Diversity, abundance and vertical distribution of methane-oxidizing bacteria (methanotrophs) in the sediments of the Xianghai wetland, Songnen Plain, northeast China

Methanotrophs in wetlands are of great importance because up to 90 % of the methane (CH4) produced in such wetlands could be oxidized by methanotrophs before reaching the atmosphere. The Xianghai wetland of Songnen Plain represents an important ecosystem in northeast China. However, methanotrophic characteristics in this ecosystem have not been studied in detail. The aim of this study is to give an overview of methanotrophic diversity and vertical distribution in the sediments of this important wetland. Sediment cores were collected from three freshwater marshes, each dominated by a particular vegetation type: Carex alata, Phragmites australis and Typha orientalis. The diversity of methanotrophs was studied by phylogenetic analysis of both the 16S rRNA gene and the particulate methane monooxygenase (pmoA) gene. Methanotroph abundance was determined by quantitative PCR (qPCR) targeting the pmoA gene; group-specific pmoA gene quantification was also used to estimate the abundance of each methanotrophic group. 16S rRNA and pmoA gene homological analysis revealed the presence of type Ia, Ib and II methanotrophs. Novel pmoA sequences distantly affiliated to cultured Methylococcus sp. were detected, implying the existence of novel methanotrophs in the wetland. Most obtained representatives of Methylobacter genus (both 16S rRNA and pmoA genes) were closely clustered in relation to sequences acquired from the Zoige wetland, Tibet and Siberia permafrost soils, therefore suggesting methanotrophs belonging to Methylobacter genus shared characteristics with methanotrophs in cold areas. The dominance of type I methanotrophs (especially the Methylobacter genus) was detected by both clone library analysis and group-specific qPCR assay. The relatively high methanotroph diversity and pmoA copy numbers measured in the T. orientalis marsh sediments indicated that vegetation type played an important role during CH4 oxidation in the wetland. We present the first data set on methanotroph diversity and vertical distribution in the sediments of the Xianghai wetland. DNA sequences information of Methylococcus-like methanotrophs in the wetland will facilitate the isolating of novel methanotrophs from the wetland. In a worldwide context, our study has enriched the database of genotypic diversity of methanotrophs, which will help in the understanding of the geographical distribution of methanotrophic communities.