Identification of the Dominant Sulfate-Reducing Bacterial Partner of Anaerobic Methanotrophs of the ANME-2 Clade

Environ Microbiol. 2010 Aug;12(8):2327-40. doi: 10.1111/j.1462-2920.2010.02275.x. Epub 2010 Jul 9.

Abstract

The anaerobic oxidation of methane (AOM) with sulfate as terminal electron acceptor is mediated by consortia of methanotrophic archaea (ANME) and sulfate-reducing bacteria (SRB). Whereas three clades of ANME have been repeatedly studied with respect to phylogeny, key genes and genomic capabilities, little is known about their sulfate-reducing partner. In order to identify the partner of anaerobic methanotrophs of the ANME-2 clade, bacterial 16S rRNA gene libraries were constructed from cultures highly enriched for ANME-2a and ANME-2c in consortia with Deltaproteobacteria of the Desulfosarcina/Desulfococcus group (DSS). Phylogenetic analysis of those and publicly available sequences from AOM sites supported the hypothesis by Knittel and colleagues that the DSS partner belongs to the diverse SEEP-SRB1 cluster. Six subclusters of SEEP-SRB1, SEEP-SRB1a to SEEP-SRB1f, were proposed and specific oligonucleotide probes were designed. Using fluorescence in situ hybridization on samples from six different AOM sites, SEEP-SRB1a was identified as sulfate-reducing partner in up to 95% of total ANME-2 consortia. SEEP-SRB1a cells exhibited a rod-shaped, vibrioid, or coccoid morphology and were found to be associated with subgroups ANME-2a and ANME-2c. Moreover, SEEP-SRB1a was also detected in 8% to 23% of ANME-3 consortia in Haakon Mosby Mud Volcano sediments, previously described to be predominantly associated with SRB of the Desulfobulbus group. SEEP-SRB1a contributed to only 0.3% to 0.7% of all single cells in almost all samples indicating that these bacteria are highly adapted to a symbiotic relationship with ANME-2.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Anaerobiosis
  • DNA, Bacterial / genetics
  • Deltaproteobacteria / classification
  • Deltaproteobacteria / genetics
  • Deltaproteobacteria / metabolism
  • Gene Library
  • Geologic Sediments / microbiology
  • In Situ Hybridization, Fluorescence
  • Methane / metabolism*
  • Microbial Consortia*
  • Molecular Sequence Data
  • Oxidation-Reduction
  • Phylogeny*
  • RNA, Ribosomal, 16S / genetics
  • Sulfur-Reducing Bacteria / classification*
  • Sulfur-Reducing Bacteria / genetics
  • Sulfur-Reducing Bacteria / metabolism
  • Symbiosis

Substances

  • DNA, Bacterial
  • RNA, Ribosomal, 16S
  • Methane

Associated data

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