KR101648719B1 - OsIT gene from Oryza sativa for improving iron availability of plant and uses thereof - Google Patents

Abstract

The present invention relates to a rice-derived iron transporter gene OsIT ( Oryza < RTI ID = 0.0 > sativa iron transporter), a method of transforming a recombinant vector containing the OsIT protein coding gene into plant cells to increase the iron utilization ability of the plant as compared to the non-transformant, a method of transforming a recombinant vector comprising the OsIT protein coding gene into plant cells A method for producing a transformed plant having enhanced iron utilization ability of a plant compared to a non-transformed plant, a method for producing a transformed plant having enhanced iron utilization ability of a plant and a seed thereof And a gene encoding an OsIT protein consisting of the amino acid sequence of SEQ ID NO: 2 as an active ingredient.

Description

A rice-derived OsIT gene which improves the ability of plants to utilize iron and uses thereof.

The present invention relates to a rice-derived OsIT gene which improves the ability of a plant to utilize iron and, more particularly, to a rice-derived iron transporter gene OsIT ( Oryza sativa iron transporter), a method of transforming a recombinant vector containing the OsIT protein coding gene into plant cells to increase the iron utilization ability of the plant as compared to the non-transformant, a method of transforming a recombinant vector comprising the OsIT protein coding gene into plant cells A method for producing a transformed plant having enhanced iron utilization ability of a plant compared to a non-transformed plant, a method for producing a transformed plant having enhanced iron utilization ability of a plant and a seed thereof And a gene encoding an OsIT protein consisting of the amino acid sequence of SEQ ID NO: 2 as an active ingredient.

Iron is a coenzyme of about 140 enzymes that catalyze important biochemical reactions and plays a key role in plant growth and development. The physiological function depends on the property of binding complexes with ligands such as proteins, amino acids and nucleic acids in vivo, and the property of electron transfer by atomic changes (Fe ^{2 +} , Fe ^{3 +} ) will be. Iron is present in heme enzymes such as cytochromes, catalase, and peroxidase, in chloroplasts, and in ferredoxin, another iron protein form, which causes oxidation and reduction reactions while acting as an electron transport. In plants, most of the chloroplasts contain phytoferritin (phytoferritin), which provides the iron required for the development of pigments needed for photosynthesis.

Most of the earth's soil is said to be bad soil. Generally, poor soil qualitatively or quantitatively deficient in elements essential for plant growth inhibits plant growth or causes growth disruption due to soil containing heavy metals or the like. Calcareous soils cause iron deficiency disorders. Because iron in soil is often insoluble or sparingly soluble, it is a form that plants can not use, so even if the iron content is high in the soil, the actual amount of plant available is very small. Therefore, the development of crops that can grow efficiently even under iron deficiency conditions is required.

The mechanism of iron acquisition of higher plants is divided into two categories, strategy-I and strategy-II. Strategy-I is an iron acquisition system for higher plants except for Gramineae. It is a system that reduces trivalent iron in soil by a trivalent fermentation enzyme present on the cell surface of plant roots, It is a device that absorbs with a transporter. Among the plants having this mechanism, there is a mechanism to increase the activity of the trivalent fermentation enzyme by lowering the pH of the rhizosphere, and that the phenolic compound is released to the rhizosphere and the formed Fe (III) -phenol compound There exists a mechanism in which a chelate feeds Fe (III) to a trivalent fermentation enzyme present on the cell membrane surface. Strategy-II is an iron-harvesting mechanism that is visible only to the germplasm of monocotyledonous plants. Glycyrrhiza radishes release myuzinic acids with trivalent iron chelating activity to the rhizosphere under iron deficiency conditions and absorb iron from roots as 'Fe (Ⅲ) - musijnate' complex.

The present invention analyzes the function of rice transporter gene, which is known to have the function of promoting iron utilization efficiency by expressing under iron deficiency conditions, and to develop a plant that can adapt to iron deficiency conditions.

On the other hand, Korean Patent Publication No. 2002-0009604 discloses a rice flour resistant to iron deficiency, but the rice-derived OsIT gene which improves the iron utilization ability of the plant of the present invention and its use has not been described.

The present invention is derived by the request as described above, the present inventors have separated a to genomic DNA of rice plants and producing a transgenic plant was cloned for the iron transporter gene OsIT using the PCR method, and overexpressing the OsIT gene The results of the comparison between the wild type and the growth state of the wild type showed that the transgenic rice plants and the Arabidopsis plants overexpressing OsIT were superior to the non-transformed wild type plants in their iron deficiency conditions and completed the present invention.

In order to solve the above problems, the present invention provides a rice-derived iron transporter gene OsIT ( Oryza sativa iron transporter.

The present invention also provides a method for transforming a recombinant vector comprising an iron transporter OsIT protein coding gene derived from rice into a plant cell to increase the iron utilization ability of the plant as compared to the non-transformant.

In addition, the present invention provides a method for producing a transgenic plant in which a recombinant vector comprising an iron transporter OsIT protein coding gene derived from rice is transformed into plant cells to thereby increase iron availability of the plant compared to the non-transformant .

In addition, the present invention provides a transformed plant having improved iron utilization ability of a plant and seeds thereof compared to the non-transformed plant produced by the above method.

In addition, the present invention provides a composition for increasing the iron utilization ability of a plant, which comprises, as an active ingredient, a gene encoding an OsIT protein consisting of the amino acid sequence of SEQ ID NO: 2.

The OsIT gene of the present invention improves the ability of the plant to utilize iron, thereby enabling plant growth even under the condition of iron deficiency. Therefore, it is possible to develop a new crop having improved iron utilization ability by biotechnological method using the gene, and thus it can be usefully used in the field of new plant variety development.

1 is to investigate the expression pattern of OsIT genes in iron deficiency conditions using dongjinbyeo, showing experimental results of the iron sufficient and deficient condition by stem and root regions as (A), iron outer OsIT in other nutrient deficiency conditions (B) the expression of the gene. Stem in S +, iron-sufficient condition; S-, stems in iron deficiency conditions; R +, roots in iron-rich conditions; R-, roots in iron deficiency conditions; Full, complete nutritional condition; -N, nitrogen deficiency conditions; -P, phosphoric acid deficiency condition; -K, potassium deficiency conditions; -Fe, iron deficiency conditions.
FIG. 2 shows the result of time- course expression of OsIT gene in stem and root in iron-deficient condition using wild-type rice, (A) showing stem and (B) showing results in root.
FIG. 3 shows PCR results (A and C) and Northern results (B and D, respectively) of rice plants and Arabidopsis plants transgenic with the OsIT gene. OsIT-OX: OsIT gene overexpressed plant, WT: wild-type plant.
FIG. 4 shows the results of confirming the growth conditions under conditions of iron sufficiency and deficiency using wild type plants, OsIT overexpressed transgenic rice (A) and Arabidopsis thaliana (C) plants. (B) is the result of the growth of stem and root length in the wild type and OsIT overgrown transgenic rice plants under iron sufficiency and depletion conditions, and (D) is the result of wild type and OsIT over transgenic transgenic plants The length of roots and the growth state through fresh weight in iron deficient and deficient conditions were measured.

In order to achieve the object of the present invention, the present invention provides a rice-derived iron transporter gene OsIT ( Oryza sativa iron transporter.

The OsIT gene of the present invention may preferably consist of the nucleotide sequence of SEQ ID NO: 1. In addition, homologues of the nucleotide sequences are included within the scope of the present invention. Specifically, the gene has a nucleotide sequence having a sequence homology of 70% or more, more preferably 80% or more, still more preferably 90% or more, and most preferably 95% or more, with the nucleotide sequence of SEQ ID NO: 1 . "% 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 >

In addition, the present invention relates to a method for producing a recombinant vector comprising the step of transforming a recombinant vector comprising an iron transporter OsIT protein coding gene derived from rice into a plant cell to overexpress an OsIT protein coding gene, / RTI >

The scope of the OsIT protein according to the present invention includes the protein having the amino acid sequence represented by SEQ ID NO: 2 and the functional equivalent of 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 bioactivity" means an activity that increases the iron utilization capacity of a plant compared to a non-transformant.

The method according to one embodiment of the present invention comprises the step of culturing a rice-derived iron transporter OsIT ( Oryza < RTI ID = 0.0 > sativa iron transporter) protein coding gene into Arabidopsis plant cells and overexpressing the OsIT protein coding gene in the presence of the recombinant vector. . In the case of Arabidopsis thaliana transformants, the growth of transformants was excellent under the iron deficiency condition compared to the rice transformants.

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.

In the present invention, the OsIT gene sequence can be inserted into a recombinant expression vector. The term "recombinant expression vector" means a bacterial plasmid, a phage, a yeast plasmid, a plant cell virus, a mammalian cell virus, or other vector. 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.

Expression vectors containing the OsIT gene sequence and appropriate transcription / translation control signals can be constructed by methods known to those skilled 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 the recombinant vector of the present invention is a Ti-plasmid vector capable of transferring a so-called T-region to a plant cell when 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.

The expression vector will preferably comprise one or more selectable markers. The marker is typically a nucleic acid sequence having a property that can be selected by a chemical method, and includes all genes capable of distinguishing a transformed cell from a non-transformed cell. Examples include herbicide resistance genes such as glyphosate or phosphinothricin, antibiotics such as kanamycin, G418, Bleomycin, hygromycin, chloramphenicol, Resistant genes, but are not limited thereto.

In the recombinant vector of the present invention, the promoter may be CaMV, 35S, actin, ubiquitin, pEMU, MAS, or histone promoter, but is not limited thereto. The term "promoter " refers to the region of DNA upstream from the structural gene and refers to a DNA molecule to which an RNA polymerase binds to initiate transcription. A "plant promoter" is a promoter capable of initiating transcription in plant cells. A "constitutive promoter" is a promoter that is active under most environmental conditions and developmental status or cell differentiation. A continuous promoter may be preferred in the present invention, since the choice of transformants can be made by various tissues at various stages. Thus, a persistent promoter does not limit selectivity.

In the recombinant vector of the present invention, conventional terminators can be used. Examples thereof include nopaline synthase (NOS), rice α-amylase RAmy1 A terminator, phaseoline terminator, Agrobacterium tumefaciens (Agrobacterium tumefaciens ) Octopine gene terminator, but the present invention is not limited thereto. Regarding the need for terminators, it is generally known that such regions increase the certainty and efficiency of transcription in plant cells. Therefore, the use of a terminator is highly desirable in the context of the present invention.

Transformation of a plant means any method of transferring DNA to a plant. Such transformation methods do not necessarily have a regeneration and / or tissue culture period. Transformation of plant species is now common for plant species, including both terminal plants as well as dicotyledonous plants. In principle, any transformation method can be used to introduce the hybrid DNA according to the present invention into suitable progenitor cells. The method is based on the calcium / polyethylene glycol method for protoplasts (Krens, FA et al., 1982, Nature 296, 72-74; Negrutiu I. et al., June 1987, Plant Mol. Biol. 8, 363-373) (Shillito RD et al., 1985 Bio / Technol. 3, 1099-1102), microinjection into plant elements (Crossway A. et al., 1986, Mol. Gen. Genet. 202,179-185 (Klein et al., 1987, Nature 327, 70), the infiltration of plants or the transformation of mature pollen or vesicles into Agrobacterium tumefaciens Infection by viruses (non-integrative) in virus-mediated gene transfer (EP 0 301 316), and the like. A preferred method according to the present invention comprises Agrobacterium mediated DNA delivery. Particularly preferred is the use of so-called binary vector techniques as described in EP A 120 516 and U.S. Pat. No. 4,940,838.

In addition,

Transforming a plant cell with a recombinant vector comprising an iron transporter OsIT protein coding gene derived from rice; And

The present invention provides a method for producing a transgenic plant having increased iron utilization ability of a plant compared to a non-transformant comprising the step of regenerating the plant from the transformed plant cell.

The scope of the OsIT protein according to the present invention is as described above.

The method of the present invention comprises transforming a plant cell with a recombinant vector according to the invention, said transformation being mediated, for example, by Agrobacterium tumefaciens. In addition, the method of the present invention comprises regenerating a transgenic plant from the transformed plant cell. Any of the methods known in the art can be used for regeneration of transgenic plants from transgenic plant cells.

Transformed plant cells must be regenerated into whole plants. Techniques for the regeneration of mature plants from callus or protoplast cultures are well known in the art for a number of different species.

In addition, the present invention provides a transformed plant having improved iron utilization ability of a plant and seeds thereof compared to the non-transformed plant produced by the above method.

In one embodiment of the present invention, the plant is selected from the group consisting of Arabidopsis, potato, eggplant, cigarette, pepper, tomato, burdock, ciliaceae, lettuce, bellflower, spinach, modern sweet potato, celery, carrot, parsley, parsley, cabbage, Rice, barley, wheat, rye, corn, sorghum, oats, onion, etc., such as rice bran, watermelon, melon, cucumber, amber, pak, strawberry, soybean, mung bean, , Preferably a dicot or a monocotyledon, more preferably a Arabidopsis or a rice plant, but is not limited thereto.

In addition, the present invention provides a composition for increasing the iron utilization ability of a plant, which comprises, as an active ingredient, a gene encoding an OsIT protein consisting of the amino acid sequence of SEQ ID NO: 2. The composition of the present invention contains a gene encoding an OsIT protein consisting of the amino acid sequence of SEQ. ID. NO. 2 as an active ingredient. The gene can be transformed into a plant to increase the iron utilization ability of the plant. The composition according to one embodiment of the present invention preferably comprises OsIT ( Oryza < RTI ID = 0.0 > but are not limited to, the ability to utilize iron in Arabidopsis plants in conditions of iron deficiency, including the gene encoding sativa iron transporter protein as an active ingredient.

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  One. OsIT  Identification and isolation of genes

Genomic DNA of Dongjinbyeon, a cultivar of Korea, was isolated, and based on the entire genome sequence of rice, iron - transporter genes ( OsIT , Oryza sativa 3) (SEQ ID NO: 3) and reverse primer 5'-TGACGCCCACTTGGCCATGA-3 '(SEQ ID NO: 4)) were prepared and PCR was carried out to prepare the oligonucleotide primer specific for the iron transporter Was amplified. The amplified product was cloned into a pEntr vector and the sequence of the gene was confirmed by sequencing.

Example  2. OsIT  Analysis of gene expression pattern

In order to confirm the expression pattern of OsIT gene, Donggin rice was hydroponically cultured for 2 weeks, and the cells were treated with Hogland solution (20 mM Ca ((20 mM) Fe-sequestrate and 20 mM Fe- NO ₃ ) ₂ .4H ₂ O, 2.5 mM K ₂ SO ₄ , 2 mM MgSO ₄ .7H ₂ O, 0.5 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 7 days, and total RNA was isolated using a trizol reagent (Invitrogen, USA) . 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 [ ³² P] dCTP labeled probes with 20% (w / v) SDS, 20X SSPE, 100g / L PEG (8,000mWt), 250mg / L heparin and 10ml / L Hering sperm DNA 10 mg / ml) was reacted overnight at 65 ° C to confirm the expression pattern of the gene. As a result, it was confirmed that expression of the OsIT gene in Dong Jin-yang was strongly induced at the stem region under iron deficiency (0.01 mM Fe-EDTA) (Fig. 1A). In addition, nitrogen (0.25 mM (NH ₄ ) ₂ SO ₄ , KNO ₃ ) phosphoric acid (0.0125 mM NaH ₂ PO ₄ .2H ₂ O), potassium (0.01 mM KNO ₃ ) and iron mM Fe-EDTA) deficiency treatment. As a result, the expression level of the OsIT gene was found to be the highest in iron deficiency conditions (FIG. 1B).

Based on the expression pattern in Dongjinbyeon, Dongjinbyeong was cultivated for 2 weeks under hydroponic conditions and treated with iron deficiency (0.01mM Fe-Sequestrate) for 7 days. The expression of OsIT gene in plant tissues was examined, The expression level increased from the early stage of the condition to the strongest expression at day 7, whereas the root level decreased at day 3 after the strong expression on day 1 of iron deficiency condition, gradually increasing to the highest level on day 7 2).

Example  3. Preparation and Expression Analysis of Transgenic Plants

In order to investigate the function of the iron transporter gene ( OsIT ) isolated from rice in the plants, Dongjinbyeong and Arabidopsis thalania ) were transfected with OsIT overexpression. The method simply cloned the coding region of the OsIT gene into a destination vector (pH7WG2D, 1) to create a structure regulated by the CaMV 35S promoter, in which the OsIT gene is always expressed strongly. The triparental mating method was used to construct Agrobacterium Agrobacterium tumefaciens ) EHA105 strain. The callus was transformed into callus by treatment with the above Agrobacterium tumefaciens EHA105 strain. In the case of Arabidopsis thaliana, wild type (Col-0) firefly was transformed with Agrobacterium tumefaciens GV3301 strain containing the above OsIT gene in the same manner. Transformants were selected after confirming the introduction of OsIT gene and the amount of gene expression in T ₀ plants by PCR analysis (FIGS. 3A and 3C) and RNA blotting method (FIGS. 3B and D).

Example  4. OsIT  Growth analysis of over-expressing transgenic plants

For the overexpressed rice plants, the harvested seeds were cultivated for 2 weeks with hydroponic culture with the control group and cultured for 7 days in iron-rich (20 mM Fe-Sequestrate) and deficient (0.01 mM Fe-Sequestrate) Respectively. Growth of both stem and root was better than that of the wild type plant where the growth of the transformant was in the control condition. Even in the iron-deficient condition, it was confirmed that the growth of the transformant was better in stem and root growth than in the wild-type plant as the control group (Fig. 4A). It was also confirmed from the quantitative investigation that stem growth was better in iron-rich and deficient conditions in the transformants compared with wild-type plants, and root growth was also good in iron-rich and deficient conditions (FIG. 4B ). Over-expressing Arabidopsis thaliana plants for the tooth shape and harvested seeds on MS medium with the control group 5 days, enough iron _{(10mM FeSO 4 · 7H 2 O} ) and absence _{(0.01mM FeSO 4 · 7H 2 O} ) MS medium conditions (200mM 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 mM NaSO 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, 55 μM Myo inositol, 3 μM Tiamin-HCl, 0.8 μM Nicotinic acid, 0.5 μM The cells were transferred to Pyridoxine HCl, 0.01 μM CuSO ₄ .5H ₂ O, 0.01 μM CoCl ₂ .6H ₂ O, 2.3 mM MES, 1.2% Sucrose, 6% Agar; pH 5.7) Respectively. The growth of the transformants in the iron-sufficient condition was similar to that of the wild-type plants in the control group, but the growth of the transformants in the depletion condition was found to be better than that of the wild-type plants in which the control group (FIG. 4C). As a result of the quantitative investigation, it was confirmed that root growth was good in the transformant as compared with the wild-type plant (Fig. 4D).

<110> Dong-A University Research Foundation For Industry-Academy Cooperation <120> OsIT gene from Oryza sativa for improving iron availability of          plant and uses thereof <130> PN14306 <160> 4 <170> Kopatentin 2.0 <210> 1 <211> 1125 <212> DNA <213> Oryza sativa <400> 1 atggcgacgc cgcggacact ggtgcccatt ctgccgcccg tcgccgcgct cctcctcctc 60 ttcgtcgccg cctcctccat ccccatcctc gccgccgcgc agccggcgga cgcgtgcggc 120 ggcgcaccgg atcaggcggc ggcggacggc gcgtgccacg acgtgccgag ggcgctgcgg 180 ctgaagctga tcgccatccc gaccatcctc gtgtcgagcg tcgtcggcgt gtgcctgccg 240 ctcctctccc gctccgtgcc ggcgctccgc cccgacggcg gcctcttcgc cgtcgtcaag 300 gcgttcgcgt cgggcgtcat cctcgccacg ggctacatgc acgtgctccc ggacgccttc 360 aacaacctca cctcgccgtg cctgcccagg aagccgtggt cggagttccc gttcgcggcg 420 ttcgtcgcca tgctcgccgc cgtgtccacg ctcatggccg actcgctcat gctcacctac 480 tacaaacgca gcaagccccg gccgtctagc ggcggcgacg tcgccgccgt cgccgaccac 540 ggcgagagcc ccgaccaggg ccaccggcac ggacacggac acggccatgg gcatggcatg 600 gcggtggcca agcccgacga cgtcgaggcc actcaggtgc agctgcgccg gaaccgcgtc 660 gtcgttcagg tcctcgagat aggcatcgtg gtgcactcgg tggtgatcgg cctcggcatg 720 ggggcgtcgc agaacgtgtg caccatccgg ccgctggtgg cggcgatgtg cttccaccaa 780 atgttcgagg gcatgggact cggtggctgc atcgtgcagg cggagtacgg ccgccggatg 840 cctcggcctc 900 gccctgacca gggtgtacag ggacaacagc ccgacggcgc tcatcgtcgt cggcctcctc 960 aacgccgcct ccgcggggct gctccactac atggcgctgg tggagctcct cgccgccgac 1020 ttcatggggc ccaagctgca gggcaacgtc cgcctccagc tcgccgcctt cctcgccgtc 1080 ctcctcggcg ccggcggcat gtccgtcatg gccaagtggg cgtga 1125 <210> 2 <211> 374 <212> PRT <213> Oryza sativa <400> 2 Met Ala Thr Pro Arg Thr Leu Val Pro Ile Leu   1 5 10 15 Leu Leu Leu Leu Phe Val Ala Ser Ser Ile Pro Ile Leu Ala Ala              20 25 30 Ala Gln Pro Ala Asp Ala Cys Gly Gly Ala Pro Asp Gln Ala Ala Ala          35 40 45 Asp Gly Ala Cys His Asp Val Pro Arg Ala Leu Arg Leu Lys Leu Ile      50 55 60 Ala Ile Pro Thr Ile Leu Val Ser Ser Val Val Gly Val Cys Leu Pro  65 70 75 80 Leu Leu Ser Arg Ser Val Pro Ala Leu Arg Pro Asp Gly Gly Leu Phe                  85 90 95 Ala Val Val Lys Ala Phe Ala Ser Gly Val Ile Leu Ala Thr Gly Tyr             100 105 110 Met His Val Leu Pro Asp Ala Phe Asn Asn Leu Thr Ser Pro Cys Leu         115 120 125 Pro Arg Lys Pro Trp Ser Glu Phe Pro Phe Ala Ala Phe Val Ala Met     130 135 140 Leu Ala Ala Val Ser Thr Leu Met Ala Asp Ser Leu Met Leu Thr Tyr 145 150 155 160 Tyr Lys Arg Ser Lys Pro Arg Pro Ser Ser Gly Gly Asp Val Ala Ala                 165 170 175 Val Ala Asp His Gly Glu Ser Pro Asp Gln Gly His Arg His Gly His             180 185 190 Gly His Gly His Gly His Gly Met Ala Val Ala Lys Pro Asp Asp Val         195 200 205 Glu Ala Thr Gln Val Gln Leu Arg Arg Asn Arg Val Val Val Gln Val     210 215 220 Leu Glu Ile Gly Ile Val Val His Ser Val Val Ile Gly Leu Gly Met 225 230 235 240 Gly Ala Ser Gln Asn Val Cys Thr Ile Arg Pro Leu Val Ala Ala Met                 245 250 255 Cys Phe His Gln Met Phe Glu Gly Met Gly Leu Gly Gly Cys Ile Val             260 265 270 Gln Ala Glu Tyr Gly Arg Arg Met Met Ser Val Leu Val Phe Phe Phe         275 280 285 Ser Thr Thr Thr Pro Phe Gly Ile Ala Leu Gly Leu Ala Leu Thr Arg     290 295 300 Val Tyr Arg Asp Asn Ser Pro Thr Ala Leu Ile Val Val Gly Leu Leu 305 310 315 320 Asn Ala Ala Ala Gly Leu Leu His Tyr Ala Leu Val Glu Leu                 325 330 335 Leu Ala Ala Asp Phe Met Gly Pro Lys Leu Gln Gly Asn Val Arg Leu             340 345 350 Gln Leu Ala Ala Phe Leu Ala Val Leu Leu Gly Ala Gly Gly Met Ser         355 360 365 Val Met Ala Lys Trp Ala     370 <210> 3 <211> 24 <212> DNA <213> Artificial Sequence <220> <223> primer <400> 3 caccatggcg acgccgcgga cact 24 <210> 4 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> primer <400> 4 tgacgcccac ttggccatga 20

Claims (11)

delete Comprising the step of over-expressing an OsIT protein coding gene by transforming a plant cell with a recombinant vector comprising a rice-derived iron-transporter OsIT ( Oryza sativa iron transporter) protein coding gene consisting of the amino acid sequence of SEQ ID NO: 2, To increase the iron utilization capacity of the plant compared to the non-transformant. delete delete Transforming a plant cell with a recombinant vector comprising a rice encoding OIT ( Oryza sativa iron transporter) protein coding gene comprising the amino acid sequence of SEQ ID NO: 2; And
And regenerating the plant from the transformed plant cell, wherein the ability of the plant to utilize iron is increased compared to the non-transformant under iron-deficient conditions. delete A transformed plant having enhanced iron utilization capacity of the plant compared to the non-transformed plant in the iron-deficient condition produced by the method of claim 5. delete A transformed seed of a plant according to claim 7. A composition for increasing the iron utilization ability of a plant in an iron-deficient condition, which comprises, as an active ingredient, a gene encoding an OsIT ( Oryza sativa iron transporter) protein consisting of the amino acid sequence of SEQ ID NO: 2. delete

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