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KMID : 0545120120220030360
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Journal of Microbiology and Biotechnology
2012 Volume.22 No. 3 p.360 ~ p.370
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Microbial Community Analysis of a Methane-Oxidizing Biofilm Using Ribosomal Tag Pyrosequencing
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Kim Tae-Gwan
Lee Eun-Hee
Cho Kyung-Suk
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Abstract
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Current ecological knowledge of methanotrophic biofilms is incomplete, although they have been broadly studied in biotechnological processes. Four individual DNA samples were prepared from a methanotrophic biofilm, and a multiplex 16S rDNA pyrosequencing was performed. A complete library (before being de-multiplexed) contained 33,639 sequences (average length, 415 nt). Interestingly, methanotrophs were not dominant, only making up 23% of the community. Methylosinus, Methylomonas, and Methylosarcina were the dominant methanotrophs. Type II methanotrophs were more abundant than type I (56 vs. 44%), but less richer and diverse. Dominant non-methanotrophic genera included Hydrogenophaga, Flavobacterium, and Hyphomicrobium. The library was de-multiplexed into four libraries, with different sequencing efforts (3,915 - 20,133 sequences). Sørrenson abundance similarity results showed that the four libraries were almost identical (indices > 0.97), and phylogenetic comparisons using UniFrac test and P-test revealed the same results. It was demonstrated that the pyrosequencing was highly reproducible. These survey results can provide an insight into the management and/or manipulation of methanotrophic biofilms.
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KEYWORD
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methanotrophs, biofilm, community analysis, pyrosequencing, quantitative real-time PCR
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