Joshua T. Ellis, Cody Tramp, Ronald C. Sims, Charles D. Miller
The microbial diversity and metabolic potential of a methanogenic consortium residing in a 3785 liter anaerobic digester, fed with wastewater algae, was analyzed using 454-pyrosequencing technology. DNA was extracted from anaerobic sludge material and used in metagenomic analysis through PCR amplification of the methyl coenzyme-M reductase α subunit (mcrA) gene using primer sets ML, MCR, and ME. The majority of annotated mcrA sequences were assigned taxonomically to the genera Methanosaeta in the order Methanosarcinales. Methanogens from the genus Methanosaeta are obligate acetotrophs, suggesting this genus plays a dominant role in methane production from the analyzed fermentation sample. Numerous analyzed sequences within the algae fed anaerobic digester were unclassified, and could not be assigned taxonomically. Relative amplicon frequencies were determined for each primer set to determine the utility of each in pyrosequencing. Primer sets ML and MCR performed better quantitatively (representing the large majority of analyzed sequences) than primer set ME. However, each of these primer sets was shown to provide a quantitatively unique community structure, and thus are of equal importance in mcrA metagenomic analysis.
Joshua T. Ellis, Cody Tramp, Ronald C. Sims, and Charles D. Miller. 2012. Metagenome analysis of a methanogenic community within an algal fed anaerobic digester. ISRN Microbiology In Press.
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