Ultra-deep pyrosequencing ofpmoAamplicons confirms the prevalence ofMethylomonasandMethylocystisinSphagnummosses from a Dutch peat bog

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Sphagnumpeatlands are important ecosystems in the methane cycle. Methanotrophs in these ecosystems have been shown to reduce methane emissions and provide additional carbon toSphagnummosses. However, little is known about the diversity and identity of the methanotrophs present in and onSphagnummosses in these peatlands. In this study, we applied apmoA microarray and high-throughput 454 pyrosequencing topmoA PCR products obtained from total DNA fromSphagnummosses from a Dutch peat bog to investigate the presence of methanotrophs and to compare the two different methods. Both techniques showed comparable results and revealed an abundance ofMethylomonasandMethylocystisspecies in theSphagnummosses. The advantage of the microarray analysis is that it is fast and cost-effective, especially when many samples have to be screened. Pyrosequencing is superior in providingpmoA sequences of many unknown or uncultivated methanotrophs present in theSphagnummosses and, thus, provided much more detailed and quantitative insight into the microbial diversity.

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