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Distribution of bacterioplankton in meromictic Lake Saelenvannet, as determined by denaturing gradient gel electrophoresis of PCR-amplified gene fragments coding for 16S rRNA

Applied and Environmental Microbiology · 1997 · Vol. 63(9) · pp. 3367–3373
Lise ØvreåsLarry J. ForneyFrida Lise DaaeVigdis Torsvik

Abstract

The community structure of bacterioplankton in meromictic Lake Saelenvannet was examined by PCR amplification of the V3 region of 16S rRNA from microbial communities recovered from various depths in the water column. Two different primer sets were used, one for amplification of DNA from the domain Bacteria and another specific for DNA from the domain Archaea. Amplified DNA fragments were resolved by denaturing gradient gel electrophoresis (DGGE), and the resulting profiles were reproducible and specific for the communities from different depths. Bacterial diversity estimated from the number and intensity of specific fragments in DGGE profiles decreased with depth. The reverse was true for the Archaea, with the diversity increasing with depth. Hybridization of DGGE profiles with oligonucleotide probes specific for phylogenetic groups of microorganisms showed the presence of both sulfate-reducing bacteria and methanogens throughout the water column, but they appeared to be most abundant below the chemocline. Several dominant fragments in the DGGE profiles were excised and sequenced. Among the dominant populations were representatives related to Chlorobium phaeovibrioides, chloroplasts from eukaryotic algae, and unidentified Archaea.

Microbial Community Ecology and PhysiologyEnvironmental DNA in Biodiversity StudiesProtist diversity and phylogenyTemperature gradient gel electrophoresisBiologyArchaeaChemoclineBacteria16S ribosomal RNABacterioplanktonRibosomal RNAGel electrophoresisRibosomal DNA

MeSH terms

AnimalsBacteriaBase SequenceDNA, BacterialDNA, RibosomalFresh WaterMarine BiologyMolecular Sequence DataNorwayPlanktonRNA, BacterialRNA, Ribosomal, 16SSpecies SpecificityTime FactorsGenetic Variation
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Bias caused by template annealing in the amplification of mixtures of 16S rRNA genes by PCR
Applied and Environmental Microbiology · 1996 · 1,890 citations
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