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Construction of filamentous fungal artificial chromosome and analysis of horizontal transfer and pathogenicity evolution mechanisms in the tomato wilt fungus.

Research Project

Project/Area Number 18K19218
Research Category

Grant-in-Aid for Challenging Research (Exploratory)

Allocation TypeMulti-year Fund
Review Section Medium-sized Section 39:Agricultural and environmental biology and related fields
Research InstitutionTokyo University of Science

Principal Investigator

ARAZOE Takayuki  東京理科大学, 理工学部応用生物科学科, 講師 (40749975)

Project Period (FY) 2018-06-29 – 2022-03-31
Project Status Completed (Fiscal Year 2021)
Budget Amount *help
¥6,370,000 (Direct Cost: ¥4,900,000、Indirect Cost: ¥1,470,000)
Fiscal Year 2020: ¥2,080,000 (Direct Cost: ¥1,600,000、Indirect Cost: ¥480,000)
Fiscal Year 2019: ¥2,600,000 (Direct Cost: ¥2,000,000、Indirect Cost: ¥600,000)
Fiscal Year 2018: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000)
Keywordsトマト萎凋病菌 / ゲノム編集 / 糸状菌人工染色体 / 小型染色体 / 病原性染色体 / アクセサリー染色体 / ゲノム進化 / 水平伝播 / 病原性変異
Outline of Final Research Achievements

We developed a genome editing technology optimized for Fusarium oxysporum to enable efficient targeted gene knock-out, knock-in, base editing, and chromosomal deletion. Using this technology, we constructed a filamentous fungal artificial chromosome by removing a large proportion of the accessory chromosome, and showed the potential of the technology to carry out intracellular cloning of chromosomal regions, chromosome reconstruction, and edited chromosome transfer. Our findings and developed technologies would enable comprehensive analysis of hyphal fusion, horizontal transfer, pathogenicity evolution mechanisms, and the function of the accessory chromosomal region in F. oxysporum

Academic Significance and Societal Importance of the Research Achievements

酵母,植物,マウスおよびヒトにおける人工染色体は確立されており,これまで導入が困難であった広域な染色体領域の導入やモデルマウスの作成等,従来の研究領域を大幅に拡大させた。本研究では,糸状菌では初となる人工染色体の作出に成功し,新たなゲノム編集手法を含む遺伝子工学・染色体工学的な利用可能性を示した。また人工染色体作製に用いたゲノム領域の広域欠失誘導は,染色体領域を標的とした新たなゲノム機能解析手法として有用である。以上より,応用微生物学や植物保護科学,遺伝子工学,染色体工学等,多様な学術的・社会的分野において重要な知見や技術を得ることができた。

Report

(3 results)
  • 2021 Final Research Report ( PDF )
  • 2019 Research-status Report
  • 2018 Research-status Report
  • Research Products

    (32 results)

All 2020 2019 2018

All Journal Article (4 results) (of which Peer Reviewed: 4 results,  Open Access: 4 results) Presentation (25 results) (of which Int'l Joint Research: 5 results,  Invited: 1 results) Book (2 results) Patent(Industrial Property Rights) (1 results) (of which Overseas: 1 results)

  • [Journal Article] Extremely Low Concentrations of Acetic Acid Stimulate Cell Differentiation in Rice Blast Fungus2020

    • Author(s)
      Kuroki Misa、Shiga Yuriko、Narukawa-Nara Megumi、Arazoe Takayuki、Kamakura Takashi
    • Journal Title

      iScience

      Volume: 23 Issue: 1 Pages: 100786-100786

    • DOI

      10.1016/j.isci.2019.100786

    • Related Report
      2019 Research-status Report
    • Peer Reviewed / Open Access
  • [Journal Article] Single crossover-mediated targeted nucleotide substitution and knock-in strategies with CRISPR/Cas9 system in the rice blast fungus2019

    • Author(s)
      Yamato, T., Handa, A., Arazoe, T., Kuroki, M., Nozaka, A., Kamakura, T., Ohsato, S., Arie, T., Kuwata, S.
    • Journal Title

      Scientific Reports

      Volume: 印刷中 Issue: 1 Pages: 7427-7427

    • DOI

      10.1038/s41598-019-43913-0

    • Related Report
      2018 Research-status Report
    • Peer Reviewed / Open Access
  • [Journal Article] Chloramphenicol inhibits eukaryotic Ser/Thr phosphatase and infection-specific cell differentiation in the rice blast fungus2019

    • Author(s)
      Nozaka, A., Nishiwaki, A., Nagashima Y., Endo, S., Kuroki, M., Nakajima, M., Narukawa, M., Kamisuki, S., Arazoe, T., Taguchi, H., Sugawara, F., Kamakura T.
    • Journal Title

      Scientific Reports

      Volume: 印刷中

    • Related Report
      2018 Research-status Report
    • Peer Reviewed / Open Access
  • [Journal Article] Targeted nucleotide editing technologies for microbial metabolic engineering2018

    • Author(s)
      Arazoe, T., Kondo, A., Nishida, K.
    • Journal Title

      Biotechnology Journal

      Volume: 13 Issue: 9 Pages: 170596-170596

    • DOI

      10.1002/biot.201700596

    • Related Report
      2018 Research-status Report
    • Peer Reviewed / Open Access
  • [Presentation] Ectopic recombination between solo-long terminal repeats triggered pathogenic changes and genome rearrangement in the rice blast fungus2020

    • Author(s)
      Arazoe, T., Murochi, S., Hiraoka, D., Kiguchi, K., Tanaka, T., Sakuma, T., Yamamoto, T., Arie, T., Kuwata, S., Chuma, I., Ohsato, S., Tosa, Y., Kamakura, T.
    • Organizer
      15th European Conference on Fungal Genetics
    • Related Report
      2019 Research-status Report
    • Int'l Joint Research
  • [Presentation] Exploration of the genetic cause of female sterility in the rice blast fungus2020

    • Author(s)
      Uchida, M., Kita, K., Fujigasaki, A., Konishi, T., Arie, T., Teraoka, T., Arazoe, T., Kamakura, T.
    • Organizer
      15th European Conference on Fungal Genetics
    • Related Report
      2019 Research-status Report
    • Int'l Joint Research
  • [Presentation] イネいもち病菌における推定カルシウムチャネル及びそ の構成因子の機能解析2020

    • Author(s)
      山内優季,平岡輪子,林平,内田百岳,荒添貴之, 鎌倉高志
    • Organizer
      日本農芸化学会2020年度大会
    • Related Report
      2019 Research-status Report
  • [Presentation] イネいもち病菌非病原力遺伝子の欠失 の規模を決める要因2020

    • Author(s)
      平岡大輝,村井俊子,荒添貴之, 曾根輝雄,土佐幸雄,中馬いづみ
    • Organizer
      令和2年度日本植物病理学会大会
    • Related Report
      2019 Research-status Report
  • [Presentation] CRISPR/Cas9系を用いたFusarium 属菌でのSIX1およびfgm4-like 遺伝子破壊株の作出2020

    • Author(s)
      齊藤大幹,荒添貴之,小寺俊丞,小松健,有江 力
    • Organizer
      令和2年度日本植物病理学会大会
    • Related Report
      2019 Research-status Report
  • [Presentation] イネいもち病菌のキチン脱アセチル化酵素タンパク質Cbp1のGPI アンカー化検証2020

    • Author(s)
      前村知佳,黒木美沙,荒添貴之,鎌倉高志
    • Organizer
      令和2年度日本植物病理学会大会
    • Related Report
      2019 Research-status Report
  • [Presentation] トマト萎凋病菌におけるCRISPR/Cas9およびゲノム編集系の確立2020

    • Author(s)
      新門想太,齊藤大幹,小寺俊丞, 荒添貴之,鎌倉高志
    • Organizer
      令和2年度日本植物病理学会大会
    • Related Report
      2019 Research-status Report
  • [Presentation] 超低濃度脂肪酸によって誘導されるイネいもち病菌の付着器形成誘導機構の解析2020

    • Author(s)
      佐藤大元,齋藤翔太,黒木美沙,荒添貴之,鎌倉高志
    • Organizer
      令和2年度日本植物病理学会大会
    • Related Report
      2019 Research-status Report
  • [Presentation] 植物病原糸状菌ゲノム編集技術の開発2020

    • Author(s)
      荒添貴之
    • Organizer
      令和2年度日本植物病理学会大会
    • Related Report
      2019 Research-status Report
    • Invited
  • [Presentation] イネいもち病菌におけるクロラムフェニコールの新規作用点の探索2019

    • Author(s)
      樋口絵莉香,野坂亮仁,田代綾子,荒添貴之,鎌倉高志
    • Organizer
      第19回糸状菌分子生物学コンファレンス
    • Related Report
      2019 Research-status Report
  • [Presentation] イネいもち病菌 (Pyricularia oryzae) における交配型決定領域の機能解析2019

    • Author(s)
      喜多光徹,内田百岳,藤ヶ崎礼夏,小西高裕,寺岡徹,有江力,荒添貴之,鎌倉高志
    • Organizer
      第19回糸状菌分子生物学コンファレンス
    • Related Report
      2019 Research-status Report
  • [Presentation] 生命の設計図を操作するゲノム編集:神の領域?悪魔の技術?2019

    • Author(s)
      荒添貴之
    • Organizer
      明治大学科学技術研究所 公開講演会 「ゲノム編集:何ができるか、その原理と活用方法」
    • Related Report
      2019 Research-status Report
  • [Presentation] イネいもち病菌における核数制御因子の探索2019

    • Author(s)
      斎藤翔太,菊澤佑斗,黒木美沙,荒添貴之,鎌倉高志
    • Organizer
      日本農芸化学会2019年度東京大会
    • Related Report
      2018 Research-status Report
  • [Presentation] Lactobacillus nodensisにおけるD-arabinose代謝経路の解析2019

    • Author(s)
      堀江絢子,奥村友梨,ジョ ミン,荒添貴之,鎌倉高志
    • Organizer
      日本農芸化学会2019年度東京大会
    • Related Report
      2018 Research-status Report
  • [Presentation] イネいもち病菌非病原力遺伝子のrepeat-poor領域における欠失2019

    • Author(s)
      平岡大輝,荒添貴之,曾根輝雄,土佐幸雄,中馬いづみ
    • Organizer
      平成31年度日本植物病理学会大会
    • Related Report
      2018 Research-status Report
  • [Presentation] Development of CRISPR/Cas9 system and genome editing technology for the non-model fungus, Moniliella polinis2019

    • Author(s)
      Marusawa, D., Inaba, A., Arazoe, T., Kamakura, T.
    • Organizer
      30th Fungal Genetics Conference
    • Related Report
      2018 Research-status Report
    • Int'l Joint Research
  • [Presentation] DNA double-strand breaks-mediated ectopic recombination between solo-long terminal repeats triggered pathogenic changes and genome rearrangement in the rice blast fungus2019

    • Author(s)
      Arazoe, T., Hiraoka, D., Murochi, S., Kiguchi, K., Tanaka, T., Sakuma, T., Yamamoto, T., Arie, T., Kuwata, S., Chuma, I., Ohsato, S., Tosa, Y., Kamakura T.
    • Organizer
      MPMI XVIII Congress
    • Related Report
      2018 Research-status Report
    • Int'l Joint Research
  • [Presentation] Chitin-deacetylase activity has important roles in inducing appressorium formation2019

    • Author(s)
      Saito, S.. Shiga, Y., Kuroki, M., Arazoe, T., Kamakura, T.
    • Organizer
      IS-MPMI XVIII Congress
    • Related Report
      2018 Research-status Report
    • Int'l Joint Research
  • [Presentation] 麹菌 ligD遺伝子がゲノム編集に起因する大規模欠失変異に与える影響2018

    • Author(s)
      上元優,林梨咲,渡嘉敷直杏,荒添貴之,山田修,外山博英,水谷治
    • Organizer
      日本ゲノム編集学会第3回大会
    • Related Report
      2018 Research-status Report
  • [Presentation] イネいもち病菌の相同組換え関連遺伝子破壊株におけるDNAリセクション反応系の構築に向けて2018

    • Author(s)
      木口歌菜,田中寿樹,荒添貴之,佐久間哲史,山本卓,桑田茂,大里修一
    • Organizer
      平成30年度日本植物病理学会九州部会
    • Related Report
      2018 Research-status Report
  • [Presentation] イネいもち病菌を利用した抗生物質Roxithromycinと標的候補タンパク質Cdc27のin vivo解析2018

    • Author(s)
      鈴木優花,林真優子,石井晶,荒添貴之,鎌倉高志
    • Organizer
      日本農芸化学会関東支部2018年度大会
    • Related Report
      2018 Research-status Report
  • [Presentation] イネいもち病菌における核数制御因子の探索2018

    • Author(s)
      斎藤翔太,菊澤佑斗,荒添貴之,鎌倉高志
    • Organizer
      日本農芸化学会関東支部2018年度大会
    • Related Report
      2018 Research-status Report
  • [Presentation] イネいもち病菌におけるキチン脱アセチル化酵素を介した細胞分化誘導2018

    • Author(s)
      黒木美沙,志賀友里子,前村知佳,荒添貴之,鎌倉高志
    • Organizer
      第18回糸状菌分子生物学コンファレンス
    • Related Report
      2018 Research-status Report
  • [Presentation] イネいもち病菌におけるクロラムフェニコールの作用点の探索2018

    • Author(s)
      田代綾子,野坂亮仁,樋口絵莉香,荒添貴之,鎌倉高志
    • Organizer
      第18回糸状菌分子生物学コンファレンス
    • Related Report
      2018 Research-status Report
  • [Presentation] CRISPR/Cas9系を用いたイネいもち病菌における標的遺伝子ノックダウン法の確立2018

    • Author(s)
      平岡輪子,林平,荒添貴之,鎌倉高志
    • Organizer
      第18回糸状菌分子生物学コンファレンス
    • Related Report
      2018 Research-status Report
  • [Book] Elsevior2020

    • Author(s)
      Arazoe, T., Mizutani, O.
    • Total Pages
      361
    • Publisher
      Genome Engineering via CRISPR-Cas9 System, Chapter5-Targeted genome editing using CRISPR/Cas9 system in fungi
    • ISBN
      9780128181409
    • Related Report
      2019 Research-status Report
  • [Book] 完全版 ゲノム編集実験スタンダード,2-9. 糸状菌でのゲノム編集2019

    • Author(s)
      荒添貴之
    • Total Pages
      386
    • Publisher
      羊土社
    • ISBN
      9784758122443
    • Related Report
      2019 Research-status Report
  • [Patent(Industrial Property Rights)] METHOD FOR CONVERTING NUCLEIC ACID SEQUENCE OF CELL SPECIFICALLY CONVERTING NUCLEIC ACID BASE OF TARGETED DNA USING CELL ENDOGENOUS DNA MODIFYING ENZYME, AND MOLECULAR COMPLEX USED THEREIN2018

    • Inventor(s)
      西田敬二,近藤昭彦,荒添貴之,吉岡伸
    • Industrial Property Rights Holder
      西田敬二,近藤昭彦,荒添貴之,吉岡伸
    • Industrial Property Rights Type
      特許
    • Filing Date
      2018
    • Acquisition Date
      2018
    • Related Report
      2018 Research-status Report
    • Overseas

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Published: 2018-07-25   Modified: 2025-01-30  

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