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PMID: 15528517 Published · ppublish English Journal Article Research Support, U.S. Gov't, Non-P.H.S.

Representative freshwater bacterioplankton isolated from Crater Lake, Oregon.

Applied and environmental microbiology ·Vol. 70 ·No. 11 ·2004-11-00 ·Pages 6542-50

Page KA, Connon SA, Giovannoni SJ

Abstract

High-throughput culturing (HTC) methods that rely on dilution to extinction in very-low-nutrient media were used to obtain bacterial isolates from Crater Lake, Oregon. 16S rRNA sequence determination and phylogenetic reconstruction were used to determine the potential ecological significance of isolated bacteria, both in Crater Lake and globally. Fifty-five Crater Lake isolates yielded 16 different 16S rRNA gene sequences. Thirty of 55 (55%) Crater Lake isolates had 16S rRNA gene sequences with 97% or greater similarity to sequences recovered previously from Crater Lake 16S rRNA gene clone libraries. Furthermore, 36 of 55 (65%) Crater Lake isolates were found to be members of widely distributed freshwater groups. These results confirm that HTC is a significant improvement over traditional isolation techniques that tend to enrich for microorganisms that do not predominate in their environment and rarely correlate with 16S rRNA gene clone library sequences. Although all isolates were obtained under dark, heterotrophic growth conditions, 2 of the 16 different groups showed evidence of photosynthetic capability as assessed by the presence of puf operon sequences, suggesting that photoheterotrophy may be a significant process in this oligotrophic, freshwater habitat.

MeSH Terms
Actinobacteria/classification,genetics,isolation & purification Animals Bacteria/classification,genetics,isolation & purification Bacteroidetes/classification,genetics,isolation & purification DNA, Ribosomal/analysis Fresh Water/microbiology Genes, rRNA Molecular Sequence Data Oregon Phylogeny Plankton/classification,genetics,isolation & purification Proteobacteria/classification,genetics,isolation & purification RNA, Ribosomal, 16S/genetics Sequence Analysis, DNA
Chemicals
DNA, Ribosomal RNA, Ribosomal, 16S
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Page Kathleen A
Department of Biology, Southern Oregon University, Ashland, Oregon 97520, USA. page@sou.edu
Connon Stephanie A
Giovannoni Stephen J
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Article Info
Journal
Applied and environmental microbiology
Abbr.
Appl Environ Microbiol
ISSN
0099-2240
DOI
10.1128/AEM.70.11.6542-6550.2004
Email
page@sou.edu
Published
2004-11-00
Pages
6542-50
Language
English
Region
United States
NLM ID
7605801
PMCID
PMC525233
Subset
IM
Databases
GENBANK
AY584571, AY584572, AY584573, AY584574, AY584575, AY584576, AY584577, AY584578, AY584579, AY584580, AY584581, AY584582, AY584583, AY584584, AY584585, AY584586, AY584587, AY584588, AY584589
External Links
PubMed source DOI: 10.1128/AEM.70.11.6542-6550.2004 PMC Full Text View journal details
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