title: Ultra-deep pyrosequencing of pmoA amplicons confirms the prevalence of Methylomonas and Methylocystis in Sphagnum mosses from a Dutch peat bog.
abstract: Sphagnum peatlands are important ecosystems in the methane cycle. Methanotrophs in these ecosystems have been shown to reduce methane emissions and provide additional carbon to Sphagnum mosses. However, little is known about the diversity and identity of the methanotrophs present in and on Sphagnum mosses in these peatlands. In this study, we applied a pmoA microarray and high-throughput 454 pyrosequencing to pmoA PCR products obtained from total DNA from Sphagnum mosses 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 of Methylomonas and Methylocystis species in the Sphagnum mosses. 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 providing pmoA sequences of many unknown or uncultivated methanotrophs present in the Sphagnum mosses and, thus, provided much more detailed and quantitative insight into the microbial diversity.
authors.affiliation: Radboud University Nijmegen, Institute for Water and Wetland Research (IWWR), Department of Microbiology, Heyendaalseweg 135, 6525 AJ Nijmegen, the Netherlands. Centre for Molecular and Biomolecular Informatics, Nijmegen Centre for Molecular Life Sciences, Radboud University Nijmegen Medical Centre, Geert Grooteplein 28, 6525 GA, Nijmegen, the Netherlands. Department of Bioresources, AIT, Austrian Institute of Technology GmbH, 2444 Seibersdorf, Austria. Department of Human Genetics, Radboud University Nijmegen Medical Centre, Geert Grooteplein 10, 6525 GA Nijmegen, the Netherlands.
authors.forename: Nardy
authors.initials: N
authors.lastname: Kip
authors.forename: Bas E
authors.initials: BE
authors.lastname: Dutilh
authors.forename: Yao
authors.initials: Y
authors.lastname: Pan
authors.forename: Levente
authors.initials: L
authors.lastname: Bodrossy
authors.forename: Kornelia
authors.initials: K
authors.lastname: Neveling
authors.forename: Michael P
authors.initials: MP
authors.lastname: Kwint
authors.forename: Mike S M
authors.initials: MS
authors.lastname: Jetten
authors.forename: Huub J M
authors.initials: HJ
authors.lastname: Op den Camp