KR100781076B1 - Osglu2 gene for increasing disease resistant and crop yield - Google Patents

Abstract

A beta-1,3-glucanase(OsGlu2) gene is provided to promote the growth of rice by increasing the resistance against diseases such as rice blast disease, thereby being effectively utilized for breeding a new species able to enlarge the quantity. A method for increasing the disease-resistance of a plant comprises a step of transforming a plant using a recombinant plant expression vector including a cDNA gene encoding beta-1,3-glucanase(OsGlu2). A plant prepared by the method is characterized in that it has the increased disease resistance, wherein the disease is a rice blast disease.

Description

OsGlu2 Gene for Increasing Disease Resistant and Crop Yield

1 is a protein after inoculation of the plaque from the rice leaves to confirm the protein expression by rice blast, Figure 1 (A) is 8 types of defense-related proteins through the 2-DE analysis, Figure 1 (B) Are two β-1,3-glucanase (OsGlu1, OsGlu2) genes identified by MALDI-TOF analysis.

Figure 2 is a cloning of the identified β-1,3-glucanase (OsGlu2) from the cDNA library, Figure 2 (A) is a specific primer combination of OsGlu2, Figure 2 (B) DNA fragments amplified by PCR It is shown.

FIG. 3 (A) shows a comparison of the amino acid sequences of the well-known β-1,3-glucanase from two types of β-1,3-glucanase (OsGlu1, OsGlu2) identified from rice and other plants (Arabidopsis and tobacco). , Figure 3 (B) shows their phylogenetic relationship.

Figure 4 shows the Genomic DNA gel blot analysis of the OsGlu2 gene.

Figure 5 is to confirm and produced the expression of the recombinant protein of OsGlu2 using a protein expression vector with histag.

Figure 6 shows the Northern blot analysis of the OsGlu2 gene according to plant hormones (JA, SA, ABA) and stress (CN, CHX, ST).

Figure 7 shows the change in the expression pattern of OsGlu2 gene by treating the blasts from rice leaves by time, Figure 7 (A) is Northern blot analysis confirmed from RNA samples treated with blasts by time from rice leaves, Figure 7 (B ) Is Western blot analysis from protein samples.

8 is an over-expression of the OsGlu2 gene, an RNAi rice transformed crop was produced by the Gateway method, and FIG. 8 (A) is a cloning method through the Gateway method of the OsGlu2 gene to produce a rice transformed crop. , Figure 8 (B) is to prepare a rice transformed crop using the over-expression vector OsCcl and RNAi vector P-Ai which is the basic vector of the present invention.

9 is a process of producing transformed rice by Agrobacterium transformation method.

10 is a constituent of the medium used in accordance with the various conditions and growth process used in the production of transformed rice.

Figure 11 is the expression pattern of each line of rice transformant by Northern blot analysis and Western blot analysis.

Figure 12 is a photograph of the comparison of the height of the transformed rice plants (OsCcl :: OsGlu2 and Ai :: OsGlu2) and control Nakdong rice (WT).

Figure 13 is a comparison of Stem of each transformed rice plant (OsCcl :: OsGlu2 and Ai :: OsGlu2) and control Nakdong rice (WT).

Figure 14 is a comparative photograph according to the deletion of the rice transformant of Ai :: OsGlu2 and control Nakdong rice (WT).

Figure 15 is a comparison table according to the quantity components of each transformed rice plants (OsCcl :: OsGlu2 and Ai :: OsGlu2) and control Nakdong rice (WT).

FIG. 16 is a comparison of resistance after 48 hours of rice blast treatment to Na: dong rice (WT) leaves of Ai :: OsGlu2 and control.

The present invention relates to the OsGlu2 gene useful for improving rice blast resistance and yield, β-1,3-glucanase, which is a rice leaf blast resistance protein that directly protects against diseases of plants (rice), It relates to a gene (OsGlu2) encoding it. The gene OsGlu2 according to the present invention may be effectively used to cultivate new varieties that can increase rice resistance by increasing resistance to diseases such as rice blasts and consequently increase yield.

As the plant grows, it is exposed to various environmental stresses, which have a great influence on the growth, development, and productivity of the plant. Plants actively combat environmental stress by operating various defenses to survive. A hydrolytic enzyme, β-1,3-glucanases, a member of pathogen-related genes (PRs), a well-known member of the plant defense system, constitutes the cell wall of most filamentous fungi. By reducing the substance β-1,3-glucan (glucan) is a protective action. In addition, β-1,3-glucanase affects the physiological or developmental stages of microsporogenesis of the crop itself, germination of pollen or seeds, and fertilization. Crazy

Control of diseases and pests is a very important factor in the cultivation of rice. Especially Pyricularia oryzae ) is an important disease that determines growth and yield. Until now, chemical fungicides (pesticides) and control methods have been developed and used, but many chemicals have to overcome such as toxicity, safety (residual problems) and control effects. It has a flaw. To date, molecular biology efforts have been attempted in some plant groups to introduce recombinant genes to make plants resistant to pests.

The present invention was able to identify two types of β-1,3-glucanase (OsGlu1, OsGlu2) during the analysis of disease-related proteins with optimal protection against rice blast. Among them, β-1,3-glucanase (OsGlu2) was cloned, and the antibody was produced to be used for its functional studies. By using this gene, we will contribute to the development of new varieties that can be expected to increase the resistance to diseases such as blast diseases and increase yields due to growth promotion and yield components.

Accordingly, an object of the present invention is to identify proteins having a protective resistance against leaf blasts of rice ( Oryza sativa ), to clone the genes and to identify their functions, among which analyzes the proteins related to defense against rice leaf blasts. The purpose is to provide the gene.

In order to achieve the above object, the present invention provides β-1,3-glucanase, a gene encoding the same (OsGlu2), which is a rice leaf blast resistant protein that directly protects against diseases of plants (rice). do.

More specifically, through the present invention, β-1,3-glucanase (OsGlu2) having an amino acid sequence represented by SEQ ID NO: 1 (see FIG. 3 (A)) and resistant to plant diseases, and this specific Provided is a cDNA gene comprising a nucleotide sequence represented by SEQ ID NO: 2 encoding an enzyme protein.

The gene is SEQ ID NO: encoding β-1,3-glucanase, characterized in that the expression is induced by the plant hormone jasmonic acid (JA) or the expression is suppressed by the plant hormone absic acid (ABS). CDNA gene comprising the nucleotide sequence represented by 2.

Hereinafter, the content of the present invention will be described in more detail with reference to Examples. However, these examples are only presented to understand the content of the present invention, the scope of the present invention should not be construed as being limited to these embodiments.

Example 1 Identification of β-1,3-glucanase in Defense Related Proteins against Rice Blast

After inoculating the plaque from the rice leaves, the proteins were extracted to identify defense-related proteins, mainly on the proteins with increased protein expression by the rice plaque.

Protein Extraction, 2-DE Assay, MALDI-TOF Identification

Protein Extraction: Samples of rice blast fungus (M. grisea) were inoculated for 48 hours and 72 hours on the leaves of Jinheung rice grown for 4-5 weeks in a greenhouse at 20-30 ° C. The promotion rice which did not treat blasting was used as a control. Peg (PEG) precipitation was used to separate proteins from this. This method is an effective method for analyzing relatively low-expression proteins by effectively eliminating rubisco proteins, which are significantly expressed in green plant protein analysis. The sampled rice leaves were ground with liquid nitrogen, and then 2 g of each sample was treated with Mg / NP-40 buffer (0.5 M Tris-HCl (pH 8.3), 2% v / v NP-40, 20 mM MgCl2, 2% v). / v b-mercaptoethanol, 1 mM phenyl methyl sulfonyl fluoride, and 1% w / v polyvinyl polypyrrolidone) were centrifuged at 12000xg for 15 minutes, the supernatant was extracted, and 50% w / v PEG was added to give a final sample supernatant concentration. 15% w / v PEG. The supernatant was left on ice for 30 minutes and then centrifugation (12000xg) for 15 minutes to precipitate supernatant protein extracts obtained from acetone.

2-DE Assay: 15% PEG supernatant protein extract was analyzed as 2-DE. IEF (1-DE) gel (4.5% w / v acrylamide solution, 9.5 M urea, 2% v / v NP-40, and 2.5% v / v ampholytes (pH 310: pH 58: pH 58: pH 46.5 = 1: 3.5: 2.5) is hardened for 6 hours Each sample (150ug for silver stained protein and 500ug for CBB stained protein) is dissolved in sample buffer and loaded into IEF gel for 12 hours at 600V, 6 hours at 800V, and 1000V at 1000V. 3 hours each gel is placed in a 20 mL screw-cap tube and 5 mL of equilibration buffer (10% v / v glycerol, 2.5% w / v SDS, 125 mM Tris-HCl (pH 6.8), 5% v / v Add β-mercaptoethanol, and 0.1 mg / mL bromophenol blue) at room temperature for 30 minutes, equilibrate the gel with Low-Melting Agarose (low-melting agarose: 0.2g, 4 × buffer5ml, H2O: 15ml, 10% SDS: 200ul) and put it on the pre-hardened 2-DE gel, SDS-PAGE (second dimension) overnight at 40 ~ 50 V. After the dye is removed the next day, fix it sufficiently (50% methanol, 10% acetic acid, 40% dd H2O). Silver stain (Pretreat, Impregnate, Develop). Remove aine protein spots from the gel, wash (50% v / v ACN in 0.1 M NH4HCO3), and vacuum dry the gel fragments in 10 mM DTT solution (55 mM iodoacetamide in 0.1 M NH4HCO3) in 55 ° C for 45 minutes. Add 0.1 M NH4HCO3, cool on ice, wash with (50% ACN in 0.1 M NH4HCO3), add (10 mL digestion buffer containing 25 mM NH4HCO3 and 12.5 ng / mL trypsin), and dry. As shown in FIG. 1 (A), eight types of defense-related proteins were identified through 2-DE analysis.

MALDI-TOF Identification: All proteins were run using a Voyager-DE STR MALDI-TOF mass spectrometer (PerSeptive Biosystems, Framingham, Mass., USA). Figure 1 (B) is to identify two types of β-1,3-glucanase (OsGlu1, OsGlu2) gene by MALDI-TOF analysis.

< Example  2: Identified  β-1,3- glucanase ( Osglu2 Genetic Identification>

Identified  β-1,3- glucanase  ( Osglu2 ) cDNA library from cloning

The identified β-1,3-glucanase (OsGlu2) was identified using a database program of the National Center for Biotechnology Information (NCBI) to identify genes and specific primers were prepared. Β-1,3-glucanase (OsGlu2) was amplified by PCR using gene-specific primers from cDNA gene library obtained by reacting mRNA prepared from rice leaves derived from blasts, and then pGEM-T Easy Vector (Promega, Madison). , WI, USA) and confirmed the sequence by cloning. Figure 2 (A) shows a specific primer combination of OsGlu2.

 OsGlu2 (AB027430) Forward 5'- ATG GGT TTC GCT CCC ATG-3 '(SEQ ID NO: 3)

 OsGlu2 (AB027430) Reverse 5'-TAA GAA ACT AAT GCT GTA CGA-3 '(SEQ ID NO: 4)

DNA markers and OsGlu2 PCR products were loaded from the agarose gel and confirmed to be 1008 bp. Figure 2 (B) shows a DNA fragment amplified by PCR.

Identified  β-1,3- glucanase ( OsGlu1 , Osglu2 Β-1,3- in other plants glucanase  Amino acid sequence Homology  compare

Cloned OsGlu2 is a phylogenetic tree using Neighbor Joining methods of Clustal X program from another identified β-1,3-glucanase (OsGlu1) and other β-1,3-glucanases present in other plants. The comparison was made using (see FIG. 3 (A)). Β-1,3-glucanase genes (AtGLU1 (NCBI Acc. No AY096465) and AtGLU2 (AY096541)) and two β-1,3-glucanase genes NtGLU1 (NCBI Acc. No M20618) ) And NtGLU2 (M20619)) had high similarities to each other, and relatively low, with 62% homology between OsGlu1 (AB027428) and OsGlu2 present in rice (see FIG. 3 (B)). This confirmed that the gene is β-1,3-glucanase (OsGlu2).

Cloning Done Osglu2 of genome On copy  Can be ok

DNA fragment of OsGlu2 cloned into pGEM-T Easy Vector (10ug) was cut for 3-6 hours at 37 ° C using restriction enzymes (EcoR I, Hind III, Xba I) and loaded onto 1% DNA agarose gel. Next, transfer to nylon membrane (GeneScreen Plus, NEN). The membrane was labeled with OsGlu2 probe using [-32P] dCTP (see FIG. 4).

Protein expression and antibodies ( antibody Production

As shown in FIG. 5, expression of the recombinant protein of OsGlu2 was confirmed and produced using a protein expression vector with histag. To construct the antibody, the OsGlu2 gene was cloned using pET28 with histag, a protein expression vector. First, primers were prepared to clone OsGlu2 into pET28 vector.

  OsGlu2 Nde I Forward 5'- GCG CAT ATG ATG GGT TTC GCT CCC ATG-3 '(SEQ ID NO: 5)

  OsGlu2 BamH I Reverse 5'- GCG GGA TCC TTA GAA ACT AAT GCT GTA CGA-3 '(SEQ ID NO: 6)

After recovering the amplified insert by PCR using the prepared primers, the plasmid DNA extracted by ligation to the pGEM-T Easy Vector was cut with Nde I / BamH I restriction enzyme to recover the insert, and the DNA was extracted from the pET28 vector. The insert recovered from OsGlu2 was cut and cloned with Nde I / BamH I restriction enzyme.

To confirm the protein expression, colonies generated by transforming pET28 vector and pET28 :: OsGlu2 into XL-1 blue cells were inoculated into LB broth and grown in IPTG at 37 ° C. The pellet obtained by centrifugation (6000xg) for 5 minutes was added to the protein loading butter, boiled for 10 minutes, centrifugation (6000xg) for 5 minutes, and only the supernatant was loaded on a 12% SDS-polyacrylamide gel. Soluble recombinant protein pET28 :: OsGlu2 and protein markers are also loaded. The gel was removed and stained with comssie brilliant blue R250 to confirm protein expression and OsGlu2 protein size.

To prepare an antibody from the recombinant protein pET28 :: OsGlu2, 1 ~ 2mg / mL protein and freund's complete adiuvants were emulsified 1: 1 or more for 30 minutes. In this way, the rabbit is injected first, secondly after 4 weeks, and thirdly after 2 weeks. Seven days after the third injection, blood is collected and tittered. Two weeks after the third injection, only four (4) injections of protein (antigen) were taken and a week later, blood was collected, and only serum was recovered from the blood.

For plant hormones and rice blast Osglu2 Expression Analysis of

Jasmonic acid (JA, 250uM), Salicylic acid (SA, 5mM), Abscisic acid (ABA, 200uM) and cantharidin (CN, 1uM), Cycolheximide (CHX, 1uM), Staurosporine (ST, 10uM) was treated for 48 hours (+), no treatment (-) RNA was isolated. In addition, RNA and protein were isolated by treating rice blasts (KJ401, incompatible and KJ101, compatible) with time (12, 18, 24, 36, 48, 60, 72 hours).

As shown in FIG. 6, Northern blot analysis of the OsGlu2 gene according to plant hormones (JA, SA, ABA) and stress (CN, CHX, ST) revealed that this gene was induced by JA but expressed by ABA. This was suppressed. FIG. 7 (A) shows Northern blot analysis confirmed from RNA samples treated with blasting from rice leaves by time, and FIG. 7 (B) shows Western blot analysis confirmed from protein samples. The amount of mRNA (Northern blot) and protein (Western blot) expressed was found to be higher in the incompatible reaction earlier than in the compatible reaction.

RNA isolation: Sampled rice leaves are ground with liquid nitrogen, followed by voltex with RNA extraction butter (0.2M tris (pH 9.0), 0.4M Licl, 25mM EDTA, 1% SDS) and phenol for RNA and β mercaptoethanol. (3000xg) 5 minutes to take only the supernatant, then PCI (phenol 25: chroloform 24: isoamine alcohol 1 for RNA) and add 10 minutes to the voltex, centrifugation (7500xg) to separate the supernatant. Add chroloform 1: 1 and centrifugation (7500xg) for 10 minutes, and then take only the supernatant, add isoprophanol (1: 1), add 3M sodium acetate (1/10) and settle for 1 hour at -20 ° C. . After 1 hour, discard the supernatant by centrifugation (7500xg) for 10 minutes, dry only the remaining pellet, add TE solution for RNA to dissolve, and add LiCl so that the final concentration becomes 2M LiCl. After 15 minutes of centrifugation (12000xg), wash with 70% EtOH, and after drying, add TE solution to dissolve it, add RNA in P.C.I solution, chroloform, 100% EtOH and 3M sodium acetate (1/10), and clean the RNA. Finally, store at -20 ° and dissolve in 10 mM tris-Hcl (pH7.5).

Northern blot analysis: RNA samples (20 uM) were loaded onto formaldehyde denaturing agarose gels and transferred to nylon membrane (GeneScreen Plus, NEN). The membrane was labeled with OsGlu2 probe using [-32P] dCTP.

Western blot analysis: Put protein loading butter, boil for 10 minutes, centrifugation (6000xg) for 5 minutes, load only the supernatant onto 12% SDS-polyacrylamide gel, blotting onto PVDF membrane, confirm with Ponceau S, wash with distilled water, and then wash with 5% v / v Cross-link with glutaraldehyde solution. The cross-linked membrane was blocked with 7% w / v nonfat dry milk dissolved in TTBs (50 mM Tris-HCl pH 8.2, 0.1% v / v Tween 20, and 150 mM NaCl) solution for 2 hours and then purified. Add rabbit antibody OsGlu2 (diluted 1: 1000). This antigen-anti-body interaction is for 2 hours and then washed 20 times with TTBs 3 times. Secondary goat anti-rabbit IgG antibody conjugated with horseradish peroxidase diluted 1: 5000 was added to TTBs and immunodetection was performed for 2 hours. The blots are again washed 20 times with TTBs solution three times. Finally, immunoblot signals are detected using ECL (PerkinElmer Life Sciences, Boston, MA, USA).

< Example  3: Osglu2  Gene Production and Growth of Rice Transformant>

Gateway  Through the way Osglu2 Rice transformant vector cloning

Gateway method (Invitrogen) was used to prepare rice transformants. First, primers were prepared using NCBI's database program to prepare adapter PCR primers (see FIG. 8 (A)).

   OsGlu2 12b attB1 5'- GGG GAC AAG TTT GTA CAA AAA AGC AGG CTT CAT GGA AAG CAT GGCGTTGCT -3 '(SEQ ID NO: 7)

   OsGlu2 12b attB2 5'- GGG GAC CAC TTT GTA CAA GAA AGC TGG GTC GAA ATT GTA GGA GTA TGT CGG CGA -3 '(SEQ ID NO: 8)

   attB1 adapter primer 5'- GGG GAC AAG TTT GTA CAA AAA AGC AGG CT-3 '(SEQ ID NO: 9)

   attB2 adapter primer 5'- GGG GAC CAC TTT GTA CAA GAA AGC TGG GT-3 '(SEQ ID NO: 10)

Using a full-length cDNA of OsGlu2, amplify and recover 30 cycles of fist PCR, and then amplify again by rotating 25 cycles of second PCR using an adapter primer to recover the PCR product of OsGlu2 with an adapter. It is BP reacted with pDONR201 (Invitrogen) vector (more than 25 ° C for 16 hours). Subsequently, incubation at 25 ° C for 8 hours using an LR reaction (OsCc1 promoter (overexpression vector), P-Ai promoter (ABA inducible vector), which is the vector to be coined with the BP reaction mixture, followed by adding proteinase K for 1 hour at 37 ° C) incubation) to transform to DH 5β. The vector to be coined contains the 3 'region of the potato proteinase inhibitor II gene (3'pinII) and the bar gene expression cassette that contains a 35S promoter, bar coding region, 3' region of the nopaline synthase gene (3'nos). have.

A rice transgenic crop was prepared using OsCcl, an over-expression vector of the present invention, and P-Ai, an RNAi vector (see FIG. 8 (B)).

Preparation of Rice Transformant Using Agrobacterium

OsGlu2 gene in each of the overexpression vector and ABA-nducible vector was cultured at 28 ° C for 3 days in ABST medium by cell mix with Agrobacterium tumefaciens strain (LBA4404) and E. coli HB101 (pRK2013). transformation to callus.

Nakdong rice seeds were disinfected with 70% EtOH, washed in 3% Chlorax for 1 hour, washed, and planted in 2N6 medium to incubate at 28 ° C for 3-4 weeks in the dark. Only good callus formed was screened and cultured at 28 ° C in 2N6 medium 4 days before transformation. The Agrobacterium concentration of the cell mix was adjusted to OD600nm to 3 at OD600nm, and then infected with the selected callus. Then, the cells were inoculated in 2N6-AS medium and cultured for 3 days at 20 ° C. After immersing the callus in a solution containing cefortaxiume in sterile water for about 1 hour, dried on filter paper and placed on a 2N6-CP medium and incubated for 3 weeks at 28 ° C. Only the callus, showing no growth, was selected and transferred to N6-70CP medium, and then cultured for 10 days at 28 ° C. Selected callus was transferred to MSCP medium and cultured in light. Root and stem-derived plants were transferred to magent medium and grown in soil (FIG. 9 shows propagation process from callus of rice transformants and FIG. 10 shows the medium and its composition used).

Osglu2 Transformation and Verification of Rice Transformant Lines

Northern blot analysis and Western blot analysis were performed to verify the To generation line of rice transformant Ai :: OsGlu2 of ABA-inducible vector grown from callus induced by Agrobacterium transformation. As shown in FIG. 11, the expression patterns of each line of rice transformants by Northern blot analysis and Western blot analysis were examined, and lines were expressed after the expression of the lines.

< Example  4: Osglu2  Functional Verification from Rice Transformants of Genes>

Osglu2 Growth of Rice Transformants in Korea

The lines of verified rice transformant Ai :: OsGlu2 were selected and propagated to T2 generations to observe their growth. Nakdong rice was used as a control. Among them, the Ai :: OsGlu2 T2 generation line 2-2 and 10-3 line were intensively observed. We also observed and compared line 1 of OsCcl :: OsGlu2 using the OsCc1 promoter (overexpression vector). Yield components such as growth growth, height, flag leaf length, stem thickness, number of grains, rice grain per ear, and grain weight were investigated.

As shown in FIG. 12, the photographs of the transgenic rice plants (OsCcl :: OsGlu2 and Ai :: OsGlu2) and the control of Nakdong rice (WT) were compared to the Ai :: OsGlu2 transformants (T2- 2-2, T2-10-3) was found to promote growth than the control (WT) line, overexpression (OsCcl :: OsGlu2 T2-2-1) line. FIG. 13 is a Stem comparison photograph of each of the transformed rice plants (OsCcl :: OsGlu2 and Ai :: OsGlu2) and the control Nakdong rice (WT), wherein the Ai :: OsGlu2 lines of the transformants were grown more strongly. Figure 14 is a comparison picture of the deletion of the Ai :: OsGlu2 rice transformant and the control Nakdong rice (WT), it can be seen that the transformant growth is large. FIG. 15 is a comparison table according to the yield components of each transformed rice plant (OsCcl :: OsGlu2 and Ai :: OsGlu2) and a control, Nakdong rice (WT), where the transformants have a large number of heights, grains and rice grains per ear. Could confirm.

Osglu2 Of rice transformants against rice blast disease

To verify the protective function of β-1,3-glucanases, a hydrolytic enzyme belonging to pathogen-related genes (PRs), OsGlu2 rice transformants were inoculated with blasts. Ai :: OsGlu2 T2 generation, which grew well for 4 ~ 4 weeks, was inoculated with a blast (KJ101) line-by-line and incubated for 3 days. Among them, the number of lesions on the leaves of lines 18 and 27 of Ai :: OsGlu2 T2 generation showed a significant decrease and resistance. Then, Northern blot analysis and Western blot analysis were performed for verification. At this time, the ABA was treated before inoculation of the blast (KJ101) to express the OsGlu2 gene.

As shown in FIG. 16, a rice transformant of Ai :: OsGlu2 and a control group of Nakdong rice (WT) leaves were treated after 48 hours of rice blasting. Figure 16 (A) was treated with ABA to the rice transformants and control Nakdong rice (WT) leaves of Ai :: OsGlu2 in advance, and treated the rice blast for 48 hours to examine the lesions of these leaves. FIG. 16 (B) shows RNA and protein separation from Ai :: OsGlu2 rice transformants treated with rice blast and Nakdong rice (WT) leaves. I looked. The lines of the two transformant rice (Ai :: OsGlu2 T2-2-2, T2-10-3) were resistant to blast disease compared to the control (WT), which increased the amount of gene expression in the plant, resulting in β-1,3- Increasing expression of glucanase also suggests that blast resistance is also increased. This shows that the disease resistance in Ai :: OsGlu2 transformants is correlated with blast resistance due to the increased expression of β-1,3-glucanase.

The present invention relates to β-1,3-glucanase, which is a rice leaf blast resistance protein that directly protects against diseases of plants (rice), and a gene encoding the same (OsGlu2). By using this gene, which is expressed specifically in diseases in rice, it is possible to increase the resistance to diseases such as rice blast diseases, thereby cultivating new varieties that promote the growth of rice and consequently increase the yield. While reducing the use of chemicals such as pesticides, which have many problems, it is expected to greatly contribute to economic efficiency by increasing the yield and producing high quality rice by dramatically increasing the environmental stress resistance to diseases.

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Claims (8)

delete delete delete delete CDNA gene encoding β-1,3-glucanase (OsGlu2) derived from rice, consisting of the nucleotide sequence of SEQ ID NO: 2 A method for increasing disease resistance of a plant, comprising the step of transforming the plant with a recombinant plant expression vector comprising. Plants characterized by increased disease resistance of plants produced by the method of claim 5. The plant of claim 6, wherein the disease is rice blast. The plant of claim 6, wherein the plant is rice.

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