stage. Since the identification of teosinte branched 1 (tb1), a stem production inhibitor that enabled maize breeding, identification of genes related to stems such as zfl (FLORICAULA / LEAFY), bif (barren inflorescence2) and pin (Lukens and Doebley 2001, Barazesh and McSteen 2008). The first known tb1 gene in maize has been identified as a key gene in rice and Arabidopsis (Takeda et al. 2003, Finlayson 2007, Aquilar-Martinez et al. 2007).

(Schumacher et al. 1999, Schmitz et al. 2002), and other genes such as sugarcane, bamboo, etc. have been identified in addition to major crops such as rice, maize and corn Research into the stem production regulatory genes to promote biomass is at an early stage (Aiteken et al. 2008). In addition, recent studies have shown that tb1 gene of maize is introduced into wheat to inhibit cleavage (Lewis 2008), indicating that these genes can be used in biotechnology.

In order to increase the yield of plants, the number of ears is not only increased but also has a proper plant form to make a lot of ears. For this purpose, it is important to control the development of the stem of the rice, flowering time, ear growth.

For example, the flowering time of rice is an important trait for recognizing environmental factors such as seasonal changes and temperature changes. Rice is a monocotyledonous plant whose flowering is promoted in a single condition, and the flowering period (water extraction period) is influenced by the photoperiod and nutrient growth period, and these factors are important agricultural traits that are involved in growing rice in various fields. Among them, photoperiod significantly influences growth by controlling the flowering time and the biological clock of rice, which directly affects productivity. In addition, recently, as the nucleotide sequence of the entire genome of rice has been completed and research on comparative genomics has progressed, rice has attracted attention as an important monocotyledonous plant.

In other words, to increase the yield, it is necessary to study each gene which regulates stem development, flowering time, ear development, or a single gene that can control all of them and a new breed using the same.

However, to date, some flowering-related genes, stem development-related genes and ear developments have been identified in rice, but the number is limited and little is known about rice-specific genes distinguished from Arabidopsis. Considering that huge agricultural benefits can be obtained by controlling stem development, flowering time, and ear development, efforts to search for genes involved in this will be urgently needed.

(US) 10-2007-7016825 (US) 10-2006-0095687

The present invention provides a LOC_Os02g05840 gene, a recombinant vector comprising the gene, a transformant transformed with the recombinant vector, and a method of preparing the same, for the purpose of increasing the yield through controlling stem development, ear growth, and flowering time of plants. I would like to.

In order to achieve the above object, the present inventors made diligent efforts to discover genes for increasing the quantitative yield, such as the ear development, stem development, flowering time of plants. In this process, the LOC_Os02g05840 gene of rice was identified as a gene for regulating ear number, stem development, and flowering time, and accordingly, this gene was secured to prepare a recombinant vector including the same, and introduced into a plant to transform it. The present invention was completed by confirming that the number of spikes, the stems, and the flowering period of the converting body increased.

More specifically, the present inventors observed mutants with changes in the number of eggs, stem development, and flowering time in the 100,000 rice mutant population produced by the transformation method using Agrobacterium. Each mutant plant is a mutant in which the gene in the region inserted by insertion of T-DNA into the genome by Agrobacterium has been rendered inoperable. The location of T-DNA insertion can be confirmed by genomic DNA PCR and sequencing, and the gene locus id can be found in the published gene database of rice.

Through the phenotypic observations of the mutant population, two lines 3A-10852 and 1B-21530 with varying flowering time were selected. 3A-10852 shows that T-DNA was inserted into the second intron of LOC_Os02g05840, and 1B-21530 was a mutant with T-DNA inserted into the third exon. Thus, the function of LOC_Os02g05840 was lost in these two lines by PCR there was.

DNA of LOC_Os02g05840 was isolated and cloned into pGA3426, pGA3428, and pGA3438 vectors based on the nucleotide sequencing information. This was introduced into Agrobacterium and used for transformation.

The phenotype of the transformed plant was analyzed to confirm that the LOC_Os02g05840 gene regulates ear development, stem development, and flowering time.

The present invention provides a LOC_Os02g05840 gene (hereinafter referred to as "gene of the present invention") for increasing the yield of rice. The increase in yield by the LOC_Os02g05840 gene is due to one or more selected from ear growth, stem development or shortening of flowering time.

The genome DNA size of LOC_Os02g05840 is 5600 bp and the size of the representative coding sequence (LOC_Os02g05840.1) is 2250 bp. It consists of 4 exons and 3 introns and is translated into 749 amino acids to make protein. This protein encodes the PHD finger domain, the fibronectin type III (FNIII) domain, and the VIN3 interacting domain (VID). The PHD finger domain binds to the histone residue of the DNA polymer, And the expression of these genes is known to play a role. The PHD finger domain that LOC_Os02g05840 encodes also binds histone H3.

The gene encodes information necessary for constructing and maintaining the cells of the organism, and can increase the yield by using the LOC_Os02g05840 gene involved in the development of ear, stem and flowering time according to the present invention.

The present invention also provides a recombinant vector for increasing the yield including the gene.

The term "vector" refers to a DNA fragment (s), nucleic acid molecule, which transfers into a cell, which can be cloned and independently reprogrammed in host cells. "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. The recombinant vector may generally be derived from a plasmid or viral DNA, or may contain both elements. Thus, a recombinant vector refers to a recombinant DNA or RNA construct, such as a plasmid, phage, recombinant virus, or other vector that, upon introduction into an appropriate host cell, results in the expression of the cloned DNA. Suitable recombinant vectors are well known to those of skill in the art and include those that replicate in eukaryotic and / or prokaryotic cells and those that remain as episomes or that are integrated into the host cell genome.

The vector may be any vector capable of introducing the gene of the present invention. Preferably, pCAMBIA or pGA family vectors such as pGA3383, pCAMBIA1381, pCAMBIA1391, pGWB3, pGA3426, pGA3428, pGA3438, pGA2715, and a vector derived from a Ti-plasmid such as pGA2717. More preferably, pGA3426, pGA3428 or pGA3438 may be used.

Preferably, the recombinant vector of the present invention is a recombinant vector comprising a promoter operably linked to the LOC_Os02g05840 gene.

For example, the promoter may be anything that is expressed by being limited to the entire body or a specific tissue of a plant, preferably a promoter that induces systemic expression, such as a CaMV virus as a promoter of the 35S RNA gene and the promoter for promoting the expression of the whole plant in a cotyledon plant are other tissue specific promoters such as rice actin, maize ubiquitin gene promoter and rice cytochrome C gene (OsOc1) promoter and leaf For example, ribosyl bisphosphate carboxylase / oxygenase small subunit (rbcS) promoter derived from rice and maize, RolD promoter inducing expression of plant roots derived from Agrobacterium, patatin promoting expression specific to potato-derived tubers, (PDS: phytoen < RTI ID = 0.0 > e synthase) promoter, the promoter of the present invention LOC_Os02g05840 gene, and the like.

More preferably, there is provided a recombinant vector comprising the promoter of LOC_Os02g05840, which is a promoter represented by SEQ ID NO: 3, which is operatively linked to the LOC_Os02g05840 gene.

Means 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. Such nucleic acid fragments are 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, compared to the nucleotide sequences set forth in SEQ ID NO: 1 With 96%, 97%, 98%, 99% or more sequence homology, such nucleic acid fragments can be readily produced by molecular biological methods well known in the art.

The recombinant vector may be one obtained by introducing the gene of the present invention into a conventional vector. The production of a recombinant vector by introducing the gene of the present invention may be easily carried out according to known methods by those skilled in the art. It is possible.

The present invention also provides a transformed cell transformed with said 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 hosts include, but are not limited to, bacterial cells such as Escherichia coli, Streptomyces and Salmonella typhimurium cells and plant cells. Preferably, the transformed cells are transformed cells that are microorganisms of the genus Agrobacterium. More preferably, Agrobacterium tumefaciens LB4404 can be used as the microorganism of the genus Agrobacterium.

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 also provides a transformed transgenic plant transfected with said vector or said transformed cell.

The transformed monocotyledonous plant exhibits an increase in yield, stem development, or shortening of flowering time by the introduced LOC_Os02g05840 gene, thereby exhibiting an increase in the yield of monocotyledonous plants.

"Transformed monocotyledonous plant" and "functionally equivalent transgenic plant" of the present invention is 60%, 65%, 70%, 75%, 80%, Transgenic monocots prepared using sequences having 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence homology As a plant, it is a transgenic plant having substantially the same characteristics as the effect of increasing the yield by increasing ear, stem development or flowering time.

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) Infections caused by (non-integrative) viruses in Agrobacterium tumefaciens mediated gene transfer (EP 0 301 316), and the like.

The plant to be transformed in the present invention can be used without limitation in a terminal plant including the rice (Oryza sativa L.) used in the present embodiment.

Preferably rice (Oryza sativa L.) is used.

Such mutants can be easily produced by molecular biological methods well known in the art.

In another aspect, the present invention provides a method for transforming a plant comprising the step of transforming the plant with a vector using a foreign promoter to express the gene LOC_Os02g05840 gene involved in the development of ear, stem and flowering time.

Preferably, a step of preparing a vector comprising the gene of SEQ ID NO: 1; Introducing the recombinant vector into rice; And it provides a plant transformation method comprising the step of selecting a rice transformant having the effect of increasing the ear, stem development, shortening the flowering time according to the introduction of the recombinant vector.

LOC_Os02g05840 gene of the present invention is a gene that improves yield by affecting ear development, stem development, and flowering time, and is used to increase the yield of various grains including rice, increase biomass of plants, and study feed crops. Can be.

1 is a diagram showing the LOC_Os02g05840 gene and T-DNA insertion positions of the 3A-10852 and 1B-21530 lines.
2 is a diagram showing the presence or absence of LOC_Os02g05840 transcript according to the flowering phenotype and genotype of the 3A-10852 and 1B-21530 lines.
Figure 3 is a diagram of a vector for the production of LOC_Os02g05840 overexpressed transgenic monocots.
FIG. 4 is a graph showing the results of confirming the LOC_Os02g05840 gene expression of transgenic rice plants overexpressing the LOC_Os02g05840 gene.
5 is a diagram showing the phenotype of transgenic rice plants overexpressing the LOC_Os02g05840 gene.
FIG. 6 is a diagram showing elongation lengths and internode lengths of transgenic rice plants transgenic overexpressing LOC_Os02g05840 gene.
7 is a diagram showing stem thickness and transverse shape of LOC_Os02g05840 gene overexpressed transgenic rice.
FIG. 8 is a graph showing the numerical value and the total number of seeds of primary and secondary sprouts of LOC_Os02g05840 gene overexpressed transgenic rice.
FIG. 9 is a diagram showing the cell shape of LOC_Os02g05840 gene overexpressed transgenic rice, centering on length and width.

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> Selection of flowering timing control mutant strains affecting yield

The plant phenotype of 100,000 T-DNA mutant populations was observed to find genes that regulate ear, stem, and flowering time, which are factors affecting the yield. Of these, two strains 3A-10852 and 1B-21530 were selected for the slow flowering period. The difference in flowering time was confirmed by comparing the two lines with the control group, Dongjin rice, which was grown in 12 light / 12 dark (single condition), 16 light / 8 dark (long day condition)

The results of the flowering time are shown in Table 1.

Table 1. Changes in Flowering Time of 3A-10852 and 1B-21530 Systems

As shown in Table 1, it was confirmed that the flowering time of each of the two lines was slowed down compared with that of the control group, Dongjin rice, under single condition, long-day condition or rice field condition.

< Example  2 > The T- DNA  Cover co - segregation  Verification

The 3A-10852 and 1B-21530 lines generated by T-DNA insertion were trapped by pGA2715 and pGA2717 vectors, respectively. T-DNA-inserted chromosomes were analyzed by inverse-PCR and sequencing of the two lines.

The selected two lines of rice seeds were cultured in MSO medium at 28 DEG C for 10 days. 100 mg of the leaves of the cultivated individuals were sampled and pulverized by rapid cooling using liquid nitrogen. DNA was extracted from the pulverized leaves using one step buffer (5 ml of 1 M Tris (pH 9.5), 1 ml of 0.5 M EDTA, and 3.73 g of KCl, followed by 50 ml of water).

3A-10852 is a system in which T-DNA is inserted into the second intron of the LOC_Os02g05840 gene. PCR was performed using primers (SEQ ID NO: 5 and SEQ ID NO: 6) and NGUS1 primer (SEQ ID NO: 9) Genotype analysis was performed.

1B-21530 was genotyped with primers (SEQ ID NO: 7 and SEQ ID NO: 8) and NGUS1 primer (SEQ ID NO: 9) prepared before and after the insertion site in which T-DNA was inserted into the third exon of the LOC_Os02g05840 gene.

3A-10852 and 1B-21530 insertion positions and corresponding sequences in the LOC_Os02g05840 gene are shown in FIG. 1 and SEQ ID NO: 1, and the primers used in the genotyping analysis are shown in Table 2.

Table 2. Primers 3A-10852 and 1B-21530

1 is the nucleotides 160 to 178 of SEQ ID NO: 4, 2 is the nucleotides 1627 to 2314 of SEQ ID NO: 4, the T-DNA insertion site of the 3A-10852 strain is 2550 and 3 is the nucleotide of SEQ ID NO: 4 3298 to 3602, the T-DNA insertion site of the 1B-21530 strain is 3591, and 4 is 3680 to 4917 of SEQ ID NO: 4. 1 to 4 in Fig. 1 are sequences corresponding to respective exons.

PCR reactions were performed after 5 minutes at 95 ° C; 30 seconds at 95 ° C, 20 seconds at 53 ° C, 1 minute at 72 ° C was repeated 35 times, and finally 7 minutes at 72 ° C. PCR products were confirmed by electrophoresis using agarose gel. This confirmed the genotype.

The results are shown in Fig. 2A.

FIG. 2A shows a comparison of the flowering time of pure (HO) experimental groups 3A-10852 and 1B-21530 with the flowering time of wild type (WT). As shown in FIG. 2A, it can be confirmed that the flower buds have already been formed in the wild type (WT) and the seeds are developed at the time when the flower buds of purebred (HO) 3A-10852 and 1B-21530 are emerging, . In addition, genotype analysis results through PCR also support the above results. The upper band corresponds to the PCR product of the gene specific primer (SEQ ID NO: 5 to SEQ ID NO: 8), and the lower band corresponds to the NGUS1 (SEQ ID NO: 9) on the RB (right border) of T-DNA and SEQ ID NO: . In both experimental groups, the wild-type (WT) was amplified only by gene specific primers, confirming that obedience (HO) was only amplified by SEQ ID NO: 5 or SEQ ID NO: 7 and NGUS1 (SEQ ID NO: 9) on the RB I could.

That is, it was confirmed that the LOC_Os02g05840 gene affects the flowering time.

In order to confirm whether the function of the LOC_Os02g05840 gene was actually lost, it was confirmed that the mRNA was produced using the primer (SEQ ID NO: 10) produced in the first exon of FIG. 1 and the primer (SEQ ID NO: 11) Respectively. As a control, primers of rice ubiquitin-5 (SEQ ID NO: 12 and SEQ ID NO: 13) were used. The primers for identifying such transcripts are shown in Table 3.

Table 3. LOC_Os02g03670 transcript confirmation primer

Two lines of seeds were grown in MSO medium for 10 days, and then 100 mg of leaves of each individual were collected 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 μg of total RNA was synthesized using Moloney murine leukemia virus reverse transcriptase (Promega, Madison, WI, USA) and the cDNA was confirmed by PCR.

PCR reactions were performed after 5 minutes at 95 ° C; 30 seconds at 95 ° C, 20 seconds at 53 ° C, 2 minutes at 72 ° C, and finally 7 minutes at 72 ° C. PCR products were confirmed by electrophoresis using agarose gel.

The results are shown in Fig. 2B.

As shown in FIG. 2B, it was confirmed by PCR that there was no mRNA production of the LOC_Os02g05840 gene in the pure (HO) experimental groups 3A-10852 and 1B-21530. The upper band of FIG. 2B corresponds to the PCR product of the mRNA primers (SEQ ID NO: 10 and SEQ ID NO: 11) of LOC_Os02g05840 and the lower band corresponds to the PCR of the specific primers of the rice ubiquitin 5 (SEQ ID NO: 12 and SEQ ID NO: 13) It corresponds to product.

That is, 3A-10852 and 1B-21530 isolated from the genotypic analysis were delayed to flowering in comparison with the control group, Dongjin, and the mRNA was not amplified when the presence of mRNA was observed by PCR.

From the above results, it was confirmed that the 3A-10852 and 1B-21530 lines were co-segregated with the LOC_Os02g05840 gene.

< Example  3> LOC _ Os02g05840  Genes for expression Cloning

In order to clone the LOC_Os02g05840 gene, PCR was performed using SEQ ID NO: 10 and SEQ ID NO: 11 shown in Table 3 with the cDNA used in FIG. 2C as a template. 20 ng of cDNA template and 5 pmol of each primer were used. PCR reactions were performed after 5 minutes at 95 ° C; 30 seconds at 95 ° C, 20 seconds at 53 ° C, 2 minutes at 72 ° C, and finally 7 minutes at 72 ° C. The PCR product was electrophoresed using an agarose gel, and the size of the PCR product was determined. Then, the PCR product was isolated and identified and cloned into a pGA3724 vector (vector for sub-cloning). The cloned DNA was digested with HindIII and SpeI, followed by ligation with pGA3426 vector (over expression vector) digested with the same restriction enzyme at 28 DEG C for 3 hours. The ligation product was mixed with 50 μl of Top10 E. coli competent cells, transferred to a 1.5 ml tube, and left on ice for 15 minutes. Subsequently, the plate was allowed to stand in a 37 ° C oven for 1 minute, and then 1 ml of LB liquid medium was further placed in the tube and left in a 37 ° C shaking incubator for 3 hours. Then, the cells were plated on kanamycin-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 a miniprep was digested with restriction enzymes HindIII and SpeI, and the agarose gel was separated by electrophoresis to confirm the band. The obtained cloning DNA was analyzed again to select DNA that did not cause an error. FIG. 3 shows a diagram of the vector for producing the overexpressed plant LOC_Os02g05840. Sequence analysis was performed using a 3730XL DNA analyzer (AB, USA) and BigDye v3.1 (AB, USA), and primers using sequences in the LOC_Os02g05840 gene were used.

Table 4. Primers for Sequencing

< Example  4> Production of transformed cells

The plant recombinant vector produced in Example 3 was extracted. 50 μl of Agrobacterium tumefaciens LB4404 and 10 μg of the plant recombinant 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  5> Production of Transgenic Plants

From the N6D solid medium, Dong - Jin - jin seeds were grown for 30 days at 28 ℃ in the growth chamber under dark conditions to produce calli of rice. The resulting callus was mixed with the cells cultured for 72 hours with Agrobacterium transformed with the plant recombinant vector obtained in Example 4, and left in a culture medium containing N6D-Acetosyringone at a temperature of 22 占 폚.

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 cleaved callus was transferred to MSR (hygromycin 40 mg / L) solid medium, which is a regeneration medium, to induce re-differentiation in the growth room at 28 ° C. for 4 weeks, and then the plants were transferred to MS solid medium and grown in 28 ° C. Transferred to grow replanted plants.

< Example  6> transgenic plants LOC _ Os02g05840  Gene expression verification

To confirm the increased expression of LOC_Os02g05840 in regenerated plants, isolated total RNA was isolated from regenerated plants and quantitative real-time RT-PCR was performed using a Rotor-Gene 6000 PCR instrument. A primer to be used for confirmation of expression was prepared at the 4'-axon and 5'UTR sites in Fig. 4A. The primers are shown in Table 5.

Table 5. Verification of expression of LOC_Os02g05840 in transgenic plants Primer

As shown in FIG. 4B, LOC_Os02g05840 expression was observed in different LOC_Os02g05840 overexpressed rice OX1 and OX2. As a result, it was confirmed that the amount of transcript was significantly increased in comparison with the control group, Dongjin rice. In other words, the transgenic rice with the LOC_Os02g05840 transcript increased using the LOC_Os02g05840 overexpression recombinant vector.

< Example  7> LOC _ Os02g05840  Over-expression transgenic plants were characterized

LOC_Os02g05840 Overexpressed transgenic rice plants were identified by phenotypic observations.

The results are shown in Figs. 5 to 8. Fig.

5 is a diagram showing a phenotype of transgenic rice. The three transgenic lines of the transgenic rice were different from those of the control group.

First, OX1 and OX2 were elongated longer than the control, and the results are shown in Fig.

As shown in Fig. 6, it can be confirmed that the elongation length is longer than that of the control group. The kidney length was measured from the stem end of the plant to the end of the ears. In FIG. 6A, the control elongation length was 122 cm, and the average elongation lengths of the OX1 and OX2 plants were 158 and 173 cm, respectively. This figure is due to the fact that OX1 and OX2 plants are 30% and 42% longer, respectively, than the control, and this increase in length is due to the longer length of each internode. The results of measuring the lengths of the major and minor intervertebral bodies of the control and two line transformants are shown in Figure 6B. As shown in FIG. 6B, it was confirmed that the lengths of all the intervertebral discs were longer than those of the control.

As shown in FIG. 6, when the expression level of LOC_Os02g05840 was increased, it was confirmed that the kidney which is a trait related to the yield was lengthened. In other words, LOC_Os02g05840 can play an important role in increasing the yield by regulating plant kidney development.

Second, the transgenic lines of both strains showed a thicker stem compared to the control.

The results are shown in Fig.

As shown in Fig. 7A, the thicknesses of the major and minor stems of the control and OX1 and OX2 plants of the control and two lines were measured at 4 cm above each node using a vernier caliper. The major stem thickness of transgenic plants was 19% larger than that of the control group, and the minor stem thickness was 20% larger on average.

A section was made to confirm this in cell units.

section, the third section of the plant was discarded after 30 days and the FAA fix solution (formalin (35%): acetic acid: alcohol (70%) = 1: 1: 18) The spiral cells were fixed. The cross-sectional tissue was cut with a LEICA RM2265 section apparatus in a transverse section within 1 μm, fixed on a slide glass, fixed with a cover glass, stained with 0.05% toluidine blue solution and observed with an optical microscope at 20-40 magnification.

The results are shown in Fig. 7B.

As shown in Fig. 7B, it was confirmed that the stem thickness of the OX1 plant was thicker than that of the control group.

In other words, it was confirmed that LOC_Os02g05840 gene is involved in whole stem development through influencing thickness development as well as stem length.

Third, total seed number of main tiller increased.

The number of primary spikes and secondary spikes increased in the transformants compared to the control group, and the total number of seeds was increased, as shown in FIG.

As shown in Fig. 8, in the transformant, the number of primary spikes was increased by 40% as compared with that of the control group. In Fig. 8B, the number of secondary spikes increased by 28% Respectively. The increase in the number of seeds resulted in an increase in the total number of seeds. As shown in FIG. 8C, the number of seeds of OX1 plants was increased by 47% as compared with the control.

The results of FIG. 8 show that the number of seeds can be increased to increase the yield, as well as controlling the height of the kidney, which is one of the factors that increase the yield.

< Example  8> LOC _ Os02g05840  Overexpressed Transforming Cytological Observation

The cytotoxicity of the LOC_Os02g05840 overexpressant was investigated.

In order to observe the size and number of the cells, the third section of the plant that had been over 30 days old was cut and FAA fix solution (formalin (35%): acetic acid: alcohol (70%) = 1: : 18). The cross-sectional tissue was cut with a LEICA RM2265 section instrument to a length of less than 1 μm, fixed on a slide glass, fixed with a cover glass, stained with 0.05% toluidine blue solution and observed at 20-40 magnification under an optical microscope.

As shown in Fig. 9A, the OX1 transformant cells were smaller in size than the control group, and the number of cells was found to be the same within the same range. As shown in FIG. 9B, when the cells were quantitatively observed, it was confirmed that the transformant cells were about 40% shorter in length than the control group.

In other words, FIG. 9 supports that the effects on the elongation and thickness increase of the stem are due to the increase in elongation and thickness as the number of cells constituting the plant increases in FIG. 6 and FIG.

Based on the above results, the LOC_Os02g05840 gene shows that one gene can control several traits related to increased quantitative properties such as flowering time, height of kidney and increase of ear yield. That is, it was confirmed that the LOC_Os02g05840 gene could increase the yield.

<110> University-Industry Cooperation Group of Kyung Hee University <120> LOC_Os02g05840 gene, recombinant vector comprising the same,          transformed plants by and method for preparation thereof          a yield-enhancing of Oryza sativa <130> P12-086-KHU <160> 22 <170> Kopatentin 2.0 <210> 1 <211> 2247 <212> DNA <213> Oryza sativa <220> <221> CDS <222> (1) (2247) <223> LOC_Os02g05840 <400> 1 atg gat cca ccc tac gca gga gta cct att gat cct gct aaa tgc cga 48 Met Asp Pro Pro Tyr Ala Gly Val Pro Ile Asp Pro Ala Lys Cys Arg   1 5 10 15 ttg atg agt gtg gat gaa aag cgg gaa ctt gtc cgt gaa tta tcg aag 96 Leu Met Ser Val Asp Glu Lys Arg Glu Leu Val Arg Glu Leu Ser Lys              20 25 30 cgg cca gaa agt gct cct gac aaa ctg cag tct tgg agt cgc cgt gaa 144 Arg Pro Glu Ser Ala Pro Asp Lys Leu Gln Ser Trp Ser Arg Arg Glu          35 40 45 att gta gag att ctt tgt gct gat tta gga agg gaa agg aag tac act 192 Ile Val Glu Ile Leu Cys Ala Asp Leu Gly Arg Glu Arg Lys Tyr Thr      50 55 60 gga tta tcg aag cag aga atg ttg gaa tat ctc ttc aga gtt gt Gly Leu Ser Lys Gln Arg Met Leu Glu Tyr Leu Phe Arg Val Val Thr  65 70 75 80 ggc aaa tca tct ggt ggt ggc gtt gtg gag cat gtg caa gag aag gag 288 Gly Lys Ser Ser Gly Gly Gly Val Val Glu His Val Gln Glu Lys Glu                  85 90 95 cct acc cct gaa ccc aac aca gcc aac cat cag tcc cct gcg aaa cgg 336 Pro Thr Pro Glu Pro Asn Thr Ala Asn His Gln Ser Pro Ala Lys Arg             100 105 110 cag cga aag agt gac aac cca tca cga cta cca att gt t gca agc agt 384 Gln Arg Lys Ser Asp Asn Pro Ser Arg Leu Pro Ile Val Ala Ser Ser         115 120 125 cca act aca gaa ata ccc agg cca gca agt aat gct cgc ttc tgc cac 432 Pro Thr Thr Glu Ile Pro Arg Pro Ala Ser Asn Ala Arg Phe Cys His     130 135 140 aat tta gct tgc aga gcg act ctt aat cca gaa gat aaa ttt tgc aga 480 Asn Leu Ala Cys Arg Ala Thr Leu Asn Pro Glu Asp Lys Phe Cys Arg 145 150 155 160 cgc tgt tca tgc tgt att tgt ttc aag tac gat gac aat aag gat cct 528 Arg Cys Ser Cys Cys Ile Cys Phe Lys Tyr Asp Asp Asn Lys Asp Pro                 165 170 175 agc ctc tgg tta ttc tgt agt tca gat caa ccc ttg cag aaa gat tct 576 Ser Leu Trp Leu Phe Cys Ser Ser Asp Gln Pro Leu Gln Lys Asp Ser             180 185 190 tgt gta ttt tcg tgc cat ctt gaa tgt gct ctt aag gat gga aga act 624 Cys Val Phe Ser Cys His Leu Glu Cys Ala Leu Lys Asp Gly Arg Thr         195 200 205 ggc atc atg cag agt ggg cag tgc aag aaa ctt gat ggt ggt tat tac 672 Gly Ile Met Gln Ser Gly Gln Cys Lys Lys Leu Asp Gly Gly Tyr Tyr     210 215 220 tgc act cgc tgt cgg aaa cag aat gat ctg ctt ggg tcc tgg aag aaa 720 Cys Thr Arg Cys Arg Lys Gln Asn Asp Leu Leu Gly Ser Trp Lys Lys 225 230 235 240 caa ctg gtg ata gct aaa gat gct cgc cgg ttg gat gta ttg tgt cat 768 Gln Leu Val Ile Ala Lys Asp Ala Arg Arg Leu Asp Val Leu Cys His                 245 250 255 cgg att ttt ttg agt cat aag att ctt gtc tcc acg gag aag tac ttg 816 Arg Ile Phe Leu Ser His Lys Ile Leu Val Ser Thr Glu Lys Tyr Leu             260 265 270 gtt ttg cat gaa att gtt gac aca gcg atg aag aaa ctg gag gct gag 864 Val Leu His Glu Ile Val Asp Thr Ala Met Lys Lys Leu Glu Ala Glu         275 280 285 gtt ggt cct ata tct gga gtt gca aat atg ggt cgt gga att gtg agc 912 Val Gly Pro Ile Ser Gly Val Ala Asn Met Gly Arg Gly Ile Val Ser     290 295 300 cgg ctt gct gtt ggt gct gaa gtt cag aaa ctt tgt gct cga gca ata 960 Arg Leu Ala Val Gly Ala Glu Val Gln Lys Leu Cys Ala Arg Ala Ile 305 310 315 320 gaa acc atg gag tct ctg ttt tgt gga tct cct tct aac ttg caa ttt 1008 Glu Thr Met Glu Ser Leu Phe Cys Gly Ser Pro Ser Asn Leu Gln Phe                 325 330 335 caa cgt tca cgg atg ata cca tca aac ttc gta aag ttt gaa gct ata 1056 Gln Arg Ser Ser Met Ile Pro Ser Asn Phe Val Lys Phe Glu Ala Ile             340 345 350 acc caa aca tct gtc act gta gtt ttg gat ttg ggt cct ata ctt gct 1104 Thr Gln Thr Ser Val Thr Val Val Leu Asp Leu Gly Pro Ile Leu Ala         355 360 365 caa gat gta aca tgc ttt aat gta tgg cac aga gtg gca gcc aca ggc 1152 Gln Asp Val Thr Cys Phe Asn Val Trp His Arg Val Ala Ala Thr Gly     370 375 380 tcg ttc tca tca agt cca act ggc atc ata ctt gca cca tta aaa acg 1200 Ser Phe Ser Ser Ser Pro Thr Gly Ile Ile Leu Ala Pro Leu Lys Thr 385 390 395 400 tta gtg gtc act caa ctt gtg cca gct aca agc tat ata ttc aag gta 1248 Leu Val Val Thr Gln Leu Val Pro Ala Thr Ser Tyr Ile Phe Lys Val                 405 410 415 gtt gcc ttc agt aac tac aag gag ttt gga tcg tgg gaa gcc aaa atg 1296 Val Ala Phe Ser Asn Tyr Lys Glu Phe Gly Ser Trp Glu Ala Lys Met             420 425 430 aag aca agc tgt cag aag gaa gtt gat ctg aag ggt ttg atg cca ggt 1344 Lys Thr Ser Cys Gln Lys Glu Val Asp Leu Lys Gly Leu Met Pro Gly         435 440 445 ggg tct ggg cta gac caa aac aat ggg agc cca aag gca aac agt ggt 1392 Gly Ser Gly Leu Asp Gln Asn Asn Gly Ser Pro Lys Ala Asn Ser Gly     450 455 460 ggt cag tct gat cct tct tca gaa ggt gtg gac tca aat aat aac act 1440 Gly Gln Ser Asp Pro Ser Ser Glu Gly Val Asp Ser Asn Asn Asn Thr 465 470 475 480 gcg gtg tat gct gat ctc aat aaa tca cca gaa agt gat ttt gaa tat 1488 Ala Val Tyr Ala Asp Leu Asn Lys Ser Pro Glu Ser Asp Phe Glu Tyr                 485 490 495 tgt gaa aat cct gag ata ctt gat tca gac aaa gca agt cat cac ccc 1536 Cys Glu Asn Pro Glu Ile Leu Asp Ser Asp Lys Ala Ser His His Pro             500 505 510 aat gaa cct aca aac aac tca cag agt atg ccg atg gtc gta gct agg 1584 Asn Glu Pro Thr Asn Asn Ser Gln Ser Met Pro Met Val Val Ala Arg         515 520 525 gtt acg gag gta tct gga ttg gag gaa gct cct gga ctc tca gca tca 1632 Val Thr Glu Val Ser Gly Leu Glu Glu Ala Pro Gly Leu Ser Ala Ser     530 535 540 gct ttg gac gag gag ccc aat tca gca gtt caa aca caa tta ctt aga 1680 Ala Leu Asp Glu Glu Pro Asn Ser Ala Val Gln Thr Gln Leu Leu Arg 545 550 555 560 gaa tcc tca aat tca atg gag cag aac cag aga gaa gtt cct gga 1728 Glu Ser Ser Asn Ser Met Glu Gln Asn Gln Arg Ser Glu Val Pro Gly                 565 570 575 tca cag gat gca tca aat gct cct gct gga aat gag gtg gtg att gtt 1776 Ser Gln Asp Ala Ser Asn Ala Pro Ala Gly Asn Glu Val Val Ile Val             580 585 590 cca cct cga tat tct ggc tct att cca cca act gca cct aga tat atg 1824 Pro Pro Arg Tyr Ser Gly Ser Ile Pro Pro Thr Ala Pro Arg Tyr Met         595 600 605 gaa aat ggt aag gat atc agt ggg agg agc ttg aaa gca aaa cct ggt 1872 Glu Asn Gly Lys Asp Ile Ser Gly Arg Ser Leu Lys Ala Lys Pro Gly     610 615 620 gat aac atc ct caa aat ggc tc tcc aag cct gaa agg gaa cca ggg 1920 Asp Asn Ile Leu Gln Asn Gly Ser Ser Lys Pro Glu Arg Glu Pro Gly 625 630 635 640 aat tct aaa aaa aga aca tca ggt aaa tgt gag gaa atc ggc cac 1968 Asn Ser Ser Asn Lys Arg Thr Ser Gly Lys Cys Glu Glu Ile Gly His                 645 650 655 aag gat gga tgc cca gaa gca tct tat gag tac tgt gtt aag gtg gtc 2016 Lys Asp Gly Cys Pro Glu Ala Ser Tyr Glu Tyr Cys Val Lys Val Val             660 665 670 agg tgg ctg gaa tgt gag ggt tac att gag acc aac ttc aga gtg aag 2064 Arg Trp Leu Glu Cys Glu Gly Tyr Ile Glu Thr Asn Phe Arg Val Lys         675 680 685 ttt ctg act tgg tat agc ctt cgt gct acc cct cat gac agg aag ata 2112 Phe Leu Thr Trp Tyr Ser Leu Arg Ala Thr Pro His Asp Arg Lys Ile     690 695 700 gtc agc gtc tac gta aac act ctt att gat gat cct gtt agc ctt tct 2160 Val Ser Tyr Val Asn Thr Leu Ile Asp Asp Pro Val Ser Leu Ser 705 710 715 720 ggc cag ctt gct gac act ttc tct gag gcc atc tac agc aaa agg cca 2208 Gly Gln Leu Ala Asp Thr Phe Ser Glu Ala Ile Tyr Ser Lys Arg Pro                 725 730 735 cct tct gtt cgc tcc ggt ttc tgc atg gaa ctt tgg cat 2247 Pro Ser Val Arg Ser Gly Phe Cys Met Glu Leu Trp His             740 745 <210> 3 <211> 3000 <212> DNA <213> Oryza sativa <220> <221> promoter &Lt; 222 > (1) .. (3000) <223> Promoter of LOC_Os02g05840 <400> 3 ttggttccta aactctcaaa atacatatcc aaagtcccaa aacttatcaa aatatgttat 60 ttaggtctca aatcgccaca acctcttcag aattttacgt ggggctcccc acatggatgt 120 ggcgtggtat catcttgact gaaaaatagg acccacttat tttttttctt ttttcttctt 180 ttccttttcc ttctcttttt tttccttttc tgttcctttc tctgtgacct gagtacccca 240 tgcggcggta gcctattttt agtgagagga aggagaagaa atgaaaaaaa taagagaagg 300 agaagaaaaa taattttttt taagtaggtc ccatttttaa gtcaagatta tgctacatca 360 gt; caagttttga gatctagatt tacatttaag agattgggga cctagatgac actctaagga 480 ctgctcatgc actttactct atttttctag aaatgtcggc accttctttg tgatcctggt 540 cgaatttgag gagtttttgg cttaaaacaa atccaaaacg acttataaac ttgacaagtt 600 tagatatttg ttacaaacga cttatattaa cgacaaatct agacaatctt ttatctaaaa 660 cctaagctct aaccatttgc ccagattagt tatattagag caagtttaat agtatagccc 720 actactagct ccaatttatc tatagccaat ctaatagcta attcatacaa tagttgttta 780 ctatactatt aatatatggt ctcacctgtc atacacatac tgtgtcttgg agtccgtgct 840 acagctggct acagatctgt agactgcttc tcttatctct catcttttat ctcattaaaa 900 tatgtttata gctgactaat agtctgctat tgtacctgct tttaggaggg gttatatttg 960 gttgagtatt tttcgggacg gatggagtac actgcaatat agctgtagaa tattcataaa 1020 attttctcta aaaaatactc ctcgtgaaat gattgttcat tagtcttaag tatactccta 1080 ggtcaaaaaa aattgccaag tgaggtctta ataaaagaaa gggagaaagg cgtttgcact 1140 gcatgcagca actgcagcag tagtggtagt taattaatta attaattaat aatggcgatc 1200 gacagctcag ctcggaagac gatgcccacc cctctctccg aggccgaggc cgcacgaatc 1260 ttgaagcgcc tagccgttcg ccgcgcgtgc cgcgggcggc ggaggcggtg gccacgtaag 1320 cgcccgcccc acgcatgcag ccgtcggatt ccgattcctt ttgccgcttt tcccgtctgc 1380 ccccctcgtt ctccgtcgcc tttaaaccta gagtaatcaa tcaattgatt tattggtaat 1440 tcacgattaa ttaattgttt tctttctcca tcaaccccaa ttaggttata aattatgctt 1500 atatacccta atgtataaag tattctttta ttctagtact ttatactagc actattactc 1560 catactattg tactccctcc attccaaatt gactacatat agtttttttt aaggttattc 1620 ctaaatgatc tacatatttg tgcttattta ttaagtctat tcgttacttg tgcattggag 1680 taaatagaca ttgatgcatg catccatgca cacaagtatt tataacacac atgcaatatc 1740 ttgatttgct attggctagg aaatagtggg gatggtgttt gagtttgttt ctagaataaa 1800 tatagtatga gagagttatt agcttttctt ggtcttggtg tacctatgaa atatgtagat 1860 caatttgaaa tggagggagt actatagtag atgtagatcc actggattat gatttgtgag 1920 gaagattatg acggtgtgta tagtatagta ttggtgttag ctgaatgcac cctttacaaa 1980 atggcatatt gtttgtttcg gtcttagata catgtcattt taattatcaa tgtcatctgc 2040 caaatacaat tcttttcatg tgttttattt tactataatc acactacagg catatttgtg 2100 acaaaaaaaa gaatgggaat tatgtggtag tatgtatcac tttatcatgt atatgtatgt 2160 cgaggagctt agctaagcga agactacatt taatattgat tagcggatct aattacacac 2220 aaacggttgg ttcaactaac cctacaactt gttacgtgcg atcctataag cctttgttta 2280 tattgtacgt aaactataca aaaatgacct aaaaataact caattgaaat atgaatcaac 2340 gcgacaaaaa tctacaagat gtttggtaca gttgaactcc agctgtgtga cacttacaac 2400 tacaatacca agagatcgaa atatacaact ttggttgtac agcgctaccg gctataccca 2460 tagcccaacc acatgtcaga cctcatccaa ccctagatcc accgcaactc gatattatca 2520 cacttctcaa cctactctta ttatcatact acaagcttgt tagcttggtt acaatgtatg 2580 ctaatttgaa gctctgaact atgaacacga actataccac tattactatt ttgcccaaca 2640 aaccccaagt agagccgcct aaccaacaac caacctcgtt gaccgctaga gccgctaatc 2700 actctggagt cgggacccat caccgaatac gcgagaaccc gaggaccaaa ccgcaaaagc 2760 ctcacctcac cctaggcttt gatatgcgtg cggacgccta aggctaggag ttaacctcgc 2820 gtagagag agagagagag agagagataa ctagcagagc actcgcggag gcgaggagag 2880 agagagagag agagagaggt tggaagagga ggggagagtc cacgtgagtg agaggaggag 2940 gaggaggagg agatccctct cccgcgcgca tccgcccctc tcgccgatcc ccaattcgcc 3000                                                                         3000 <210> 4 <211> 5619 <212> DNA <213> Oryza sativa <220> <221> gene &Lt; 222 > (1) .. (5619) <223> LOC_Os02g05840 <400> 4 gagagataac tagcagagca ctcgcggagg cgaggagaga gagagagaga gagagaggtt 60 ggaagaggag gggagagtcc acgtgagtga gaggaggagg aggaggagga gatccctctc 120 ccgcgcgcat ccgcccctct cgccgatccc caattcgcca tggatccacc ctacgcaggt 180 cggtcagcgc cgcccccgca tccgcggttt tcgccttgtt tttttttttt gttgttgttg 240 ttgcgtgtgt gtggtggtgg tgatggctca ggtctcgcgg cggcgtcggg gggagctcga 300 tctgtttgtt ccggcgacga atccgctgtt ccggcggtgt aattgcgggc ttggtggtgg 360 aggaggagga atttgtgggc gtggatttta gggtgggagg cggttttttt ttttttgggg 420 ggggggtggt gtagtgttcg tcgagattta gactttttgt gtggatggat ggatgggtta 480 gattatgcgc gcgtggatta gattgcggcg gtagagggtt tgtactgttc attgattggt 540 ttgttgggtt gatttggagc tgtagctgat gcgtggtaga ggggtggagg tggaagcgcg 600 ttggggaggg tggatggcga tgccgctggt gtcagattcg ccgctgcatg cggtgatggg 660 tactcacatt gcgccccctc tctaccgggg gaggtggcag cggagtggtt cctgcggctt 720 tacgtcgttt tcgtgggatt tacgtccgcg ggcgttggta gcagcagcct atcctctttt 780 gggacgggaa aataatatgg cgttaggctg tttgaggcct cctgttcgtg cgttgggtgt 840 tgcattggtg ctgatttggt gggatcacgg ctgccgaagg cgggtttgtt tgctgttagt 900 tgatggtaag gattggctga tacgtcgatt atatcttcta gaggagctgc gtctacttgc 960 atgtcaaggt taccacgttt aggggttata tgattgatga tagtttcttt gtttagtggt 1020 ccaaaattag aaagactcat ttctttttag ttttctcgtt tcccttagac agtctgtaga 1080 gagttgactt tctgcggctc gtgccagctt cgatagctct agttaacatt gcgccttgtg 1140 gtatcacaac tgtgctttgt agttcacatt tagtcctata caatatggta catgttgatg 1200 agcctttaat tcttagatta ttgaccgaat catctttttt ttccgaggtt aagacttatt 1260 ctgttgcaat gtgctataat tataatttga cttctactcc agtaagacag tgtgctactc 1320 acattaattt gattcagcat gtcgagaatt taatttcctt tggaacgtca ttcattgagt 1380 ttttccttgc acgctaagtt actcagtacc gtctcatagt tgttctgaag aagagaaaga 1440 atgatgcctt caaaatatgt attcttctac tggatctaat gaactttaaa cctttccatt 1500 ctggactgta aatatgaatg aacgtctaaa tttccttcgt tgtttgcttc ttgtccgact 1560 ggattttgta actatttctg actttatcaa attgataata atagtttcca gtgttctctt 1620 gtttaggagt acctattgat cctgctaaat gccgattgat gagtgtggat gaaaagcggg 1680 aacttgtccg tgaattatcg aagcggccag aaagtgctcc tgacaaactg cagtcttgga 1740 gtcgccgtga aattgtagag attctttgtg ctgatttagg aagggaaagg aagtacactg 1800 gattatcgaa gcagagaatg ttggaatatc tcttcagagt tgtgactggc aaatcatctg 1860 gtggtggcgt tgtggagcat gtgcaagaga aggagcctac ccctgaaccc aacacagcca 1920 accatcagtc ccctgcgaaa cggcagcgaa agagtgacaa cccatcacga ctaccaattg 1980 ttgcaagcag tccaactaca gaaataccca ggccagcaag taatgctcgc ttctgccaca 2040 atttagcttg cagagcgact cttaatccag aagataaatt ttgcagacgc tgttcatgct 2100 gtatttgttt caagtacgat gacaataagg atcctagcct ctggttattc tgtagttcag 2160 atcaaccctt gcagaaagat tcttgtgtat tttcgtgcca tcttgaatgt gctcttaagg 2220 atggaagaac tggcatcatg cagagtgggc agtgcaagaa acttgatggt ggttattact 2280 gcactcgctg tcggaaacag aatgatctgc ttgggtattc tccctaaaca actttattat 2340 ttgcacagtt ttttttcctg ttctgtaact atacgaaatt atgttttaat tattcctatg 2400 cctccagttt ttactgcctt aagtaatgtt accattattt attgcaacga taactactag 2460 aaatatgtta gtttgacttt tgtgagagtt agctttgata atttgcatgc attcttgctt 2520 tccagccact ttgattacct tactgattct gtttttagga ttcttcggtg aagatccttg 2580 tagtgtcatc atcattggtc caatcatatg atgcatattt ctcaattgtg tcttaagaag 2640 tgggttggct gcttttgccc ttcagcttca tgctttgagc ttgtggtctg tgtgcttact 2700 tctgcattaa gctgaattag tgttggattc ttccttctcc atctggagta aatttgtttg 2760 ctgggataac cctattttca taggttgcga gctgcgaaaa caggctttcg actgcatttt 2820 ggtgtctttg tgttgcattc catagggttc ttgtagttca agtttcaggg tatgaaccaa 2880 gttactaagc cagtaaaact tggtgaggca ctgaggccag gtggattcaa tattttcaag 2940 gatgaataga tcatggagaa atgcttagac tagtatatgc aaggagtctc taatttgtct 3000 gtgcatgttt ataagccatt tgagtaacct gggaaatagg attcattttg aagttttgaa 3060 ctacaagaca agtgattaat tgtctttctc aaaagaatct gcttggctat tagtatcttg 3120 tagtagtaag ttttgaacca gttatatgtt atttagagtt caagattctt tggccagtcc 3180 agcacaatgc acttaagtaa ttcaatcaag tttactggcc cattttccaa aactatgctc 3240 ctttcaagag tttggatatt gaaacagtaa attagcttgc cgagcaatac ttgactgttt 3300 aatgccttgt tcaggtcctg gaagaaacaa ctggtgatag ctaaagatgc tcgccggttg 3360 gatgtattgt gtcatcggat ttttttgagt cataagattc ttgtctccac ggagaagtac 3420 ttggttttgc atgaaattgt tgacacagcg atgaagaaac tggaggctga ggttggtcct 3480 atatctggag ttgcaaatat gggtcgtgga attgtgagcc ggcttgctgt tggtgctgaa 3540 gttcagaaac tttgtgctcg agcaatagaa accatggagt ctctgttttg tggatctcct 3600 tctaacttgc aatttcaacg taagtttcca cgtgcttcac gttagaccac atacatacaa 3660 atagtgttga attctcatgc atatgtgcat tttcaggttc acggatgata ccatcaaact 3720 tcgtaaagtt tgaagctata acccaaacat ctgtcactgt agttttggat ttgggtccta 3780 tacttgctca agatgtaaca tgctttaatg tatggcacag agtggcagcc acaggctcgt 3840 tctcatcaag tccaactggc atcatacttg caccattaaa aacgttagtg gtcactcaac 3900 ttgtgccagc tacaagctat atattcaagg tagttgcctt cagtaactac aaggagtttg 3960 gatcgtggga agccaaaatg aagacaagct gtcagaagga agttgatctg aagggtttga 4020 tgccaggtgg gtctgggcta gaccaaaaca atgggagccc aaaggcaaac agtggtggtc 4080 agtctgatcc ttcttcagaa ggtgtggact caaataataa cactgcggtg tatgctgatc 4140 tcaataaatc accagaaagt gattttgaat attgtgaaaa tcctgagata cttgattcag 4200 acaaagcaag tcatcacccc aatgaaccta caaacaactc acagagtatg ccgatggtcg 4260 tagctagggt tacggaggta tctggattgg aggaagctcc tggactctca gcatcagctt 4320 tggacgagga gcccaattca gcagttcaaa cacaattact tagagaatcc tcaaattcaa 4380 tggagcagaa ccagagaagc gaagttcctg gatcakhga tgcatcaaat gctcctgctg 4440 gaaatgaggt ggtgattgtt ccacctcgat attctggctc tattccacca actgcaccta 4500 gatatatgga aaatggtaag gatatcagtg ggaggagctt gaaagcaaaa cctggtgata 4560 acatccttca aaatggctct tccaagcctg aaagggaacc agggaattct tcaaataaaa 4620 gaacatcagg taaatgtgag gaaatcggcc acaaggatgg atgcccagaa gcatcttatg 4680 agtactgtgt taaggtggtc aggtggctgg aatgtgaggg ttacattgag accaacttca 4740 gagtgaagtt tctgacttgg tatagccttc gtgctacccc tcatgacagg aagatagtca 4800 gcgtctacgt aaacactctt attgatgatc ctgttagcct ttctggccag cttgctgaca 4860 ctttctctga ggccatctac agcaaaaggc caccttctgt tcgctccggt ttctgcatgg 4920 aactttggca ttaaagagag gggagctcgt accgttctct ttgtagggga gcatattttg 4980 gtcccattct ttcgtagcag atcgttgcat tgattcttca ttggaaattt cattgaagcg 5040 gttaacttat acaatctgtt cattgctcta ctttatttgc tatgacagta ggtttaattt 5100 atccagcaac ttatttctgt tttttcactt aggagtccat agtttacagt tctgttatcc 5160 gtaccgccat gttggtacag agcatcaaga gctgtgctct tttggtcagc tttaggaaca 5220 ttctgaaatt gccacaagtg tgtgggtacc tgaagctctg ttgctagctg ctgtgagagg 5280 gagggatgat tctcaggatg taccgggaaa cattgggatg aagcaggctt cgtgtcgaca 5340 ctgacgaatc cgagctaacc atgccttgtc aggataaatg atgatgtggt gctgtatctg 5400 gt; tttttccttt tcttaagaca atagatgcaa aatgtctgca ctaatgttat gttcggttta 5520 gacccttgct tttactgtct cctactgcac tttcttgcct tctagttcta ccgtttgaga 5580 agttaaattt atgtgcacat gcaaaaaggt ttggacctt 5619 <210> 5 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> Forward primer of 3A-10852 <400> 5 gtgccatctt gaatgtgctc 20 <210> 6 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> Reverse primer of 3A-10852 <400> 6 ggcgagcatc tttagctatc 20 <210> 7 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> Forward primer of 1B-21530 <400> 7 gccgagcaat acttgactgt 20 <210> 8 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> Reverse primer of 1B-21530 <400> 8 tccttctgac agcttgtctt 20 <210> 9 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> Primer of NGUS1 <400> 9 aacgctgatc aattccacag 20 <210> 10 <211> 14 <212> DNA <213> Artificial Sequence <220> <223> Forward Primer of LOC_Os02g05840 <400> 10 atggatccac ccta 14 <210> 11 <211> 19 <212> DNA <213> Artificial Sequence <220> <223> Reverse Primer of LOC_Os02g05840 <400> 11 atgccaaagt tccatgcag 19 <210> 12 <211> 19 <212> DNA <213> Artificial Sequence <220> <223> Forward Primer of Ubiquitine-5 <400> 12 aaccagctga ggcccaaga 19 <210> 13 <211> 24 <212> DNA <213> Artificial Sequence <220> <223> Reverse Primer of Ubiquitine-5 <400> 13 acgattgatt taaccagtcc atga 24 <210> 14 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> Primer of LOC_Os02g05840-1 <400> 14 cgcttctgcc acaatttagc 20 <210> 15 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> Primer of LOC_Os02g05840-2 <400> 15 tgttggtgct gaagttcaga 20 <210> 16 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> Primer of LOC_Os02g05840-3 <400> 16 tggactcaaa taataacact 20 <210> 17 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> Primer of LOC_Os02g05840-4 <400> 17 ccattctttc gtagcagatc 20 <210> 18 <211> 23 <212> DNA <213> Artificial Sequence <220> <223> Primer of Tnos R <400> 18 ccatctcata aataacgtca tgc 23 <210> 19 <211> 24 <212> DNA <213> Artificial Sequence <220> <223> Primer of pUbi F3 <400> 19 cagctatatg tggatttttt tagc 24 <210> 20 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> Foward primer of LOC_Os02g05840 EN + EX <400> 20 actgtgttaa ggtggtcagg 20 <210> 21 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> Reverse primer of LOC_Os02g05840 EN <400> 21 atcaatgcaa cgatctgcta c 21 <210> 22 <211> 19 <212> DNA <213> Artificial Sequence <220> <223> Reverse primer of LOC_Os02g05840 EX <400> 22 atgccaaagt tccatgcag 19

Claims (11)

Gene for increasing the yield of monocotyledonous plants represented by SEQ ID NO: 1. The gene for increasing the yield of monocotyledonous plants according to claim 1, wherein the increase in yield is at least one selected from an increase in the number of ear, stem development, or shortening of flowering time in comparison with wild type. A recombinant vector comprising a gene of claim 1 or 2 and an operably linked promoter. The recombinant vector of claim 3, wherein the promoter is a promoter represented by SEQ ID NO: 3. 5. A transformed cell transformed with the recombinant vector according to claim 3. The transformed cell of claim 5, wherein the cell is a microorganism of the genus Agrobacterium. The transformed cell of claim 6, wherein the microorganism of the genus Agrobacterium is Agrobacterium tumefaciens LB4404. A transformed monocotyledonous plant transformed by the recombinant vector according to claim 3. The transgenic plant of claim 8, wherein the monocotyledonous plant is Oryza sativa. A method of transforming a monocotyledonous plant comprising the step of transforming the monocotyledonous plant with the recombinant vector of claim 3 to express the LOC_Os02g05840 gene in the plant. The method of claim 10, wherein the LOC_Os02g05840 gene is prepared, comprising the following steps:
Preparing a recombinant vector comprising a LOC_Os02g05840 gene;
Introducing the recombinant vector into rice; And
Selecting a rice transformant in which the LOC_Os02g05840 gene is expressed as the recombinant vector is introduced.

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