Insights into the fate of a 13C labelled phenol pulse for stable isotope probing (SIP) experiments
Allbwn ymchwil: Cyfraniad at gyfnodolyn › Erthygl › adolygiad gan gymheiriaid
StandardStandard
Yn: Journal of Microbiological Methods, Cyfrol 69, Rhif 2, 05.2007, t. 340-344.
Allbwn ymchwil: Cyfraniad at gyfnodolyn › Erthygl › adolygiad gan gymheiriaid
HarvardHarvard
APA
CBE
MLA
VancouverVancouver
Author
RIS
TY - JOUR
T1 - Insights into the fate of a 13C labelled phenol pulse for stable isotope probing (SIP) experiments
AU - Manefield, Mike
AU - Griffiths, Rob
AU - McNamara, Niall P.
AU - Sleep, Darren
AU - Ostle, Nick
AU - Whiteley, Andrew
PY - 2007/5
Y1 - 2007/5
N2 - 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.
AB - 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.
KW - Microbial biomass
KW - DNA
KW - RNA
KW - Carbon
KW - Respiration
KW - CO
U2 - 10.1016/j.mimet.2007.01.019
DO - 10.1016/j.mimet.2007.01.019
M3 - Article
VL - 69
SP - 340
EP - 344
JO - Journal of Microbiological Methods
JF - Journal of Microbiological Methods
SN - 0167-7012
IS - 2
ER -