1997 年度実績報告書

宿主特異的毒素が介在する植物-微生物相互作用の分子遺伝学に関する日米共同研究

研究課題

研究課題/領域番号	08044207
研究機関	鳥取大学
研究代表者	甲元啓介鳥取大学, 農学部, 教授 (80032093)
研究分担者	伊藤靖夫信州大学, 理学部, 助手 (70283231) 秋光和也香川大学, 農学部, 助教授 (80263888) 柘植尚志名古屋大学, 大学院・生命農学研究科, 助教授 (30192644) 児玉基一朗鳥取大学, 農学部, 講師 (00183343) 尾谷浩鳥取大学, 大学院・連合農学研究科, 教授 (50032305)
キーワード	REMI / CPS遺伝子 / PKS遺伝子 / 宿主特異的毒素 / AM毒素 / AK毒素 / AAL毒素 / 毒素感受性因子
研究概要	1)リンゴ斑点落葉病菌の宿主特異的AM毒素の生合成に関与する遺伝子:(1)REMIによる遺伝子タギング法を用いてリンゴ斑点落葉病菌の形質転換体557株を得た。これらについて毒素生産能を生物検定法及びHPLC分析法で調査し、6菌株の毒素非生産変異体を得た。そのうち5菌株は培地上の菌糸成長など野生菌株と差はないが、病原性を喪失していた。これらについてサザンハイブリダイゼーションしたところ、1菌株(71HR-97)においてはゲノム中1箇所に1コピーの組込みがみられた。しかし、その他の毒素非生産変異体では複数部位に複数のプラスミドの組込みが生じていた。(2)CPS遺伝子のユニバーサルPCRプライマーを利用して得た約300bpのPCR産物をサブクローニングし、塩基配列をそれぞれ決定したところ、4種類の配列が得られた。これらすべては、他のCPS遺伝子と相同性が認められ、さらにサザン解析の結果、AM毒素生産菌に特異的に存在する配列であることが明らかとなった。また、本遺伝子断片を用いた相同的組込みによる遺伝子破壊(ジーンターゲッティング)により、毒素非生産形質転換体が得られた。以上の結果は、本CPS遺伝子断片がAM毒素生合成遺伝子の一部であることを示唆するものである。 2)トマトアルターナリア茎枯病菌の宿主特異的AAL毒素生合成遺伝子:REMI法に茎枯病菌生株よりAAL毒素非生産変異体を得た。これら変異体はトマトに対する病原性を同時に失活していたことから、本菌の病原性発現におけるAAL毒素の重要性が確認された。 3)ナシ黒斑病菌のAK毒素生合成遺伝子:遺伝子タギング法を用いて毒素生産菌に特異的に存在する染色体断片が得られた。これら一連の遺伝子群には、毒素生産に関与すると考えられるポリケタイド合成酵素(PKS)遺伝子が含まれていた。また、AK毒素と類似の化学構造を有するAF及びACT毒素の生産菌も、本遺伝子を保有することが明らかとなった。 4)ACR毒素に対するラフレモン特異的感受性因子を支配しているmtDNAで大腸菌を形質転換した。 5)AM毒素感受性リンゴに特異的に発現しているタンパク質を検出した。

研究成果
(21件)

すべてその他

すべて文献書誌 (21件)

[文献書誌] Kusaba, M.and Tsuge, T.: "Mitochondrial DNA variation in host-specific toxin-producing pathogens in the genus Alternaria" Ann.Phytopathol.Soc.Jpn. 63・6. 463-469 (1997)
[文献書誌] Akamatsu, H., Itoh, Y., Kodama, M.et al: "AAL-toxin-deficient mutants of Alternaria alternata tomato pathotype by restriction enzyme-mediated integration" Phytopathology. 87・9. 967-972 (1997)
[文献書誌] Yoder, O.C.: "A mechanistic view of the fungal/plant interaction based on hostspecific toxin studies" Proceedings of 3rd Tottori International Symposium on Molecular Genetics of Host-Specific Toxins. 3-15 (1998)
[文献書誌] Walton, J.D.et al: "Enzymology,Molecular Genetics,and regulation of biosynthesis of the host-selective toxin HC-toxin" Proceedings of 3rd Tottori International Symposium on Molecular Genetics of Host-Specific Toxins. 25-34 (1998)
[文献書誌] Kodama, M., Akamatsu, H., et al: "Host-specific toxin cleficient mutants of the tomato pathotype of Alternaria alternata obtained by restriction engyme-mediated integration" Proceedings of 3rd Tottori International Symposium on Molecular Genetics of Host-Specific Toxins. 35-42 (1998)
[文献書誌] Tsuge, T., et al: "Molecular analysis of AK-toxin biosynthesis in the Japanese pear pathotype of Alternaria alternata" Proceedings of 3rd Tottori International Symposium on Molecular Genetics of Host-Specific Toxins. 43-52 (1998)
[文献書誌] Itoh, Y., et al: "A catalytic domain of a cyclic peptide synthetase that is specific for the apple pathotype of Alternaria alternata and its possible involvement in host-specific AM-toxin production." Proceedings of 3rd Tottori International Symposium on Molecular Genetics of Host-Specific Toxins. 53-61 (1998)
[文献書誌] Otani, H., et al: "Involvement of host factors in the production of a protein host-specific toxin produced by Alternaria brassicicola" Proceedings of 3rd Tottori International Symposium on Molecular Genetics of Host-Specific Toxins. 63-69 (1998)
[文献書誌] Nakajima, H.: "Structures and biosynthesis of phytotoxins in Cochliobolus spicifer and Bipolaris sorokiniana" Proceedings of 3rd Tottori International Symposium on Molecular Genetics of Host-Specific Toxins. 71-79 (1998)
[文献書誌] Wolpert.T.J.: "Victorin-induced oat cell death" Proceedings of 3rd Tottori International Symposium on Molecular Genetics of Host-Specific Toxins. 105-114 (1998)
[文献書誌] Arase, S.et al: "Primary effect of a host-selective toxin from Magraporthe Grisela to mitochondria of rice leaves" Proceedings of 3rd Tottori International Symposium on Molecular Genetics of Host-Specific Toxins. 115-124 (1998)
[文献書誌] Park, P.: "The accelerated effects of AK-toxin I on exocytosis and endocytosis of susceptible Japanese pear leaves" Proceedings of 3rd Tottori International Symposium on Molecular Genetics of Host-Specific Toxins. 125-135 (1998)
[文献書誌] G.lehrist, D., et al: "Lethal logic in apoptosis : Toxins trigger programmed cell death cluring disoase in eukaryotic cells" Proceedings of 3rd Tottori International Symposium on Molecular Genetics of Host-Specific Toxins. 141-150 (1998)
[文献書誌] Targeon, B.G.et al: "Evolution of pathogenic and reproductive strategies in Cochliobolus and related genera" Proceedings of 3rd Tottori International Symposium on Molecular Genetics of Host-Specific Toxins. 153-163 (1998)
[文献書誌] Bronson, C.R.: "The genetics of pathogenicity in Cochliobolus heterostrophus" Proceedings of 3rd Tottori International Symposium on Molecular Genetics of Host-Specific Toxins. 187-193 (1998)
[文献書誌] Panaccione, D.G.: "Ergopeptine toxins and peptide synthotase genes in clavicipitaceous pathogens and symbionts of plants" Proceedings of 3rd Tottori International Symposium on Molecular Genetics of Host-Specific Toxins. 199-209 (1998)
[文献書誌] Akimitsu, K., et al: "Biochemical and molecular roles of HST and enzymes produced by pathogen of citrus brown spot disease" Proceedings of 3rd Tottori International Symposium on Molecular Genetics of Host-Specific Toxins. 281-290 (1998)
[文献書誌] Rhoads D.M.and Siedow, J.N.: "Characterization of the interaction between fungal pathotoxins and URF-13,the cms-Tmaize mitochondrial T-toxin receptor" Proceedings of 3rd Tottori International Symposium on Molecular Genetics of Host-Specific Toxins. 355-365 (1998)
[文献書誌] Tabira, H., et al: "Selection of a resistant mutant to Alternaria blotch in apple and analysis of proteins associated with the susceptibility by 2-D gel electrophoresis" Proceedings of 3rd Tottori International Symposium on Molecular Genetics of Host-Specific Toxins. 387-397 (1998)
[文献書誌] Kobayashi, H.et al: "Several strategies for dissecting and controlling functions in plant cells" Proceedings of 3rd Tottori International Symposium on Molecular Genetics of Host-Specific Toxins. 399-400 (1998)
[文献書誌] K.Kohmoto and O.C.Yoder(eds): "Moleaclar Gemetics of Host-Specific Tokins in Plant Disease" Kluwer Academic Publishers Dardrecht, 413 (1998)

1997 年度 実績報告書

宿主特異的毒素が介在する植物-微生物相互作用の分子遺伝学に関する日米共同研究

研究代表者

甲元 啓介 鳥取大学, 農学部, 教授 (80032093)

研究成果

[文献書誌] Kusaba, M.and Tsuge, T.: "Mitochondrial DNA variation in host-specific toxin-producing pathogens in the genus Alternaria" Ann.Phytopathol.Soc.Jpn. 63・6. 463-469 (1997)

[文献書誌] Akamatsu, H., Itoh, Y., Kodama, M.et al: "AAL-toxin-deficient mutants of Alternaria alternata tomato pathotype by restriction enzyme-mediated integration" Phytopathology. 87・9. 967-972 (1997)

[文献書誌] Yoder, O.C.: "A mechanistic view of the fungal/plant interaction based on hostspecific toxin studies" Proceedings of 3rd Tottori International Symposium on Molecular Genetics of Host-Specific Toxins. 3-15 (1998)

[文献書誌] Walton, J.D.et al: "Enzymology,Molecular Genetics,and regulation of biosynthesis of the host-selective toxin HC-toxin" Proceedings of 3rd Tottori International Symposium on Molecular Genetics of Host-Specific Toxins. 25-34 (1998)

[文献書誌] Kodama, M., Akamatsu, H., et al: "Host-specific toxin cleficient mutants of the tomato pathotype of Alternaria alternata obtained by restriction engyme-mediated integration" Proceedings of 3rd Tottori International Symposium on Molecular Genetics of Host-Specific Toxins. 35-42 (1998)

[文献書誌] Tsuge, T., et al: "Molecular analysis of AK-toxin biosynthesis in the Japanese pear pathotype of Alternaria alternata" Proceedings of 3rd Tottori International Symposium on Molecular Genetics of Host-Specific Toxins. 43-52 (1998)

[文献書誌] Otani, H., et al: "Involvement of host factors in the production of a protein host-specific toxin produced by Alternaria brassicicola" Proceedings of 3rd Tottori International Symposium on Molecular Genetics of Host-Specific Toxins. 63-69 (1998)

[文献書誌] Nakajima, H.: "Structures and biosynthesis of phytotoxins in Cochliobolus spicifer and Bipolaris sorokiniana" Proceedings of 3rd Tottori International Symposium on Molecular Genetics of Host-Specific Toxins. 71-79 (1998)

[文献書誌] Wolpert.T.J.: "Victorin-induced oat cell death" Proceedings of 3rd Tottori International Symposium on Molecular Genetics of Host-Specific Toxins. 105-114 (1998)

[文献書誌] Arase, S.et al: "Primary effect of a host-selective toxin from Magraporthe Grisela to mitochondria of rice leaves" Proceedings of 3rd Tottori International Symposium on Molecular Genetics of Host-Specific Toxins. 115-124 (1998)

[文献書誌] Park, P.: "The accelerated effects of AK-toxin I on exocytosis and endocytosis of susceptible Japanese pear leaves" Proceedings of 3rd Tottori International Symposium on Molecular Genetics of Host-Specific Toxins. 125-135 (1998)

[文献書誌] G.lehrist, D., et al: "Lethal logic in apoptosis : Toxins trigger programmed cell death cluring disoase in eukaryotic cells" Proceedings of 3rd Tottori International Symposium on Molecular Genetics of Host-Specific Toxins. 141-150 (1998)

[文献書誌] Targeon, B.G.et al: "Evolution of pathogenic and reproductive strategies in Cochliobolus and related genera" Proceedings of 3rd Tottori International Symposium on Molecular Genetics of Host-Specific Toxins. 153-163 (1998)

[文献書誌] Bronson, C.R.: "The genetics of pathogenicity in Cochliobolus heterostrophus" Proceedings of 3rd Tottori International Symposium on Molecular Genetics of Host-Specific Toxins. 187-193 (1998)

[文献書誌] Panaccione, D.G.: "Ergopeptine toxins and peptide synthotase genes in clavicipitaceous pathogens and symbionts of plants" Proceedings of 3rd Tottori International Symposium on Molecular Genetics of Host-Specific Toxins. 199-209 (1998)

[文献書誌] Akimitsu, K., et al: "Biochemical and molecular roles of HST and enzymes produced by pathogen of citrus brown spot disease" Proceedings of 3rd Tottori International Symposium on Molecular Genetics of Host-Specific Toxins. 281-290 (1998)

[文献書誌] Rhoads D.M.and Siedow, J.N.: "Characterization of the interaction between fungal pathotoxins and URF-13,the cms-Tmaize mitochondrial T-toxin receptor" Proceedings of 3rd Tottori International Symposium on Molecular Genetics of Host-Specific Toxins. 355-365 (1998)

[文献書誌] Tabira, H., et al: "Selection of a resistant mutant to Alternaria blotch in apple and analysis of proteins associated with the susceptibility by 2-D gel electrophoresis" Proceedings of 3rd Tottori International Symposium on Molecular Genetics of Host-Specific Toxins. 387-397 (1998)

[文献書誌] Kobayashi, H.et al: "Several strategies for dissecting and controlling functions in plant cells" Proceedings of 3rd Tottori International Symposium on Molecular Genetics of Host-Specific Toxins. 399-400 (1998)

[文献書誌] K.Kohmoto and O.C.Yoder(eds): "Moleaclar Gemetics of Host-Specific Tokins in Plant Disease" Kluwer Academic Publishers Dardrecht, 413 (1998)

1997 年度実績報告書

甲元啓介鳥取大学, 農学部, 教授 (80032093)