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Viable microbes in ice: application of molecular assays to McMurdo Dry Valley lake ice communities

Published online by Cambridge University Press:  23 June 2010

Markus Dieser
Affiliation:
Montana State University, Center for Biofilm Engineering, Bozeman, MT 59717, USA Montana State University, Department of Land Resources and Environmental Sciences, Bozeman, MT 59717, USA
Andreas Nocker
Affiliation:
Montana State University, Center for Biofilm Engineering, Bozeman, MT 59717, USA Netherlands Organisation for Applied Scientific Research (TNO), 3704 HE Zeist, The Netherlands
John C. Priscu
Affiliation:
Montana State University, Department of Land Resources and Environmental Sciences, Bozeman, MT 59717, USA
Christine M. Foreman*
Affiliation:
Montana State University, Center for Biofilm Engineering, Bozeman, MT 59717, USA Montana State University, Department of Land Resources and Environmental Sciences, Bozeman, MT 59717, USA

Abstract

The permanent ice covers of the McMurdo Dry Valley lakes, Antarctica, are colonized by a diverse microbial assemblage. We collected ice cores from Lakes Fryxell, Hoare and Bonney. Propidium monoazide (PMA) was used in combination with quantitative PCR (qPCR) and denaturing gradient gel electrophoresis (DGGE) to examine membrane integrity of prokaryotes in these extreme environments. PMA selectively penetrates cells with compromised membranes and modifies their DNA resulting in the suppression of PCR amplification. Our results based on analysis of 16S rRNA genes demonstrate that despite the hostile conditions of the Dry Valleys, the permanent ice covers of the lakes support a ‘potentially viable’ microbial community. The level of membrane integrity, as well as diversity, was higher in samples where sediment was entrapped in the ice cover. Pronounced differences in the fraction of cells with intact and compromised cell membranes were found for Lake Fryxell and east lobe of Lake Bonney, both expressed in differences in DGGE banding patterns and qPCR signal reductions. Limitations in the ability to distinguish between intact or compromised cells occurred in samples from Lake Hoare and west lobe of Lake Bonney due to low DNA template concentrations recovered from the samples.

Type
Biological Sciences
Copyright
Copyright © Antarctic Science Ltd 2010

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