Phylogenetic diversity of methanogenic, sulfate-reducing and methanotrophic prokaryotes from deep-sea hydrothermal vents and cold seeps

Microbial communities of methanogenic, sulfate-reducing and methanotrophic prokaryotes from deepsea environments were investigated by molecular phylogenetic analysis of the deduced amino acid sequences of the genes encoding for the methyl coenzyme M reductase (mcrA), dissimilatory sulfite reductase (dsrAB) and particulate methane monoxygenase (pmoA), respectively. Clone libraries of PCR amplified genes were constructed using DNA extracted from deep-sea vent chimneys (Rainbow and Logatchev hydrothermal vent fields, Mid-Atlantic Ridge, Atlantic Ocean; 91N East Pacific Rise, Pacific Ocean) and from vertically subsampled sediment cores from cold-seep areas (Blake Ridge, western Atlantic Ocean; Florida Escarpment, Gulf of Mexico). Recombinant clones were screened by RFLP and representative dsrAB, mcrA and pmoA genes were sequenced. The dsrAB sequences grouped primarily within the orders Desulfobacterales, Syntrophobacterales and the Gram-positive order Clostridales. Coldseep mcrA sequences were distributed among the ANME-2c, -2d and -2e groups, which were previously shown to be associated with the anaerobic oxidation of methane. This study also reports the first mcrA sequences from a high-temperature, black smoker chimney (Logatchev) to group within the ANME-2e subgroup. The majority of the remaining hydrothermal vent mcrA sequences were primarily related to thermophilic members of the anaerobic, methanogenic order Methanococcales. A shift in the dominant ANME-2 group with depth in the sediment for both Florida Escarpment and Blake Ridge mcrA libraries was detected. ANME-2d related clones were detected in the top zones of both cores, with the frequency of ANME-2e related clones increasing with depth. All pmoA sequences retrieved from the cold-seep sites were found to be related to Type I methanotrophic members of the g-proteobacteria, and were primarily distributed among three major clusters of sequences. No Type II pmoA sequences related to methanotrophic members of the a-proteobacteria were detected, suggesting that the methanotrophic communities in these cold-seep areas are dominated by Type I g-proteobacteria. & 2009 Elsevier Ltd. All rights reserved.