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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 ,
Andreas Nocker ,
John C. Priscu  and
Christine M. Foreman
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
*
*[email protected]
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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.

Keywords

Antarcticabacterialive/deadPropidium monoazide (PMA)
Type
Biological Sciences
Information
Antarctic Science , Volume 22 , Issue 5 , October 2010 , pp. 470 - 476
Copyright
Copyright © Antarctic Science Ltd 2010

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References

Campbell, I.B., Claridge, G.G.C., Campbell, D.I.Balks, M.R. 1998. The soil environment of the McMurdo Dry Valleys, Antarctica. Antarctic Research Series, 72, 297–322.Google Scholar
Choi, J.W., Sherr, E.B.Sherr, B.F. 1996. Relation between presence-absence of a visible nucleoid and metabolic activity in bacterioplankton cells. Limnology and Oceanography, 41, 1161–1168.CrossRefGoogle Scholar
Christner, B.C., Kvitko, B.H.Reeve, J. 2003. Molecular identification of bacteria and eukarya inhabiting an Antarctic cryconite hole. Extremophiles, 7, 177–183.CrossRefGoogle ScholarPubMed
Davidson, A.T., Thomson, P.G., Westwood, K.van den Enden, R. 2004. Estimation of bacterioplankton activity in Tasmanian coastal waters and between Tasmania and Antarctica using stains. Aquatic Microbial Ecology, 37, 33–45.CrossRefGoogle Scholar
Foreman, C.M., Sattler, B., Mikucki, J.A., Porazinska, D.L.Priscu, J.C. 2007. Metabolic activity and diversity of cryoconites in the Taylor Valley, Antarctica. Journal of Geophysical Research, 112, 10.1029/2006JG000358.CrossRefGoogle Scholar
Fritsen, C.H., Adams, E.E., McKay, C.P.Priscu, J.P. 1998. Permanent ice covers of the McMudo Dry Valley lakes, Antarctica: liquid water contents. Antarctic Research Series, 72, 269–280.Google Scholar
Fritsen, C.H.Priscu, J.C. 1998. Cyanobacterial assemblages in permanent ice covers on Antarctic lakes: distribution, growth rate, and temperature response of photosynthesis. Journal of Phycology, 34, 587–597.CrossRefGoogle Scholar
Gafan, G.P., Lucas, V.S., Roberts, G.J., Petric, A., Wilson, M.Spratt, D.A. 2005. Statistical analyses of complex denaturing gradient gel electrophoresis profiles. Journal of Clinical Microbiology, 43, 3971–3978.CrossRefGoogle ScholarPubMed
Glatz, R.E., Lepp, P.W., Ward, B.B.Francis, C.A. 2006. Planktonic microbial community composition across steep physical/chemical gradients in permanently ice-covered Lake Bonney, Antarctica. Geobiology, 4, 53–67.CrossRefGoogle Scholar
Goldman, C.R., Mason, D.T.Hobbie, J.E. 1967. Two Antarctic desert lakes. Limnology and Oceanography, 12, 295–310.CrossRefGoogle Scholar
Gordon, D.A., Priscu, J.Giovannoni, S. 2000. Origin and phylogeny of microbes living in permanent Antarctic lake ice. Microbial Ecology, 39, 197–202.Google ScholarPubMed
Lancaster, N. 2002. Flux of eolian sediment in the McMurdo Dry Valleys, Antarctica: a preliminary assessment. Arctic Antarctic and Alpine Research, 34, 318–323.CrossRefGoogle Scholar
Lee, J.L.Levin, R.E. 2009. A comparative study of the ability of EMA and PMA to distinguish viable from heat killed mixed bacteria from fish fillets. Journal of Microbiological Methods, 76, 93–96.CrossRefGoogle ScholarPubMed
Luna, G.M., Manini, E.Danovaro, R. 2002. Large fraction of dead and inactive bacteria in coastal marine sediments: comparison of protocols for determination and ecological significance. Applied and Environmental Microbiology, 68, 3509–3513.CrossRefGoogle ScholarPubMed
Mosier, A.C., Murray, A.E.Fritsen, C.H. 2007. Microbiota within the perennial ice cover of Lake Vida, Antarctica. FEMS Microbiology. Ecology, 59, 274–288.CrossRefGoogle ScholarPubMed
Murray, A.E., Hollibaugh, J.T.Orrego, C. 1996. Phylogenetic compositions of bacterioplankton from two California estuaries compared by denaturing gradient gel electrophoresis of 16S rDNA fragments. Applied and Environmental Microbiology, 62, 2676–2680.CrossRefGoogle ScholarPubMed
Muyzer, G., Hottentrager, S., Teske, A.Waver, C. 1996. Denaturing gradient gel electrophoresis of PCR-amplified 16S rDNA. A new molecular approach to analyze the genetic diversity of mixed microbial communities. In Akkermans, A.D.L., van Elsas, J.D. & de Bruijn F.J., eds. Molecular microbial ecology manual 3.4.4. Dordrecht: Kluwer, 1–23.Google Scholar
Niederberger, T.D., Mcdonald, I.R., Hacker, A.L., Soo, R.M., Barrett, J.E., Wall, D.H.Cary, S.C. 2008. Microbial community composition in soils of northern Victoria Land, Antarctica. Environmental Microbiology, 10, 1713–1724.CrossRefGoogle Scholar
Nocker, A., Cheung, C.Y.Camper, A.K. 2006. Comparison of propidium monoazide with ethidium monoazide for differentiation of live vs. dead bacteria by selective removal of DNA from the cells. Journal of Microbiological Methods, 67, 310–320.CrossRefGoogle Scholar
Nocker, A., Sossa, K.E.Camper, A.K. 2007a. Molecular monitoring of disinfection efficacy using propidium monoazide in combination with quantitative PCR. Journal of Microbiological Methods, 70, 252–260.CrossRefGoogle ScholarPubMed
Nocker, A., Sossa-Fernandez, P., Burr, M.D.Camper, A.K. 2007b. Use of propidium monoazide for live/dead distinction in microbial ecology. Applied and Environmental Microbiology, 73, 5111–5117.CrossRefGoogle ScholarPubMed
Olson, J.B., Steppe, T.F., Litaker, R.W.Paerl, H.W. 1998. N2-fixing microbial consortia associated with the ice cover of Lake Bonney, Antarctica. Microbial Ecology, 36, 231–238.CrossRefGoogle ScholarPubMed
Paerl, H.W.Pinckney, J.L. 1996. A mini-review of microbial consortia: their roles in aquatic production and biogeochemical cycling. Microbial Ecology, 31, 225–247.CrossRefGoogle ScholarPubMed
Paerl, H.W.Priscu, J.C. 1998. Microbial phototrophic, heterotrophic and diazotrophic activities associated with aggregates in the permanent ice cover of the Lake Bonney, Antarctica. Microbial Ecology, 36, 221–230.CrossRefGoogle ScholarPubMed
Porazinska, D.L., Fountain, A.G., Nylen, T.H., Tranter, M., Virginia, R.A.Wall, D.H. 2004. The biodiversity and biogeochemistry of cryoconite holes from McMurdo Dry Valley glaciers, Antarctica. Arctic, Antarctic and Alpine Research, 36, 84–91.CrossRefGoogle Scholar
Price, B.P. 2000. A habitat for psychrophiles in deep Antarctic ice. PNAS, 97, 1247–1251.CrossRefGoogle ScholarPubMed
Priscu, J.C., Adams, E.E., Paerl, H.W., Fritsen, C.H., Dore, J.E., Lisle, J.T., Wolf, C.F.Mikucki, J.A. 2005. Perennial Antarctic lake ice: a refuge for cyanobacteria in an extreme environment. In Castello, J.D. & Rogers, S.O., eds. Life in ancient ice. Princeton, NJ: Princeton University Press, 22–49.CrossRefGoogle Scholar
Priscu, J.C., Fritsen, C.H., Adams, E.E., Giovannoni, S.J., Paerl, H.W., McKay, C.P., Doran, P.T., Gordon, D.A., Lanoil, B.D.Pinckney, J.L. 1998. Perennial Antarctic lake ice: an oasis for life in a polar desert. Science, 280, 2095–2098.CrossRefGoogle Scholar
Psenner, R.Sattler, B. 1998. Life at the freezing point. Science, 280, 2073–2074.CrossRefGoogle ScholarPubMed
Rieder, A., Schwartz, T., Schön-Hölz, K., Marten, S.M., Süss, J., Gusbeth, C.Kohnen, W. 2008. Molecular monitoring of inactivation efficiencies of bacteria during pulsed electric field treatment of clinical wastewater. Journal of Applied Microbiology, 105, 2035–2045.CrossRefGoogle ScholarPubMed
Roberts, E.C., Priscu, J.C.Laybourn-Parry, J. 2004. Microplankton dynamics in a perennially ice-covered Antarctic lake - Lake Hoare. Freshwater Biology, 49, 853–869.CrossRefGoogle Scholar
Roberts, E.C., Laybourn-Parry, J., McKnight, D.Novarino, G. 2000. Stratification and dynamics of microbial loop communities in Lake Fryxell, Antarctica. Freshwater Biology, 44, 649–661.CrossRefGoogle Scholar
Sikorskya, J.A., Primeranoa, D.A., Fengera, T.W.Denvir, J. 2004. Effect of DNA damage on PCR amplification efficiency with the relative threshold cycle method. Biochemical and Biophysical Research Communications, 323, 823–830.CrossRefGoogle Scholar
Smith, J.J., Tow, L.A., Stafford, W., Cary, C.Cowan, D.A. 2006. Bacterial diversity in three different Antarctic cold desert mineral soils. Microbial Ecology, 51, 413–421.CrossRefGoogle ScholarPubMed
Suzuki, M.T.Giovannoni, S.J. 1996. Bias caused by template annealing in the amplification of mixtures of 16S rRNA genes by PCR. Applied and Environmental Microbiology, 62, 625–630.CrossRefGoogle ScholarPubMed
Takacs, C.T.Priscu, J.C. 1998. Bacterioplankton dynamics in the McMurdo Dry Valley lakes: production and biomass loss over four seasons. Microbial Ecology, 36, 239–250.CrossRefGoogle ScholarPubMed
Van Trappen, S., Mergaert, J., van Eygen, S., Dawyndt, P., Cnockaert, M.C.Swings, J. 2002. Diversity of 746 heterotrophic bacteria isolated from microbial mats from ten Antarctic lakes. Systematic and Applied Microbiology, 25, 603–610.CrossRefGoogle ScholarPubMed
Wahman, D.G., Wulfeck-Kleier, K.A.Pressman, J.G. 2009. Monochloramine disinfection kinetics of Nitrosomonas europaea using propidium monoazide quantitative PCR (PMA-qPCR) and LIVE/DEAD(R) BacLight methods. Applied and Environmental Microbiology, 75, 5555–5562.CrossRefGoogle Scholar