KR101546728B1 - OgPT gene improving phosphaste uptake efficiency in plant and uses thererof - Google Patents

Abstract

The present invention relates to an OgPT gene for promoting phosphoric acid absorption efficiency of a plant and a use thereof. Since a gene encoding a wild rice-derived phosphate transporter protein OgPT ( Oryza gandiglumis phosphate transporter) promotes phosphoric acid absorption efficiency of a plant, Can be used to develop plants that can enhance phosphorus uptake and overcome phosphorylation stress through modulation of OgPT gene expression.

Description

OgPT gene promoting phosphoric acid uptake efficiency of plants and its use {OgPT gene improving phosphaste uptake efficiency in plant and uses thererof}

The present invention relates to an OgPT gene for promoting phosphoric acid absorption efficiency of a plant and a use thereof, and more particularly, to an OgPT gene for OgPT gene ( Oryza gandiglumis phosphate transporter) derived from wild rice, a gene encoding the protein, And a step of overexpressing the OgPT gene by transforming the plant cell with a recombinant vector comprising the OgPT gene, an antibody against the OgPT protein, and a recombinant vector containing the recombinant vector, the host cell transformed with the recombinant vector, the antibody against the OgPT protein, , A method for producing a transgenic plant in which the efficiency of absorption of phosphoric acid is enhanced as compared with a wild type using a recombinant vector containing the OgPT gene, a method for producing transgenic plants having increased phosphorus absorption efficiency and a recombinant containing the gene OgPT of its seeds, stems and wild rice It relates to a composition for acid absorption efficiency increase of the plant containing an emitter.

Due to the rapid increase in population, food shortage is becoming a big problem all over the world, and it is required to cultivate multi-sex food crops to overcome this problem. In order to cultivate multiple food crops, it is necessary to supply sufficient nutrients to the crops. One of the main ingredients of fertilizer is phosphoric acid. 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. It is also an important component of phospholipids and nucleic acids, and is the nutrient element most needed by plants along with nitrogen.

Although there are a lot of phosphoric acid in the soil, most of the phosphoric acid in the soil exists in a form that is combined with an organic matter or a metal ion such as iron or aluminum which is not available to plants. Therefore, the amount of effective phosphoric acid available in actual plants is very small It is present in quantity. In addition, even when phosphoric acid is supplied as fertilizer, it is bonded to an organic matter or metal ion present in the soil and immobilized in a form that plants can not use. Particularly, in acidic soil, active immobilization occurs due to excessive accumulation of aluminum ions. The effectiveness of the phosphoric acid is further lowered, which causes an excessive introduction of fertilizer.

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.

Recently, wild rice ( Oryza gandiglumis ) has been attracting attention as a research crop for securing various kinds of useful genes that do not exist in existing cultivated rice. Various useful genes possessed by wild rice have been reported, but no transgenic gene has been reported to date in the case of phosphate deficiency. Accordingly, the present invention aims to analyze a transgenic transgenic gene derived from wild rice which is expressed under the conditions of phosphoric acid deficiency and enhance the utilization efficiency of phosphoric acid, and to develop a plant which can adapt to the phosphoric acid deficiency condition using the gene.

On the other hand, Korean Patent No. 0654958 discloses a high-affinity phosphate carrier gene (OsPT8) and a phosphate-deficient stress-resistant plant transformed with the gene. In Korean Patent Publication No. 2008-0074887, Plants and their use ". However, as in the present invention, the OgPT gene which promotes the phosphoric acid absorption efficiency of plants and its use has not been disclosed.

The present invention is derived by the request as described above, the inventors of the wild rice-derived phosphoric acid transporter protein OgPT (Oryza gandiglumis phosphate transporter) was cultured under a phosphate-deficient condition, the present inventors completed the present invention by confirming that the phosphoric acid content in the plant was increased compared to the wild type.

In order to solve the above problems, the present invention provides a wild rice-derived phosphate transporter protein OgPT ( Oryza gandiglumis phosphate transporter).

The present invention also provides a gene encoding the transporter protein OgPT.

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

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

The present invention also provides an antibody against the OgPT protein.

The present invention also provides a method for increasing the phosphoric acid uptake efficiency of a plant as compared to a wild type, comprising the step of overexpressing an OgPT gene by transforming a plant cell with a recombinant vector comprising a gene encoding an OgPT 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 an OgPT 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 recombinant vector comprising a gene encoding a wild-type rice-derived OgPT protein.

Since the gene encoding the wild rice-derived transgenic phosphate protein OgPT ( Oryza gandiglumis phosphate transporter) of the present invention promotes the phosphorus absorption efficiency of the plant, it is possible to develop a plant in which the absorption rate of phosphoric acid is increased by controlling the expression of the OgPT gene in plants Can be used. 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 shows the nucleotide sequence of the OgPT gene which is a transporter gene derived from wild rice ( Oryza gandiglumis ) whose expression level increases under phosphoric acid-deficient conditions.
Figure 2 shows the amino acid sequence deduced from the cDNA of the OgPT gene of wild rice ( Oryza gandiglumis ) whose expression level increases under phosphate-deficient conditions.
FIG. 3 shows the results of RACE (rapid amplification of cDNA end) -PCR results (A) and OgPT gene transcription ( OgPT ) (B) obtained by comparing the homology.
FIG. 4 shows the results of investigation of the expression pattern of OgPT gene of wild rice in the phosphate sufficiency and deficiency conditions by the parts (leaf, stem and root). The expression level of OgPT gene was increased in leaves, stems and roots in the absence of phosphate.
FIG. 5 shows morphological characteristics (A) and northern blot results (B) of the rice plants transformed with the OgPT gene. Transgenic plants were selected by comparing morphological characteristics and gene expression patterns of overexpressed OgPT gene.
FIG. 6 shows the results of confirming the growth characteristics of wild-type plants and OgPT- overexpressing transformants under phosphate-sufficiency conditions and deficient conditions. (A) Photographs of wild type and OgPT overexpressing transformants. (B) Results of phosphoric acid content measurement. Plant length and root length of wild type and OgPT overexpressing transgenic plants were not significantly different under phosphate - rich condition and deficient condition. However, phosphorus content in plant was significantly increased in transgenic plants under phosphate - depleted conditions.

In order to accomplish the object of the present invention, the present invention provides a wild rice-derived phosphate transporter protein OgPT ( Oryza gandiglumis phosphate transporter) which promotes the phosphoric acid absorption efficiency of plants.

The scope of the OgPT protein according to the present invention includes proteins having the amino acid sequence shown in SEQ ID NO: 2 isolated from rice ( Oryza gandiglumis ) and functional equivalents of the proteins. 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 OgPT protein. As used herein, the terms "fragments", "derivatives" and "analogs" refer to polypeptides having substantially the same biological function or activity as the OgPT polypeptides of the invention.

The present invention also provides a gene encoding the transporter protein OgPT. The gene of the present invention is characterized by enhancing the phosphoric acid uptake efficiency of a plant, and includes all the genomic DNA, cDNA, and synthetic DNA encoding the OgPT 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 OgPT gene according to the present invention. In the recombinant vector of the present invention, the OgPT 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.

Also provided is a host cell transformed with the recombinant vector of the present invention. 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.

The present invention also provides an antibody against the transporter protein OgPT.

The term "antibody" refers to a monoclonal antibody, a multispecific antibody, a human antibody, a humanized antibody, a camelised antibody, a chimeric antibody, a single chain Fvs (scFv) Fab fragments, F (ab) fragments, disulfide-linked Fvs (sdFv), and antiidiotypic (anti-Id) antibodies, and any of the epitope binding fragments. In particular, the antibody comprises an immunoglobulin molecule and an immunologically active immunoglobulin molecule fragment, i.e., a molecule containing an antigen binding site. The immunoglobulin molecule may be of any kind (e.g., IgG, IgE, IgM, IgD, IgA, and IgY), classes (eg, IgG1, IgG2, IgG3, IgG4, IgA1, and IgA2)

The antibody of the present invention can be prepared by cloning the gene of the present invention into an expression vector according to a conventional method to obtain an OgPT protein and then obtaining the protein from the obtained protein by a conventional method. Also included are partial peptides that can be made from the protein, and the partial peptides of the invention include at least 7 amino acids, preferably 9 amino acids, more preferably 12 or more amino acids. The form of the antibody of the present invention is not particularly limited, and a polyclonal antibody, a monoclonal antibody or a part thereof having antigen binding ability is included in the antibody of the present invention and includes all immunoglobulin antibodies. Furthermore, the antibodies of the present invention include special antibodies such as humanized antibodies.

The present invention also relates to a method for increasing the phosphorus uptake efficiency of a plant as compared to a wild type comprising overexpressing a OgPT gene by transforming a plant cell with a recombinant vector containing a gene encoding a transgenic phosphate transporter protein OgPT from wild rice ≪ / RTI >

In one embodiment of the present invention, the OgPT protein may comprise 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 provides a method for producing a plant cell, which comprises transforming a plant cell with a recombinant vector comprising a gene encoding wild-type rice-derived transporter protein OgPT; 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 OgPT protein may comprise 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.

In addition, the present invention provides a transgenic plant having increased 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, It may be a monocot plant, preferably a monocot plant, more preferably a Poaceae plant, more preferably rice ( Oryza sativa ) or wild rice ( Oryza gandiglumis ) It is not.

The present invention also provides a composition for increasing the phosphoric acid uptake efficiency of a plant comprising a recombinant vector comprising a gene encoding the wild-type rice transporter protein OgPT comprising the amino acid sequence of SEQ ID NO: 2. The composition includes a recombinant vector containing the gene encoding the amino acid sequence of SEQ ID NO: 2 as an active ingredient, and can increase the phosphoric acid absorption efficiency of the plant by transforming the gene into a plant.

Hereinafter, the present invention will be described in detail with reference to examples. However, the following examples are illustrative of the present invention, and the present invention is not limited to the following examples.

Example 1: Transgenic rice transgenic gene ( OgPT )

The present invention was carried out using wild rice ( Oryza gandiglumis ) distributed in South America. RNA was isolated from leaves of wild rice and specific oligonucleotide primers (forward primer F1: 5'-TTCCTCATCGATGTCGTCGGC-3 ') were prepared based on the conserved base sequence of phosphate transporter genes ( OsPT ) (SEQ ID NO: 3), F2: 5'-CTGCTCGGATTCTTCATGATG-3 '(SEQ ID NO: 4) and reverse primer R1: 5'-GGTCGACCTCGTACGCCT- 6)) and RACE (rapid amplification of cDNA end) -PCR was performed. The OgPT gene, which is a transgenic transgenic gene derived from wild rice, was obtained by RACE-PCR using SMART ™ RACE cDNA Amplification Kit (Clontech) (FIG. 3A), and the amplified product was cloned into pEntr-vector to confirm the nucleotide sequence. The phosphate transporter gene, OgPT, which is expressed in phosphate-deficient conditions, is a new gene that has not been identified yet. It has a total nucleotide sequence of 1623 bp and codes for 540 amino acids. The cDNA nucleotide sequence and the amino acid sequence of the OgPT gene are shown in detail in FIGS. 1 and 2, respectively. Next, the homology between the OgPT gene isolated in the present invention and the previously identified rice transporter genes from rice was compared. As a result, these genes were found to have about 70% homology, and the phylogenetic tree construction result is shown in FIG. 3B.

Example 2. In the presence of phosphoric acid OgPT  Expression pattern of gene

The OgPT gene isolated in the present invention is expected to be strongly expressed in the absence of phosphoric acid, like other transporter genes, and the gene expression pattern during phosphorylation is examined by northern blot. A primer having a gene-specific base sequence (forward direction: 5'-TACTGGCGGATGAAGATGC-3 '(SEQ ID NO: 7) and reverse direction: 5'-CGATGAGGAAGACGGTGAAC-3' (SEQ ID NO: 8)) was used as a probe.

Wild rice 1 after hydroponics in weekly distilled water and phosphate deficiency conditions (0.02 mM KH ₂ PO ₄₎ or hogeul land solution of phosphoric acid and sufficient conditions _{(2.0 mM KH 2 PO 4)} (Hogland solution) [Ca (NO 3) 2 _{· 4H 2 O, K 2 SO} 4, MgSO 4 · 7H 2 O, KH 2 PO 4, KCl, MnSO 4, H 3 BO 3, ZnSO 4, CuSO 4, Na 2 MoO 4, Fe-Sequestrate, K 2 SO ₄ ; pH 5.5] at 28 [deg.] C, 16 hours light and 8 hours dark cycle for 7 days.

The wild-type RNA was isolated using Trizol reagent (Invitrogen), and 5 μg of the separated RNA was electrophoresed on a 1.2% (w / v) denaturing formaldehyde agarose gel and transferred to a nylon membrane (Amersham) RNA was attached. The nylon membrane RNA is attached to ^[32 P] 20% with a probe labeled with dCTP (w / v) SDS, 20 × SSPE, 100 g / L PEG (8,000 mwt), 250 mg / L heparin, and 10 ml / L Hybridization was carried out overnight at 65 ° C with a solution containing Herring sperm DNA (10 mg / ml).

OgPT gene expression patterns of leaves, stems, and roots of rice cultivated for 7 days under phosphate-depletion and phosphorus-containing conditions were slightly different from each other, but it was confirmed that OgPT gene was strongly expressed at phosphate deficiency 4).

Example 3. OgPT  Generation of overexpressed transgenic plants

To investigate the function of the OgPT transporter gene isolated from wild rice in the plant, the OgPT gene was transformed into Donggin - Over-expressing transformants. The production process of rice transgenic plants is as follows. A coding region of the OgPT gene was cloned into a destination vector (pH7WG2D, 1) whose expression was regulated by the CaMV 35S promoter, which is a strong allosteric promoter. The cloned construct was introduced into Agrobacterium tumefaciens EHA105 by a triparental mating method and transformed with Agrobacterium tumefaciens EHA105. The introduction of the OgPT gene into the transformed T0 plant was confirmed by PCR (data not shown). The morphological characteristics of wild type rice and OgPT overexpressing transformants were compared, showing no growth or morphological differences of the plants (FIG. 5A). The expression level of the OgPT gene was measured by Northern blot (FIG. 5B), and a transformant having a high expression level was selected and used for further experiments.

Example 4. OgPT  Growth Characteristics of Transgenic Transgenic Plants

Seed and wild type seeds (control) harvested from transgenic plants transfected with the OgPT gene were overgrown and cultivated in distilled water for 2 weeks. The cells were grown under phosphate-free conditions (0.02 mM KH ₂ PO ₄ ) KH hogeul land solution of _{2 PO 4) [Ca (NO} 3) 2 · 4H 2 O, K 2 SO 4, MgSO 4 · 7H 2 O, KH 2 PO 4, KCl, MnSO 4, H 3 BO 3, ZnSO 4 _{, CuSO 4, Na 2 MoO 4} , Fe-Sequestrate, K 2 SO 4; pH 5.5] for 3 weeks, and the growth state and phosphate content of the transformants were compared with the wild type. The analysis method of phosphoric acid content is as follows. The plant tissue was pulverized using liquid nitrogen and 10 μl of extraction buffer (100 mM NaCl, 10 mM Tris (pH 8.0), 1 mM EDTA, 0.1% mercaptoethanol) was added per 1 mg of the pulverized sample, 1% glacial acetic acid was added and reacted at 42 ° C for 30 minutes. Then, the supernatant was separated by centrifugation at 13,000 rpm for 5 minutes, and 60 μl of the separated sample was added to 140 μl of a mixed solution [0.35% (NH ₄ ) ₆ Mo ₇ O ₂₄ .4H ₂ O, 0.86 NH ₂ SO ₄ , 1.43 % Ascorbic acid] and reacted at 45 캜 for 20 minutes. The phosphoric acid content of the culture broth was determined by measuring absorbance at A ₈₂₀ using KH ₂ PO ₄ solution as a phosphate standard solution.

As a result, the plant height and root length of the acid sufficient and deficient conditions, wild-type plants and OgPT over-expressing transformant showed no significant differences (FIG. 6A), plant in phosphate content of phosphate deficiency conditions are noted in OgPT overexpressing transformant (Fig. 6B).

<110> Dong-A University Research Foundation For Industry-Academy Cooperation <120> OgPT gene improving phosphaste uptake efficiency in plant and          uses thererof <130> PN13328 <160> 8 <170> Kopatentin 2.0 <210> 1 <211> 1623 <212> DNA <213> Oryza gandiglumis <400> 1 atggcccggc gggagcagca gcacctgcag gtgctgagcg cgctggacgc ggcgaagacg 60 cagtggtacc acttcacggc catcgtcgtc gccggcatgg gcttcttcac cgacgcctac 120 gacctcttct gcatctccct cgtcaccaag ctgctcggcc gcatctacta caccgacctc 180 gccaaggaga accccggcag cctgccgccc aacgtcgcgg cggcggtgag cggcgtagcg 240 ttctgtggga cgctggcggg gcagctcttc ttcgggtggc tcggcgacaa gctcgggcgg 300 aagagcgtct acgggatgac gctgctgctg atggtgatct gctccatcgc gtcggggctg 360 tcgttctcga acacgcccac cagcgtcatg gcgacgctct gcttcttccg cttctggctc 420 ggcttcggca tcggcggcga ctacccgctg tcggcgacca tcatgtcgga gtacgccaac 480 aagaagaccc gcggcgcgtt catcgccgcc gtgttcgcga tgcaggggtt cggcatcctc 540 gccggcggca tcgtcaccct catcatctca tccgcgttcc gcgcggggtt cccggcgcag 600 gcgtacgagg tcgaccgcgc gggctccacc gtccgccagg ccgactacgt gtggcgtatc 660 atcctcatgc tcggcgccgc gccggcgctg ctcacctact actggcggat gaagatgccg 720 gagacggcac gctacaccgc gctggtcgcc aagaacgcga agcaggcggc ggccgacatg 780 tccaaggttc tccaggtgga gatccaggag gagcaagaca agctggagga gatggtgacc 840 cggaacagca gcagcttcgg gctcttctcc cgccagttcg cacgccgaca tggcctccac 900 ctcgtcggca ccgccacaac gtggttcctc ctcgacatcg ccttctacag ccagaacctg 960 ttccagaagg acattttcac cagcatcaac tggatcccca aggccaagac catgtcggcg 1020 ctggaggagg tgttccgcat cgcgcgcgcg cagacgctca tcgcgctgtg cggcaccgtg 1080 ccgggctact ggttcaccgt cttcctcatc gatgtcgtcg gccgcttcgc catccagctg 1140 ctcggattct tcatgatgac cgtcttcatg ctcggcctcg ccgtgccgta ccaccactgg 1200 acaacgaagg ggaaccacat cggcttcgtc gtcatgtacg cgttcacctt cttcttcgcc 1260 aacttcgggc caaactccac caccttcatc gtgccggcgg agatcttccc ggcgaggctg 1320 cgttccacgt gccacggcat ctccgccgcc gccgggaagg ccggcgccat cgtcggatcg 1380 ttcgggttcc tctacgcggc gcaggacccg cacaagcccg acgccgggta caagcctggc 1440 atcggcgtgc gcaactcact gttcgtgctc gccggatgca acatgctcgg cttcatctgc 1500 accttcctcg tgccggagtc gaaggggaag tcgctggagg agatgtccgg cgaggtggag 1560 gaggacgacg aggtggccgg cggcggcggc ggtggcgccg ccgtgcggcc tcagacggcg 1620 tag 1623 <210> 2 <211> 540 <212> PRT <213> Oryza gandiglumis <400> 2 Met Ala Arg Arg Glu Gln Gln His Leu Gln Val Leu Ser Ala Leu Asp   1 5 10 15 Ala Ala Lys Thr Gln Trp Tyr His Phe Thr Ala Ile Val Val Ala Gly              20 25 30 Met Gly Phe Phe Thr Asp Ala Tyr Asp Leu Phe Cys Ile Ser Leu Val          35 40 45 Thr Lys Leu Leu Gly Arg Ile Tyr Tyr Thr Asp Leu Ala Lys Glu Asn      50 55 60 Pro Gly Ser Leu Pro Pro Asn Val Ala Ala Ala Val Ser Gly Val Ala  65 70 75 80 Phe Cys Gly Thr Leu Ala Gly Gln Leu Phe Phe Gly Trp Leu Gly Asp                  85 90 95 Lys Leu Gly Arg Lys Ser Val Tyr Gly Met Thr Leu Leu Leu Met Val             100 105 110 Ile Cys Ser Ile Ala Ser Gly Leu Ser Phe Ser Asn Thr Pro Thr Ser         115 120 125 Val Met Ala Thr Leu Cys Phe Phe Arg Phe Trp Leu Gly Phe Gly Ile     130 135 140 Gly Gly Asp Tyr Pro Leu Ser Ala Thr Ile Met Ser Glu Tyr Ala Asn 145 150 155 160 Lys Lys Thr Arg Gly Ala Phe Ile Ala Ala Val Phe Ala Met Gln Gly                 165 170 175 Phe Gly Ile Leu Ala Gly Gly Ile Val Thr Leu Ile Ile Ser Ser Ala             180 185 190 Phe Arg Ala Gly Phe Pro Ala Gln Ala Tyr Glu Val Asp Arg Ala Gly         195 200 205 Ser Thr Val Arg Gln Ala Asp Tyr Val Trp Arg Ile Ile Leu Met Leu     210 215 220 Gly Ala Ala Pro Ala Leu Leu Thr Tyr Tyr Trp Arg Met Lys Met Pro 225 230 235 240 Glu Thr Ala Arg Tyr Thr Ala Leu Val Ala Lys Asn Ala Lys Gln Ala                 245 250 255 Ala Ala Asp Met Ser Lys Val Leu Gln Val Glu Ile Gln Glu Glu Gln             260 265 270 Asp Lys Leu Glu Glu Met Val Thr Arg Asn Ser Ser Ser Phe Gly Leu         275 280 285 Phe Ser Arg Gln Phe Ala Arg Arg His Gly Leu His Leu Val Gly Thr     290 295 300 Ala Thr Thr Trp Phe Leu Leu Asp Ile Ala Phe Tyr Ser Gln Asn Leu 305 310 315 320 Phe Gln Lys Asp Ile Phe Thr Ser Ile Asn Trp Ile Pro Lys Ala Lys                 325 330 335 Thr Met Ser Ala Leu Glu Glu Val Phe Arg Ile Ala Arg Ala Gln Thr             340 345 350 Leu Ile Ala Leu Cys Gly Thr Val Pro Gly Tyr Trp Phe Thr Val Phe         355 360 365 Leu Ile Asp Val Val Gly Arg Phe Ala Ile Gln Leu Leu Gly Phe Phe     370 375 380 Met Met Thr Val Phe Met Leu Gly Leu Ala Val Pro Tyr His His Trp 385 390 395 400 Thr Thr Lys Gly Asn His Ile Gly Phe Val Val Met Tyr Ala Phe Thr                 405 410 415 Phe Phe Phe Ala Asn Phe Gly Pro Asn Ser Thr Thr Phe Ile Val Pro             420 425 430 Ala Glu Ile Phe Pro Ala Arg Leu Arg Ser Thr Cys His Gly Ile Ser         435 440 445 Ala Ala Aly Gly Aly Gly Aly Ile Val Gly Ser Phe Gly Phe Leu     450 455 460 Tyr Ala Gln Asp Pro His Lys Pro Asp Ala Gly Tyr Lys Pro Gly 465 470 475 480 Ile Gly Val Arg Asn Ser Leu Phe Val Leu Ala Gly Cys Asn Met Leu                 485 490 495 Gly Phe Ile Cys Thr Phe Leu Val Pro Glu Ser Lys Gly Lys Ser Leu             500 505 510 Glu Glu Met Ser Gly Glu Val Glu Glu Asp Asp Glu Val Ala Gly Gly         515 520 525 Gly Gly Gly Gly Ala Ala Val Arg Pro Gln Thr Ala     530 535 540 <210> 3 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> Primer <400> 3 ttcctcatcg atgtcgtcgg c 21 <210> 4 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> Primer <400> 4 ctgctcggat tcttcatgat g 21 <210> 5 <211> 18 <212> DNA <213> Artificial Sequence <220> <223> Primer <400> 5 ggtcgacctc gtacgcct 18 <210> 6 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> Primer <400> 6 gcggatgaga tgatgagggt g 21 <210> 7 <211> 19 <212> DNA <213> Artificial Sequence <220> <223> Primer <400> 7 tactggcgga tgaagatgc 19 <210> 8 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> Primer <400> 8 cgatgaggaa gacggtgaac 20

Claims (14)

delete delete delete delete delete delete Comprising the step of transfecting a plant cell with a recombinant vector comprising a gene encoding a wild-type rice transporter protein OgPT comprising the amino acid sequence of SEQ ID NO: 2 to overexpress the OgPT gene, Of phosphoric acid. delete Transforming a plant cell with a recombinant vector comprising a gene encoding a wild-type rice transporter protein OgPT consisting of the amino acid sequence of SEQ ID NO: 2; And
A method for producing a transgenic plant having increased phosphoric acid uptake efficiency under a phosphorous-depletion condition compared to a wild-type plant, comprising regenerating a transgenic plant from the transgenic plant cell. delete 9. A transgenic plant having increased phosphorus uptake efficiency under phosphate-depleted conditions produced by the method of claim 9. 12. The transgenic plant according to claim 11, wherein the plant is a Poaceae plant. 12. Seeds of transgenic plants according to claim 11. A composition for increasing the phosphoric acid uptake efficiency of a plant in a phosphate-depleted condition containing a recombinant vector comprising a gene encoding the wild-rice-derived phosphate transporter protein OgPT, the amino acid sequence of SEQ ID NO: 2.

Images