Molecular characterization of methanotrophic isolates from freshwater lake sediment

A. J. Auman, S. Stolyar, Andria Costello Staniec, M. E. Lidstrom

Research output: Contribution to journalArticle

158 Citations (Scopus)

Abstract

Profiles of dissolved O2 and methane with increasing depth were generated for Lake Washington sediment, which suggested the zone of methane oxidation is limited to the top 0.8 cm of the sediment. Methane oxidation potentials were measured for 0.5-cm layers down to 1.5 cm and found to be relatively constant at 270 to 350 υmol/liter of sediment/h. Approximately 65% of the methane was oxidized to cell material or metabolites, a signature suggestive of type I methanotrophs. Eleven methanotroph strains were isolated from the lake sediment and analyzed. Five of these strains classed as type I, while six were classed as type II strains by 16S rRNA gene sequence analysis. Southern hybridization analysis with oligonucleotide probes detected, on average, one to two copies of pmoA and one to three copies of 16S rRNA genes. Only one restriction length polymorphism pattern was shown for pmoA genes in each isolate, and in cases where, sequencing was done, the pmoA copies were found to be almost identical. PCR primers were developed for mmoX which amplified 1.2-kb regions from all six strains that tested positive for cytoplasmic soluble methane mono-oxygenase (sMMO) activity. Phylogenetic analysis of the translated PCR products with published mmoX sequences showed that MmoX falls into two distinct clusters, one containing the orthologs from type I strains and another containing the orthologs from type II strains. The presence of sMMO-containing Methylomonas strains in a pristine freshwater lake environment suggests that these methanotrophs are more widespread than has been previously thought.

Original languageEnglish (US)
Pages (from-to)5259-5266
Number of pages8
JournalApplied and Environmental Microbiology
Volume66
Issue number12
DOIs
StatePublished - 2000
Externally publishedYes

Fingerprint

freshwater sediment
Methane
Lakes
Fresh Water
lacustrine deposit
methane
lakes
sediments
methanotrophs
Oxygenases
rRNA Genes
gene
Methylomonas
oxidation
Polymerase Chain Reaction
ribosomal RNA
freshwater environment
Oligonucleotide Probes
lacustrine environment
oligonucleotide probes

ASJC Scopus subject areas

  • Environmental Science(all)
  • Biotechnology
  • Microbiology

Cite this

Molecular characterization of methanotrophic isolates from freshwater lake sediment. / Auman, A. J.; Stolyar, S.; Staniec, Andria Costello; Lidstrom, M. E.

In: Applied and Environmental Microbiology, Vol. 66, No. 12, 2000, p. 5259-5266.

Research output: Contribution to journalArticle

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