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Development of isolation and cultivation methods for uncultivable microbes

Research Project

Project/Area Number 17K19218
Research Category

Grant-in-Aid for Challenging Research (Exploratory)

Allocation TypeMulti-year Fund
Research Field Agricultural chemistry and related fields
Research InstitutionUniversity of Tsukuba

Principal Investigator

Aoyagi Hideki  筑波大学, 生命環境系, 教授 (00251025)

Project Period (FY) 2017-06-30 – 2019-03-31
Project Status Completed (Fiscal Year 2018)
Budget Amount *help
¥6,500,000 (Direct Cost: ¥5,000,000、Indirect Cost: ¥1,500,000)
Fiscal Year 2018: ¥3,250,000 (Direct Cost: ¥2,500,000、Indirect Cost: ¥750,000)
Fiscal Year 2017: ¥3,250,000 (Direct Cost: ¥2,500,000、Indirect Cost: ¥750,000)
Keywords未培養微生物 / 微生物ダークマター / 寒天平板培養法 / 特殊セルロースシート / 培養基材 / 寒天平板培地 / 特殊セルロース / 振盪培養 / 模擬微小重力 / 環境試料 / フィルター
Outline of Final Research Achievements

A feasible cultivation method for uncultivable microorganisms was developed. This culture method aimed at solving the limitations of agar plate culture method. After cultivation from environmental samples such as soil by the developed culture method and agar plate culture method, the grown microbial communities were compared by the denaturing agent gradient gel electrophoresis method. Results showed that the uncultivable microbes in the agar plate method could be successfully grown with the developed cultivation method. In contrast to the conventional culture methods, this method was able to cultivate a higher ratio of new microbes. Moreover, the developed cultivation method maintained a more diverse microbial community as compared to the agar plate culture method. Further investigations on the cultivation method and applications of pretreatment of samples, enhance the diversity and growth of microbes.

Academic Significance and Societal Importance of the Research Achievements

本研究で開発した培養基材は、寒天などのゲル化剤を使用せず、経済性、素材の品質や均一性、操作の簡便性、実用性に優れ、多くの未培養微生物の単離培養が期待できる、閉塞状態にある微生物関連産業界の活性化への貢献や、国内外の産官学の様々な微生物分野(生物工学、応用微生物学、海洋微生物学、病原微生物学、食品微生物学、環境微生物学、極限環境微生物学、腸内細菌学など)で有用性が高い。また、簡便性も高く、未培養微生物資源が豊富なアジアやアフリカなどの現地でも使用可能である。本培養基材の利用性、応用性は拡がりがある。

Report

(3 results)
  • 2018 Annual Research Report   Final Research Report ( PDF )
  • 2017 Research-status Report
  • Research Products

    (22 results)

All 2019 2018 2017 Other

All Int'l Joint Research (1 results) Journal Article (5 results) (of which Int'l Joint Research: 2 results,  Peer Reviewed: 4 results) Presentation (16 results) (of which Invited: 3 results)

  • [Int'l Joint Research] 国立台湾大学(その他の国・地域)

    • Related Report
      2018 Annual Research Report
  • [Journal Article] Adsorption preference for divalent metal ions by Lactobacillus casei JCM1134.2019

    • Author(s)
      Rin Endo and Hideki Aoyagi
    • Journal Title

      Applied Microbiology and Biotechnology

      Volume: 102 Issue: 14 Pages: 6155-6162

    • DOI

      10.1007/s00253-018-9050-1

    • Related Report
      2018 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Monitoring of CO2 and O2 concentrations in the headspace of Sakaguchi flasks during liquid culture of microorganism.2018

    • Author(s)
      Masato Takahashi and Hideki Aoyagi
    • Journal Title

      Applied Microbiology and Biotechnology

      Volume: 102 Issue: 15 Pages: 6637-6645

    • DOI

      10.1007/s00253-018-9076-4

    • Related Report
      2018 Annual Research Report
    • Peer Reviewed
  • [Journal Article] 従来の微生物培養法の生物化学工学的解析とその利用2018

    • Author(s)
      高橋将人、青柳秀紀
    • Journal Title

      日本放線菌学会誌

      Volume: 32 Pages: 25-26

    • Related Report
      2018 Annual Research Report
  • [Journal Article] Effect of intermittent opening of breathable culture plugs and aeration of headspace on the structure of microbial communities in shake-flask culture2018

    • Author(s)
      Takahashi Masato、Aoyagi Hideki
    • Journal Title

      Journal of Bioscience and Bioengineering

      Volume: in press Issue: 1 Pages: 1-6

    • DOI

      10.1016/j.jbiosc.2018.01.009

    • Related Report
      2017 Research-status Report
    • Peer Reviewed / Int'l Joint Research
  • [Journal Article] Practices of shake-flask culture and advances in monitoring CO2 and O22018

    • Author(s)
      Masato Takahashi, Hideki Aoyagi
    • Journal Title

      Applied Microbiology and Biotechnology

      Volume: in press Issue: 10 Pages: 1-11

    • DOI

      10.1007/s00253-018-8922-8

    • Related Report
      2017 Research-status Report
    • Peer Reviewed / Int'l Joint Research
  • [Presentation] セルロースメンブレンを用いた植物成長促進細菌の評価と培養法の開発2019

    • Author(s)
      大矢裕之、青柳秀紀、林乃君
    • Organizer
      化学工学会 第84回年会
    • Related Report
      2018 Annual Research Report
  • [Presentation] 発酵微生物のスクリーニングを考える2018

    • Author(s)
      青柳秀紀、高橋将人、松﨑大典、長谷川智弘
    • Organizer
      第70回日本生物工学会大会
    • Related Report
      2018 Annual Research Report
    • Invited
  • [Presentation] 微生物を用いた液体培養におけるフラスコ気相環境の解析2018

    • Author(s)
      高橋将人、青柳秀紀
    • Organizer
      第70回日本生物工学会大会
    • Related Report
      2018 Annual Research Report
  • [Presentation] pH変動培養法を活用した耐酸性微生物や好酸性微生物の高効率なスクリーニング系の開発2018

    • Author(s)
      高野力、村山晃一、青柳秀紀
    • Organizer
      第70回日本生物工学会大会
    • Related Report
      2018 Annual Research Report
  • [Presentation] 固定化細胞培養法を活用した生物細胞の選択的取得法の開発2018

    • Author(s)
      吉田真梨、青柳秀紀
    • Organizer
      第70回日本生物工学会大会
    • Related Report
      2018 Annual Research Report
  • [Presentation] 植物試料の凍結前処理が培養微生物群集に与える影響の解析2018

    • Author(s)
      小林和輝、青柳秀紀
    • Organizer
      第70回日本生物工学会大会
    • Related Report
      2018 Annual Research Report
  • [Presentation] 増殖の速い微生物が環境試料からの微生物の培養化に及ぼす影響の解析と利用2018

    • Author(s)
      長谷川智弘、原田玲奈、青柳秀紀
    • Organizer
      第70回日本生物工学会大会
    • Related Report
      2018 Annual Research Report
  • [Presentation] マイクロ波(非熱効果)が微生物の生理活性に及ぼす影響の解析2018

    • Author(s)
      松田将典、青柳秀紀
    • Organizer
      第70回日本生物工学会大会
    • Related Report
      2018 Annual Research Report
  • [Presentation] 従来の微生物培養法の生物化学工学的解析とその利用2018

    • Author(s)
      高橋将人、青柳秀紀
    • Organizer
      日本放線菌学会 学術講演会
    • Related Report
      2018 Annual Research Report
    • Invited
  • [Presentation] 振盪フラスコ培養中のフラスコ気相部が培養微生物群集に及ぼす影響の解析2018

    • Author(s)
      高橋将人、青柳秀紀
    • Organizer
      第69回日本生物工学会大会
    • Related Report
      2017 Research-status Report
  • [Presentation] 物理的な前処理を活用した樹皮試料からの新規微生物の培養化2018

    • Author(s)
      小林和輝、村山晃一、青柳秀紀
    • Organizer
      第69回日本生物工学会大会
    • Related Report
      2017 Research-status Report
  • [Presentation] 従来の微生物培養法の特性:問題点と解決策の提案2018

    • Author(s)
      青柳秀紀
    • Organizer
      第69回日本生物工学会大会
    • Related Report
      2017 Research-status Report
    • Invited
  • [Presentation] ストロンチウムがツバキ花粉の生理活性に及ぼす影響の解析2018

    • Author(s)
      中村真由、青柳秀紀
    • Organizer
      第69回日本生物工学会大会
    • Related Report
      2017 Research-status Report
  • [Presentation] 固定化培養を活用したシロアリ腸内の未培養微生物の培養化2018

    • Author(s)
      森峻一、青柳秀紀
    • Organizer
      第69回日本生物工学会大会
    • Related Report
      2017 Research-status Report
  • [Presentation] SCF法を用いた強酸性条件下における未培養微生物の培養化2017

    • Author(s)
      高野力、村山晃一、青柳秀紀
    • Organizer
      第69回日本生物工学会大会
    • Related Report
      2017 Research-status Report
  • [Presentation] セシウムが花粉の生理活性に及ぼす影響の解析(第2 報)2017

    • Author(s)
      青柳秀紀
    • Organizer
      日本花粉学会第58回(2017年)大会
    • Related Report
      2017 Research-status Report

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Published: 2017-07-21   Modified: 2024-01-30  

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