Mitotic homologous recombination and pathogenic changes in the fungus Pyricularia oryzae

• Project/Area Number: 24580070
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Plant pathology
• Research Institution: Meiji University
• Principal Investigator: SHUICHI Ohsato 明治大学, 農学部, 講師 (30533228)
• Project Period (FY): 2012-04-01 – 2015-03-31
• Project Status: Completed (Fiscal Year 2014)
• Budget Amount: ¥5,460,000 (Direct Cost: ¥4,200,000、Indirect Cost: ¥1,260,000); Fiscal Year 2014: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000); Fiscal Year 2013: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000); Fiscal Year 2012: ¥2,470,000 (Direct Cost: ¥1,900,000、Indirect Cost: ¥570,000)
• Keywords: イネいもち病菌 / 相同組換え / DNA二本鎖切断 / 病原性変異 / 準有性生殖 / I-Sce I

Outline of Final Research Achievements

To study somatic homologous recombination (HR) in Pyricularia oryzae, we created an HR detection system, using which we showed that somatic HR occurs at high levels in P. oryzae after treatment with several chemical agents. Somatic HR was also seen to be involved in the instability and diversification of the avirulence gene Avr-Pita in P. oryzae. We found that DNA double-strand breaks (DSBs) generated by the endonuclease I-SceI increased the frequency of ectopic HR in the genome of P. oryzae. To generate DSBs at a specific site in the desired locus within the P. oryzae genome, we designed and engineered nucleases called Platinum-Fungal TALENs. To summarize, genetic mutations caused by somatic HR in P. oryzae are responsible for the diversification of its virulence.

Research Products

• [Journal Article] Tailor-made TALEN system for highly efficient targeted gene replacement in the rice blast fungus (2015)
  • Author(s): Arazoe T, Ogawa T, Miyoshi K, Yamato T, Ohsato S, Sakuma T, Yamamoto T, Arie T, Kuwata S
  • Journal Title: Biotechnology and Bioengineering Volume: - Issue: 7 Pages: 1335-1342
  • DOI: 10.1002/bit.25559
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] The effect of chemicals on somatic homologous recombination in the rice blast fungus: its possible application for detection of mycotoxins (2014)
  • Author(s): Arazoe T, Ohsato S, Maeda K, Arie T, Kuwata S
  • Journal Title: JSM Mycotoxins Volume: 64 Issue: 2 Pages: 141-146
  • DOI: 10.2520/myco.64.141
  • NAID: 130004699346
  • ISSN: 0285-1466, 1881-0128
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Construction and characterization of a copy number-inducible fosmid library of Xanthomonas oryzae pathovar oryzae MAFF311018 (2014)
  • Author(s): Ichida H, Sun X, Imanaga S, Ito Y, Yoneyama K, Kuwata S, Ohsato S
  • Journal Title: Gene Volume: 546 Issue: 1 Pages: 68-72
  • DOI: 10.1016/j.gene.2014.05.035
  • Peer Reviewed
• [Journal Article] Site-specific DNA double-strand break generated by I-SceI endonuclease enhances ectopic homologous recombination in Pyricularia oryzae. (2014)
  • Author(s): Arazoe T, Younomaru T, Ohsato S, Kimura M, Arie T, Kuwata S
  • Journal Title: FEMS Microbiol. Lett. Volume: 352 Issue: 2 Pages: 221-229
  • DOI: 10.1111/1574-6968.12396
  • Peer Reviewed
• [Journal Article] Experimental evidence of a pathogenic change caused by homologo us recombination between endogenous and introduced-dysfunctional Avr-Pita genes in Pyricularia oryzae (2014)
  • Author(s): Arazoe, T., Kuwata, S., Arie, T., Ohsato, S.
  • Journal Title: Journal of General Plant Pathology Volume: 80 Issue: 2 Pages: 153-157
  • DOI: 10.1007/s10327-013-0502-2
  • Peer Reviewed
• [Journal Article] Construction of a system for exploring mitotic homologous recombination in the genome of Pyricularia oryzae (2013)
  • Author(s): Arazoe, T., Younomaru, T., Ohsato, S., Arie, T., Yoneyama, K. and Kuwata, S.
  • Journal Title: Journal of General Plant Pathology Volume: 79 Issue: 6 Pages: 422-430
  • DOI: 10.1007/s10327-013-0474-2
  • Peer Reviewed
• [Presentation] イネいもち病菌Srs2DNAヘリカーゼの機能解析 (2015)
  • Author(s): 小川哲央，荒添貴之，佐久間哲史，山本卓，桑田茂，草野好司，大里修一
  • Organizer: 日本植物病理学会創立100周年記念大会
  • Place of Presentation: 明治大学駿河台キャンパス
  • Year and Date: 2015-03-29 – 2015-03-31
• [Presentation] 糸状菌型人工ヌクレアーゼPlatinum Fungal TALENsを用いたイネいもち病菌における新規遺伝子ノックインおよび塩基置換導入法 (2015)
  • Author(s): 三好健之介，荒添貴之，大和澄，小川哲央，佐久間哲史，山本卓，大里修一，有江力，桑田茂
  • Organizer: 日本植物病理学会創立100周年記念大会
  • Place of Presentation: 明治大学駿河台キャンパス
  • Year and Date: 2015-03-29 – 2015-03-31
• [Presentation] DNA二本鎖切断とその修復過程において生じるイネいもち病菌の病原性変異 (2015)
  • Author(s): 荒添貴之，田中寿樹，小川哲央，三好健之介，大和澄，佐久間哲史，山本卓，有江力，中馬いづみ，大里修一，土佐幸雄，桑田茂
  • Organizer: 日本植物病理学会創立100周年記念大会
  • Place of Presentation: 明治大学駿河台キャンパス
  • Year and Date: 2015-03-29 – 2015-03-31
• [Presentation] 人工ヌクレアーゼTALENsを用いたイネいもち病菌における高効率遺伝子改変法 (2014)
  • Author(s): 荒添貴之，小川哲央，大里修一，佐久間哲史，山本卓，有江力，桑田茂
  • Organizer: 第14回糸状菌分子生物コンファレンス
  • Place of Presentation: 東北大学川内北キャンパス
  • Year and Date: 2014-11-15 – 2014-11-16
• [Presentation] 植物病原糸状菌 (イネいもち病菌) のゲノム編集および変異機構解析に向けて (2014)
  • Author(s): 荒添貴之，大里修一，佐久間哲史，山本卓，有江力，桑田茂
  • Organizer: 植物ゲノム編集ワークショップ
  • Place of Presentation: 岡山大学資源植物科学研究所
  • Year and Date: 2014-11-04
• [Presentation] 糸状菌型Platinum TALENの作製とイネいもち病菌 (糸状菌) におけるゲノム編集 (2014)
  • Author(s): 荒添貴之，小川哲央，大里修一，佐久間哲史，山本卓，有江力，桑田茂
  • Organizer: 第4回ゲノム編集研究会
  • Place of Presentation: 広島国際会議場ダリア
  • Year and Date: 2014-10-06 – 2014-10-07
• [Presentation] TALENを用いた麹菌Aspergillus oryzae におけるゲノム編集 (2014)
  • Author(s): 水谷治，荒添貴之，利田賢次，林梨咲，大里修一，佐久間哲史，山本卓，桑田茂，山田修
  • Organizer: 第4回ゲノム編集研究会
  • Place of Presentation: 広島国際会議場ダリア
  • Year and Date: 2014-10-06 – 2014-10-07
• [Presentation] Highly efficient gene targeting in Pyricularia oryzae by Zinc Finger Nuclease (2014)
  • Author(s): Arazoe T, Ohsato S, Arie T, Kuwata S
  • Organizer: XVI International Congress on Molecular Plant-Microbe Interactions
  • Place of Presentation: Rhodes, Greece
  • Year and Date: 2014-07-06 – 2014-07-10
• [Presentation] Platinum Gate TALENシステムを用いたイネいもち病菌における高効率遺伝子ターゲッティング (2014)
  • Author(s): 荒添貴之，用之丸哲也，大里修一，佐久間哲史，山本卓，有江力，桑田茂
  • Organizer: 平成26年度日本植物病理学会
  • Place of Presentation: 札幌コンベンションセンター
  • Year and Date: 2014-06-02 – 2014-06-04
• [Presentation] イネいもち病菌Srs2DNAヘリカーゼの単離と機能解析 (2014)
  • Author(s): 小川哲央，荒添貴之，桑田茂，草野好司，大里修一
  • Organizer: 平成26年度日本植物病理学会
  • Place of Presentation: 札幌コンベンションセンター
  • Year and Date: 2014-06-02 – 2014-06-04
• [Presentation] DNA二本鎖切断によるイネいもち病菌の標的遺伝子改変法 (2013)
  • Author(s): 用之丸哲也，荒添貴之，大里修一，有江力， 桑田茂
  • Organizer: 第13回糸状菌分子生物コンファレンス
  • Place of Presentation: つくば国際会議場
• [Presentation] Development of a novel reporter system for analyzing gene expressions in rhizobia (2013)
  • Author(s): Yasuhiro Ito, Hiroyuki Ichida, Shuichi Ohsato, Shigeru Kuwata
  • Organizer: 18th International Congress on Nitrogen Fixation
  • Place of Presentation: Miyazaki Prefecture, Phoneix Seagaia
• [Presentation] DNA二本鎖切断を介したイネいもち病菌の遺伝子ターゲティング法 (2013)
  • Author(s): 用之丸哲也，荒添貴之，大里修一，有江力， 桑田茂
  • Organizer: 日本植物病理学会関東部会
  • Place of Presentation: 法政大学市ヶ谷キャンパス
• [Presentation] Zinc finger nucleaseおよびtranscription activator-like effector nucleaseを用いたイネいもち病菌ゲノムDNAの配列特異的二本鎖切断導入の試み (2013)
  • Author(s): 荒添貴之，用之丸哲也，大里修一，有江力，米山勝美，桑田茂
  • Organizer: 平成25年度日本植物病理学会大会
  • Place of Presentation: 岐阜大学
• [Presentation] DNA double-strand breaks generated by yeast endonuclease I-Sce I induce ectopic homologous recombination and targeted gene replacement in Magnaporthe oryzae (2013)
  • Author(s): T. Arazoe, T. Younomaru, S. Ohsato, T. Arie, S. Kuwata
  • Organizer: 27th Fungal Genetics Conference
  • Place of Presentation: Asilomar Conference Grounds CA, USA
• [Presentation] イネいもち病菌におけるDNA二本鎖切断誘導系を用いた遺伝的変異機構の解析と遺伝子ターゲッティング法への応用 (2012)
  • Author(s): 荒添貴之，用之丸哲也，大里修一，有江力，桑田茂
  • Organizer: 第12回糸状菌分子生物コンファレンス
  • Place of Presentation: ウインクあいち(愛知県名古屋市)
• [Presentation] イネいもち病菌感染誘導性を示すジベレリン2-酸化酵素遺伝子 (2012)
  • Author(s): 真籠洋、藤枝俊介、花田篤志、大里修一、神谷勇治、山口信次郎
  • Organizer: 第47回植物化学調節学会大会
  • Place of Presentation: 山形大学農学部(鶴岡)
• [Presentation] Various stress conditions induce somatic homologous recombination in Magnaporthe oryzae (2012)
  • Author(s): T. Arazoe, S. Ohsato, T. Arie, K. Yoneyama, S. Kuwata
  • Organizer: XV International Congress of Molecular Plant-Microbe Interactions
  • Place of Presentation: Kyoto International Conference Center