Insights into the fate of a 13C labelled phenol pulse for stable isotope probing (SIP) experiments

Manefield, Mike; Griffiths, Rob ORCID: https://orcid.org/0000-0002-3341-4547; McNamara, Niall P. ORCID: https://orcid.org/0000-0002-5143-5819; Sleep, Darren ORCID: https://orcid.org/0009-0002-1128-1883; Ostle, Nick; Whiteley, Andrew. 2007 Insights into the fate of a 13C labelled phenol pulse for stable isotope probing (SIP) experiments. Journal of Microbiological Methods, 69 (2). 340-344. 10.1016/j.mimet.2007.01.019

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Official URL: http://dx.doi.org/10.1016/j.mimet.2007.01.019

Abstract/Summary

Stable isotope probing (SIP) using DNA or RNA as a biomarker has proven to be a useful method for attributing substrate utilisation to specific microbial taxa. In this study we followed the transfer of a 13C6-phenol pulse in an activated sludge micro-reactor to examine the resulting distribution of labelled carbon in the context of SIP. Most of the added phenol was metabolically converted within the first 100 min after 13C6-phenol addition, with 49% incorporated into microbial biomass and 6% respired as CO2. Less than 1% of the total 13C labelled carbon supplied was incorporated into microbial RNA and DNA, with RNA labelling 6.5 times faster than DNA. The remainder of the added 13C was adsorbed and/or complexed to suspended solids within the sludge. The 13C content of nucleic acids increased beyond the initial consumption of the 13C-phenol pulse. This study confirms that RNA labels more efficiently than DNA and reveals that only a small proportion of a pulse is incorporated into nucleic acids. Evidence of continued 13C incorporation into nucleic acids suggests that cross-feeding of the SIP substrate was rapid. This highlights both the benefits of using a biomarker that is rapidly labelled and the importance of sampling within appropriate timescales to avoid or capture the effects of cross-feeding, depending on the goal of the study.

Item Type:	Publication - Article
Digital Object Identifier (DOI):	10.1016/j.mimet.2007.01.019
Programmes:	CEH Programmes pre-2009 publications > Biogeochemistry CEH Programmes pre-2009 publications > Biodiversity
UKCEH and CEH Sections/Science Areas:	Shore Hails
ISSN:	0167-7012
Additional Keywords:	Microbial biomass, DNA, RNA, Carbon, Respiration, CO2
NORA Subject Terms:	Agriculture and Soil Science Biology and Microbiology Ecology and Environment
Date made live:	11 Feb 2008 13:59 +0 (UTC)
URI:	https://nora.nerc.ac.uk/id/eprint/2282

Item Type:

Publication - Article

Digital Object Identifier (DOI):

10.1016/j.mimet.2007.01.019

Programmes:

CEH Programmes pre-2009 publications > Biogeochemistry
CEH Programmes pre-2009 publications > Biodiversity

UKCEH and CEH Sections/Science Areas:

Shore
Hails

ISSN:

0167-7012

Additional Keywords:

Microbial biomass, DNA, RNA, Carbon, Respiration, CO2