Detection of putatively thermophilic anaerobic methanotrophs in diffuse hydrothermal vent fluids

Appl Environ Microbiol. 2013 Feb;79(3):915-23. doi: 10.1128/AEM.03034-12. Epub 2012 Nov 26.

Abstract

The anaerobic oxidation of methane (AOM) is carried out by a globally distributed group of uncultivated Euryarchaeota, the anaerobic methanotrophic arachaea (ANME). In this work, we used G+C analysis of 16S rRNA genes to identify a putatively thermophilic ANME group and applied newly designed primers to study its distribution in low-temperature diffuse vent fluids from deep-sea hydrothermal vents. We found that the G+C content of the 16S rRNA genes (P(GC)) is significantly higher in the ANME-1GBa group than in other ANME groups. Based on the positive correlation between the P(GC) and optimal growth temperatures (T(opt)) of archaea, we hypothesize that the ANME-1GBa group is adapted to thrive at high temperatures. We designed specific 16S rRNA gene-targeted primers for the ANME-1 cluster to detect all phylogenetic groups within this cluster, including the deeply branching ANME-1GBa group. The primers were successfully tested both in silico and in experiments with sediment samples where ANME-1 phylotypes had previously been detected. The primers were further used to screen for the ANME-1 microorganisms in diffuse vent fluid samples from deep-sea hydrothermal vents in the Pacific Ocean, and sequences belonging to the ANME-1 cluster were detected in four individual vents. Phylotypes belonging to the ANME-1GBa group dominated in clone libraries from three of these vents. Our findings provide evidence of existence of a putatively extremely thermophilic group of methanotrophic archaea that occur in geographically and geologically distinct marine hydrothermal habitats.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Anaerobiosis
  • Archaea / classification*
  • Archaea / genetics
  • Archaea / metabolism*
  • Base Composition
  • Cluster Analysis
  • DNA Primers / genetics
  • DNA, Archaeal / chemistry
  • DNA, Archaeal / genetics
  • DNA, Ribosomal / chemistry
  • DNA, Ribosomal / genetics
  • Hot Temperature
  • Hydrothermal Vents / microbiology*
  • Methane / metabolism*
  • Molecular Sequence Data
  • Pacific Ocean
  • Phylogeny
  • RNA, Ribosomal, 16S / genetics
  • Sequence Analysis, DNA

Substances

  • DNA Primers
  • DNA, Archaeal
  • DNA, Ribosomal
  • RNA, Ribosomal, 16S
  • Methane

Associated data

  • GENBANK/JQ740748
  • GENBANK/JQ740749
  • GENBANK/JQ740750
  • GENBANK/JQ740751
  • GENBANK/JQ740752
  • GENBANK/JQ740753
  • GENBANK/JQ740754
  • GENBANK/JQ740755
  • GENBANK/JQ740756
  • GENBANK/JQ740757
  • GENBANK/JQ740758
  • GENBANK/JQ740759
  • GENBANK/JQ740760
  • GENBANK/JQ740761
  • GENBANK/JQ740762