KR101707032B1 - OsMYB5P gene improving phosphate uptake efficiency in plant and uses thereof - Google Patents

Abstract

A gene coding rice-derived transcriptional regulatory protein, Oryza sativa MYB5P (OsMYB5P), improves phosphoric acid absorption efficiency in plants, thereby being used for developing plants with improved phosphoric acid absorption rates through the expression regulation of OsMYB5P genes. Also, plants with improved phosphoric acid absorption abilities can overcome phosphoric acid deficiency stress, thereby being capable of obtaining effects of reducing agriculture production costs and preventing environmental pollution.

Description

OsMYB5P gene and its use for enhancing the phosphoric acid uptake efficiency of plants (OsMYB5P gene improving phosphate uptake efficiency in plant and uses thereof)

The present invention relates to OsMYB5P gene and its use for promoting phosphoric acid absorption efficiency of plants.

Phosphoric acid is one of the essential nutrients needed by plants and plays an important role in all metabolism processes occurring in plant cells such as energy transfer, signal transduction, regulation of enzyme activity, biosynthesis of metabolites, photosynthesis and respiration. Since phosphoric acid is also an important component of phospholipids and nucleic acids, it is the nutrient element most needed by plants with nitrogen.

Although there are a lot of phosphoric acid in the earth's lithosphere, most of the phosphoric acid in the soil exists in a form that is combined with an organic matter that is not available to plants or a metal ion such as iron or aluminum. Therefore, the amount of available phosphoric acid It is present in small quantities. Therefore, phosphoric acid acts as a factor limiting crop productivity. However, most of the supplied phosphoric acid is bound to the organic matter or metal ion present in the soil, and is fixed in a form that the plant can not use. In particular, in the case of acidic soil, Phosphoric acid availability in the soil is even lower because the accumulated aluminum ions cause more active phosphate immobilization.

Thus, the fertilizer treated to increase the effective phosphoric acid concentration of the soil required for crop growth causes the increase in agricultural production costs. In addition, fertilizers that have not been absorbed due to excessive use of fertilizer are accumulated in the soil and acidify the soil, adversely affecting the growth of crops, causing groundwater pollution and eutrophication of surface water. The pollution of the agricultural environment not only leads to quantitative and qualitative deterioration in the production of agricultural products, but also pollutes the living environment and threatens the public health. This problem can be solved by reducing the amount of chemical fertilizer added. Currently, various organic fertilizers have been developed and used to replace chemical fertilizers, but many problems still remain. A fundamental solution to this problem is to reduce the use of fertilizer by improving the plant's ability to absorb phosphoric acid. Therefore, it is required to develop a crop that can grow efficiently even under the condition of phosphoric acid deficiency.

In the meantime, studies for improving the nutrient utilization efficiency of crops have been performed on rice, one of the most important food crops in the world, and many efforts have been made to obtain useful genes and to preoccupy them after the whole gene sequence of rice has been revealed . Studies on the inorganic nutrient uptake of plants have been carried out for a long time in the research of cultivated aspects and plant physiological aspects. Recently, based on the basic study on the molecular level of inorganic nutrient uptake, Studies have been actively conducted around the world to increase the growth and yield of crops and to breed crops capable of growing under extremely limited nutrient conditions.

Under the conditions of phosphorus deficiency, the plant promotes root growth and root hair formation to broaden the surface area, and induces the expression of genes involved in the biosynthesis and secretion of highly phosphorylated phosphoric acid carriers, dephosphorylated enzymes and organic acids, , Thereby improving the phosphate availability. The present invention analyzes the function of the rice-derived R2R3-MYB transcriptional regulatory gene ( OsMYB5P ) expressed in a phosphate-deficient condition and improves the phosphorus absorption efficiency of the plant using the gene to develop a plant capable of adapting to phosphorus deficiency conditions I want to.

Korean Patent No. 0640284 discloses a method for producing a high-affinity phosphate carrier gene, a promoter of the gene, and a plant transformed using the gene, which are always expressed in all tissues of rice regardless of changes in the phosphoric acid concentration Korean Patent No. 0770203 discloses a method of increasing the absorption of phosphoric acid by a plant using a gene of a tobacco-derived phosphoric acid transporter, and a transgenic plant in which the rate of absorption of phosphoric acid is increased by the same method. However, as in the present invention, the OsMYB5P gene which promotes the phosphoric acid absorption efficiency of a plant and its use has not been disclosed.

SUMMARY OF THE INVENTION The present invention has been made in view of the above-mentioned needs, and the present inventors have found that rice transcription control protein OsMYB5P ( Oryza sativa MYB5P), the present inventors have completed the present invention by confirming that the transgenic rice plant overexpressing the OsMYB5P gene is cultivated and the phosphoric acid content in the plant is increased compared to the wild type.

In order to solve the above problems, the present invention provides rice transcription control protein OsMYB5P ( Oryza sativa MYB5P).

The present invention also relates to the above transcription regulatory protein OsMYB5P ( Oryza sativa MYB5P).

The present invention also provides a recombinant vector comprising the gene.

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

In addition, the present invention provides a method for increasing the phosphoric acid uptake efficiency of a plant as compared to a wild type comprising overexpressing OsMYB5P gene by transforming a plant cell with a recombinant vector comprising a gene encoding OsMYB5P protein.

In addition, the present invention provides a method for producing a transgenic plant having increased phosphorus absorption efficiency as compared to a wild type using a recombinant vector containing a gene encoding OsMYB5P protein.

Also, the present invention provides a transgenic plant having enhanced phosphorus absorption efficiency produced by the above method and seeds thereof.

The present invention also provides a composition for promoting phosphoric acid absorption efficiency of a plant containing a gene encoding OsMYB5P protein derived from rice.

The rice-derived electron control protein OsMYB5P ( Oryza sativa MYB5P) promotes the phosphorus uptake efficiency of plants, and thus can be used for the development of plants with enhanced absorption of phosphoric acid through regulation of OsMYB5P gene expression in plants. In addition, plants with improved phosphate absorption capacity can overcome phosphorus deficiency stress, and therefore, it is expected that reduction of agricultural production cost and prevention of environmental pollution due to reduction of fertilizer input amount can be obtained.

1 compares the amino acid sequences of the OsMYB5P gene of the present invention with MYB2P- 1 and OsMYB4P known as rice transcriptional regulatory genes.
FIG. 2 shows the expression pattern ( Os ) of OsMYB5P gene (A) and the expression level of OsmYB5P gene ( Oryza ) in the conditions of nitrogen, phosphoric acid, potassium and iron (B) of the OsMYB5P gene according to the treatment time of phosphate deficiency in the stem and root of S. sativa . EtBr is the loading control.
FIG. 3 is a graph showing the activity of OsMYB5P Whether the overexpressed transformants and RNAi mutants were transfected with OsMYB5P gene and OsMYB5P Gene expression pattern. A and B are PCR analysis, and C and D are Northern blot results.
FIG. 4 is a photograph (A) of a wild-type plant, OsMYB5P overexpressing transformant and RNAi mutant, a stem length measurement result (B), and a main length measurement result (C) under phosphate sufficiency and deficiency conditions.
FIG. 5 is a photograph showing the main muscle photograph (A) of the wild type plant, the OsMYB5P overexpressing transformant and the RNAi mutant at the phosphate sufficiency and depletion conditions, the result of measuring the anisometrical density (B), the result of measuring the phosphate content of the stem and root (C) And the measurement result (D).
FIG. 6 shows photographs (A), (B), (C), (C), (D) and (D) the phosphorus in the stem and root of the wild type Arabidopsis thaliana and OsMYB5P overexpressing transformants under phosphate sufficient conditions (E).

In order to achieve the object of the present invention, the present invention provides rice transcriptional regulatory protein OsMYB5P ( Oryza sativa MYB5P).

The range of the OsMYB5P protein according to the present invention is rice ( Oryza comprising the amino acid sequence shown in the SEQ ID NO: 2 isolated from sativa) comprises the functional equivalent of a protein and the protein. Is at least 70% or more, preferably 80% or more, more preferably 90% or more, still more preferably 90% or more, more preferably 90% or more, Quot; refers to a protein having a homology of at least 95% with a physiological activity substantially equivalent to that of the protein represented by SEQ ID NO: 2. "Substantially homogenous physiological activity" means a phosphoric acid absorption efficiency enhancing activity. The invention also includes fragments, derivatives and analogues of the OsMYB5P protein. As used herein, the terms "fragments", "derivatives" and "analogs" refer to polypeptides that possess substantially the same biological function or activity as the OsMYB5P polypeptides of the present invention.

The present invention also relates to the above transcription regulatory protein OsMYB5P ( Oryza sativa MYB5P).

The gene of the present invention is characterized by enhancing the phosphoric acid uptake efficiency of plants, and includes genomic DNA, cDNA, and synthetic DNA encoding OsMYB5P protein. Preferably, the gene of the present invention may include the nucleotide sequence shown in SEQ ID NO: 1. In addition, homologues of the nucleotide sequences are included within the scope of the present invention. Specifically, the gene includes a nucleotide sequence having a sequence homology of 70% or more, more preferably 80% or more, more preferably 90% or more, and most preferably 95% or more, with the nucleotide sequence of SEQ ID NO: 1 can do. "% Of sequence homology to polynucleotides" is ascertained by comparing the comparison region with two optimally aligned sequences, and a portion of the polynucleotide sequence in the comparison region is the reference sequence for the optimal alignment of the two sequences (I. E., A gap) relative to the < / RTI >

The present invention also provides a recombinant vector comprising the OsMYB5P gene according to the present invention. In the recombinant vector of the present invention, the OsMYB5P The gene may preferably consist of the nucleotide sequence of SEQ ID NO: 1.

The term "recombinant" refers to a cell in which a cell replicates a heterologous nucleic acid, expresses the nucleic acid, or expresses a protein encoded by a peptide, heterologous peptide or heterologous nucleic acid. The recombinant cell can express a gene or a gene fragment that is not found in the natural form of the cell in one of the sense or antisense form. In addition, the recombinant cell can express a gene found in a cell in its natural state, but the gene has been modified and reintroduced intracellularly by an artificial means.

The term "vector" is used to refer to a DNA fragment (s), nucleic acid molecule, which is transferred into a cell. The vector replicates the DNA and can be independently regenerated in the host cell. The term "carrier" is often used interchangeably with "vector ". The term "expression 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. In principle, any plasmid and vector can be used if it can replicate and stabilize within the host. An important characteristic of the expression vector is that it has a replication origin, a promoter, a marker gene and a translation control element. Enhancers, ribosome binding sites, termination signals, polyadenylation signals, and promoters, which are available in eukaryotic cells, are well known in the art. The expression vector can be constructed by a method known in the art. Such methods include in vitro recombinant DNA technology, DNA synthesis techniques, and in vivo recombination techniques. The DNA sequence can be effectively linked to appropriate promoters in the expression vector to drive mRNA synthesis. The expression vector may also include a ribosome binding site and a transcription terminator as a translation initiation site.

A preferred example of a recombinant vector is a Ti-plasmid vector which is capable of transferring a so-called T-region to a plant cell when it is present in a suitable host such as Agrobacterium tumefaciens. Other types of Ti-plasmid vectors (see EP 0 116 718 B1) are currently used to transfer hybrid DNA sequences to plant cells or protoplasts in which new plants capable of properly inserting hybrid DNA into the plant's genome can be produced have. A particularly preferred form of the Ti-plasmid vector is a so-called binary vector as claimed in EP 0 120 516 B1 and U.S. Patent No. 4,940,838. Other suitable vectors that can be used to introduce the DNA according to the invention into the plant host include viral vectors such as those that can be derived from double-stranded plant viruses (e. G., CaMV) and single- For example, from non -complete plant virus vectors. The use of such vectors may be particularly advantageous when it is difficult to transform the plant host properly.

In the recombinant vector of the present invention, the promoter may be a promoter suitable for transformation, preferably a CaMV 35S promoter, an actin promoter, a ubiquitin promoter, a pEMU promoter, a MAS promoter or a histone promoter, preferably a CaMV 35S promoter But is not limited thereto. The term "promoter " refers to the region of DNA upstream from the structural gene and refers to a DNA molecule to which an RNA polymerase binds to initiate transcription. A "plant promoter" is a promoter capable of initiating transcription in plant cells. A "constitutive promoter" is a promoter that is active under most environmental conditions and developmental conditions or cell differentiation. Since the selection of the transformant can be carried out by various tissues at various stages, a constant promoter may be preferable in the present invention. Therefore, the constant promoter does not limit the selectivity.

In the recombinant vector of the present invention, the terminator can be a conventional terminator, such as nopaline synthase (NOS), rice α-amylase RAmy1 A terminator, phaseoline terminator, Agrobacterium tumefaciens Agrobacterium tumefaciens , Octopine gene terminator of Agrobacterium tumefaciens , and the like. Regarding the need for a terminator, it is generally known that the terminator region increases the certainty and efficiency of gene transcription in plant cells. Therefore, the use of a terminator is highly desirable in the context of the present invention.

The recombinant vector of the present invention may preferably comprise one or more selectable markers. The marker is typically a nucleic acid sequence having a property that can be selected by a chemical method, and includes all genes capable of distinguishing a transformed cell from a non-transformed cell. Examples include herbicide resistance genes such as glyphosate or phosphinothricin, antibiotic resistant genes such as kanamycin, G418, Bleomycin, hygromycin, chloramphenicol, However, the present invention is not limited thereto.

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

Host cells capable of continuously cloning and expressing the vector of the present invention in a stable manner can be any host cell known in the art. Examples of prokaryotic cells include E. coli JM109, E. coli BL21, E. coli Bacillus sp. Strains such as RR1, E. coli LE392, E. coli B, E. coli X 1776, E. coli W3110, Bacillus subtilis, and Bacillus thuringiensis, and Salmonella typhimurium, Serratia marcesensis And enterobacteria and strains such as Pseudomonas spp.

When the vector of the present invention is transformed into eukaryotic cells, yeast ( Saccharomyce cerevisiae ), insect cells, human cells (e.g., Chinese hamster ovary, W138, BHK, COS-7, 293, HepG2 , 3T3, RIN and MDCK cell lines) and plant cells. The host cell is preferably a plant cell.

The recombinant vector of the present invention can be produced by the CaCl ₂ method, Hanahan, D., J. Mol. Biol., 166: 557-580 (1983) And can be delivered into cells. When the host cell is a eukaryotic cell, the vector may be injected into the host cell by microinjection, calcium phosphate precipitation, electroporation, liposome-mediated transfection, DEAE-dextran treatment, and gene bombardment have.

In addition, the present invention provides a method for increasing the phosphoric acid uptake efficiency of a plant as compared to a wild type comprising overexpressing OsMYB5P gene by transforming a plant cell with a recombinant vector comprising a gene encoding OsMYB5P protein.

In one embodiment of the present invention, the OsMYB5P protein comprises the amino acid sequence of SEQ ID NO: 2, but is not limited thereto.

When transforming a vector of the present invention into a plant cell, the host cell may preferably be a Poaceae plant cell, and more preferably, it may be a rice cell, but is not limited thereto. The method of transforming the plant cell is as described above.

The present invention also relates to a method for transforming a plant cell with a recombinant vector comprising a gene encoding a wild-type rice transporter protein OsMYB5P; And

And regenerating the transformed plant from the transformed plant cell. The present invention also provides a method for producing a transgenic plant having increased phosphorus absorption efficiency as compared to the wild type.

In one embodiment of the present invention, the OsMYB5P protein comprises the amino acid sequence of SEQ ID NO: 2, but is not limited thereto.

The method of transforming plant cells is as described above. Regeneration of transgenic plants from the transformed plant cells can be carried out by any method known in the art.

Also, the present invention provides a transgenic plant having enhanced phosphorus absorption efficiency produced by the above method and seeds thereof.

In one embodiment of the present invention, the plants used in the present invention are selected from the group consisting of Arabidopsis, potato, eggplant, cigarette, pepper, tomato, burdock, ciliaceae, lettuce, bellflower, spinach, modern sweet potato, celery, carrot, Such as rice, barley, wheat, rye, corn, sugar cane, oats, onions, etc., such as Chinese cabbage, cabbage, gangbu, watermelon, melon, cucumber, amber, pak, strawberry, soybean, mung bean, May be monocotyledons, preferably monocotyledons, more preferably Poaceae plants, more preferably rice ( Oryza < RTI ID = 0.0 > sativa , but is not limited thereto.

The present invention also provides a composition for promoting phosphoric acid absorption efficiency of a plant containing a gene encoding OsMYB5P protein derived from rice.

The composition includes an effective component as a gene encoding the amino acid sequence of SEQ ID NO: 2 or a recombinant vector containing the gene. The gene can be transformed into a plant to increase the phosphoric acid absorption efficiency of the plant.

Hereinafter, the present invention will be described in more detail with reference to Examples. It is to be understood by those skilled in the art that these embodiments are merely illustrative of the present invention and that the scope of the present invention is not limited thereto.

Example  1. Rice-derived transcriptional control genes ( OsMYB5P )

In this example, the cultivated varieties of Oryza sativa cv. Dongjin ) was used. Genomic DNA was isolated from Dongjin-jin, and oligonucleotide primers (forward primer 5'-CACCATGGGGAGGGCGCCGTGCTG-3 '(SEQ ID NO: 3)) specific for R2R3-MYB transcriptional regulatory gene ( OsMYB5P ) derived from rice were selected based on the entire genome sequence of rice. Reverse primer 5'-TTAAATCTGGGGCAAATTCT-3 '(SEQ ID NO: 4)). After amplifying the coding region of the gene using the primer using the PCR method, the amplified product was cloned into the pEntr-vector and the nucleotide sequence of the gene was confirmed by sequencing. OsMYB5P , a transcriptional regulatory gene expressed in phosphate-deficient conditions, is a novel gene that has not been previously identified and has the nucleotide sequence of SEQ ID NO: 1 (777 bp) and encodes 258 amino acid sequences of SEQ ID NO: 2. In addition, the homology of the isolated OsMYB5P gene to the amino acids of the previously identified MYB2P- 1 and OsMYB4P rice transcriptional control genes was compared with that of the CLASTAL W program. As a result, there was about 70% amino acid sequence identity between these genes One).

Example  2. In the absence of phosphoric acid, OsMYB5P  Expression pattern of gene

In general, the expression of a gene having a similar function is specifically regulated by a tissue - specific or inducible factor. Therefore, the expression pattern of the isolated transcription regulatory gene was examined using an RNA blotting method. At this time, a primer having an OsMYB5p transcription regulatory gene-specific base sequence was used as a probe. By using distilled water for two weeks by using the hydroponic seed dongjinbyeo to affect the growth of the rice plant nitrogen _{(0.25mM (NH 4) 2 SO} 4, KNO 3), phosphoric acid (0.0125mM KH ₂ PO _4), potassium (0.01 mM KNO _3), and iron (Fe-EDTA 0.01mM) a component-deficient hogeul land solution (Hogland solution; 200mM Ca (NO 3) 2 · 4H 2 O, 2.5mM K 2 SO 4, 2mM MgSO 4 · 7H 2 O , 1.25 mM KH ₂ PO ₄ , 9 mM KCl, 0.25 mM MnSO ₄ , 0.25 mM H ₃ BO ₃ , 0.25 mM ZnSO ₄ , 0.25 mM CuSO ₄ , 0.25 mM Na ₂ MoO ₄ , 20 mM Fe-Sequestrate and 0.5 mM K ₂ SO ₄ , pH 5.5) for one week. In addition, phosphoric acid sufficient to confirm that the gene expression profiles in phosphate deficiency over time for the stem and root (1.25mM KH ₂ PO ₄₎ and phosphoric acid-deficient condition, respectively 7 in hogeul land solution was treated with (0.0125mM KH ₂ PO ₄₎ (Invitrogen, USA), and the total RNA was isolated. 20 μg of RNA was electrophoresed on 1.2% (w / v) denatured formaldehyde agarose gel to attach RNA to a nylon membrane (Amersham, UK). RNA-attached nylon membranes were prepared by incubating probes labeled with [ ³² P] dCTP in 20% w / v SDS, 20 x SSPE, 100 g / L PEG (8,000 mwt), 250 mg / L heparin and 10 mL / (Hering sperm) DNA (10 mg / ml) overnight at 65 ° C to confirm the expression pattern of the gene. RNA blots were performed to obtain OsMYB5P Expression of the gene showed that the expression of OsMYB5P gene was high only when phosphorus was deficient without expression in the case of deficiency of nitrogen, potassium or iron (Fig. 2A). In addition, OsMYB5P gene was expressed at the early stage of deficiency treatment in the stem and gradually decreased over time, and it was decreased from the root It was confirmed that it was not expressed at the early stage and was highly expressed at the 7th day (Fig. 2B).

Example  3. OsMYB5P  Overexpressing transformants of the genes and RNAi  Production of Mutants

In order to investigate the function of MYB transcriptional control genes isolated from rice in plants, transformants and RNAi mutants overexpressing OsMYB5P were constructed in Dongjinbil. The production process of rice transgenic plants is as follows. The coding region of the OsMYB5P gene was cloned into a destination vector (pH7WG2D, 1) whose expression was regulated by the CaMV 35S promoter, which is a strong allosteric promoter, and a fragment of the OsMYB5p gene was cloned into a specific oligonucleotide primer Was cloned into a destination vector (pB7GWIWG2 (II)) using the forward primer '5-CACC CGTCGTCCGGCCGGGAAGCC-3' (SEQ ID NO: 5) and the reverse primer '5- CCGAGGTGAAACTGTCCGTG-3' (SEQ ID NO: Was knocked out and introduced into Agrobacterium tumefaciens EHA105 as a triparental mating method. The callus was transformed with Agrobacterium tumefaciens EHA105 containing the construct. After confirming the introduction of OsMYB5P gene and the expression level of the gene in the transformed T0 plants by PCR analysis (FIGS. 3A and 3B) and RNA blotting method (FIG. 3C and 3D), overexpressing transformants (# 2, # 3 and # 9) and gene expression knock-out RNAi mutants (# 3, # 5 and # 8) were selected. In the case of Arabidopsis, the wild-type (Col-0) firefly was transformed in the same manner with the Agrobacterium tumefaciens GV3301 strain containing the OsNMYB5P gene.

Example  4. OsMYB5P  Growth characteristics of transgenic plants overexpressing genes

The harvested over-expressing transformants and the RNAi mutant seeds were hydroponically cultured for 2 weeks with the control group, and then treated with the hogleland solution for 7 days under the conditions of phosphate deficiency (0.0125 mM KH ₂ PO ₄ ) and phosphoric acid sufficiency (1.25 mM KH ₂ PO ₄ ) Were cultured and cultured, and the growth condition of the plants was compared with the wild type. The growth of overexpressed transgenic plants was found to be better than that of the control wild type plants in the conditions of phosphate sufficiency and deficiency. On the other hand, in the case of RNAi transformants, it was confirmed that the growth was worse than that of wild-type plants as the control group (Fig. 4A). As a result of quantitative investigation, it was confirmed that stem and main muscle growth was good in the overexpressed transformant as compared with the wild type plant, and stem and main muscle growth was reduced in the mutant (FIGS. 4B and 4C).

Phosphorylation-related genes have been shown to cause structural changes in roots and regulate phosphorus uptake. To confirm the function of the OsMYB5P gene, the development of the lateral root and root hair was observed using a main muscle treated for 7 days under the conditions of phosphate deficiency (0.0125 mM KH ₂ PO ₄ ) and phosphoric acid (1.25 mM KH ₂ PO ₄ ) 5A). In the case of the overexpressed transformants, the anterior muscle density and length were increased in comparison with wild type rice, which was the control, whereas the anterior muscle density and length were decreased in the RNAi mutants (FIGS. 5B and 5D). Phosphorus contents of stem and root were increased in both phosphate-rich (1.25 mM KH ₂ PO ₄ ) and deficient (0.0125 mM KH ₂ PO ₄ ) conditions compared with wild-type rice in control, and RNAi mutation The sieve was found to decrease in both phosphate sufficiency and deficiency conditions (Fig. 5C).

In the overexpressed Arabidopsis thaliana plants, the harvested seeds were placed on MS medium together with the control group, and after 5 days, the MS medium (0.0125 mM KH ₂ PO ₄ ) and phosphate-sufficient (1.25 mM KH ₂ PO ₄ ) 200 mM NH ₄ NO ₃ , 200 mM KNO ₃ , 3 mM MgSO ₄ .7H ₂ O, 1.25 mM KH ₂ PO ₄ , 6 mM CaCl ₂ .2H ₂ O, 10 mM Na ₂ -EDTA, 10 mM FeSO ₄ .7H ₂ O, 1 mM H ₃ BO ₃ , 1.5 mM MnSO ₄ .4H ₂ O, 0.3 mM ZnSO ₄ .7H ₂ O, 0.5 mM KI, 10 uM Na ₂ MoO ₄ .2H ₂ O, 55M Myo inositol, 3M Tiamin-HCl, 0.8M Nicotinic acid, 0.5 The cells were transferred to M Pyridoxine HCl, 0.01 M CuSO ₄ .5H ₂ O, 0.01 M CoCl ₂ .6H ₂ O, 2.3 mM MES, 1.2% Sucrose, 6% Agar; pH 5.7) and cultured for 7 days. (Fig. 6A). Quantitative investigations showed that the growth of the transgenic plants was better in the phosphate-rich and deficient conditions than in the wild-type plants in which the growth of the transformants was the control (Fig. 6B). Comparing the development of root hair in phosphate sufficient and deficient conditions, it was confirmed that the root development of the overexpressed transformant was increased in the absence of phosphoric acid (FIGS. 6C and 6D). The phosphoric acid content was found to be increased in both the phosphoric acid sufficiency and the deficiency condition in the transformant compared to the wild type plant (Fig. 6E). OsMYB5P , a rice transcriptional regulatory gene that increases phosphorus uptake, also plays a role in increasing phosphorus uptake in Arabidopsis thaliana. It also suggests that it is possible to improve the phosphoric acid uptake efficiency by overexpression of the OsMYB5P gene in all plants transcriptionally regulated by the R2R3-MYB transcriptional regulatory protein.

<110> Dong-A University Research Foundation For Industry-Academy Cooperation <120> OsMYB5P gene improving phosphate uptake efficiency in plant and          uses thereof <130> PN15350 <160> 6 <170> Kopatentin 2.0 <210> 1 <211> 777 <212> DNA <213> Oryza sativa <400> 1 atggggaggg cgccgtgctg cgagaagatg gggctgaaga gggggccgtg gacggcggag 60 gaggacagga tcctggtggc gcacatcgag cggcacgggc acagcaactg gcgcgcgctg 120 ccgaggcagg ccggccttct ccgctgcggc aagagctgcc gcctccggtg gatcaactac 180 ctccgccccg acatcaagcg cggcaacttc acccgcgagg aggaggacgc catcatccac 240 ctccacgacc ttctcggcaa ccgatggtcc gcgattgcag cgaggctgcc ggggaggacg 300 gacaacgaga tcaagaatgt gtggcacact cacctcaaga agcggctgga gccgaagccg 360 tcgtccggcc gggaagccgc cgcgcccaag cgaaaggcga ccaagaaggc tgcggcggtg 420 gcggtggcga tcgacgttcc gaccaccgtg ccggtgtcgc cggagcagtc gctctcgacc 480 acgacgacgt cggccgccac caccgaggag tactcgtact cgatggcctc ctccgcggat 540 cacaacacca cggacagttt cacctcggag gaggagttcc agatcgacga cagcttctgg 600 tcggagacgc tggcaatgac ggtggacagc accgactccg ggatggagat gagcggcggc 660 gatcctctcg gcgcgggcgg tgcctcgccg tcgtcgagca acgacgacga catggacgac 720 ttctggctca agctgttcat ccaggccggt ggcatgcaga atttgcccca gatttaa 777 <210> 2 <211> 258 <212> PRT <213> Oryza sativa <400> 2 Met Gly Arg Ala Pro Cys Cys Glu Lys Met Gly Leu Lys Arg Gly Pro   1 5 10 15 Trp Thr Ala Glu Glu Asp Arg Ile Leu Val Ala His Ile Glu Arg His              20 25 30 Gly His Ser Asn Trp Arg Ala Leu Pro Arg Gln Ala Gly Leu Leu Arg          35 40 45 Cys Gly Lys Ser Cys Arg Leu Arg Trp Ile Asn Tyr Leu Arg Pro Asp      50 55 60 Ile Lys Arg Gly Asn Phe Thr Arg Glu Glu Glu Asp Ala Ile Ile His  65 70 75 80 Leu His Asp Leu Leu Gly Asn Arg Trp Ser Ala Ile Ala Ala Arg Leu                  85 90 95 Pro Gly Arg Thr Asp Asn Glu Ile Lys Asn Val Trp His Thr His Leu             100 105 110 Lys Lys Arg Leu Glu Pro Lys Pro Ser Ser Gly Arg Glu Ala Ala Ala         115 120 125 Pro Lys Arg Lys Ala Thr Lys Lys Ala Ala Ala Val Ala Val Ala Ile     130 135 140 Asp Val Pro Thr Thr Val Ser Ser Pro Glu Gln Ser Leu Ser Thr 145 150 155 160 Thr Thr Ser Ala Ala Thr Thr Glu Glu Tyr Ser Tyr Ser Ala                 165 170 175 Ser Ser Ala Asp His Asn Thr Thr Asp Ser Phe Thr Ser Glu Glu Glu             180 185 190 Phe Gln Ile Asp Asp Ser Phe Trp Ser Glu Thr Leu Ala Met Thr Val         195 200 205 Asp Ser Thr Asp Ser Gly Met Glu Met Ser Gly Gly Asp Pro Leu Gly     210 215 220 Ala Gly Gly Ala Ser Ser Ser Ser Asn Asp Asp Asp Met Asp Asp 225 230 235 240 Phe Trp Leu Lys Leu Phe Ile Gln Ala Gly Gly Met Gln Asn Leu Pro                 245 250 255 Gln Ile         <210> 3 <211> 24 <212> DNA <213> Artificial Sequence <220> <223> primer <400> 3 caccatgggg agggcgccgt gctg 24 <210> 4 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> primer <400> 4 ttaaatctgg ggcaaattct 20 <210> 5 <211> 24 <212> DNA <213> Artificial Sequence <220> <223> primer <400> 5 cacccgtcgt ccggccggga agcc 24 <210> 6 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> primer <400> 6 ccgaggtgaa actgtccgtg 20

Claims (12)

delete delete delete delete delete Comprising transfecting a plant cell with a recombinant vector comprising a gene encoding a rice-derived transcription regulatory protein OsMYB5P ( Oryza sativa MYB5P) comprising the amino acid sequence of SEQ ID NO: 2 and overexpressing the OsMYB5P gene, A method for increasing phosphoric acid absorption efficiency. delete Transforming a plant cell with a recombinant vector comprising a gene encoding a rice-derived transcription regulatory protein OsMYB5P ( Oryza sativa MYB5P) comprising the amino acid sequence of SEQ ID NO: 2; And
And regenerating the transformed plant from the transformed plant cell, wherein the efficiency of absorption of phosphoric acid is increased compared to the wild type. 9. A transgenic plant having increased phosphorus absorption efficiency produced by the method of claim 8. 10. The transgenic plant according to claim 9, wherein the plant is a monocotyledonous plant or a dicotyledonous plant. 10. A transformed seed of a plant according to claim 9. A composition for increasing the phosphoric acid uptake efficiency of a plant comprising the gene encoding the rice-derived transcription control protein OsMYB5P ( Oryza sativa MYB5P), which comprises the amino acid sequence of SEQ ID NO: 2.

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