Dilution-to-Extinction Culturing of Psychrotolerant Planktonic Bacteria from Permanently Ice-covered Lakes in the McMurdo Dry Valleys,Antarctica |
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Authors: | U Stingl J-C Cho W Foo K L Vergin B Lanoil S J Giovannoni |
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Institution: | (1) Department of Microbiology, Oregon State University, Corvallis, OR 97331, USA;(2) Division of Biology and Ocean Sciences, Inha University, Incheon, 402-751, Republic of Korea;(3) Department of Environmental Sciences, University of California, Riverside, CA 92521, USA |
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Abstract: | Lakes in the McMurdo Dry Valleys of Antarctica are characterized by a permanent ice cover and little or no anthropogenic influence.
Although bacterial cultures have been obtained from these habitats, recent culture-independent studies indicate that the most
abundant microbes in these systems are not yet cultivated. By using dilution-to-extinction cultivation methods with sterilized
and nutrient-amended lake water as media, we isolated 148 chemotrophic psychrotolerant bacterial cultures from fresh surface
water of Lake Fryxell and the east lobe of Lake Bonney and the hypersaline, suboxic bottom water from the west lobes of Lake
Bonney. Screening of the 16S ribosomal ribonucleic acid (rRNA) genes of the cultures by restriction fragment length polymorphism
(RFLP) yielded 57 putatively pure psychrotolerant, slow growing cultures grouped into 18 clusters. The sequencing of 16S rRNA
genes of randomly selected representatives of each RFLP cluster revealed that the corresponding isolates belong to the Alphaproteobacteria (six RFLP patterns), Betaproteobacteria (six RFLP patterns), Bacteroidetes (four RFLP patterns), and Actinobacteria (two RFLP patterns). Phylogenetic analysis of the sequences showed that the vast majority of the isolates were not closely
related to previously described species. Thirteen of 18 RFLP patterns shared a 16S ribosomal deoxyribonucleic acid sequence
similarity of 97% or less with the closest described species, and four isolates had a sequence similarity of 93% or less with
the nearest described species. Phylogenetic analysis showed that these sequences were representatives of deeply branching
organisms in the respective phylum. A comparison of the isolates with 16S rRNA clone libraries prepared from the same environments
showed substantial overlap, indicating that dilution-to-extinction culturing in natural lake water media can help isolate
some of the most abundant organisms in these perennially ice-covered lakes. |
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