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Estimating high-affinity methanotrophic bacterial biomass, growth, and turnover in soil by phospholipid fatty acid 13C labeling

Estimating high-affinity methanotrophic bacterial biomass, growth, and turnover in soil by phospholipid fatty acid 13C labeling

Applied and Environmental Microbiology 72(6): 3901-3907

A time series phospholipid fatty acid (PLFA) [superscript 13]C-labeling study was undertaken to determine methanotrophic taxon, calculate methanotrophic biomass, and assess carbon recycling in an upland brown earth soil from Bronydd Mawr (Wales, United Kingdom). Laboratory incubations of soils were performed at ambient CH[subscript 4] concentrations using synthetic air containing 2 parts per million of volume of [superscript 13]CH[subscript 4]. Flowthrough chambers maintained a stable CH[subscript 4] concentration throughout the 11-week incubation. Soils were analyzed at weekly intervals by gas chromatography (GC), GC-mass spectrometry, and GC-combustion-isotope ratio mass spectrometry to identify and quantify individual PLFAs and trace the incorporation of [superscript 13]C label into the microbial biomass. Incorporation of the [superscript 13]C label was seen throughout the experiment, with the rate of incorporation decreasing after 9 weeks. The [delta][superscript 13]C values of individual PLFAs showed that [superscript 13]C label was incorporated into different components to various extents and at various rates, reflecting the diversity of PLFA sources. Quantitative assessments of [superscript 13]C-labeled PLFAs showed that the methanotrophic population was of constant structure throughout the experiment. The dominant [superscript 13]C-labeled PLFA was 18:1[chi][omicron][beta][rho][beta][omega]7c, with 16:1[chi][omicron][beta][rho][beta][omega]5 present at lower abundance, suggesting the presence of novel type II methanotrophs. The biomass of methane-oxidizing bacteria at optimum labeling was estimated to be about 7.2 x 10[superscript 6] cells g[superscript -1] of soil (dry weight). While recycling of [superscript 13]C label from the methanotrophic biomass must occur, it is a slower process than initial [superscript 13]CH[subscript 4] incorporation, with only about 5 to 10% of [superscript 13]C-labeled PLFAs reflecting this process. Thus, [superscript 13]C-labeled PLFA distributions determined at any time point during [superscript 13]CH[subscript 4] incubation can be used for chemotaxonomic assessments, although extended incubations are required to achieve optimum [superscript 13]C labeling for methanotrophic biomass determinations.

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Accession: 012786484

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PMID: 16751495

DOI: 10.1128/aem.02779-05

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