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Cultivation of microbial dark matter using novel isolation strategies

Research Project

Project/Area Number 16H04833
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section一般
Research Field Biodiversity/Systematics
Research InstitutionHiroshima University

Principal Investigator

Kindaichi Tomonori  広島大学, 工学研究科, 助教 (10379901)

Co-Investigator(Kenkyū-buntansha) 青井 議輝  広島大学, 先端物質科学研究科, 准教授 (40386636)
Project Period (FY) 2016-04-01 – 2019-03-31
Project Status Completed (Fiscal Year 2018)
Budget Amount *help
¥17,550,000 (Direct Cost: ¥13,500,000、Indirect Cost: ¥4,050,000)
Fiscal Year 2018: ¥4,810,000 (Direct Cost: ¥3,700,000、Indirect Cost: ¥1,110,000)
Fiscal Year 2017: ¥5,200,000 (Direct Cost: ¥4,000,000、Indirect Cost: ¥1,200,000)
Fiscal Year 2016: ¥7,540,000 (Direct Cost: ¥5,800,000、Indirect Cost: ¥1,740,000)
Keywords微生物ダークマター / Candidate division微生物群 / 16S rRNA遺伝子 / MAR-FISH法 / 新規分離培養手法 / Candidate division細菌群 / 群集・生態系多様性 / 分離培養
Outline of Final Research Achievements

In this study, Candidate division microorganisms, such as TM7, BD1-5, SR1, and OD1 were selected from activated sludge samples. Specific probes for the fluorescence in situ hybridization were designed for above four groups, then probes specific for TM7 and BD1-5 were optimized. Metagenomic analysis were performed for the activated sludge sample and revealed that BD1-5 have a potential for uptake of amino acids. MAR-FISH analysis for BD1-5 also revealed the uptake of amino acids under anaerobic conditions. Specific primer set for TM7 for quantitative PCR was established. We demonstrated the optimal condition for the growth of TM7. Isolation technique using cell sorting showed separation of anammox bacteria, as model uncultivable bacteria, from the complex microbial community. We also demonstrated the concept of an innovative cultivation method that enable isolating pure culture automatically using model microorganisms.

Academic Significance and Societal Importance of the Research Achievements

これまでに機能が未知であった未培養微生物群の機能が明らかになる。従来の微生物分離培養手法を凌駕する高性能で汎用性の高い分離培養手法の実用化が達成できる。未培養細菌の純粋培養系が得られ、新規単離菌株の学名提案ができる。これまで培養可能な微生物からのみ得られてきた物質・情報が飛躍的に高まるため、本研究成果の波及効果は極めて大きく、多分野(学術面・医学面・産業面など)に及ぶ。環境中の微生物の99%は未培養であり、未利用バイオリソースの開拓に貢献できる。生物界の3 つのドメインの境界または共通祖先微生物などの生物の進化に関連する理解が深まる。

Report

(4 results)
  • 2018 Annual Research Report   Final Research Report ( PDF )
  • 2017 Annual Research Report
  • 2016 Annual Research Report
  • Research Products

    (14 results)

All 2018 2017 2016 Other

All Int'l Joint Research (1 results) Journal Article (1 results) (of which Peer Reviewed: 1 results,  Open Access: 1 results) Presentation (12 results) (of which Int'l Joint Research: 2 results)

  • [Int'l Joint Research] Aalborg University(Denmark)

    • Related Report
      2016 Annual Research Report
  • [Journal Article] Specificities and Efficiencies of Primers Targeting Candidatus Phylum Saccharibacteria in Activated Sludge2018

    • Author(s)
      Takenaka Ryota、Aoi Yoshiteru、Ozaki Noriatsu、Ohashi Akiyoshi、Kindaichi Tomonori
    • Journal Title

      Materials

      Volume: 11 Issue: 7 Pages: 1129-1129

    • DOI

      10.3390/ma11071129

    • Related Report
      2018 Annual Research Report
    • Peer Reviewed / Open Access
  • [Presentation] 活性汚泥内Saccharibacteriaの集積培養条件の検討.2018

    • Author(s)
      竹中 亮太, 青井 議輝, 大橋 晶良, 金田一 智規.
    • Organizer
      第53回日本水環境学会年会
    • Related Report
      2018 Annual Research Report
  • [Presentation] 活性汚泥内に存在するGracilibacteria(BD1-5)門の機能解析.2018

    • Author(s)
      赤木 秀至, 青井 議輝, 大橋 晶良, 金田一 智規.
    • Organizer
      第53回日本水環境学会年会
    • Related Report
      2018 Annual Research Report
  • [Presentation] Investigation of growth conditions of Candidatus Saccharibacteria in activated sludge.2018

    • Author(s)
      Ryota Takenaka, Yoshiteru Aoi, Noriatsu Ozaki, Akiyoshi Ohashi, Tomonori Kindaichi.
    • Organizer
      日本微生物生態学会第32回大会
    • Related Report
      2018 Annual Research Report
  • [Presentation] 活性汚泥内の Candidatus Saccharibacteria の集積培養の試み.2018

    • Author(s)
      竹中 亮太, 尾崎 則篤, 大橋 晶良, 金田一 智規.
    • Organizer
      第52回日本水環境学会年会
    • Related Report
      2017 Annual Research Report
  • [Presentation] 環境中の微生物を自動的に“捕え”て“分離”する革新的分離培養手法の開発.2018

    • Author(s)
      植田 雄人, 金田一 智規, 大橋 晶良, 青井 議輝.
    • Organizer
      第52回日本水環境学会年会
    • Related Report
      2017 Annual Research Report
  • [Presentation] 活性汚泥内の Candidatus Saccharibacteria(TM7)を標的とするプライマーの特異性の検証2017

    • Author(s)
      竹中亮太, 金田一智規, 尾崎則篤, 大橋晶良
    • Organizer
      第51回日本水環境学会年会
    • Place of Presentation
      熊本市
    • Year and Date
      2017-03-15
    • Related Report
      2016 Annual Research Report
  • [Presentation] Validation of the specificity of primers targeting Canditatus Saccharibacteria (TM7) in activated sludge.2017

    • Author(s)
      Ryota Takenaka, Tomonori Kindaichi, Yoshiteru Aoi, Noriatsu Ozaki, Akiyoshi Ohashi.
    • Organizer
      International Conference on Civil and Environmental Engineering ICCEE 2017
    • Related Report
      2017 Annual Research Report
    • Int'l Joint Research
  • [Presentation] 活性汚泥内のCandidatus Saccharibacteria (TM7)に対するプライマーの特異性の検証.2017

    • Author(s)
      竹中 亮太, 金田一 智規, 大橋 晶良, 尾崎 則篤, 青井 議輝.
    • Organizer
      環境微生物系学会合同大会2017
    • Related Report
      2017 Annual Research Report
  • [Presentation] 活性汚泥内に存在する Candidatus Saccharibacteria(TM7)に対するプライマーの特異性の検証.2017

    • Author(s)
      竹中 亮太, 金田一 智規, 大橋 晶良, 尾崎 則篤, 青井 議輝.
    • Organizer
      第69回土木学会中国支部研究発表会
    • Related Report
      2017 Annual Research Report
  • [Presentation] 活性汚泥内に存在するCandidate divisio 細菌群のFISH法を用いた可視化.2016

    • Author(s)
      西村 恭平, 金田一 智規, 大橋 晶良, 尾崎 則篤, 青井 義輝.
    • Organizer
      第31回日本微生物生態学会大会
    • Place of Presentation
      横須賀市
    • Year and Date
      2016-10-22
    • Related Report
      2016 Annual Research Report
  • [Presentation] 微生物を自動的に“捕え”て“分離”する革新的分離培養手法.2016

    • Author(s)
      植田 雄人, Dawoon Jung, Nil Tandogan, Edger D. Goluch, 金田一 智規, 大橋 晶良, 青井 議輝.
    • Organizer
      第31回日本微生物生態学会大会
    • Place of Presentation
      横須賀市
    • Year and Date
      2016-10-22
    • Related Report
      2016 Annual Research Report
  • [Presentation] An innovative cultivation method that enable isolating pure culture automatically.2016

    • Author(s)
      Yuto Ueda, Nil Tandogan, Edger D. Goluch, Tomonori Kindaichi, Akiyoshi Ohashi, Yoshiteru Aoi.
    • Organizer
      16th International Symposium on Microbial Ecology (isme16)
    • Place of Presentation
      Montreal, Canada
    • Year and Date
      2016-08-21
    • Related Report
      2016 Annual Research Report
    • Int'l Joint Research

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Published: 2016-04-21   Modified: 2020-03-30  

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