Identification of methanogenic archaea in the hyporheic sediment of Sitka stream

Abstract

Methanogenic archaea produce methane as a metabolic product under anoxic conditions and they play a crucial role in the global methane cycle. In this study molecular diversity of methanogenic archaea in the hyporheic sediment of the lowland stream Sitka (Olomouc, Czech Republic) was analyzed by PCR amplification, cloning and sequencing analysis of the methyl coenzyme M reductase alpha subunit (mcrA) gene. Sequencing analysis of 60 clones revealed 24 different mcrA phylotypes from hyporheic sedimentary layers to a depth of 50 cm. Phylotypes were affiliated with Methanomicrobiales, Methanosarcinales and Methanobacteriales orders. Only one phylotype remains unclassified. The majority of the phylotypes showed higher affiliation with uncultured methanogens than with known methanogenic species. The presence of relatively rich assemblage of methanogenic archaea confirmed that methanogens may be an important component of hyporheic microbial communities and may affect CH4 cycling in rivers.

Figure 1. Phylogenetic tree of mcrA gene clone (phylotype) sequences retrieved from hyporheic river sediment.

The clones come from upper sediment layer (0-25 cm depth) are described as „P“, clones come from deeper sediment layer (25-50 cm depth) are described as „H“. The numbers in parenthesis indicate the number of clones. The phylogenetic tree is rooted with Methanopyrus kandleri.