title: Stable isotope probing analysis of the diversity and activity of methanotrophic bacteria in soils from the Canadian high Arctic.
abstract: The melting of permafrost and its potential impact on CH(4) emissions are major concerns in the context of global warming. Methanotrophic bacteria have the capacity to mitigate CH(4) emissions from melting permafrost. Here, we used quantitative PCR (qPCR), stable isotope probing (SIP) of DNA, denaturing gradient gel electrophoresis (DGGE) fingerprinting, and sequencing of the 16S rRNA and pmoA genes to study the activity and diversity of methanotrophic bacteria in active-layer soils from Ellesmere Island in the Canadian high Arctic. Results showed that most of the soils had the capacity to oxidize CH(4) at 4 degrees C and at room temperature (RT), but the oxidation rates were greater at RT than at 4 degrees C and were significantly enhanced by nutrient amendment. The DGGE banding patterns associated with active methanotrophic bacterial populations were also different depending on the temperature of incubation and the addition of nutrients. Sequencing of the 16S rRNA and pmoA genes indicated a low diversity of the active methanotrophic bacteria, with all methanotroph 16S rRNA and pmoA gene sequences being related to type I methanotrophs from Methylobacter and Methylosarcina. The dominance of type I methanotrophs over type II methanotrophs in the native soil samples was confirmed by qPCR of the 16S rRNA gene with primers specific for these two groups of bacteria. The 16S rRNA and pmoA gene sequences related to those of Methylobacter tundripaludum were found in all soils, regardless of the incubation conditions, and they might therefore play a role in CH(4) degradation in situ. This work is providing new information supporting the potential importance of Methylobacter spp. in Arctic soils found in previous studies and contributes to the limited body of knowledge on methanotrophic activity and diversity in this extreme environment.
authors.affiliation: National Research Council Canada, Biotechnology Research Institute, 6100 Royalmount Ave., Montréal, QC H4P 2R2, Canada.
authors.forename: Christine
authors.initials: C
authors.lastname: Martineau
authors.forename: Lyle G
authors.initials: LG
authors.lastname: Whyte
authors.forename: Charles W
authors.initials: CW
authors.lastname: Greer
mesh_terms: D001110:Arctic Regions; D001419:Bacteria / Q000145:classification* / Q000378:metabolism*; D044822:Biodiversity; D002170:Canada; D016000:Cluster Analysis; D016172:DNA Fingerprinting; D004269:DNA, Bacterial / Q000737:chemistry / Q000235:genetics; D004275:DNA, Ribosomal / Q000737:chemistry / Q000235:genetics; D004591:Electrophoresis, Polyacrylamide Gel; D007554:Isotopes / Q000378:metabolism*; D008697:Methane / Q000378:metabolism*; D008969:Molecular Sequence Data; D009691:Nucleic Acid Denaturation; D010084:Oxidation-Reduction; D010802:Phylogeny; D016133:Polymerase Chain Reaction; D012336:RNA, Ribosomal, 16S / Q000235:genetics; D017422:Sequence Analysis, DNA; D012988:Soil Microbiology; D013696:Temperature
chemical_list: D004269:DNA, Bacterial; D004275:DNA, Ribosomal; D007554:Isotopes; D012336:RNA, Ribosomal, 16S; D008697:Methane