Isolation of functional single cells from environments using a micromanipulator: application to study denitrifying bacteria

Applied Microbiology and Biotechnology
Naoaki AshidaKeishi Senoo

Abstract

We developed a novel method to isolate functionally active single cells from environmental samples and named it the functional single-cell (FSC) isolation method. This method is based on a combination of substrate-responsive direct viable counts, live-cell staining with 5-carboxyfluorescein diacetate acetoxymethyl ester, and micromanipulation followed by cultivation in a medium. To evaluate this method, we applied it to study a denitrifying community in rice paddy soil. Similar denitrifier counts were obtained by the conventional most probable number analysis and our FSC isolation method. Using the FSC isolation method, 37 denitrifying bacteria were isolated, some of which harbored copper-containing nitrite reductase gene (nirK). The 16S rRNA gene analysis showed that members belonging to the genera Azospirillum and Ochrobactrum may be the major denitrifiers in the rice paddy soil. These results indicate that the FSC isolation method is a useful tool to obtain functionally active single cells from environmental samples.

References

Mar 1, 1979·Canadian Journal of Microbiology·K KogureN Taga
Apr 1, 1977·Applied and Environmental Microbiology·T N GambleJ M Tiedje
Nov 15, 2000·FEMS Microbiology Reviews·J Fröhlich, H König
Aug 19, 2003·Molecular Imaging·Vasilis NtziachristosRalph Weissleder
Sep 9, 2004·Microbiology and Molecular Biology Reviews : MMBR·Byron F Brehm-Stecher, Eric A Johnson
Sep 11, 2004·Journal of Applied Microbiology·G Wang, H D Skipper
Feb 5, 2005·Applied and Environmental Microbiology·Kathryn E R DavisPeter H Janssen
Sep 1, 1995·Applied and Environmental Microbiology·T TsujiS Tanaka
Mar 15, 2006·Applied and Environmental Microbiology·F Joux, P Lebaron
Apr 7, 2006·Applied and Environmental Microbiology·Kim HeylenPaul De Vos
Nov 17, 2006·Applied Microbiology and Biotechnology·Thomas KvistPeter Westermann
Jun 26, 2007·Applied and Environmental Microbiology·Qiong WangJames R Cole
Jun 24, 2009·Extremophiles : Life Under Extreme Conditions·Karine Alain, Joël Querellou

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Citations

Jun 17, 2011·The ISME Journal·Satoshi IshiiKeishi Senoo
Nov 16, 2010·Applied and Environmental Microbiology·Satoshi IshiiKeishi Senoo
Oct 20, 2011·Microbes and Environments·Satoshi IshiiKeishi Senoo
Nov 26, 2010·The Journal of General and Applied Microbiology·Hoan Thi VuKeishi Senoo
Mar 24, 2010·Applied Microbiology and Biotechnology·Satoshi IshiiKeishi Senoo
Apr 9, 2014·Critical Reviews in Biotechnology·Xu ShiDeirdre R Meldrum
Jul 10, 2012·Trends in Biotechnology·Van H T Pham, Jaisoo Kim
Mar 8, 2011·Microbial Biotechnology·Koki MaedaTakashi Osada
May 23, 2015·FEMS Microbiology Reviews·Mark A LeverBo Barker Jørgensen
Oct 9, 2012·International Journal of Systematic and Evolutionary Microbiology·Shigeto OtsukaKeishi Senoo
May 20, 2014·Microbial Biotechnology·Jackson Z LeeJohn R Spear
Nov 11, 2018·Applied and Environmental Microbiology·Jeonghwan JangSatoshi Ishii
Sep 14, 2012·Microbes and Environments·Megumi YoshidaKeishi Senoo
Jul 20, 2016·Microbes and Environments·Satoshi IshiiSatoshi Okabe
Apr 20, 2019·Frontiers in Microbiology·Jeonghwan JangSatoshi Ishii

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