Ecophysiological analysis on subseafloor microorganisms by using NanoSIMS

• Project/Area Number: 24687004
• Research Category: Grant-in-Aid for Young Scientists (A)
• Allocation Type: Partial Multi-year Fund
• Research Field: Ecology/Environment
• Research Institution: Japan Agency for Marine-Earth Science and Technology
• Principal Investigator: Morono Yuki 国立研究開発法人海洋研究開発機構, 高知コア研究所, グループリーダー代理 (30421845)
• Project Period (FY): 2012-04-01 – 2016-03-31
• Project Status: Completed (Fiscal Year 2015)
• Budget Amount: ¥27,560,000 (Direct Cost: ¥21,200,000、Indirect Cost: ¥6,360,000); Fiscal Year 2014: ¥8,190,000 (Direct Cost: ¥6,300,000、Indirect Cost: ¥1,890,000); Fiscal Year 2013: ¥8,450,000 (Direct Cost: ¥6,500,000、Indirect Cost: ¥1,950,000); Fiscal Year 2012: ¥10,920,000 (Direct Cost: ¥8,400,000、Indirect Cost: ¥2,520,000)
• Keywords: 海底下生命圏 / NanoSIMS / 安定同位体 / ＮａｎｏＳＩＭＳ / 代謝活性 / 二次イオン質量分析計

Outline of Final Research Achievements

We employed NanoSIMS(Nano-scale Secondary Ion Mass Spectroscopy) for studying eco-physiology of microorganism in subseafloor biosphere, which is largest microbial biosphere on the Earth. Through developments of techniques to detect, separate, and purify microbial cells from the matrix of subseafloor samples (e.g. sediment grains), we successfully detected substrate incorporation activity of subeseafloor microbes.

Research Products

• [Int'l Joint Research] カリフォルニア工科大学(米国)
• [Int'l Joint Research] ドイツ地球科学研究センター （GFZ）/ブレーメン大学(ドイツ)
• [Int'l Joint Research] オーフス大学(デンマーク)
• [Journal Article] Variance and potential niche separation of microbial communities in subseafloor sediments off Shimokita Peninsula, Japan (2016)
  • Author(s): Takuro Nunoura, Yoshihiro Takaki, Shigeru Shimamura, Jungo Kakuta, Hiromi Kazama, Miho Hirai, Noriaki Masui, Hitoshi Tomaru, Yuki Morono, Hiroyuki Imachi, Fumio Inagaki, Ken Takai
  • Journal Title: Environmental Microbiology Volume: 18 Issue: 6 Pages: 1-16
  • DOI: 10.1111/1462-2920.13096
  • Peer Reviewed
• [Journal Article] Exploring deep microbial life in coal-bearing sediments down to ~2.5km below the seafloor (2015)
  • Author(s): INAGAKI FUMIO, Kai-Uwe Hinrichs,KUBO YUSUKE,Marshall W. Bowles,Verena B. Heuer,Wei-Li Hong,HOSHINO TATSUHIKO,IJIRI AKIRA,IMACHI HIROYUKI,ITO MOTOO,KANEKO MASANORI,Mark A. Lever,Lin Yu-Shin,Barbara A. Methe,Morita Sumito,MORONO YUKI,et al.
  • Journal Title: SCIENCE/AAAS/ Volume: 349 Issue: 6246 Pages: 420-424
  • DOI: 10.1126/science.aaa6882
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Ecophysiology of Zetaproteobacteria Associated with Shallow Hydrothermal Iron-Oxyhydroxide Deposits in Nagahama Bay of Satsuma Iwo-Jima, Japan (2015)
  • Author(s): HOSHINO TATSUHIKO, Kuratomi Takashi,MORONO YUKI,Hori Tomoyuki,Oiwane Hisashi,Kiyokawa Shoichi,INAGAKI FUMIO
  • Journal Title: Frontiers in Microbiology Volume: 6 Pages: 1554-1554
  • DOI: 10.3389/fmicb.2015.01554
  • Peer Reviewed / Open Access
• [Journal Article] NanoSIMSにより基質の流れを可視化するー未培養微生物のシングルセルレベルでの生理生態解析 (2015)
  • Author(s): 諸野 祐樹、伊藤 元雄
  • Journal Title: 水環境学会誌 Volume: 38 Pages: 375-379
  • Acknowledgement Compliant
• [Journal Article] Presence of oxygen and aerobic communities from sea floor to basement in deep-sea sediments (2015)
  • Author(s): D’Hondt, S., Inagaki, F., Zarikian, C.A., Yamaguchi, Y.T., et al.
  • Journal Title: Nature Geoscience Volume: 8 Issue: 4 Pages: 299-304
  • DOI: 10.1038/ngeo2387
  • Peer Reviewed
• [Journal Article] Intact preservation of environmental samples by freezing under an alternating magnetic field (2015)
  • Author(s): Morono, Y., Terada, T., Yamamoto, Y., Xiao, N., Hirose, T., Sugeno, M., Ohwada, N., Inagaki F
  • Journal Title: Environ. Microbiol. Rep. Volume: 7 Issue: 2 Pages: 243-251
  • DOI: 10.1111/1758-2229.12238
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Origins of lithium in submarine mud volcano fluid in the Nankai accretionary wedge (2015)
  • Author(s): Nishio, Y., Ijiri, A., Toki, T., Morono, Y., Tanimizu, M., Nagaishi, K., and Inagaki, F.
  • Journal Title: Earth Plant. Sci. Lett. Volume: 414 Pages: 144-155
  • DOI: 10.1016/j.epsl.2015.01.018
  • Peer Reviewed
• [Journal Article] Gold-ISH: A nano-size gold particle-based phylogenetic identification compatible with NanoSIMS. (2014)
  • Author(s): Kubota, K., Morono, Y. , Ito, M., Terada, T., Itezono, S., Harada, H., and Inagaki, F.
  • Journal Title: Syst. Appl. Microbiol. Volume: 37 Issue: 4 Pages: 261-266
  • DOI: 10.1016/j.syapm.2014.02.003
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Variability of subseafloor viral abundance at the geographically and geologically distinct continental margins. (2014)
  • Author(s): Yanagawa, K., Morono, Y., Yoshida-Takashima, Y., Eitoku, M., Sunamura, M., Inagaki, F., Imachi, H., Takai, K., and Nunoura, T.
  • Journal Title: FEMS Microbiol. Ecol. Volume: 12269 Issue: 1 Pages: 60-68
  • DOI: 10.1111/1574-6941.12269
  • Peer Reviewed
• [Journal Article] 海底下環境における微生物生命圏とその生態 (2014)
  • Author(s): 諸野祐樹
  • Journal Title: Journal of Japanese Society for Extremophiles Volume: 13 Pages: 4-10
  • Acknowledgement Compliant
• [Journal Article] Hot-alkaline DNA extraction method for deep subseafloor archaeal communities. (2014)
  • Author(s): Morono, Y., Terada, T., Hoshino, T., and Inagaki, F.
  • Journal Title: Appl. Environ. Microbiol. Volume: 80 Issue: 6 Pages: 1985-1994
  • DOI: 10.1128/aem.04150-13
  • Peer Reviewed
• [Journal Article] Gold-ISH: A nano-size gold particle-based phylogenetic identification compatible with NanoSIMS. (2014)
  • Author(s): Kubota, K., Morono, Y., Ito, M., Terada, T., Itezono, S., Harada, H., and Inagaki, F.
  • Journal Title: Syst. Appl. Microbiol Volume: 37 Pages: 261-266
  • Peer Reviewed
• [Journal Article] Distribution of dehalogenation activity and characterization of organohalide-responsive genes in marine sediments of the Nankai Trough subduction zone (2013)
  • Author(s): Futagami, T., Morono, Y., Terada, T., Kaksonen, A. H., and Inagaki, F.
  • Journal Title: Phil. Trans. R. Soc. B Volume: 368 Issue: 1616 Pages: 20120249-20120249
  • DOI: 10.1098/rstb.2012.0249
  • Peer Reviewed
• [Journal Article] An improved cell separation technique for marine subsurface sediments: Applications for high-throughput analysis using flow cytometry and cell sorting (2013)
  • Author(s): Morono, Y., Terada, T., Kallmeyer, J., and Inagaki, F.
  • Journal Title: Envion. Microbiol. Volume: 15 Issue: 10 Pages: 2841-2849
  • DOI: 10.1111/1462-2920.12153
  • Peer Reviewed
• [Journal Article] 超高空間分解能二次イオン質量分析法による微小領域イメージングと環境微生物学―微小領域の組成イメージ分析から見えてくる環境微生物個々の代謝活性と機能 (2013)
  • Author(s): 諸野祐樹, 伊藤元雄, 稲垣史生
  • Journal Title: 化学と生物 Volume: 51 Pages: 205-207
  • NAID: 10031164803
• [Journal Article] 海底下における微生物とその代謝・活性－未知の海底下生命圏を紐解くテクニックとその応用 (2013)
  • Author(s): 諸野祐樹
  • Journal Title: 日本極限環境生物学会誌 Volume: 12 Pages: 65-70
  • NAID: 40020070807
• [Journal Article] An Improved Cell separation Technique for Marine Subsurface Sediments : Applications for High-throughput Analysis Using Flow Cytometry and Cell Sorting (2013)
  • Author(s): Yuki Morono, Takeshi Terada, Jens Kallmeyer, Fumio Inagaki
  • Journal Title: Environmental Microbiology Volume: (accepted)
  • Peer Reviewed
• [Journal Article] Cell-Specific Thioautotrophic Productivity of Epsilon-Proteobacterial Epibionts Associated with Shinkaia crosnieri. (2012)
  • Author(s): Watsuji, T., Nishizawa, M., Morono, Y. et al.,
  • Journal Title: PLoS ONE Volume: - Issue: 10 Pages: e46282-e46282
  • DOI: 10.1371/journal.pone.0046282
  • Peer Reviewed
• [Presentation] OXIC AND ANOXIC REGIONS OF SUBSEAFLOOR SEDIMENT (2015)
  • Author(s): D'Hondt Steven, Pockalny Robert,Spivack Arthur,INAGAKI FUMIO,Murray RICHARD,Adhikari Rishi Ram,Britta Gribsholt,Kallmeyer Jens,McKinley Claire,MORONO YUKI,Roy Hans,Justine Sauvage,Wiebke Ziebis
  • Organizer: 2015 AGU Fall Meeting
  • Place of Presentation: Moscone Center (San Francisco, USA)
  • Year and Date: 2015-12-17
  • Int'l Joint Research
• [Presentation] Size and Carbon Content of Sub-seafloor Microbial Cells (2015)
  • Author(s): Stefan Braun, MORONO YUKI,Sten Littmann,Jorgensen Bo Barker,Lomstein Bente Aa.
  • Organizer: 2015 AGU Fall Meeting
  • Place of Presentation: Moscone Center (San Francisco, USA)
  • Year and Date: 2015-12-17
  • Int'l Joint Research
• [Presentation] 海底下生命のセンサス：海底下微生物群集の空間的分布と地球環境との相互作用の解明 (2015)
  • Author(s): 星野 辰彦, 諸野 祐樹,稲垣 史生
  • Organizer: 第16回極限環境生物学会
  • Place of Presentation: 東京海洋大学 品川キャンパス （東京都港区）
  • Year and Date: 2015-11-07
• [Presentation] Analysis of biomolecules of sub-seafloor microbial cells separated from the sediment matrix (2015)
  • Author(s): Braun Stefan, MORONO YUKI,Kevin Becker,Hinrichs Kai-Uwe,Jorgensen Bo Barker,Lomstein Bente Aa.
  • Organizer: 3rd International Workshop on Microbial Life under Extreme Energy Limitation
  • Place of Presentation: Aarhus (Denmark)
  • Year and Date: 2015-09-22
  • Int'l Joint Research
• [Presentation] Technical developments to explore sedimentary biosphere: current status, limitation, and future perspectives (2015)
  • Author(s): MORONO YUKI
  • Organizer: CDEBI Sediment Workshop
  • Place of Presentation: University of Southern California （USA)
  • Year and Date: 2015-06-25
  • Int'l Joint Research / Invited
• [Presentation] 超貧栄養海域のマンガン団塊表面に密集する微生物群集 (2015)
  • Author(s): 白石 史人、光延 聖,諸野 祐樹,鈴木 勝彦,稲垣 史生
  • Organizer: 日本地球惑星科学連合2015年大会
  • Place of Presentation: 幕張メッセ(千葉県千葉市）
  • Year and Date: 2015-05-26
• [Presentation] 掘削コア試料の高品質、長期間保管を可能とする磁場振動型凍結とその評価 (2015)
  • Author(s): 諸野 祐樹, 寺田 武志,山本 裕二,肖 楠,廣瀬 丈洋,菅野 正也,大和田 哲男,稲垣 史生
  • Organizer: 日本地球惑星科学連合2015年大会
  • Place of Presentation: 幕張メッセ (千葉県千葉市）
  • Year and Date: 2015-05-26
• [Presentation] Probing metabolic activity of aerobic microbial cells in ultraoligotrophic sediments of the South Pacific Gyre (2015)
  • Author(s): MORONO YUKI, Terada Takeshi,ITO MOTOO,INAGAKI FUMIO
  • Organizer: Deep Carbon Observatory International Science Meeting
  • Place of Presentation: Lisbon （Portugal)
  • Year and Date: 2015-05-08
  • Int'l Joint Research / Invited
• [Presentation] Probing Metabolic Activity of Deep Subseafloor Life with NanoSIMS. (2014)
  • Author(s): Morono, Y., Terada, T., Ito, M., and Inagaki, F.
  • Organizer: AGU Fall Meeting 2014
  • Place of Presentation: Moscone Center, San Francisco, USA
  • Year and Date: 2014-12-15
  • Invited
• [Presentation] 海底下で太古の生命を探せー最先端技術で迫る海底下深部生命とその姿 (2014)
  • Author(s): 諸野祐樹
  • Organizer: 高知大学自主セミナー
  • Place of Presentation: 高知大学農学部、高知県南国市
  • Year and Date: 2014-11-05
  • Invited
• [Presentation] Development of Analytical Technologies for Low-Biomass Microbial Communities in the Subseafloor Sedimentary Biosphere (2014)
  • Author(s): Morono, Y., Terada, T., Ito, M., Hoshino, T., and Inagaki, F.
  • Organizer: Ninth International Symposium on Subsurface Microbiology
  • Place of Presentation: Pacific Grove, California, USA
  • Year and Date: 2014-10-06
• [Presentation] Intact preservation of environmental samples by freezing under an alternating magnetic field (2014)
  • Author(s): Morono Y
  • Organizer: Fourth C-DEBI Annual Meeting
  • Place of Presentation: Marina, California, USA
  • Year and Date: 2014-10-03
  • Invited
• [Presentation] 超高空間分解能質量分析計（NanoSIMS）によるサブミクロンレベルでの安定同位体追跡 (2014)
  • Author(s): 諸野祐樹
  • Organizer: 第32回 「重窒素圃場利用研究会」討論会
  • Place of Presentation: 東京農工大学、東京都府中市
  • Year and Date: 2014-09-08
  • Invited
• [Presentation] Metabolic activity of subseafloor microbes in the South Pacific Gyre (2014)
  • Author(s): Morono, Y., Ito, M., Terada, T., and Inagaki, F.
  • Organizer: 15th International Symposium on Microbial Ecology
  • Place of Presentation: Seoul, Korea
  • Year and Date: 2014-08-25
• [Presentation] 海底下に広がる生命圏 (2014)
  • Author(s): 諸野祐樹
  • Organizer: 統合国際深海掘削計画10年の成果一般公開シンポジウム、深海を掘る
  • Place of Presentation: 国立科学博物館、東京都台東区
  • Year and Date: 2014-04-06
  • Invited
• [Presentation] 海底下堆積物からの細胞分離手法、およびフローサイトメトリーによる細胞計数手法の確立 (2013)
  • Author(s): 寺田武志、諸野祐樹、稲垣史生
  • Organizer: 農芸化学会2013年度大会
  • Place of Presentation: 東北大学
  • Year and Date: 2013-03-26
• [Presentation] NanoSIMSおよびセルソーティングによる南太平洋環流域堆積物試料中の微生物代謝活性解析 (2013)
  • Author(s): 諸野祐樹， 伊藤元雄，寺田武志， 稲垣史生
  • Organizer: 日本地球惑星科学連合2013年大会
  • Place of Presentation: 幕張メッセ、千葉県
• [Presentation] A new method of geobiological sample storage by snap freezing under alternating magnetic field (2012)
  • Author(s): Morono y, Terada T., Yamamoto Y., Hirose T., Xiao N., Sugeno M., Ohwada N., Inagaki F.
  • Organizer: AGU Fall meeting
  • Place of Presentation: Moscone center, USA
  • Year and Date: 2012-12-06
• [Presentation] Mineral-Association and Activity of Bacteria and Archaea in the Deep Subsurface South Pacific Gyre Sediment (2012)
  • Author(s): Steele JA, Dekas AE, Harrison BK, Morono Y, Inagaki F, Xievis W, Orphan VJ
  • Organizer: AGU Fall meeting
  • Place of Presentation: Moscone center, USA
  • Year and Date: 2012-12-06
• [Presentation] Exploration of serpentine seamount, South Chamorro seamount (2012)
  • Author(s): Miyazaki J., Morono Y., Hirayama H., Inagaki F., Wheat CG, Takai K.
  • Organizer: AGU Fall meeting
  • Place of Presentation: Moscone center, USA
  • Year and Date: 2012-12-06
• [Presentation] Imaging microstructure of fine-grained marine sediments : employing resin embedding method for biological specimens and comparison with conventional technique using a combined observation by microfocus X-ray CT and SEM (2012)
  • Author(s): Uramoto, G.I., Morono, Y., Uematsu, K, and Inagaki, F
  • Organizer: AGU Fall meeting
  • Place of Presentation: Moscone center, USA
  • Year and Date: 2012-12-03
• [Presentation] 海底下堆積物からの細胞分離法の確立、およびフローサイトメトリーによる菌数カウントと選択的細胞濃縮 (2012)
  • Author(s): 諸野祐樹・寺田武志・稲垣史生
  • Organizer: 極限環境生物学会
  • Place of Presentation: 日本大学文理学部
  • Year and Date: 2012-12-02
• [Presentation] Cell abundance, metabolic activity, and genetic functions of subseafloor microbes in the South Pacific Gyre (2012)
  • Author(s): Morono Y, Ito M., Inagaki F.
  • Organizer: IODP Expedition 329, 2^<nd> post cruise meeting
  • Place of Presentation: King Kamehameha Hotel, USA
  • Year and Date: 2012-11-13
• [Presentation] 微生物は一億年生きる?-最新の技術で探る海底下生命圏とその生存戦略- (2012)
  • Author(s): 諸野祐樹
  • Organizer: 酵素工学研究会第68回講演会
  • Place of Presentation: 東京大学山上会館(招待講演)
  • Year and Date: 2012-10-05
• [Presentation] Carbon and nitrogen assimilation in deep subseafloor microbial cells (2012)
  • Author(s): Morono y, Terada T., Nishizawa M., Ito M., Hillion F., Takahata N., Sano Y., and Inagaki F
  • Organizer: Microenergy 2012
  • Place of Presentation: Aarhus University, Denmark
  • Year and Date: 2012-05-07
• [Presentation] The Challenge of Contamination in Deep Life Exploration
  • Author(s): Morono Y., Lever M.A., Hoshino T., and Inagaki F.
  • Organizer: The CHIKYU+10 Workshop
  • Place of Presentation: 一ツ橋会館、東京都
• [Presentation] Technological challenges for the advanced study of subseafloor life
  • Author(s): Morono, Y., Terada, T., and Inagaki, F
  • Organizer: Goldschmidt 2013
  • Place of Presentation: Firenze Fiera Congress and Exhibition Centre, Florence, Italy
  • Invited
• [Presentation] Technological challenges for the advanced study of deep subseafloor life
  • Author(s): Morono, Y., Ito, M., Terada, T., and Inagaki, F.
  • Organizer: International Astrobiology Workshop 2013
  • Place of Presentation: JAXA/ISAS, Sagamihara, Kanagawa, Japan
• [Presentation] Metabolic activity of subseafloor microbes in the South Pacific Gyre
  • Author(s): Morono, Y., Ito, M., Terada, T., and Inagaki, F.
  • Organizer: AGU fall meeting
  • Place of Presentation: Moscone center, San Francisco, USA
• [Presentation] 生命の生息限界、代謝速度の限界を探る―海底下における微生物とその活性―
  • Author(s): 諸野祐樹
  • Organizer: 日本極限環境生物学会 第14回シンポジウム
  • Place of Presentation: 東洋大学、東京都
  • Invited
• [Presentation] 海底下環境における微生物生命圏とその生態に関する研究
  • Author(s): 諸野祐樹
  • Organizer: 日本極限環境生物学会、奨励賞受賞講演
  • Place of Presentation: 明治大学、川崎市、神奈川県
• [Presentation] NanoSIMSによる環境微生物の基質同化活性測定―海底下生命圏を例として―
  • Author(s): 諸野祐樹，伊藤元雄，寺田武志，稲垣史生
  • Organizer: 第29回日本微生物生態学会年会シンポジウム
  • Place of Presentation: 鹿児島大学、鹿児島県
  • Invited
• [Book] Earth and Life Processes Discovered from Subseafloor Environments: A Decade of Science Achieved by the Integrated Ocean Drilling Program (IODP) (2014)
  • Author(s): Morono, Y., Kallmeyer, J.
  • Total Pages: 325
  • Publisher: Elsevier
• [Book] “Life in Extreme Environments: Microbial Life in the Deep Biosphere” (2014)
  • Author(s): Morono, Y., Ito, M., and Inagaki, F.
  • Total Pages: 822
  • Publisher: DeGruyter
• [Book] Chapter 5: Detecting slow metabolism in the subseafloor: analysis of single cells using NanoSIMS. In Kallmeyer, J. (ed.), “Life in Extreme Environments: Microbial Life in the Deep Biosphere” (2014)
  • Author(s): Morono, Y., Ito, M., and Inagaki, F.
  • Total Pages: 326
  • Publisher: De Gruyter