KR101636157B1 - Methyltrasnferase gene resistant to a bacterial blight, and the transgenic plant using the same - Google Patents

Abstract

The present invention relates to a plant transformant in which the activity of the LOC_Os06g20790 gene comprising the nucleotide sequence of SEQ ID NO: 2 encoding a methyltransferase is enhanced, an expression inducing sequence for enhancing the activity of the gene, And a method for producing the plant transformant. Further, the present invention relates to a method for enhancing the disease resistance of a plant, comprising the step of enhancing the activity of the LOC_Os06g20790 gene.
The present invention relates to a transgenic rice plant which is intrinsically present in rice and has a strong resistance to blight of blight by inserting an expression inducing sequence into the expression regulatory region of LOC_Os06g20790, a resistance gene against blight of blight, Can be widely used because it can reduce the use of pesticides, improve productivity, and provide safe agricultural products through disease resistance.

Description

{Methyltrasniferase gene resistant to a bacterial blight, and a transgenic plant using the same}

The present invention relates to a plant transformant in which the activity of the LOC_Os06g20790 gene comprising the nucleotide sequence of SEQ ID NO: 2 encoding a methyltransferase is enhanced, an expression inducing sequence for enhancing the activity of the gene, And a method for producing the plant transformant. Further, the present invention relates to a method for enhancing the disease resistance of a plant, comprising the step of enhancing the activity of the LOC_Os06g20790 gene.

Rice blight caused by Xanthomonas oryzae pv. It is a bacterial tuberculosis caused by oryzae . Its onset occurs from early July to harvesting season, mainly from early July to mid-August. Bacterial wintering is possible in underground stalks, roots, or diseased rice straw and rice stubble, which are intermediate host plants. Bacteria spread along the water by flagella and invade into the water holes, air holes and cut roots of the leaves. Disease occurs mainly in the area of frequent occurrence of the disease every year, but it also occurs in areas not in the area of the pelvic area.

Usually it appears before and after the heading period, but the occurrence of the rash or the occurrence of the rash occurs at the beginning of the reply. The first occurrence of the disease in Korea was first reported in 1930 in Haenam, Jeollanam-do. Since 1960, a limited occurrence has been observed in some parts of the southern part of the country. The cultivation of Gyeongnam cultivar showed a nationwide occurrence. 23, and it is considered to be one of the three major diseases of rice, showing not only the onset of leaf in later stage during cultivation period but also the symptom of gold mold in which the whole body occurs until the post-transplantation cleavage stage.

The rice blotting bacteria are classified as Gram negative bacteria and are widely distributed throughout the world because of bacterial fungi causing bacterial fake disease of rice. Although they cause great damage to rice, pathogens propagate in the ducts of rice, And there is no specific fungicide to date.

In addition, the conventional method of imparting resistance to crops is a technique in which plants are given new traits through the introduction of foreign genes.

Meanwhile, Dongjinbyeong was developed in 1975 as a cross breed between gold southern wind, Nakdong rice and Sadominori in 1975 with the goal of raising the disease, quality and diversity of new varieties. It is resistant to streak blight in case of disease resistance, , And it is weak against blight of white blotch and other pests. Therefore, it is required to study resistance to blight of white blossom and blast disease in Dongjinbyeong, and to cultivate improved varieties.

Accordingly, the present inventors have made intensive researches to identify a resistance gene against the blight of rice blight that has not yet been overcome, and as a result, a methyltransferase gene (LOC_Os06g20790) that confers resistance to blight of blight in the wild type rice paddy gene is identified , And that the transformant activated by the transformant was resistant to blight of blight.

One object of the present invention is to provide a plant transformant in which the activity of the LOC_Os06g20790 gene comprising the nucleotide sequence of SEQ ID NO: 2 encoding the methyltransferase is enhanced.

Another object of the present invention is to provide a recombinant vector for promoting the resistance to diseases of plants, comprising an expression inducing sequence for enhancing the activity of the LOC_Os06g20790 gene comprising the nucleotide sequence of SEQ ID NO: 2 encoding the methyltransferase.

Yet another object of the present invention is to provide a method for producing a recombinant vector comprising: (a) infecting a plant with Agrobacterium to which the recombinant vector is introduced; And (b) callusing the cells of the infected plant in step (a) to obtain cells of the transformed plant. will be.

It is still another object of the present invention to provide a method for enhancing plant disease resistance, comprising the step of enhancing the activity of the LOC_Os06g20790 gene comprising the nucleotide sequence of SEQ ID NO: 2 encoding a methyltransferase.

In one aspect of the present invention, there is provided a plant transformant in which the activity of the LOC_Os06g20790 gene comprising the nucleotide sequence of SEQ ID NO: 2 encoding a methyltransferase is enhanced.

In the present invention, an expression induction sequence is inserted into the expression control region of LOC_Os06g20790 so as to enhance the activity of the LOC_Os06g20790 gene having the nucleotide sequence of SEQ ID NO: 1 or SEQ ID NO: 2, which is a methyltransferase, Were prepared.

More specifically, the present inventors have observed plant transformants resistant to blight of blight in a 100,000 rice mutant population produced by the transformation method using Agrobacterium. The plant transformant is a plant in which a gene function is activated in the region where T-DNA including an expression inducing sequence for expressing a gene by Agrobacterium is inserted into the genome, and insertion of T-DNA The location can be confirmed by genome DNA PCR and sequencing, and the gene locus id can be found in the database.

The term "methyltransferase" in the present invention is a generic term for an enzyme that catalyzes the methyltransferase, and may be referred to as a methylation enzyme by methylating an amino group, a hydroxyl group or a thiol group, which is a substrate,

The term in the invention, "LOC_Os06g20790" is Oryza Sativa is a gene located in the sixth chromosome, and its expression is a protein methyltransferase. The LOC_Os06g20790 gene has the nucleotide sequence of SEQ ID NO: 2, and the nucleotide sequence is encoded by the amino acid sequence of SEQ ID NO: 3.

In the present invention, the LOC_Os06g20790 gene used for preparing the plant transformant may be a gene having the nucleotide sequence of SEQ ID NO: 1 or SEQ ID NO: 2, preferably the nucleotide sequence of SEQ ID NO: 2, Lt; RTI ID = 0.0 > 3 < / RTI >

The genome DNA size of LOC_Os06g20790 (TIGR, Os06g0313440 (RAP-DB)) is 8025 bp (SEQ ID NO: 1) and the size of the representative coding sequence (LOC_Os06g20790.1) is 1113 bp intron, and translated into 370 amino acids to make a protein (SEQ ID NO: 3). This protein encodes the plant methyltransferase domain (Methyltransf_7) (Fig. 1B).

A gene the gene LOC_Os06g20790 is implicitly present in the rice, to mean that the term "intrinsic" is a state of natural organisms have in the present invention, in the present invention, a plant, preferably from rice (Oriza sp . It is used to denote genes that are inherently chromosomes.

As used herein, the term "enhanced activity" means enhancing the activity of a particular gene and may be used interchangeably with the term "activation ". It also includes activating inactivated genes or increasing the activity of genes with low activity. 1) introducing more than one copy of the gene of interest, 2) inserting a sequence that induces expression in the regulatory region of the gene of interest, 3) enhancing by transforming the activity into enhanced activity, or 4) The activity can be modified to enhance. In the present invention, it is preferable to insert a sequence or a promoter inducing expression in the expression control region of LOC_Os06g20790, more preferably, a sequence including an expression induction sequence of SEQ ID NO: 4 .

In the present invention, the term "low-activity gene" means that DNA is present in the gene, that is, in the gene, but the rate of transcription into RNA is low or that the protein is not translated. In the present invention, it is meant that the gene in which rice is inherently possessed by the rice is low in transcription or translation, and thus can not express its function properly. In particular, LOC_Os06g20790 gene does not express its function properly.

In a specific embodiment of the present invention, the expression of the LOC_Os06g20790 gene was increased by inserting an expression inducing sequence (SEQ ID NO: 4) in the expression control region of the LOC_Os06g20790 gene (FIG. 4). In addition, in a specific example of the present invention, it was confirmed that the plant transformant transformed to enhance the activity of the LOC_Os06g20790 gene has disease resistance against blight of blight (FIG. 5).

The term "expression inducing sequence" as used herein includes, without limitation, a sequence which functions to promote expression when it is present in a gene expression regulatory region and includes a transcription factor binding site, an enhancer, a co- binding region). In the present invention, it is preferable that the sequence inserted into the expression control region of the gene comprises the expression induction sequence of SEQ ID NO: 4.

The term "transformed" in the present invention means all the activities in which a specific trait occurs, disappears or is controlled through mutation such as insertion, deletion or substitution on the DNA base sequence. In particular, in the present invention, Which means that the genetic properties of In the present invention, the term "transformant" refers to a transgenic plant produced by transformation, and includes genetic recombinants produced by mutation of a specific gene or activation of an activity using genetic recombination technology.

In the present invention, the term "plant transformant" includes all plants that can be subjected to transformation, and in particular, all plants in which the LOC_Os06g20790 gene or a gene having the inactivated or inactivated gene are included without limitation. Preferably, the plant transformant is rice ( Oryza < RTI ID = 0.0 > sp . ), And more preferably, it may be a Dongjin rice.

In the present invention, the plant transformant may have resistance to blight of blight.

In a specific embodiment of the present invention, a transformant was prepared by inserting an expression-inducing sequence of SEQ ID NO: 4 into the regulatory region of LOC_Os06g20790, which is an endogenous gene of the present invention, in Dongjinbyeong to enhance the activity of the gene corresponding to LOC_Os06g20790 , And named it SAMT 14.

In another aspect, the present invention provides a recombinant vector for enhancing disease resistance of a plant, comprising an expression inducing sequence for enhancing the activity of the LOC_Os06g20790 gene comprising the nucleotide sequence of SEQ ID NO: 2 encoding a methyltransferase.

The methyltransferase, LOC_Os06g20790 and the expression inducing sequence are as described above.

In the present invention, the term "disease resistance" means that the onset of disease does not occur well, or that the disease progression rate is significantly slower or recoverable. In the present invention, disease resistance may mean resistance to blight of blight.

In the present invention, the term "recombinant vector" means a vector that has undergone a gene recombination process in which a specific sequence is mutated, inserted or deleted, preferably a vector into which an expression inducing sequence is inserted, Means a recombinant vector capable of inserting the sequence into the expression control region of LOC_Os06g20790.

The recombinant vector is functionally linked to an expression control sequence so that the LOC_Os06g20790 gene can be expressed. For example, the vector may comprise a signal sequence or leader sequence for membrane targeting or secretion in addition to an expression regulatory element such as a promoter, an operator, an initiation codon, a stop codon, a polyadenylation signal, an enhancer, . In addition, the vector may comprise a selectable marker, and the vector may be self-replicating or integrated into the host DNA. The vector of the present invention can be prepared using gene recombination techniques well known in the art, and site-specific DNA cleavage and linkage are performed using enzymes generally known in the art

In one embodiment of the present invention, a recombinant vector comprising the expression-inducing sequence of SEQ ID NO: 4 was prepared at the LOC_Os06g20790 gene expression regulatory region and introduced into rice cells through Agrobacterium to express the LOC_Os06g20790 gene, Respectively.

In another embodiment, the present invention provides a method for producing a recombinant vector comprising: (a) infecting a plant with Agrobacterium to which the recombinant vector is introduced; And (b) callusing cells of the infected plant in step (a) to obtain cells of the transformed plant, wherein the method comprises the steps of: .

The term "Agrobacterium" in the present invention refers to a gram-positive bacterium, which is infected with plant cells through scars of the roots or stems of plants, and inserts the gene of interest into the genome of the plant cell to transform the plant cell Is a kind of carcinoma bacteria that causes cancer. In the present invention, the Agrobacterium may be Agrobacterium tumefaciens.

The term "callus " in the present invention means an undifferentiated amorphous cell mass produced when a plant is injured, that is, undifferentiated cell. Particularly, in the present invention, Agrobacterium When the transforming carcinoma bacterium is infected, it means that the infected cells are undifferentiated and lump on the wound site. That is, in the present invention, transformed undifferentiated cells can be obtained using only the cells transformed by infection with carcinoma bacteria to produce a mass of carcinoma, which is produced as a callus.

The method for producing a transformant having enhanced disease resistance may further include the step of differentiating the obtained callus into root and adult. Since the callus is only a cell mass in an undifferentiated state as described above, it may be necessary to induce differentiation into roots and adults in order to obtain it as a whole seed.

In one specific embodiment of the present invention, the obtained callus was cultured in root-induction medium to induce differentiation into root and adult (Example 2).

In another aspect, the present invention provides a method for enhancing plant disease resistance comprising the step of enhancing the activity of the LOC_Os06g20790 gene comprising the nucleotide sequence of SEQ ID NO: 2 encoding a methyltransferase.

The LOC_Os06g20790 gene, activity enhancement and disease resistance of the present invention are as described above.

In the present invention, the term " promotion "is used herein to refer to at least 5% or 10%, preferably at least 20% or 40%, more preferably 50%, 60%, 70% Means that the disease resistance is increased by introducing the gene as compared to the inherent disease resistance of the rice.

The present invention relates to transgenic rice plants having strong resistance against blight of blight by inserting an expression inducing sequence into the expression regulatory region of LOC_Os06g20790, a resistance gene against blight of blight present intrinsically in rice, Can be widely used because it can reduce the use of pesticides, improve productivity, and provide safe agricultural products through disease resistance.

Fig. 1 is an outline of the structure of LOC_Os06g20790 selected as a gene related to resistance to blight of blight. Fig. 1 (A) shows the structure of the gene phase. Fig. 1 (B) shows the structure of the gene-expressing protein. Methyltransf_7, as shown, is a methyltransferase domain.
FIG. 2 is a structural diagram showing a part of genomic DNA of SAMT 14 transgenic rice in which a gene expression induction sequence including T-DNA is inserted in a region regulating LOC_Os06g20790 expression.
FIG. 3 is a diagram showing PCR-confirmed positions of T-DNA insertion in SAMT 14 transgenic rice.
FIG. 4 is a graph showing the results of RT-PCR to confirm the expression of LOC_Os06g20790 in SAMT 14 transgenic rice.
FIG. 5 is a graph showing the results of an experiment for resistance to blight of blight of SAMT 14 genetically modified rice, and FIG. 5A shows the results of inoculation of blotch blight of native-type rice and SAMT 14 genetically modified rice with scissors And FIG. 5B is a graph showing the results of measuring the lengths of rice leaves that were destroyed by infection thereof.

Hereinafter, the present invention will be described in more detail with reference to the following examples. However, the following examples are intended to illustrate the present invention, but the scope of the present invention is not limited thereto.

Example 1. Screening of Transgenic Target Gene through Bioinformatics

We identified 230 methyltransferases that are expected to be closely related to the disease resistance of the plants possessed by the genomes of Dongjinbara. Based on the microarray data, 108 genes were selected for MeV v2.0 program. In the process of isolating genes having disease resistance by preparing transformants for selected genes, LOC_Os06g20790 of the present invention was selected as a gene resistant to blight of blight.

Example 2. Preparation of transgenic rice with expression derivatives

2-1. Production of recombinant vector for rice transformation

A vector was constructed to insert an expression inducing sequence into the nucleotide sequence of the expression control region of the target gene. First, the T-DNA for inserting the genome, N-GUS for identifying the left gene sequence at the insertion site, and the recombinant vector containing the expression induction sequence of the gene of SEQ ID NO: 4 were obtained from Tobacco Leaf Mosaic Virus 35S promoter DNA and a vector having L-0.5 / 1.5 to identify the right gene sequence at the insertion site. The primers for identification and amplification of the target gene from the inserted genome are as follows.

Enhancer-F (forward, SEQ ID NO: 9)

Gt;

Enhancer-R (reverse, SEQ ID NO: 10)

5'-ggatctagatatacacatca-3 '

The PCR amplification conditions were 1 cycle at 94 ° C for 3 minutes, 1 cycle at 94 ° C for 1 minute, 1 minute at 58 ° C and 2 minutes at 72 ° C for thermal denaturation of DNA for the first time, Respectively.

The amplified PCR product was electrophoresed with 1.0% agarose gel, stained with EtBr (ethidium bromide) solution, and the band corresponding to the base size was cut under UV lamp to obtain agarose gel purified ≪ / RTI >

The extracted PCR product was digested with the restriction enzyme BamHI / KpnI. Further, the recombinant vector pGA2772 vector was digested with restriction enzyme BamHI / KpnI, and the digested vector was purified by the above agarose gel purification method. The DNA fragment from the PCR product was inserted into the cleaved recombinant vector to prepare a recombinant vector pGA2772 vector into which the enhancer sequence (Enhance sequence, SEQ ID NO: 4) was inserted.

2-2. Transformant production

The prepared vector was introduced into Agrobacterium tumefaciens using electroporation. The cells were transformed into the aseptic state of Dongjinbyeon in the culture room by leaf disc method. Specifically, the rice leaves were cut into approximately 5 x 5 mm size and placed in an MSO liquid medium (4.3 g of MS salt, 30 g / L of sucrose, pH 5.6 to 5.8) containing 200 μl of Agrobacterium, The cells were cultured for 3 days and the cultured leaf slices were treated with callus-shoot induction medium (100 쨉 g NAA, 1 mg BA, 500 쨉 g cell suspension, 1 mg phospinothricin, 0.7% (phyto agar, pH 5.6) and transferred to fresh medium. Induced germination was transferred to root-induction medium (250 μg of cell suspension, 10 mg phosphinotricin, 0.7% phyto-agar, pH 5.6) in MSO medium if it grew to about 1.5 to 2 cm.

The gene transgenic rice in which the expression derivative was inserted was named SAMT 14, and its gene structure is as shown in Fig. Specifically, a T-DNA sequence (SEQ ID NO: 5) was inserted into an upstream region predicted to be an expression-regulating region of LOC_Os06g20790 present in the first derivative of Dongjin rice, To prepare a conversion body.

2-3. Identification of rice embryo insertion position of recombinant vector

T-DNA inserted chromosomes were analyzed by inverse-PCR and sequencing of SAMT 14 made by T-DNA insertion.

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

SAMT 14 is a system in which T-DNA is inserted upstream of the LOC_Os06g20790 gene expression regulatory gene. The primer (SEQ ID NO: 6 and SEQ ID NO: 7) created at the T-DNA insertion site and the NGUS1 primer 8) were used for genotyping analysis.

PCR reactions were performed after 5 minutes at 95 ° C; Repeating 38 times at 95 ° C for 1 minute, at 57 ° C for 1 minute, and 72 ° C for 1 minute, and finally at 72 ° C for 7 minutes. PCR products were confirmed by electrophoresis using agarose gel (Fig. 3). The genotype was selected through this.

The primer sequences used here are as follows.

SAMT 14_F (SEQ ID NO: 6)

5'-GTCAACATTCGCAGTCAACA- 3 '

SAMT 14_R (SEQ ID NO: 7)

Gt;

NGUS1 (SEQ ID NO: 8)

5'-AACGCTGATCAATTCCACAG-3 '

Example  3. Verification of genetic activity of transgenic rice

LOC_Os06g20790 gene activity was confirmed by reverse transcriptase-PCR (RT-PCR) in transgenic rice plants (SAMT 14) in which expression derivatives were inserted into the expression regulatory region of LOC_Os06g20790 gene.

Specifically, total RNA was extracted from 0.1 g leaf of transformed rice (SAMT 14). Reverse transcriptase PCR was performed using reverse transcription primers and reverse transcriptase DNA polymerase, which can confirm the expression of LOC_Os06g20790 using this as a template.

As can be seen from FIG. 3, it was confirmed that the expression of the gene LOC_Os06g20790 was increased in the transgenic rice of the present invention, as compared with the wild type Dongjin pine.

RT-SAMT 14_F (forward, SEQ ID NO: 11)

5'-CAGCTTGCATTGGTTATCCG-3 '

RT-SAMT 14_R (reverse, SEQ ID NO: 12)

5'-CAACTCACCCTTAATGCCA -3 '

Example  4. Transgenic rice ( SAMT  14) rice To blight of blight  Resistance test for

In order to confirm whether the transgenic rice produced in Example 2 showed resistance to blight of blight, SAMT 14, a transgenic rice in which an expression derivative was inserted into the LOC_Os06g20790 expression regulatory region, was grown as a control, Gt ; Xanthomonas < / RTI > oryzae pv . oryzae (strain: PXO99Az) scissors were inoculated at 2 weeks after inoculation (Fig. 5). Lesion lengths were measured and compared with the wild type Dongjinbyeo.

As can be seen from FIG. 5B, the average pathogenicity of SAMT 14 transformants was only about half of that of wild-type Dong Jinbin (natural type vs. SAMT 14: 24.2 cm vs. 10.6 cm) As shown in Fig.

From these results, it was confirmed that the transgenic rice SAMT 14 of the present invention can be resistant to blight of leaf blight by activating the inherent gene LOC_Os06g20790.

From the above description, it will be understood by those skilled in the art that the present invention may be embodied in other specific forms without departing from the spirit or essential characteristics thereof. In this regard, it should be understood that the above-described embodiments are to be considered in all respects as illustrative and not restrictive. The scope of the present invention should be construed as being included in the scope of the present invention without departing from the scope of the present invention as defined by the appended claims.

<110> University-Industry Cooperation Group of Kyung Hee University <120> Methyltrasniferase gene resistant to a bacterial blight, and the          transgenic plant using the same <130> KPA141024-KR <160> 12 <170> Kopatentin 2.0 <210> 1 <211> 8025 <212> DNA <213> Oryza sativa <400> 1 aaactgaatc atggccacca actccaaaca gagtgtgcat atgaatccag ggcaaggtga 60 aacgagctat gctcagaact cagcacttca ggtatgcttg tgtttaactc ttaaatgcac 120 aaacttaggc tatgcatgat cacctaattt ctcttatgtg gaatttattg gtgtactgac 180 tagatgaaga aaccatatgg tcgccttaga tgcgtcggta cggtgttgta aggcaccggt 240 tatgaagatc accgtgcggc gacggcgatg ggaattcgcc attcgttccc cttcaacaca 300 ccttaagatc gaaagtctga gagttttggt tttggctaag ccgtaaatcg atcgtcgcca 360 atgaacgggt aacgttccct ttatatgttt gtgcattgag gggatcgtgg gcgagtccgt 420 tacgtagata gcgcgtccga tcatcgcaca agagagagag caatgtgcac tcgagaccga 480 gggaatcggt tctcatcgtt aggcgcgagt gggagtgggg tggaccaagc ccattggcgc 540 aaatctctga ctcggtggcc agagatgctt gctctttctt caaagtgcta taccggcttc 600 ttatgattgt cgaaagttga gggcgttgcc tcatttctga gtattacatg ccttgtacta 660 attcgtcatc tgaaggaact gacagtagca gattataaat gaattgaatc attgaaataa 720 gaaccagtca aggcagaatt tgtactgatc agatattttt gtttttataa aaggtctatt 780 gtacaaaaat tgatgacagt ttgatatttg ccaaaaaaca gagaaatgga tttattttgg 840 tcataatgca atcttattca cctgtttttg acaaaataag gaaaggaatt gccattaata 900 tggtggaagc taattttaat ggcctaaatg tttatataaa aatgatcggc tgaatagcta 960 atatggtctc tgaagttttg cttgggctca gtttagtcct cggagtttca aatcattcaa 1020 aaaagttctc caaattaatc atttgggtta atatagtcct tgaactcaaa aaagtgtccc 1080 ttgagcatgc catgccaagt catcctgcca tgcagcggta tgaatgaaat gaccattcta 1140 tccttatata ccatataaaa aagaaagaaa agaaaagaag acaaatgttt cacaggtgta 1200 tgatcgtttt agaattgtca cttacgattg ctgtagtcaa gcaccgatcg ttgggcctcg 1260 gctagcctgt tatttttttc tatttgtgtt ttttctttct tttctatata ttatataagg 1320 ttagaatggt cattttattc ataccgctac atgcgaggat aacttggcat gctcacgtga 1380 tatttttttg gtccaaggat tatattaacc caaataatta atttggagga ctcgtttgaa 1440 cgatttgaaa cattgaggac taaattgagt ccgaagtgaa acttaaggga ccgtattggc 1500 tatttaccca aaatgatcat ataactatgt ttacttggga caattttttt aagaactagg 1560 tgaaaccccg cgcattgctg cgggaattta gtatgaagat atacattgaa acattatgtg 1620 aattatgtta gatgataatt taacatattt gtgtttgaaa agctcttaaa ttataaaaac 1680 tataaagata tagcatgctt gcatgatgtt taaatgtgat gattgttagt ggatgatgat 1740 gtggcatctt gttaattagt ggatgattat atggcatctt gttagtggat gattatgtgg 1800 catctttgca tattaagctt aaggagttag tgggggataa gtttatagta agctaggata 1860 tgctcacttg ttcaaattaa atactccaat tttaatgtaa cggaatattg caccattttt 1920 tcgaactatg atactagatg catcgaggta tgttcgagat aggagaattt ttacaatact 1980 taagagatac caagagatat taaaattttt agttgaaatt tgatacctct tgctactgga 2040 agtattgaga ggaccaaatt ttatataaaa aatatgatac ttgcaggtac cttctcaagt 2100 attataaaat cactcgctaa atagagtcag cttgaaggtt gtgtaaaaaa aatgggtttc 2160 acatggaacc aatctaccta gttgtctgac ttgagcgcat ataaatttta taattttgat 2220 gtttgacagt ttaacttggt gcattcatat tttgggtgtg tttagttcac gctaaaattg 2280 aaagtttggt tgaaattgaa gcgatgtgac ggaaaagttg gaagtttgtg tgtgtagaaa 2340 agttttgatg tgatagaaaa gttggaagtt tgaagaaaac ttttggaact aaactcggcc 2400 ttcatcatat tcccagttgg taactctatc cattgacggc tgtgttttgt tccacgctaa 2460 aattggaagt ttgactaaaa ttagaagttt aaaggaaaaa gttggaagtt tatgtgtgta 2520 ggaaagtttt gatgtgatgg aaaagttgca agtttgaaga aaagttttgg aactaaactc 2580 tgccttcatc atattcccag ctggtaactc tatccattga cggctgtgtt tagtttcaca 2640 ttaaaattag aagtttgaca aaaattagaa gtttgaagga aaaagttgga agtttatgag 2700 tgtagaaaag ttttgatgtg atggaaaagt tgaaagtttg aagtaaactc ggccttcatc 2760 atattcccag ctggtaactc tatccattga cgattgtgtt tagttccaca ccaaaattag 2820 aagtttgact aaaattagaa tttcagagaa aaaagttgaa agtttaatta tgtgtgtaaa 2880 aaaagtttta atctgatgga aaaattggaa gtttgaagaa aaaattgaaa actaaacccc 2940 gccgtaatct gaaaatcatc atactacacc caccctgcat tgtgtggcca aaaaagccaa 3000 ctatttcaat gtttactttg cttgcttgca agatctgatt gagacatatc tgaaaataat 3060 ttgcacgcag aaaacagccc aggataggat gaagactctg atagaggagg cagtcacagg 3120 cttatgcaca agctcatgtc ctcacccaaa gaacatggtg atcgcggact tgggctgctc 3180 ctccggacca aacgcgctca cgctcgtctc tgccgccgtc gacgccatcc accgctactg 3240 cgcgcagcac gagcagctgc cgccggagat gtgcgtgctc ctgaacgatc tgcctgacaa 3300 cgatttcaac gccgtggcga agagcttgga cacactgaag cacagtggtg atgaagcgct 3360 cgctcgccct actgctgtga tcaccggtat ggtacctggg tcgttctacg agaggctatt 3420 cgctcgtggc tccttgcatt tggtttgctc ggctaacagc ttgcattggt tatccgaggt 3480 aagtactgcc tccgtttttt aaatatatga tactgttgac tttttaacaa acgttcgatc 3540 attcgtctta tttaaaattt ttatgtaaat ataaaaaaaa ttaagtaata cttaaaaaaa 3600 catttaatga taaatcaagt cacaatataa taaatgataa ttacataatt ttttttaata 3660 agatgaattg tcaaatgaat ctcaaaagtc aaatggcgtc gtatattaaa aaatggagag 3720 agtaactaat tgtgttgatt acccatccat gtatggaggg ggtatgtttt gatttctgta 3780 agtagtgtgc attgtaactg caggcacctg aagatctcaa gaagagtaga atcccgatgc 3840 atgatagtga tgaacagttg agatcatcaa ggcatcaaat tgtcgcggat tcttatgctc 3900 ggcagtttag gaaggatttc atgcgattct tgagcttgag ggcacaggag attgtccctg 3960 gggccgaat ggttgtctca ctgttggtca agcgctcgga caaacctgac actgagttaa 4020 ttcaaccatg gacgccggcg gtgacggctt taagtgacat ggcattaagg gtgagttgac 4080 caatttcggc acataccaat tcagtcttat gttcgtgttt tccaatcaca tatgacacaa 4140 gcacacacat aatctctttg gactgctgtt cccttgttgc ttctgattca tctacctttt 4200 tcttcaagct cttattcgca aaaaaacaaa aacaaaaaca aaaacaaaag gacacaagca 4260 cacacaagat gtctttgcac acgtcaactc tttaattacg acctacatga ttaattaaag 4320 aggtttttac ctgtatgcca ttaagaaaga tgatgcatgt ctctgtacca cttaaaagtt 4380 cgagccttcc tatatgccat cggcgagctt tattgcgccc tccacaccat tctgtcgcta 4440 ttctgtgtgg gtttaaccgt ttggcaaatc tgacatatgg gcccgggtag gaaaaatgtc 4500 gctcttgccc tttcaaaaga ggacaggcgg cacggcggac aagcggccag gcgcggcgct 4560 gagtcaggcg accggcgcgg cggaaaagcg gccagccaca atgctgaggc aggcgaccgc 4620 gcgaggcggc caggcggcgc cagcggccag gcggcgtcgg cagccagccg aggcggcgcg 4680 gcgcggcgac caggaggggt ggggtggcgc ggcggcaagg ccgccgccgt agggaatgat 4740 ggccaccgcc acctccatcc acgccgccca ccacactcaa tccactccac agccaaatcg 4800 aaatctcacc ccgtcctcca agccgacgac tccctccccg tccgcccgtg gcatcggcgg 4860 catccgccac ctacaactcc ctccgcgcca ccatcgagct ccacttccgc aagctcaacg 4920 ccatgaaggc cggctcctcc tcctcctcct ttgccggcga cggcgcctcc ggatcatccc 4980 agcccacggg ctcctcctcg tcgcggacca agaagatgga cgaggagcgg cggcgtccgg 5040 ggagggatcc ctcctcccgc cgatccgggt gtcagagtgg gctggcgggt aagagatgag 5100 gaggagcagc ggtggcggcg gcggccgcac ggcagctacc tcctcctctc cttctccgtg 5160 ctcgccatcg tggaccggct cgtcgacttc ccactgctct cgttcactat ccagacggac 5220 gccctcgagc acgccaccat gtacctccac ctagccatgt acgcgtccgt ggcgctcgcc 5280 gccgacagtg ttgcgtcctc ccacaacatc ggcgcgcagg cgctgcttgt ggacgtggtg 5340 gccgcgctcg tcacgtcggt gctcggccag gagctgttcc cctcctcccc cttcctcggc 5400 gccttgggga cgaacgaacg aacacgttgg caagaacgcc taatggcaac taagtcattt 5460 agatgcgctg ttttctaccg ttaggacaaa aaggacggaa tggtgtgtag ggcgctataa 5520 agttcgacga tggcatagag ggaggctcga acttttcagt ggcacagagg ccagcatcat 5580 ctttcttaat ggcatacggg taataacctc ttaattaaat tctgaaaata attttcatat 5640 gtacaacgtg cgttcatcta ctgtcccaca aaatttccag tccagtgaca gttggatgtt 5700 gagtgacaga acatgtgctg caaaactttt caggcgtcct ccctttaggg attaaaagag 5760 tttcgaaata aatcagctca taggggtact tgcacaagtg catatgtagg caccaaacta 5820 ggttaattta ttacctgaat tactctacaa cttatatgta ttttatacaa agcagctaca 5880 ctgatacaaa tctactctgt tatactgaat tttcttcaat tcaatgccag ggtgttatca 5940 gcaaagaaaa gcttgactcc ttctacattc ccttgtgctg ccctatggac agtgaagtta 6000 acaacatcat agaggaggag ggttcatttg aggtcaacaa gatgatgatg cacgatccgt 6060 atgacggcac gggcaaagcc ttgctcgatc tgaagatggt agccttgcgc gtaagagctg 6120 tatttgagcc aataatagtg cagcattttg cagcgtcaga tgaaatcatg gacgacttcg 6180 tcagagcagt ggagcggcac ttgatcagtt caggggctct ggaagctagg ctgtccggcc 6240 aacacccatt tgccttcttg tgtttgtctc ttactagagc gatgtgatga gacagacggg 6300 tgatagataa tacggtgata ccctgcacct tacggtagga ttgctatgta atgtactact 6360 aacttttact ttttccatct cacaaagact ctatttttta gttttttaat ataaggtttg 6420 actcttcatc ttatttaaaa ttttatatgg ttagcacttt tattattatt agatgataaa 6480 acattaatag tactttgttt gtgtgtattt attttttatt tttttaaata agacgtgcag 6540 tcaaaaattg gacataaaac tcaaaggtaa agtctttgtg ggacggaggg tgtaatacga 6600 aactctctgt gaacaatgga gcaagagaaa aagggatatt aattattgac aactatataa 6660 gatcacgcaa caggtttgcc aataaatcag tagtggagag atcaagtaat caagtttcag 6720 tggagagata aatctgatgg acaatgtaaa gaacagataa aattattggc caagatttac 6780 agtgtcaaaa gtacaaggct gcaggataat tgatcgaaaa gccgggttta ttattacata 6840 tttatcaagg aacaaaattg ttggttatta ttatcaaacg agaggaaggg aatcgagaag 6900 aggaaggagc tccccatcca tggttcgata gactcgagtg agtcaaggaa atagaactgg 6960 ttaattaatt ggattcatgc cattataaat ttgctagttc gaaaaaatac cattattatt 7020 atttgtacaa ctgtaacatt tttaaaattg gatccataac atgccgtaca cttgatacac 7080 attttgaatc atttcttatc cactgtattt atatatggac ccaaatatcc gtatcttcta 7140 cagctcaaaa actgtggtta aatttcggta aacatagtta tacatacaag ggaaaaaggg 7200 ggaaattggt aaaagaaatt tagaaaatgg aaaaataata aaaaagaaag ggcggtggag 7260 cggtcgctat ccatgatgac tacccgagcc ggccatccca tccccaccgt cgttggcatc 7320 gacggctttt aactatttac cacttttaga tttggcaaac aactatttgc catctctatc 7380 tcaatgacat gtgggtccac atgtatttat tacatgtggc ttcagtggca aaaagttaat 7440 tgtcacatct aaatgtggca aatggttaaa tatttctctc tgctaagctc attgttgttg 7500 tcggaccatg cagtgcagct acgcttcttg tcaccgttgg ccatgtcacc tctgtcagac 7560 cgcacgtcgt agctcaacac cagacccgat accgtcgtct ccactagatc tgcttgagct 7620 gagctcaccg ccaccggtgc tggatcagct cggatagcac gcaagttggc gctaaggtca 7680 tcgtcgtcgc caggccgcgc atcgctactg ccgggatttg gggaatttgg agcttcggtt 7740 gctgggattt gggtctatgg ttaagagagg attggcagga gtacaaaaaa aaatattaca 7800 caaaaaagat tcaaaacatg tatcaagtgt ataacatggc atagatctaa tttcacgagt 7860 gttgcaatgg aatggttata aatagagaat tgtaactagt aagctatatt ttccaaagta 7920 gtaaatttat tgtggcatgt atccaataat cttttttttt tgagacgatg tatccaataa 7980 tctgattcac ttgagatggc caaaactgtc ctgattatgt tgctc 8025 <210> 2 <211> 1113 <212> DNA <213> Oryza sativa <400> 2 atggccacca actccaaaca gagtgtgcat atgaatccag ggcaaggtga aacgagctat 60 gctcagaact cagcacttca gaaaacagcc caggatagga tgaagactct gatagaggag 120 gcagtcacag gcttatgcac aagctcatgt cctcacccaa agaacatggt gatcgcggac 180 ttgggctgct cctccggacc aaacgcgctc acgctcgtct ctgccgccgt cgacgccatc 240 caccgctact gcgcgcagca cgagcagctg ccgccggaga tgtgcgtgct cctgaacgat 300 ctgcctgaca acgatttcaa cgccgtggcg aagagcttgg acacactgaa gcacagtggt 360 gatgaagcgc tcgctcgccc tactgctgtg atcaccggta tggtacctgg gtcgttctac 420 gagaggctat tcgctcgtgg ctccttgcat ttggtttgct cggctaacag cttgcattgg 480 ttatccgagg cacctgaaga tctcaagaag agtagaatcc cgatgcatga tagtgatgaa 540 cagttgagat catcaaggca tcaaattgtc gcggattctt atgctcggca gtttaggaag 600 gatttcatgc gattcttgag cttgagggca caggagattg tccctggagg ccgaatggtt 660 gtctcactgt tggtcaagcg ctcggacaaa cctgacactg agttaattca accatggacg 720 ccggcggtga cggctttaag tgacatggca ttaaggggtg ttatcagcaa agaaaagctt 780 gactccttct acattccctt gtgctgccct atggacagtg aagttaacaa catcatagag 840 gaggagggtt catttgaggt caacaagatg atgatgcacg atccgtatga cggcacgggc 900 aaagccttgc tcgatctgaa gatggtagcc ttgcgcgtaa gagctgtatt tgagccaata 960 atagtgcagc attttgcagc gtcagatgaa atcatggacg acttcgtcag agcagtggag 1020 cggcacttga tcagttcagg ggctctggaa gctaggctgt ccggccaaca cccatttgcc 1080 ttcttgtgtt tgtctcttac tagagcgatg tga 1113 <210> 3 <211> 370 <212> PRT <213> Oryza sativa <400> 3 Met Ala Thr Asn Ser Lys Gln Ser Val His Met Asn Pro Gly Gln Gly   1 5 10 15 Glu Thr Ser Tyr Ala Gln Asn Ser Ala Leu Gln Lys Thr Ala Gln Asp              20 25 30 Arg Met Lys Thr Leu Ile Glu Glu Ala Val Thr Gly Leu Cys Thr Ser          35 40 45 Ser Cys Pro His Pro Lys Asn Met Val Ile Ala Asp Leu Gly Cys Ser      50 55 60 Ser Gly Pro Asn Ala Leu Thr Leu Val Ser Ala Ala Val Asp Ala Ile  65 70 75 80 His Arg Tyr Cys Ala Gln His Glu Gln Leu Pro Pro Glu Met Cys Val                  85 90 95 Leu Leu Asn Asp Leu Pro Asp Asn Asp Phe Asn Ala Val Ala Lys Ser             100 105 110 Leu Asp Thr Leu Lys His Ser Gly Asp Glu Ala Leu Ala Arg Pro Thr         115 120 125 Ala Val Ile Thr Gly Met Val Pro Gly Ser Phe Tyr Glu Arg Leu Phe     130 135 140 Ala Arg Gly Ser Leu His Leu Val Cys Ser Ala Asn Ser Leu His Trp 145 150 155 160 Leu Ser Glu Ala Pro Glu Asp Leu Lys Lys Ser Arg Ile Pro Met His                 165 170 175 Asp Ser Asp Glu Gln Leu Arg Ser Ser Arg His Gln Ile Val Ala Asp             180 185 190 Ser Tyr Ala Arg Gln Phe Arg Lys Asp Phe Met Arg Phe Leu Ser Leu         195 200 205 Arg Ala Gln Glu Ile Val Gly Gly Arg Met Val Val Ser Leu Leu     210 215 220 Val Lys Arg Ser Asp Lys Pro Asp Thr Glu Leu Ile Gln Pro Trp Thr 225 230 235 240 Pro Ala Val Thr Ala Leu Ser Asp Met Ala Leu Arg Gly Val Ile Ser                 245 250 255 Lys Glu Lys Leu Asp Ser Phe Tyr Ile Pro Leu Cys Cys Pro Met Asp             260 265 270 Ser Glu Val Asn Asn Ile Ile Glu Glu Glu Gly Ser Ser Phe Glu Val Asn         275 280 285 Lys Met Met Met His Asp Pro Tyr Asp Gly Thr Gly Lys Ala Leu Leu     290 295 300 Asp Leu Lys Met Val Ala Leu Arg Val Val Ala Val Phe Glu Pro Ile 305 310 315 320 Ile Val Gln His Phe Ala Ala Ser Asp Glu Ile Met Asp Asp Phe Val                 325 330 335 Arg Ala Val Glu Arg His Leu Ile Ser Ser Gly Ala Leu Glu Ala Arg             340 345 350 Leu Ser Gly Gln His Pro Phe Ala Phe Leu Cys Leu Ser Leu Thr Arg         355 360 365 Ala Met     370 <210> 4 <211> 1361 <212> DNA <213> Artificial Sequence <220> <223> promoter <400> 4 gatccccaac atggtggagc acgacactct cgtctactcc aagaatatca aagatacagt 60 ctcagaagac cagagggcta ttgagacttt tcaacaaagg gtaatatcgg gaaacctcct 120 cggattccat tgcccagcta tctgtcactt catcgaaagg acagtagaaa aggaagatgg 180 cttctacaaa tgccatcatt gcgataaagg aaaggctatc gttcaagatg cctctaccga 240 cagtggtccc aaagatggac ccccacccac gaggaacatc gtggaaaaag aagacgttcc 300 aaccacgtct tcaaagcaag tggattgatg tgatatctag atccccaaca tggtggagca 360 cgacactctc gtctactcca agaatatcaa agatacagtc tcagaagacc agagggctat 420 tgagactttt caacaaaggg taatatcggg aaacctcctc ggattccatt gcccagctat 480 ctgtcacttc atcgaaagga cagtagaaaa ggaagatggc ttctacaaat gccatcattg 540 cgataaagga aaggctatcg ttcaagatgc ctctaccgac agtggtccca aagatggacc 600 cccacccacg aggaacatcg tggaaaaaga agacgttcca accacgtctt caaagcaagt 660 ggattgatgt gatatctaga tccccaacat ggtggagcac gacactctcg tctactccaa 720 gaatatcaaa gatacagtct cagaagacca gagggctatt gagacttttc aacaaagggt 780 aatatcggga aacctcctcg gattccattg cccagctatc tgtcacttca tcgaaaggac 840 agtagaaaag gaagatggct tctacaaatg ccatcattgc gataaaggaa aggctatcgt 900 tcaagatgcc tctaccgaca gtggtcccaa agatggaccc ccacccacga ggaacatcgt 960 ggaaaaagaa gacgttccaa ccacgtcttc aaagcaagtg gattgatgtg atatctagat 1020 ccccaacatg gtggagcacg acactctcgt ctactccaag aatatcaaag atacagtctc 1080 agaagaccag agggctattg agacttttca acaaagggta atatcgggaa acctcctcgg 1140 attccattgc ccagctatct gtcacttcat cgaaaggaca gtagaaaagg aagatggctt 1200 ctacaaatgc catcattgcg ataaaggaaa ggctatcgtt caagatgcct ctaccgacag 1260 tggtcccaaa gatggacccc cacccacgag gaacatcgtg gaaaaagaag acgttccaac 1320 cacgtcttca aagcaagtgg attgatgtga tatctagatc c 1361 <210> 5 <211> 8275 <212> DNA <213> Artificial Sequence <220> <223> T-DNA <400> 5 gtttacccgc caatatatcc tgtcaaacac ggatccgagg taccaggtac caggtgagtt 60 ccattcttac taccacggtg ctattttttt tgctatgtgg ctaattacat gactaacttg 120 gggtgctaaa tcttacaggt tatatgcagg ttatatgcag gtcccgggta ggtcagtccc 180 ttatgttacg tcctgtagaa accccaaccc gtgaaatcaa aaaactcgac ggcctgtggg 240 cattcagtct ggatcgcgaa aactgtggaa ttgatcagcg ttggtgggaa agcgcgttac 300 aagaaagccg ggcaattgct gtgccaggca gttttaacga tcagttcgcc gatgcagata 360 ttcgtaatta tgcgggcaac gtctggtatc agcgcgaagt ctttataccg aaaggttggg 420 caggccagcg tatcgtgctg cgtttcgatg cggtcactca ttacggcaaa gtgtgggtca 480 ataatcagga agtgatggag catcagggcg gctatacgcc atttgaagcc gatgtcacgc 540 cgtatgttat tgccgggaaa agtgtacgta tcaccgtttg tgtgaacaac gaactgaact 600 ggcagactat cccgccggga atggtgatta ccgacgaaaa cggcaagaaa aagcagtctt 660 acttccatga tttctttaac tatgccggaa tccatcgcag cgtaatgctc tacaccacgc 720 cgaacacctg ggtggacgat atcaccgtgg tgacgcatgt cgcgcaagac tgtaaccacg 780 cgtctgttga ctggcaggtg gtggccaatg gtgatgtcag cgttgaactg cgtgatgcgg 840 atcaacaggt ggttgcaact ggacaaggca ctagcgggac tttgcaagtg gtgaatccgc 900 acctctggca accgggtgaa ggttatctct atgaactgtg cgtcacagcc aaaagccaga 960 cagagtgtga tatctacccg cttcgcgtcg gcatccggtc agtggcagtg aagggccaac 1020 agttcctgat taaccacaaa ccgttctact ttactggctt tggtcgtcat gaagatgcgg 1080 acttacgtgg caaaggattc gataacgtgc tgatggtgca cgaccacgca ttaatggact 1140 ggattggggc caactcctac cgtacctcgc attaccctta cgctgaagag atgctcgact 1200 gggcagatga acatggcatc gtggtgattg atgaaactgc tgctgtcggc tttaacctct 1260 ctttaggcat tggtttcgaa gcgggcaaca agccgaaaga actgtacagc gaagaggcag 1320 tcaacgggga aactcagcaa gcgcacttac aggcgattaa agagctgata gcgcgtgaca 1380 aaaaccaccc aagcgtggtg atgtggagta ttgccaacga accggatacc cgtccgcaag 1440 tgcacgggaa tatttcgcca ctggcggaag caacgcgtaa actcgacccg acgcgtccga 1500 tcacctgcgt caatgtaatg ttctgcgacg ctcacaccga taccatcagc gatctctttg 1560 atgtgctgtg cctgaaccgt tattacggat ggtatgtcca aagcggcgat ttggaaacgg 1620 caggaaggt actggaaaaa gaacttctgg cctggcagga gaaactgcat cagccgatta 1680 tcatcaccga atacggcgtg gatacgttag ccgggctgca ctcaatgtac accgacatgt 1740 ggagtgaaga gtatcagtgt gcatggctgg atatgtatca ccgcgtcttt gatcgcgtca 1800 gcgccgtcgt cggtgaacag gtatggaatt tcgccgattt tgcgacctcg caaggcatat 1860 tgcgcgttgg cggtaacaag aaagggatct tcactcgcga ccgcaaaccg aagtcggcgg 1920 cttttctgct gcaaaaacgc tggactggca tgaacttcgg tgaaaaaccg cagcagggag 1980 gcaaacaatg aatcaacaac tctcctggcg caccatcgtc ggctacagcc tcgggaattg 2040 ctaccgagct cgaatttccc cgatcgttca aacatttggc aataaagttt cttaagattg 2100 aatcctgttg ccggtcttgc gatgattatc atataatttc tgttgaatta cgttaagcat 2160 gtaataatta acatgtaatg catgacgtta tttatgagat gggtttttat gattagagtc 2220 ccgcaattat acatttaata cgcgatagaa aacaaaatat agcgcgcaaa ctaggataaa 2280 ttatcgcgcg cggtgtcatc tatgttacta gatcgggaat taattcatcg ataggctagt 2340 catggtgact gtacgttgta agtgcagcaa actgccgacg cgatgcaaac tgtacacgtt 2400 aacatgccac tcacctggaa cgcacaatgg ccactaggtg cggccgtagt gtggatttca 2460 aagagagaga gagagagaga gagctaatca cgtaaacgta aacacagcag atagcagaga 2520 tgttgattag gcaaaacagt ataaaagcca atccaataaa ctacatttag cgaagtgcta 2580 tactaatgca ctaataacga actgttcttt tcttaagatc ggagccagta atgggttgtc 2640 agcaggagaa gcacgtaaac cttgaaacat actaagttcc acagtcgaga gtaaaccgta 2700 atcaacacaa gaaacaaaca taaaattgaa caaacgcgca tattataagt gacgaagcgg 2760 tctcacataa aacagggcac acaggttaca acaacgaggg ttgtaagccc attaagcccc 2820 aaacatcaga tcaccacaag caaatgtctc gaagacacac gcacacggca acaggataac 2880 tccacactgg cagatcatgg gatagcagca gttatcaatc aggccttgac acacagaaca 2940 tcaagccccc agacgacgac gactcctcta gatcccggtc ggcatctact ctattccttt 3000 gccctcggac gagtgctggg gcgtcggttt ccactatcgg cgagtacttc tacacagcca 3060 tcggtccaga cggccgcgct tctgcgggcg atttgtgtac gcccgacagt cccggctccg 3120 gatcggacga ttgcgtcgca tcgaccctgc gcccaagctg catcatcgaa attgccgtca 3180 accaagctct gatagagttg gtcaagacca atgcggagca tatacgcccg gagccgcggc 3240 gatcctgcaa gctccggatg cctccgctcg aagtagcgcg tctgctgctc catacaagcc 3300 aaccacggcc tccagaagaa gatgttggcg acctcgtatt gggaatcccc gaacatcgcc 3360 tcgctccagt caatgaccgc tgttatgcgg ccattgtccg tcaggacatt gttggagccg 3420 aaatccgcgt gcacgaggtg ccggacttcg gggcagtcct cggcccaaag catcagctca 3480 tcgagagcct gcgcgacgga cgcactgacg gtgtcgtcca tcacagtttg ccagtgatac 3540 acatggggat cagcaatcgc gcatatgaaa tcacgccatg tagtgtattg accgattcct 3600 tgcggtccga atgggccgaa cccgctcgtc tggctaagat cggccgcagc gatcgcatcc 3660 atggcctccg cgaccggctg cagaacagcg ggcagttcgg tttcaggcag gtcttgcaac 3720 gtgacaccct gtgcacggcg ggagatgcaa taggtcaggc tctcgctgaa ttccccaatg 3780 tcaagcactt ccggaatcgg gagcgcggcc gatgcaaagt gccgataaac ataacgatct 3840 ttgtagaaac catcggcgca gctatttacc cgcaggacat atccacgccc tcctacatcg 3900 aagctgaaag cacgagattc ttcgccctcc gagagctgca tcaggtcgga gacgctgtcg 3960 aacttttcga tcagaaactt ctcgacagac gtcgcggtga gttcaggctt tttcatatct 4020 cattgccccc cgggatccgt cgagtcagcc tgaaaggaca aaatacatgt tagcgcctta 4080 gtggtacatt attatttcag tacagcaaga taacacaatt caaaagactg acccataata 4140 aataactagt cctcaattta aaatttgagt tcctaaatag acatctatga atatgctgta 4200 catcggcact acagaaaata cgattcccaa taattgaaca attgtacttt atttagttgt 4260 tactacaaca atggaagata caagatcgtt tcaaaactac catacatgca tggagtattt 4320 gttccacaga tctggaaaaa acagatctga cgggcagtgt caccaaacac tagacatatg 4380 tatttgtatt aggtggatga cgtgtacaaa catgactacc agatctagaa ttagaacgcg 4440 gcgtgctttg gaaatgttaa gtagatccaa atacatcggt aacaaatgaa catattcata 4500 tgacatagct gtaaaacatc atgatctatc atatcaacta gaggggatct cggcatgtat 4560 cattctattg catctagaac catagcattt ccatgtgaca aacaattttg aaacatacat 4620 tgctcagatc taccataaga acaccatgtt catgacagca tcgaccatga tttccacatt 4680 taacagcatc ccgaggtcag atccagtgtt tagaactatc atttcagcaa aattcacaaa 4740 ataaatcctc caattcgcct acatatatcc attccacgca tcctaggacc agatccacca 4800 taccagtata cacgaacact aaacatttca tcagatccgc taactacgac agagaaaacg 4860 cagatccaga caaccagatc caccgacaaa taaacacagc cccccacatc caacaaccgc 4920 gaatccaccc agatctgacc cagaagcagg cacgaagaac acggggggtg agggagatgg 4980 gacgcggatc caagcttggc ggcggattgg gttgatgctg cgacggcggc ggagaaggga 5040 gagaggggag aggaggaaaa gcgcggaggc gcggagagag gcgagtacga agacgccttt 5100 ctctgcgttg tctcggctta gggtttgcga tccccgcact ccgcccgttt tatagggcca 5160 gacagcctgg acctctcagg agcggaagcg aagggatggg ggagtttttc gttttacccc 5220 ctttgacgtc ttcaaaattc actcgcatct cacatcgtgc ctgcaaaggt cgggtggttg 5280 gatcatatcg agaatattta ataagttagg gagctttctg ataatatcca ccgcaaggag 5340 gccatttcgt ttgcactcgg ttgaagtcca tacccgcctc tcgttagttt cactgacagg 5400 tgggtccaga ggcttccttc gtgtgcggtt agcgagtgca cgcgtcgctc caagaaaaag 5460 cagtaggtga cctggccacc tcgttgtagg tgcgaccgat gcagacgtcc cgcttgccgg 5520 tgggcccacg ttacagctgg gtcccacata tcggtgggtg cgatgagttg ttcggtgcgt 5580 aggagtagat tggatcccaa cgggtggttg cgtcgtcggc ggccctgccg attttatttt 5640 cgatttttga aatgcgagag cgggagaacg gcaccgttgg cttggctgtg gatgccgtta 5700 gccgcaccgg actattcggc ccagttccat tttggcccaa cttgaaacag cgcggagcac 5760 aagacggggg agcccattta ggcccgtaat ctcagccctg tagaaagtcc gtgtcgttgc 5820 gatggaccag aaagcccata aaccttggag gcgttctctg ctgggaataa aataagttca 5880 cactctgtcc ctcaacttta cgtcgagttt gtttgacatc gctaatgccc agtaccagaa 5940 atcttggata caaataaggt ttgatataaa gtggcacgtg gtgaatggca tcgtttgaaa 6000 aatcttcaga tcgagcgggc ttcacaagcc gtaagtgcaa gtgccaaaat ttgtgaagaa 6060 aattacttcc ttcatttcac aatgtaagtt attttagcat tttccacatt catattgata 6120 tcaataaatt tagatagata tatatgtctg aattcgtacc gttaacgagc tctagaagct 6180 tcagtactac gcgtctcgag gcctattatt gaagcattta tcagggttat tgtctcatga 6240 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc 6300 cccgaaaagt gccacctgac gtctaagaaa ccattattat catgacatta acctataaaa 6360 ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct 6420 gacacatgca gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac 6480 aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg 6540 catcagagca gattgtactg agagtgcacc atatgcggtg tgaaataccg cacagatgcg 6600 taaggagaaa ataccgcatc aggcgccatt cgccattcag gctgcgcaac tgttgggaag 6660 ggcgatcggt gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa 6720 ggcgattaag ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa acgacggcca 6780 gtgaattcac tagtgattgc tctagacact gatagtttcg gatctagata tcacatcaat 6840 ccacttgctt tgaagacgtg gttggaacgt cttctttttc cacgatgttc ctcgtgggtg 6900 ggggtccatc tttgggacca ctgtcggtag aggcatcttg aacgatagcc tttcctttat 6960 cgcaatgatg gcatttgtag aagccatctt ccttttctac tgtcctttcg atgaagtgac 7020 agatagctgg gcaatggaat ccgaggaggt ttcccgatat taccctttgt tgaaaagtct 7080 caatagccct ctggtcttct gagactgtat ctttgatatt cttggagtag acgagagtgt 7140 cgtgctccac catgttgggg atctagatat cacatcaatc cacttgcttt gaagacgtgg 7200 ttggaacgtc ttctttttcc acgatgttcc tcgtgggtgg gggtccatct ttgggaccac 7260 tgtcggtaga ggcatcttga acgatagcct ttcctttatc gcaatgatgg catttgtaga 7320 agccatcttc cttttctact gtcctttcga tgaagtgaca gatagctggg caatggaatc 7380 cgaggaggtt tcccgatatt accctttgtt gaaaagtctc aatagccctc tggtcttctg 7440 agactgtatc tttgatattc ttggagtaga cgagagtgtc gtgctccacc atgttgggga 7500 tctagatatc acatcaatcc acttgctttg aagacgtggt tggaacgtct tctttttcca 7560 cgatgttcct cgtgggtggg ggtccatctt tgggaccact gtcggtagag gcatcttgaa 7620 cgatagcctt tcctttatcg caatgatggc atttgtagaa gccatcttcc ttttctactg 7680 tcctttcgat gaagtgacag atagctgggc aatggaatcc gaggaggttt cccgatatta 7740 ccctttgttg aaaagtctca atagccctct ggtcttctga gactgtatct ttgatattct 7800 tggagtagac gagagtgtcg tgctccacca tgttggggat ctagatatca catcaatcca 7860 cttgctttga agacgtggtt ggaacgtctt ctttttccac gatgttcctc gtgggtgggg 7920 gtccatcttt gggaccactg tcggtagagg catcttgaac gatagccttt cctttatcgc 7980 aatgatggca tttgtagaag ccatcttcct tttctactgt cctttcgatg aagtgacaga 8040 tagctgggca atggaatccg aggaggtttc ccgatattac cctttgttga aaagtctcaa 8100 tagccctctg gtcttctgag actgtatctt tgatattctt ggagtagacg agagtgtcgt 8160 gctccaccat gttggggatc ctctagagtc gagaattcag tacattaaaa acgtccgcaa 8220 tgtgttatta agttgtctaa gcgtcaattt gtttacacca caatatatcc tgcca 8275 <210> 6 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> SAMT 14_F primer <400> 6 gtcaacattc gcagtcaaca 20 <210> 7 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> SAMT 14_R primer <400> 7 gtgggtatgt gcagggatta 20 <210> 8 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> NGUS1 primer <400> 8 aacgctgatc aattccacag 20 <210> 9 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> Enhancer-F <400> 9 gatccccaac atggtggagc 20 <210> 10 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> Enhancer-R <400> 10 ggatctagat atcacatcaa 20 <210> 11 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> RT-SAMT 14_F <400> 11 cagcttgcat tggttatccg 20 <210> 12 <211> 19 <212> DNA <213> Artificial Sequence <220> <223> RT-SAMT 14_R <400> 12 caactcaccc ttaatgcca 19

Claims (19)

As a transgenic rice containing a recombinant vector for promoting blight resistance to rice blast,
Wherein the recombinant vector comprises an expression inducing sequence that enhances the activity of the LOC_Os06g20790 gene comprising the nucleotide sequence of SEQ ID NO: 2 encoding a methyltransferase.
Transgenic rice.
The transgenic rice of claim 1, wherein the LOC_Os06g20790 gene comprises the nucleotide sequence of SEQ ID NO: 1.
2. The transgenic rice of claim 1, wherein the LOC_Os06g20790 gene is encoded by a methyltransferase comprising the amino acid sequence of SEQ ID NO: 3.
delete The transgenic rice plant according to claim 1, wherein the rice is Dongjin rice.
delete 2. The transformed rice according to claim 1, wherein the expression inducing sequence consists of the nucleotide sequence of SEQ ID NO: 4.
delete A recombinant vector for promoting resistance to blight of blight of rice blast in rice comprising an expression inducing sequence for enhancing the activity of LOC_Os06g20790 gene comprising the nucleotide sequence of SEQ ID NO: 2 encoding a methyltransferase.
10. The recombinant vector according to claim 9, wherein the LOC_Os06g20790 gene comprises the nucleotide sequence of SEQ ID NO: 1.
10. The recombinant vector according to claim 9, wherein the LOC_Os06g20790 gene is encoded by a methyltransferase comprising the amino acid sequence of SEQ ID NO: 3.
10. The recombinant vector according to claim 9, wherein the expression inducing sequence comprises the nucleotide sequence of SEQ ID NO: 4.
delete delete (a) infecting rice with Agrobacterium to which the recombinant vector of any one of claims 9 to 12 has been introduced; And
(b) callusing cells of infected rice in step (a) to obtain cells of transformed rice,
A method for producing transgenic rice with improved blight resistance to blight.
The method according to claim 15, further comprising (c) inducing differentiation of the callus produced in step (b) into root and adult.
delete delete 1. A method for promoting resistance to blight of blight of rice blast, comprising the step of enhancing the activity of the LOC_Os06g20790 gene comprising the nucleotide sequence of SEQ ID NO: 2 encoding a methyltransferase.

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