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Applied and Environmental Microbiology, January 2005, p. 467-479, Vol. 71, No. 1
0099-2240/05/$08.00+0     doi:10.1128/AEM.71.1.467-479.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Diversity and Distribution of Methanotrophic Archaea at Cold Seeps{dagger}

Katrin Knittel,1* Tina Lösekann,1 Antje Boetius,1,2 Renate Kort,3 and Rudolf Amann1

Max Planck Institute for Marine Microbiology,1 International University of Bremen, Bremen,2 ICBM-Geomicrobiology Group, University of Oldenburg, Oldenburg, Germany3

Received 16 April 2004/ Accepted 10 August 2004

In this study we investigated by using 16S rRNA-based methods the distribution and biomass of archaea in samples from (i) sediments above outcropping methane hydrate at Hydrate Ridge (Cascadia margin off Oregon) and (ii) massive microbial mats enclosing carbonate reefs (Crimea area, Black Sea). The archaeal diversity was low in both locations; there were only four (Hydrate Ridge) and five (Black Sea) different phylogenetic clusters of sequences, most of which belonged to the methanotrophic archaea (ANME). ANME group 2 (ANME-2) sequences were the most abundant and diverse sequences at Hydrate Ridge, whereas ANME-1 sequences dominated the Black Sea mats. Other seep-specific sequences belonged to the newly defined group ANME-3 (related to Methanococcoides spp.) and to the Crenarchaeota of marine benthic group B. Quantitative analysis of the samples by fluorescence in situ hybridization (FISH) showed that ANME-1 and ANME-2 co-occurred at the cold seep sites investigated. At Hydrate Ridge the surface sediments were dominated by aggregates consisting of ANME-2 and members of the Desulfosarcina-Desulfococcus branch (DSS) (ANME-2/DSS aggregates), which accounted for >90% of the total cell biomass. The numbers of ANME-1 cells increased strongly with depth; these cells accounted 1% of all single cells at the surface and more than 30% of all single cells (5% of the total cells) in 7- to 10-cm sediment horizons that were directly above layers of gas hydrate. In the Black Sea microbial mats ANME-1 accounted for about 50% of all cells. ANME-2/DSS aggregates occurred in microenvironments within the mat but accounted for only 1% of the total cells. FISH probes for the ANME-2a and ANME-2c subclusters were designed based on a comparative 16S rRNA analysis. In Hydrate Ridge sediments ANME-2a/DSS and ANME-2c/DSS aggregates differed significantly in morphology and abundance. The relative abundance values for these subgroups were remarkably different at Beggiatoa sites (80% ANME-2a, 20% ANME-2c) and Calyptogena sites (20% ANME-2a, 80% ANME-2c), indicating that there was preferential selection of the groups in the two habitats. These variations in the distribution, diversity, and morphology of methanotrophic consortia are discussed with respect to the presence of microbial ecotypes, niche formation, and biogeography.

* Corresponding author. Mailing address: Max Planck Institute for Marine Microbiology, Department of Molecular Ecology, Celsiusstrasse 1, 28359 Bremen, Germany. Phone: 49-421-2028936. Fax: 49-421-2028580. E-mail: kknittel{at}mpi-bremen.de.

{dagger} Publication GEOTECH-85 of the GEOTECHNOLOGIEN program and no. 10 of the research program GHOSTDABS of the Bundesministerium für Bildung und Forschung and the DFG.

Applied and Environmental Microbiology, January 2005, p. 467-479, Vol. 71, No. 1
0099-2240/05/$08.00+0     doi:10.1128/AEM.71.1.467-479.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.



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