WO2010023974A1 - 広範な病害抵抗性を付与するイネ遺伝子 - Google Patents
広範な病害抵抗性を付与するイネ遺伝子 Download PDFInfo
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Definitions
- the present invention relates to a method for producing a plant that provides resistance to either or both of a pathogenic bacterium and a pathogenic filamentous fungus, a plant obtained using the same, and use thereof.
- SA salicylic acid
- Non-patent Document 2 Non-patent Document 2
- SA npr1 mutant of Arabidopsis thaliana no longer induces the expression of disease resistance genes and PR genes by SA, INA, etc., and at the same time cannot grow on a medium containing SA.
- rice knowledge about the SA signal transduction pathway and signal transduction factors involved in this pathway is extremely poor.
- the present invention has been made in view of such a situation, and an object of the present invention is to isolate a novel gene imparting a wide range of disease resistance.
- the present inventors have identified genes that are resistant to pathogenic bacteria and pathogenic fungi by screening based on gain-of-function (function acquisition) as an index instead of loss-of-function. Tried. Specifically, the present inventors selected pathogenic bacteria infection resistance, pathogenic fungal infection resistance in rice full-length cDNA high-expressing Arabidopsis thaliana (rice-Family FOX line) prepared using FOX hunting system. Three-stage screening of sex selection and salicylic acid sensitivity selection was attempted. As a result, the present inventors succeeded in finding one Arabidopsis thaliana line (K15424 line) selected in any of these three types of screening.
- the K15424 line has a rice full-length cDNA (AK070024, a novel protein kinase gene).
- the present inventors produced rice overexpressing AK070024, and conducted rice white leaf blight resistance test in the T1 generation. As a result, it was confirmed that AK070024 overexpressing rice is resistant to leaf blight and blast.
- the present invention relates to an unprecedented gene that is resistant to pathogenic bacteria and pathogenic fungi, and a plant into which the gene has been introduced. More specifically, the following [1] to [7] are provided. To do. [1] A method for imparting resistance to a pathogenic bacterium and / or a pathogenic fungus to a plant, including the following steps (a) and (b): (A) a step of introducing a DNA selected from the group consisting of (i) to (iv) below or a vector containing the DNA into a plant cell; and (b) a plant into which the DNA or vector has been introduced in step (a).
- a process of regenerating plant bodies from cells (I) DNA encoding a protein comprising the amino acid sequence set forth in SEQ ID NO: 2; (Ii) DNA containing the coding region of the base sequence set forth in SEQ ID NO: 1, (Iii) DNA encoding a protein comprising an amino acid sequence in which one or more amino acids are substituted, deleted, added, and / or inserted in the amino acid sequence of SEQ ID NO: 2, and (iv) SEQ ID NO: 1 DNA that hybridizes under stringent conditions with DNA comprising the base sequence described in 1.
- SEQ ID NO: 1 DNA that hybridizes under stringent conditions with DNA comprising the base sequence described in 1.
- [3] A plant body regenerated from the plant cell according to [2], which is resistant to either or both of a pathogenic bacterium and a pathogenic filamentous fungus
- [4] A plant body that is a progeny or clone of the plant body according to [3], which is resistant to either or both of a pathogenic bacterium and a pathogenic fungus
- [5] A plant propagation material that is resistant to either or both of the pathogenic bacterium and the pathogenic fungus described in [3] or [4]
- [6] A method for producing a plant that is resistant to either or both of pathogenic bacteria and pathogenic fungi, comprising the following steps (a) and (b): (A) a step of introducing a DNA selected from the group consisting of (i) to (iv) below or a vector containing the DNA into a plant cell; and (b) a plant into which the DNA or vector has been introduced in step (a).
- a process of regenerating plant bodies from cells (I) DNA encoding a protein comprising the amino acid sequence set forth in SEQ ID NO: 2; (Ii) DNA containing the coding region of the base sequence set forth in SEQ ID NO: 1, (Iii) DNA encoding a protein comprising an amino acid sequence in which one or more amino acids are substituted, deleted, added, and / or inserted in the amino acid sequence of SEQ ID NO: 2, and (iv) SEQ ID NO: 1 DNA that hybridizes under stringent conditions with DNA comprising the base sequence described in 1.
- a method for screening a candidate compound for a drug that confers resistance to either or both of a pathogenic bacterium and a pathogenic fungus on a plant including the following steps (a) to (c): (A) contacting a test compound with a cell or cell extract containing DNA having a structure in which all or part of the base sequence set forth in SEQ ID NO: 3 and a reporter gene are functionally linked; (B) measuring the expression level of the reporter gene, and (c) selecting a compound that increases the expression level as compared to the measurement in the absence of the test compound.
- OsPSR5 OX is rice that overexpresses OsPSR5. It is a photograph showing blast resistance in OsPSR5 overexpressing rice. Photo 8th day after spray inoculation with blast fungus.
- OsPSR5 OX is rice that overexpresses OsPSR5. Nipponbare is Wt. It is a graph which shows the blast disease resistance in OsPSR5 overexpression rice.
- Koganenishiki is a slightly more resistant system than Nipponbare (Wt). Bacteria were inoculated when the 6th leaf was developed. The number of lesions indicates the total value of the diseased lesions of 5 leaves and 6 leaves 6 days after inoculation.
- the present invention provides a method for imparting resistance to a pathogenic bacterium and / or a pathogenic fungus to a plant using a protein kinase gene (DDBJ Accession No: AK070024).
- the pathogenic bacteria in the present invention include, but are not limited to, tomato spotted bacterial disease bacteria, rice white leaf blight fungus, rice withering fungus, and withering bacterial blight.
- pathogenic filamentous fungi in the present invention include, but are not limited to, the cruciferous vegetable anthracnose fungus, blast fungus, and powdery mildew.
- Whether or not a plant is resistant to pathogenic bacteria or pathogenic fungi can be determined by culturing the pathogenic bacteria or pathogenic fungi and then immersing the plant in a bacterial solution or spraying the bacterial solution on the plant. Can be evaluated.
- genomic DNA used in the present invention is not particularly limited, and may be cDNA or genomic DNA.
- Preparation of genomic DNA and cDNA can be performed by those skilled in the art using conventional means.
- genomic DNA is designed using an appropriate primer pair based on the known base sequence information (SEQ ID NO: 1) of the protein kinase gene (DDBJ Accession No: AK070024), and using the genomic DNA prepared from the target plant as a template. It can be prepared by performing PCR and screening a genomic library using the resulting amplified DNA fragment as a probe.
- the protein of the present invention A cDNA encoding a phosphorylase can be prepared. Furthermore, if a commercially available DNA synthesizer is used, the target DNA can be prepared by synthesis.
- the DNA of the present invention encodes a protein-derived protein kinase (SEQ ID NO: 2) as long as it has a function of imparting resistance to one or both of pathogenic bacteria and pathogenic fungi to plants.
- DNAs eg, mutants, derivatives, alleles, variants, and homologs
- Such DNA includes, for example, DNA encoding a protein comprising an amino acid sequence in which one or more amino acids are substituted, deleted, added, and / or inserted in the amino acid sequence set forth in SEQ ID NO: 2. .
- Methods well known to those skilled in the art for preparing DNA encoding proteins with altered amino acid sequences include, for example, the site-directed mutagenesis method (Kramer, W. and Fritz, HJ Oligonucleotide-directed construction of mutagenesis via gapped duplex DNA. Methods in Enzymology. 154, 1987, 350-367.).
- the amino acid sequence of the encoded protein may be mutated in nature due to the mutation of the base sequence.
- SEQ ID NO: 2 amino acid sequence of kinase
- pathogens may occur in plants. As long as it has a function of imparting resistance to either or both of bacteria and pathogenic fungi, it is included in the DNA of the present invention.
- the number of amino acids to be modified is not particularly limited, but is generally within 50 amino acids, preferably within 30 amino acids, more preferably within 10 amino acids (for example, within 5 amino acids, within 3 amino acids).
- the amino acid modification is preferably a conservative substitution.
- the hydropathic index (Kyte, J. and Doolittle, RF J Mol Biol. 157 (1), 1982, 105-132.) And Hydrophilicity value (US Pat. No. 4,554,101) for each amino acid before and after modification are , Preferably within ⁇ 2, more preferably within ⁇ 1, and most preferably within ⁇ 0.5.
- DNA encoding proteins structurally similar to rice protein kinase include hybridization techniques (Southern, EM Journal of Molecular Biology. 98, 1975, 503.) and polymerase chain reaction. (PCR) technology (Saiki, RK et al. Science. 230, 1985, 1350-1354 .; Saiki, RK et al. Science, 239, 1988, 487-491.) Is possible. That is, the DNA of the present invention includes DNA that hybridizes under stringent conditions with the DNA consisting of the nucleotide sequence set forth in SEQ ID NO: 1. In order to isolate such DNA, the hybridization reaction is preferably carried out under stringent conditions.
- stringent conditions refers to conditions of 6M urea, 0.4% SDS, 0.5 ⁇ SSC, or hybridization conditions of stringency equivalent thereto, but are not particularly limited to these conditions. Isolation of DNA with higher homology can be expected by using conditions with higher stringency, for example, 6M urea, 0.4% SDS, and 0.1xSSC.
- the DNA thus isolated is considered to have high homology with the amino acid sequence (SEQ ID NO: 2) of the protein phosphorylase derived from rice at the amino acid level.
- High homology means a sequence of at least 50% or more, more preferably 70% or more, more preferably 90% or more (for example, 95%, 96%, 97%, 98%, 99% or more) in the entire amino acid sequence. Refers to the identity of The identity of the amino acid sequence and nucleotide sequence is determined by the algorithm BLAST (Karlin, S.
- the DNA of the present invention may be inserted into a vector.
- the vector is not particularly limited as long as it can express the inserted gene in plant cells.
- a vector having a promoter for constitutive gene expression in plant cells for example, potato chitinase gene SK2 promoter, cauliflower mosaic virus 35S promoter, etc.
- inducibly activated by external stimulation It is also possible to use a vector having a promoter.
- the coding region of the protein in the base sequence described in SEQ ID NO: 1 is from the 34th base to the 1251st base.
- the amino acid sequence generated from the 34th base to the 1251st base in the base sequence described in SEQ ID NO: 1 is shown in SEQ ID NO: 2.
- the present invention provides a method for producing a plant that is resistant to either or both of pathogenic bacteria and pathogenic fungi.
- plant cells into which the DNA or vector is introduced include monocotyledonous plants such as rice, wheat, barley, corn, and sorghum, and dicotyledonous plants such as Arabidopsis, rapeseed, tomato, soybean, and potato. It is not limited to these.
- the form of the plant cell into which the DNA or vector is introduced is not particularly limited as long as it can regenerate the plant body, and includes, for example, suspension culture cells, protoplasts, leaf sections, and callus.
- the introduction of the DNA or vector into plant cells can be carried out by methods known to those skilled in the art such as polyethylene glycol method, electroporation method, Agrobacterium-mediated method, particle gun method and the like. it can.
- Agrobacterium for example, according to the method of Nagel et al. (Nagel, R. et al. FEMS Microbiol Lett. 67, 1990, 325-328.)
- the DNA can be introduced into a plant cell by infecting the plant cell with the Agrobacterium by direct infection method or leaf disk method.
- the regeneration of plant bodies from plant cells can be performed by methods known to those skilled in the art depending on the type of plant.
- the method of Fujimura et al. (Fujimura. Et al. Tissue Culture Lett. 2, 1995, 74.) can be mentioned, and for wheat, Harris et al. (Harris, R. et al. Plant Cell Reports. 7 1988, 337-340) and Ozgen et al. (Ozgen, M. et al. Plant Cell Reports. 18, 1998, 331-335).
- Kihara and Funatsuki Kihara, M. and Funatsuki, H. Breeding Sci. 44, 1994, 157-160.
- Lurs and Lorz (Lurs, R.
- offspring or clones can be obtained from the plant by sexual or asexual reproduction. It is also possible to obtain a propagation material (for example, seeds, fruits, cuttings, tubers, tuberous roots, strains, callus, protoplasts, etc.) from the plant body, its descendants or clones, and mass-produce the plant body based on them. Is possible.
- a propagation material for example, seeds, fruits, cuttings, tubers, tuberous roots, strains, callus, protoplasts, etc.
- the control means a plant body of the same species as the plant body of the present invention, in which the DNA of the present invention is not overexpressed.
- the control in the present invention is not limited at all as long as it is a plant of the same species as the plant of the present invention and the DNA of the present invention is not overexpressed. Therefore, the control of the present invention includes, for example, a plant into which DNA other than the DNA encoding the protein phosphorylase derived from the rice of the present invention has been introduced.
- Examples of such a plant include, for example, a plant of the same species as the transformed plant of the present invention, a plant transformed with a DNA other than the DNA of the present invention, and the function of the DNA of the present invention.
- a plant transformed with a DNA introduced with a deletion mutation, a plant transformed with a DNA converted to a function-inhibited form of the DNA of the present invention, a DNA fragment of a region insufficient for functional expression of the DNA of the present invention The plant body transformed with is mentioned, but it is not limited to these.
- the present invention provides a plant exhibiting resistance to one or both of pathogenic bacteria and pathogenic fungi, plant cells capable of regenerating the plant, plants that are descendants or clones of the plant, and the above It also provides plant propagation material.
- the plant body, plant cell, offspring or clone plant body and propagation material of the present invention may further have sensitivity to salicylic acid.
- the present invention also provides a method for screening a candidate compound for a drug that confers resistance to either or both of a pathogenic bacterium and a pathogenic fungus on a plant.
- a cell or a cell extract containing DNA having a structure in which a DNA comprising all or part of the 2000 bp upstream of the AK070024 gene including the transcriptional regulatory region of the AK070024 gene and a reporter gene are functionally linked to each other;
- the test compound is brought into contact.
- “functionally linked” means upstream of the AK070024 gene containing the transcriptional regulatory region of the AK070024 gene so that expression of the reporter gene is induced by binding of a transcription factor to the transcriptional regulatory region of the AK070024 gene. It means that a DNA comprising all or part of 2000 bp is bound to a reporter gene. Therefore, even when the reporter gene is bound to another gene and forms a fusion protein with another gene product, the transcription factor of the fusion protein binds to the transcriptional regulatory region of the AK070024 gene. Any expression that is induced is included in the meaning of “functionally linked”.
- the base sequence of 2000 bp upstream of the AK070024 gene is shown in SEQ ID NO: 3.
- the reporter gene used in the present method is not particularly limited as long as its expression can be detected, and examples thereof include CAT gene, lacZ gene, luciferase gene, and GFP gene.
- Examples of “cells containing DNA having a structure in which the transcriptional regulatory region of the AK070024 gene and a reporter gene are functionally linked” include cells into which a vector having such a structure inserted is introduced. Such vectors can be prepared by methods well known to those skilled in the art. Introduction of the vector into the cells can be performed by a general method such as calcium phosphate precipitation, electric pulse perforation, lipofectamine method, microinjection method and the like.
- Cells containing DNA having a structure in which the transcriptional regulatory region of the AK070024 gene and a reporter gene are functionally linked include cells in which the structure is inserted into the chromosome.
- the DNA structure can be inserted into the chromosome by a method generally used by those skilled in the art, for example, a gene introduction method via Agrobacterium.
- a cell extract containing DNA having a structure in which a transcriptional regulatory region of the AK070024 gene and a reporter gene are functionally linked refers to, for example, a cell extract contained in a commercially available in vitro transcription translation kit containing AK070024 gene Examples include those to which DNA having a structure in which a transcriptional regulatory region and a reporter gene are functionally linked is added.
- test compound used in this method is not particularly limited.
- natural compounds, organic compounds, inorganic compounds, proteins, peptides and other single compounds as well as compound libraries, gene library expression products, cell extracts, cell culture supernatants, fermented microorganism products, marine organism extracts Products, plant extracts and the like, but are not limited thereto.
- Contact in this method means that the test compound is added to the culture solution of “the cell containing DNA having a structure in which the transcriptional regulatory region of AK070024 gene and the reporter gene are functionally linked”, or the DNA containing the DNA.
- the test compound can be added to a commercially available cell extract.
- the test compound is a protein, for example, it can be performed by introducing a DNA vector expressing the protein into the cell.
- the expression level of the reporter gene is then measured.
- the expression level of the reporter gene can be measured by methods known to those skilled in the art depending on the type of the reporter gene. For example, when the reporter gene is a CAT gene, the expression level of the reporter gene can be measured by detecting acetylation of chloramphenicol by the gene product.
- the reporter gene is a lacZ gene, by detecting the color development of the dye compound catalyzed by the gene expression product, and when the reporter gene is a luciferase gene, the fluorescent compound catalyzed by the gene expression product By detecting fluorescence, and in the case of a GFP gene, the expression level of the reporter gene can be measured by detecting fluorescence due to the GFP protein.
- a compound that increases the expression level of the measured reporter gene is then selected as compared to the measurement in the absence of the test compound.
- the compound thus selected becomes a candidate compound for a drug that confers resistance to one or both of pathogenic bacteria and pathogenic fungi on plants.
- a cDNA library was prepared with Agrobacterium (Agrobacterium GV3101) using the cDNA standardized as described above. Using these Agrobacterium, Arabidopsis thaliana Columbia (Col-0) was transformed by the flower infection method. A rice-plant FOX line consisting of plant bodies was produced. T2 seed was used for screening.
- the cruciferous vegetable anthracnose fungus ( Colletotrichum higginsianum , Ch ) is a filamentous fungus that infects by using an attachment device and penetrating mycelia like blast fungus, and can infect Arabidopsis thaliana. This bacterium was used for selection of resistance to infection with pathogenic filamentous fungi. Infection was performed by the improved dip method used for selection with Pst 3000. The conidia concentration of the infectious solution was 10 5 to 10 6 conidia / ml, and selection was made based on the presence or absence of survival 6 days after infection.
- SA Salicylic acid
- Rice White Leaf Blight Resistance Test Using rice white leaf blight fungus race T7174, the method of Mori et al. (Mori et al. 2007) was modified and tested. The main modification was that approximately 5cm was cut from the tip of the young leaf blade of rice, and inoculation was performed by immersing the cut surface in a bacterial solution.
- Salicylic acid-sensitive selection rice-Nazuna FOX strains were first selected from the lines that killed in + SA medium (primary screening). For the selected lines, secondary screening was performed to select lines that died after germination in the + SA medium and grew to the same extent as the wild type in the -SA medium (FIG. 2), and 95 lines were selected. For these lines, the partial rice sequence was determined after PCR amplification of the inserted rice full-length cDNA.
- K15424 Lines Selected in Three Types of Screening and Their Causal Genes
- This line is likely to have acquired resistance to pathogenic bacteria and pathogenic fungi by enhancing the defense response mechanism involving the SA signaling system.
- Rice full-length cDNA was inserted into the K15424 line.
- a phylogenetic analysis of AK070024 revealed that it was a novel protein kinase gene, closely related to the NAK (Novel Arabisopsis protein Kinase) gene.
- AK070024 a novel gene (AK070024) was found by screening of rice FOX strain Arabidopsis thaliana, which gave three traits of pathogenic bacteria ( Pst 3000) resistance, pathogenic fungus (Ch) resistance, and salicylic acid hypersensitivity. Rice that overexpressed this gene was resistant to leaf blight and blast. From the above results, it was shown that plants overexpressing this gene are resistant to multiple diseases regardless of whether they are dicotyledonous or monocotyledonous plants. This gene can be a material for imparting combined disease resistance to various crops by genetic recombination technology.
- a plant body resistant to pathogenic bacteria and pathogenic filamentous fungi is provided.
- a plant overexpressed with the gene identified in the present invention is resistant to a plurality of diseases regardless of whether it is a dicotyledonous plant or a monocotyledonous plant. Therefore, the gene identified in the present invention is useful as a material for imparting combined disease resistance to various crops.
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Description
Becker, D et al., (1990) Nucleic Acid Res., 18(1): 203 Cao, H et al (1997) Cell 88:57-63 Ichikawa, T et al., (2006) Plant J. 48: 974-985 Kikuchi, S et al., (2003) Science 301: 376-379 Mori, M et al., (2007) Plant Mol. Biol., 63:847-860 Nakamura, H et al., (2007) Plant Mol Biol. 65:357-371 Taji, T et al., (2002) Plant J., 29(4): 417-426, Toki, S et al., (2006) Plant J. 47:969-76.
〔1〕下記(a)及び(b)の工程を含む、植物に病原細菌及び病原糸状菌のいずれか又は両方に対する抵抗性を付与する方法;
(a)下記(i)から(iv)からなる群より選択されるDNA又は該DNAを含むベクターを植物細胞に導入する工程、及び
(b)工程(a)においてDNA又はベクターが導入された植物細胞から植物体を再生する工程、
(i)配列番号:2に記載のアミノ酸配列を含むタンパク質をコードするDNA、
(ii)配列番号:1に記載の塩基配列のコード領域を含むDNA、
(iii)配列番号:2に記載のアミノ酸配列において1もしくは複数のアミノ酸が置換、欠失、付加、及び/又は挿入されたアミノ酸配列を含むタンパク質をコードするDNA、及び
(iv)配列番号:1に記載の塩基配列を含むDNAとストリンジェントな条件下でハイブリダイズするDNA、
〔2〕下記(a)から(d)のいずれかに記載のDNAまたは該DNAを含むベクターが導入された植物細胞であって、病原細菌及び病原糸状菌のいずれか又は両方に抵抗性である植物体を再生しうる植物細胞;
(a)配列番号:2に記載のアミノ酸配列を含むタンパク質をコードするDNA、
(b)配列番号:1に記載の塩基配列のコード領域を含むDNA、
(c)配列番号:2に記載のアミノ酸配列において1もしくは複数のアミノ酸が置換、欠失、付加、及び/又は挿入されたアミノ酸配列を含むタンパク質をコードするDNA、及び
(d)配列番号:1に記載の塩基配列を含むDNAとストリンジェントな条件下でハイブリダイズするDNA、
〔3〕〔2〕に記載の植物細胞から再生された、病原細菌及び病原糸状菌のいずれか又は両方に抵抗性である植物体、
〔4〕〔3〕に記載の植物体の子孫またはクローンである、病原細菌及び病原糸状菌のいずれか又は両方に抵抗性である植物体、
〔5〕〔3〕または〔4〕に記載の病原細菌及び病原糸状菌のいずれか又は両方に抵抗性である植物体の繁殖材料、
〔6〕下記(a)及び(b)の工程を含む、病原細菌及び病原糸状菌のいずれか又は両方に抵抗性である植物体の製造方法;
(a)下記(i)から(iv)からなる群より選択されるDNA又は該DNAを含むベクターを植物細胞に導入する工程、及び
(b)工程(a)においてDNA又はベクターが導入された植物細胞から植物体を再生する工程、
(i)配列番号:2に記載のアミノ酸配列を含むタンパク質をコードするDNA、
(ii)配列番号:1に記載の塩基配列のコード領域を含むDNA、
(iii)配列番号:2に記載のアミノ酸配列において1もしくは複数のアミノ酸が置換、欠失、付加、及び/又は挿入されたアミノ酸配列を含むタンパク質をコードするDNA、及び
(iv)配列番号:1に記載の塩基配列を含むDNAとストリンジェントな条件下でハイブリダイズするDNA、
〔7〕下記(a)から(c)の工程を含む、植物に病原細菌及び病原糸状菌のいずれか又は両方に対する抵抗性を付与する薬剤の候補化合物のスクリーニング方法;
(a)配列番号:3に記載の塩基配列の全部又は一部とレポーター遺伝子とが機能的に結合した構造を有するDNAを含む細胞または細胞抽出液と、被検化合物を接触させる工程、
(b)該レポーター遺伝子の発現レベルを測定する工程、及び
(c)被検化合物の非存在下において測定した場合と比較して、該発現レベルを増加させる化合物を選択する工程。
本発明は、タンパク質リン酸化酵素遺伝子(DDBJ Accession No: AK070024)を用いた植物に病原細菌及び病原糸状菌のいずれか又は両方に対する抵抗性を付与する方法を提供する。
本発明における病原細菌としてはトマト斑葉細菌病細菌、イネ白葉枯病菌、イネ立枯れ病菌、籾枯れ細菌病菌などが挙げられるがこれらに限定されない。また本発明における病原糸状菌としてはアブラナ科野菜類炭そ病菌、いもち病菌、うどんこ病菌などが挙げられるがこれらに限定されない。
またシロイヌナズナであればAkamaら(Akama. et al. Plant Cell Reports. 12, 1992, 7-11.)の方法が挙げられ、ナタネであればWangら(Wang, Y.P. et al. Plant Breeding. 124, 2005, 1-4.)の方法が挙げられ、トマトであればKoblitzとKoblitz(Koblitz, H and Koblitz, D. Plant Cell Reports. 1, 1982, 143-146.)の方法やMorganとCocking(Morgan, A. and Cocking, E.C. Z.Pflanzenpysiol. 106, 1982, 97-104.)の方法が挙げられ、大豆であればLazzeriら(Lazzeri, P.A. et al., Plant Mol. Biol. Rep. 3, 1985, 160-167.)の方法やRanchら(Ranch, J.P. et al., In Vitro Cell Dev. Biol. 21, 1985, 653-658.)の方法が挙げられ、ジャガイモであればVisserら(Visser, R.G.F. et al. Theor. Appl. Genet. 78, 1989, 594-600.)の方法が挙げられるが、これらに限定されない。
なお本発明の植物体、植物細胞、子孫あるいはクローンである植物体、繁殖材料は、さらにサリチル酸に対する感受性を有していてもよい。
本方法においては、まず、AK070024遺伝子の転写調節領域を含むAK070024遺伝子の上流2000bpの全部又は一部を含むDNAとレポーター遺伝子とが機能的に結合した構造を有するDNAを含む細胞または細胞抽出液と、被検化合物を接触させる。ここで「機能的に結合した」とは、AK070024遺伝子の転写調節領域に転写因子が結合することにより、レポーター遺伝子の発現が誘導されるように、AK070024遺伝子の転写調節領域を含むAK070024遺伝子の上流2000bpの全部又は一部を含むDNAとレポーター遺伝子とが結合していることをいう。従って、レポーター遺伝子が他の遺伝子と結合しており、他の遺伝子産物との融合タンパク質を形成する場合であっても、AK070024遺伝子の転写調節領域に転写因子が結合することによって、該融合タンパク質の発現が誘導されるものであれば、上記「機能的に結合した」の意に含まれる。
なおAK070024遺伝子の上流2000bpの塩基配列を配列番号:3に示す。
なお本明細書において引用された全ての先行技術文献は、参照として本明細書に組み入れられる。
1-1.イネ-ナズナFOX系統
スクリーニングには約20,000系統(理研15,000系統+岡山5,000系統)のイネ-ナズナFOX系統を用いた。イネ-ナズナFOX系統は以下のようにして作製された。独立したイネ完全長cDNA(Kikuchi et al. 2003)約13,000種類をcDNAの等量比のプールとして調製し(標準化という)、cDNAをシロイヌナズナの発現ベクターに組み込んだ。発現ベクターには、pBIG2113N (Taji et al. 2002, Becker et al. 1990)にSfiIクローニングサイトを導入したpBIG2113SFを用いた。上記のように標準化したcDNAを用いてcDNAライブラリーをアグロバクテリア(Agrobacterium GV3101)で作成し、これらのアグロバクテリアを用いて花感染法によってシロイヌナズナColumbia(Col-0)の形質転換を行い、独立した植物体から成るイネ-ナズナFOX系統が作製された。スクリーニングにはT2種子を使用した。
トマト斑葉細菌病の病原細菌であるPseudomonas syringae pv. tomato strain DC3000(Pst3000)を用いて、改良型Dip法(図1)により(0.5~2)×108 CFU/mlの菌濃度で接種した。播種後3週間後のFOX系統を上記の菌液に浸し、6日後に生存の有無で選抜した。
アブラナ科野菜類炭そ病菌(Colletotrichum higginsianum, Ch)はいもち病菌同様に付着器、貫入菌糸を用いて感染する糸状菌で、シロイヌナズナにも感染できるため、この菌を病原糸状菌感染抵抗性選抜に用いた。感染はPst3000による選抜にも用いた改良型dip法で行った。感染液の分生子濃度は105~106 conidia/mlで、感染6日後に生存の有無で選抜した。
0.05mM SAを含むMS培地(+SA培地)および含まないMS培地(-SA培地)にFOX系統を播種し、-SA培地上では生育するが+SA培地上では発芽後死滅する系統を選抜した。
イネの品種は日本晴を用いた。イネでの発現ベクターはpRiceFOX (Nakamura et al. 2007)を用いた。イネの形質転換はアグロバクテリウムEH105株を用いて高速形質転換法(Toki et al. 2006)で行った。
イネ白葉枯病菌レースT7174を用いて、森らの方法(Mori et al. 2007)を改変して検定した。主な改変点はイネの若い葉身の先端から約5cmを切断した点、接種は切断面を菌液に浸すことにより行った点である。
イネいもち病菌Kyu89-246(MAFF101506,レース003.0)を用いて噴霧接種法(Mori et al. 2007)で行った。菌濃度は2 x 105 spore/mlで行った。
2-1. 病原性細菌感染抵抗性選抜
イネ-ナズナFOX系統約2万系統を対象に1次、2次、3次のスクリーニングを行い、最終的にPst3000抵抗性を示す72系統を選抜した。選抜した全系統についてゲノムDNAを抽出し、挿入されているイネ完全長cDNAをPCRで増幅して末端の塩基配列を決定することにより遺伝子を同定した。
上記の72系統について更にCh抵抗性検定を行い、両菌に対して複合抵抗性を示す系統があるかどうか調べた結果、21系統(29%)で複合抵抗性を示した。
イネ-ナズナFOX系統約2万系統を対象にまず+SA培地で死滅する系統を選抜(1次スクリーニング)した。選抜された系統について、+SA培地では発芽後死滅し、-SA培地では野生型とほぼ同程度の生育をする系統を選抜する二次スクリーニングを行い(図2)、95系統を選抜した。これらの系統について、挿入されているイネ完全長cDNAをPCR増幅後、部分塩基配列を決定した。
上記3種類のスクリーニングのいずれにおいても選抜された系統が1系統(K15424)存在した。この系統は、SAシグナル伝達系が関わる防御応答機構の増強により病原性細菌および病原性糸状菌に対する抵抗性を獲得した可能性が高い。K15424系統にはイネの完全長cDNA(AK070024)が挿入されていた。AK070024の系統樹解析を行うと、NAK(Novel Arabisopsis protein Kinase)遺伝子と近縁で、新規のタンパク質リン酸化酵素遺伝子であることが明らかになった。更にAK070024を有する他の独立のFOX系統やcDNA再導入シロイヌナズナの解析から、AK070024のシロイヌナズナでの過剰発現がPst3000抵抗性、Ch抵抗性、及びSA感受性を付与することを確認した。
AK070024遺伝子(別名OsPSR5)を過剰発現するイネを作製し、T1世代でイネ白葉枯病抵抗性検定を行った。その結果、OsPSR5過剰発現イネは、白葉枯病に対して高度抵抗性を有する品種あそみのりと同等の、強い白葉枯病抵抗性を示した(図3)。同様にOsPSR5過剰発現イネのいもち病抵抗性検定を行ったところ、接種後、WTの日本晴では病斑が拡大していくのに対し(進展型病斑)、過剰発現体では褐点型病斑が認められたが、一定の大きさ以上に病斑が拡大しなかったことから、いもち病にも抵抗性であることが示された(図4)。抵抗性の程度は、日本晴よりも抵抗性の黄金錦よりも更に強い抵抗性を示した(図5)。
Claims (7)
- 下記(a)及び(b)の工程を含む、植物に病原細菌及び病原糸状菌のいずれか又は両方に対する抵抗性を付与する方法;
(a)下記(i)から(iv)からなる群より選択されるDNA又は該DNAを含むベクターを植物細胞に導入する工程、及び
(b)工程(a)においてDNA又はベクターが導入された植物細胞から植物体を再生する工程、
(i)配列番号:2に記載のアミノ酸配列を含むタンパク質をコードするDNA、
(ii)配列番号:1に記載の塩基配列のコード領域を含むDNA、
(iii)配列番号:2に記載のアミノ酸配列において1もしくは複数のアミノ酸が置換、欠失、付加、及び/又は挿入されたアミノ酸配列を含むタンパク質をコードするDNA、及び
(iv)配列番号:1に記載の塩基配列を含むDNAとストリンジェントな条件下でハイブリダイズするDNA。 - 下記(a)から(d)のいずれかに記載のDNAまたは該DNAを含むベクターが導入された植物細胞であって、病原細菌及び病原糸状菌のいずれか又は両方に抵抗性である植物体を再生しうる植物細胞;
(a)配列番号:2に記載のアミノ酸配列を含むタンパク質をコードするDNA、
(b)配列番号:1に記載の塩基配列のコード領域を含むDNA、
(c)配列番号:2に記載のアミノ酸配列において1もしくは複数のアミノ酸が置換、欠失、付加、及び/又は挿入されたアミノ酸配列を含むタンパク質をコードするDNA、及び
(d)配列番号:1に記載の塩基配列を含むDNAとストリンジェントな条件下でハイブリダイズするDNA。 - 請求項2に記載の植物細胞から再生された、病原細菌及び病原糸状菌のいずれか又は両方に抵抗性である植物体。
- 請求項3に記載の植物体の子孫またはクローンである、病原細菌及び病原糸状菌のいずれか又は両方に抵抗性である植物体。
- 請求項3または4に記載の病原細菌及び病原糸状菌のいずれか又は両方に抵抗性である植物体の繁殖材料。
- 下記(a)及び(b)の工程を含む、病原細菌及び病原糸状菌のいずれか又は両方に抵抗性である植物体の製造方法;
(a)下記(i)から(iv)からなる群より選択されるDNA又は該DNAを含むベクターを植物細胞に導入する工程、及び
(b)工程(a)においてDNA又はベクターが導入された植物細胞から植物体を再生する工程、
(i)配列番号:2に記載のアミノ酸配列を含むタンパク質をコードするDNA、
(ii)配列番号:1に記載の塩基配列のコード領域を含むDNA、
(iii)配列番号:2に記載のアミノ酸配列において1もしくは複数のアミノ酸が置換、欠失、付加、及び/又は挿入されたアミノ酸配列を含むタンパク質をコードするDNA、及び
(iv)配列番号:1に記載の塩基配列を含むDNAとストリンジェントな条件下でハイブリダイズするDNA。 - 下記(a)から(c)の工程を含む、植物に病原細菌及び病原糸状菌のいずれか又は両方に対する抵抗性を付与する薬剤の候補化合物のスクリーニング方法;
(a)配列番号:3に記載の塩基配列の全部又は一部とレポーター遺伝子とが機能的に結合した構造を有するDNAを含む細胞または細胞抽出液と、被検化合物を接触させる工程、
(b)該レポーター遺伝子の発現レベルを測定する工程、及び
(c)被検化合物の非存在下において測定した場合と比較して、該発現レベルを増加させる化合物を選択する工程。
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Cited By (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN102888399A (zh) * | 2012-09-30 | 2013-01-23 | 浙江师范大学 | 鉴定水稻高抗白叶枯病基因的sts分子标记及其应用 |
US20140337602A1 (en) * | 2008-01-11 | 2014-11-13 | International Business Machines Corporation | Execution Of An Instruction For Performing a Configuration Virtual Topology Change |
Families Citing this family (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN102899726A (zh) * | 2012-11-09 | 2013-01-30 | 东北林业大学 | 一种FOX hunting蒙古柳农杆菌cDNA文库的构建方法 |
CN112369314B (zh) * | 2021-01-06 | 2023-06-27 | 湖南杂交水稻研究中心 | 一种鉴定苗期水稻纹枯病表型的方法 |
Citations (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2003000898A1 (en) * | 2001-06-22 | 2003-01-03 | Syngenta Participations Ag | Plant genes involved in defense against pathogens |
US20040123343A1 (en) * | 2000-04-19 | 2004-06-24 | La Rosa Thomas J. | Rice nucleic acid molecules and other molecules associated with plants and uses thereof for plant improvement |
US20060123505A1 (en) * | 2002-05-30 | 2006-06-08 | National Institute Of Agrobiological Sciences | Full-length plant cDNA and uses thereof |
WO2008070179A2 (en) * | 2006-12-06 | 2008-06-12 | Monsanto Technology, Llc | Genes and uses for plant improvement |
-
2009
- 2009-03-04 US US13/061,396 patent/US9127290B2/en not_active Expired - Fee Related
- 2009-03-04 WO PCT/JP2009/054081 patent/WO2010023974A1/ja active Application Filing
- 2009-03-04 JP JP2010526583A patent/JP5591703B2/ja not_active Expired - Fee Related
Patent Citations (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US20040123343A1 (en) * | 2000-04-19 | 2004-06-24 | La Rosa Thomas J. | Rice nucleic acid molecules and other molecules associated with plants and uses thereof for plant improvement |
WO2003000898A1 (en) * | 2001-06-22 | 2003-01-03 | Syngenta Participations Ag | Plant genes involved in defense against pathogens |
US20060123505A1 (en) * | 2002-05-30 | 2006-06-08 | National Institute Of Agrobiological Sciences | Full-length plant cDNA and uses thereof |
WO2008070179A2 (en) * | 2006-12-06 | 2008-06-12 | Monsanto Technology, Llc | Genes and uses for plant improvement |
Non-Patent Citations (5)
Title |
---|
DATABASE GENBANK [online] 14 February 2008 (2008-02-14), OHYANAGI H ET AL.: "Oryza sativa (japonica cultivar-group) Os09g0533600 (Os09g0533600) mRNA, complete cds.", retrieved from http://www.ncbi. nlm.nih.gov/nuccore/115480352?report=genbank Database accession no. NM-001070305 * |
DUBOUZET J G ET AL.: "Ine-Nazuna FOX Keito 2 Man Keito yori Senbatsu sareta Byogen Saikin Pst3000 Kansen Teikosei Idenshi no Kaiseki", DAI 49 KAI PROCEEDINGS OF THE ANNUAL MEETING OF THE JAP, - 15 March 2008 (2008-03-15), pages 336 * |
MASAHARU KANNO ET AL.: "Ine-Shiroinunazuna FOX Keito o Mochiita Ine no Yudo Teikosei ni Kan'yo suru Shinki Inshi no Tansaku", DAI 49 KAI PROCEEDINGS OF THE ANNUAL MEETING OF THE JAPANESE SOCIETY OF PLANT PHYSIOLOGISTS, - 15 March 2008 (2008-03-15), pages 335 * |
MINAMI MATSUI ET AL.: "Ine-Nazuna FOX hunting-kei: Yuyo Keishitsu no Kosoku Tansaku no Tameno Model System", DAI 49 KAI PROCEEDINGS OF THE ANNUAL MEETING OF THE JAPANESE SOCIETY OF PLANT PHYSIOLOGISTS, - 15 March 2008 (2008-03-15), pages 89 * |
SATORU MAEDA ET AL.: "Ine Kanzencho cDNA Ko Hatsugen Shiroinunazuna Keito o Mochiita Byogen Shijokin C. higginsianum Kansen Teikosei Keito no Senbatsu to Kaiseki", JAPAN SOCIETY FOR BIOSCIENCE, BIOTECHNOLOGY, AND AGROCHEMISTRY 2008 NENDO (HEISEI 20 NENDO) TAIKAI KOEN YOSHISHU, 5 March 2008 (2008-03-05), pages 65 * |
Cited By (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US20140337602A1 (en) * | 2008-01-11 | 2014-11-13 | International Business Machines Corporation | Execution Of An Instruction For Performing a Configuration Virtual Topology Change |
CN102888399A (zh) * | 2012-09-30 | 2013-01-23 | 浙江师范大学 | 鉴定水稻高抗白叶枯病基因的sts分子标记及其应用 |
CN102888399B (zh) * | 2012-09-30 | 2014-02-05 | 浙江师范大学 | 鉴定水稻高抗白叶枯病基因的sts分子标记及其应用 |
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JPWO2010023974A1 (ja) | 2012-01-26 |
JP5591703B2 (ja) | 2014-09-17 |
US20110258737A1 (en) | 2011-10-20 |
US9127290B2 (en) | 2015-09-08 |
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