Analysis of the microbial functional diversity within water-stressed soil communities by flow cytometric analysis and CTC+ cell sorting

Whiteley, Andrew S.; Griffiths, Robert I. ORCID: https://orcid.org/0000-0002-3341-4547; Bailey, Mark J.. 2003 Analysis of the microbial functional diversity within water-stressed soil communities by flow cytometric analysis and CTC+ cell sorting. Journal of Microbiological Methods, 54 (2). 257-267. 10.1016/S0167-7012(03)00066-6

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Abstract/Summary

Total and active cell counts within soil samples were determined by culture-independent methods using flow cytometry and preparative Nycodenz gradient centrifugation. Whole cells were purified from soil cores and total extractable cell counts assessed by SYBR Green II fluorescence, while active cell counts were determined by 5-cyano-2,3-ditolyl tetrazolium chloride reduction (CTC+ cells). Parallel microcosms, maintained at either field water capacity or subjected to drying, indicated that the total extractable cell count remained between 108 and 109 g−1 (dry weight). In contrast, the CTC+ active count fell threefold in dried microcosms (6% of total cell count) when compared to wetted microcosms (18% of total cell count). Specifically, these data highlighted an overall deactivation of microbial biomass during water stress, with 16S rDNA analyses of flow-sorted CTC+ cells demonstrating shifts within the active diversity. Flow cytometry coupled with cell purification techniques represents a significant tool for operationally defining an active and redundant microbial component within soilcommunities and is demonstrated during water stress.

Item Type:	Publication - Article
Digital Object Identifier (DOI):	10.1016/S0167-7012(03)00066-6
Programmes:	CEH Programmes pre-2009 publications > Biodiversity
UKCEH and CEH Sections/Science Areas:	_ Molecular Microbial Ecology
ISSN:	0167-7012
Additional Keywords:	flow cytometry, 5-cyano-2,3-ditolyl tetrazolium chloride, CTC, soil microbes
NORA Subject Terms:	Biology and Microbiology
Date made live:	26 Apr 2012 11:33 +0 (UTC)
URI:	https://nora.nerc.ac.uk/id/eprint/13815

Item Type:

Publication - Article

Digital Object Identifier (DOI):

10.1016/S0167-7012(03)00066-6

Programmes:

CEH Programmes pre-2009 publications > Biodiversity

UKCEH and CEH Sections/Science Areas:

_ Molecular Microbial Ecology

ISSN:

0167-7012

Additional Keywords:

flow cytometry, 5-cyano-2,3-ditolyl tetrazolium chloride, CTC, soil microbes