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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Altschul SF, Madden TL, Schaffer AA, Zhang J, Zhang Z, Miller W, Lipman DJ (1997) Gapped BLAST and PSI-BLAST: a new generation of protein database search programs. Nucleic Acids Res 25:3389–3402
Ashbolt NJ (1990) Antarctic biotechnology—what is the potential? Aust J Biotechnol 4:103–105, 108
Bowman JP, Rea SM, McCammon SA, McMeekin TA (2000) Diversity and community structure within anoxic sediment from marine salinity meromictic lakes and a coastal meromictic marine basin, Vestfold Hills, Eastern Antarctica. Environ Microbiol 2:227–237
Brambilla E, Hippe H, Hagelstein A, Tindall BJ, Stackebrandt E (2001) 16S rDNA diversity of cultured and uncultured prokaryotes of a mat sample from Lake Fryxell, McMurdo Dry Valleys, Antarctica. Extremophiles 5:23–33
Brinkmeyer R, Glockner FO, Helmke E, Amann R (2004) Predominance of beta-proteobacteria in summer melt pools on Arctic pack ice. Limnol Oceanogr 49:1013–1021
Button DK (1993) Nutrient-limited microbial growth kinetics: overview and recent advances. Antonie van Leeuwenhoek 63:225–235
Cho JC, Giovannoni SJ (2004) Cultivation and growth characteristics of a diverse group of oligotrophic marine Gammaproteobacteria. Appl Environ Microbiol 70:432–440
Cho JC, Vergin KL, Morris RM, Giovannoni SJ (2004) Lentisphaera araneosa gen nov, sp nov, a transparent exopolymer producing marine bacterium, and the description of a novel bacterial phylum, Lentisphaerae. Environ Microbiol 6:611–621
Connon SA, Giovannoni SJ (2002) High-throughput methods for culturing microorganisms in very-low-nutrient media yield diverse new marine isolates. Appl Environ Microbiol 68:3878–3885
Felsenstein J (1989) PHYLIP—Phylogeny Inference Package (Version 3.2). Cladistics 5:164–166
Foght J, Aislabie J, Turner S, Brown CE, Ryburn J, Saul DJ, Lawson W (2004) Culturable bacteria in subglacial sediments and ice from two Southern Hemisphere glaciers. Microb Ecol 47:329–340
Fry JC (1990) Oligotrophs. In: Edwards C (ed) Microbiology of extreme environments. Open Univ. Press, Milton Keynes, UK, pp 93–116
Gaidos E, Lanoil B, Thorsteinsson T, Graham A, Skidmore M, Han SK, Rust T, Popp B (2004) A viable microbial community in a subglacial volcanic crater lake, Iceland. Astrobiology 4:327–344
Gich F, Schubert K, Bruns A, Hoffelner H, Overmann J (2005) Specific detection, isolation, and characterization of selected, previously uncultured members of the freshwater bacterioplankton community. Appl Environ Microbiol 71:5908–5919
Glatz RE, Lepp PW, Ward BB, Francis CA (2006) Planktonic microbial community composition across steep physical/chemical gradients in permanently ice-covered Lake Bonney, Antarctica. Geobiology 4:53–67
Green WJ, Gardner TJ, Ferdelman TG, Angle MP, Varner LC, Nixon P (1989) Geochemical process in the Lake Fryxell Basin (Victoria Land, Antarctica). Hydrobiologia 172:129–148
Irgens RL, Gosink JJ, Staley JT (1996) Polaromonas vacuolata gen nov, sp nov, a psychrophilic, marine, gas vacuolate bacterium from Antarctica. Int J Syst Evol Microbiol 46:822–826
Jung DO, Achenbach LA, Karr EA, Takaichi S, Madigan MT (2004) A gas vesiculate planktonic strain of the purple non-sulfur bacterium Rhodoferax antarcticus isolated from Lake Fryxell, Dry Valleys, Antarctica. Arch Microbiol 182:236–243
Karr EA, Sattley WM, Rice MR, Jung DO, Madigan MT, Achenbach LA (2005) Diversity and distribution of sulfate-reducing bacteria in permanently frozen Lake Fryxell, McMurdo Dry Valleys, Antarctica. Appl Environ Microbiol 71:6353–6359
Ludwig W, Strunk O, Klugbauer S, Klugbauer N, Weizenegger M, Neumaier J, Bachleitner M, Schleifer KH (1998) Bacterial phylogeny based on comparative sequence analysis. Electrophoresis 19:554–568
Lyons WB, Fountain A, Doran P, Priscu JC, Neumann K, Welch KA (2000) Importance of landscape position and legacy: the evolution of the lakes in Taylor Valley, Antarctica. Freshw Biol 43:355–367
Madigan MT (1998) Isolation and characterization of psychrophilic purple bacteria from Antarctica. In: Peschek GA, Loffelhardt W, Schmetterer G (eds) The phototrophic prokaryotes. Plenum, New York, pp 699–706
Madigan MT, Jung DO, Woese CR, Achenbach LA (2000) Rhodoferax antarcticus sp. nov., a moderately psychrophilic purple nonsulfur bacterium isolated from an Antarctic microbial mat. Arch Microbiol 173:269–277
Morris RM, Longnecker K, Giovannoni SJ (2006) Pirellula and OM43 are among the dominant lineages identified in an Oregon coast diatom bloom. Environ Microbiol 8:1361–1370
Nedashkovskaya OI, Suzuki M, Lysenko AM, Snauwaert C, Vancanneyt M, Swings J, Vysotskii MV, Mikhailov VV (2004) Cellullophaga pacifica sp nov. Int J Syst Evol Microbiol 54:609–613
Olsen GJ, Matsuda H, Hagstrom R, Overbeek R (1994) fastDNAml: A tool for construction of phylogenetic trees of DNA sequences using maximum likelihood. Comput Appl Biosci 10:41–48
Page KA, Connon SA, Giovannoni SJ (2004) Representative freshwater bacterioplankton isolated from Crater Lake, Oregon. Appl Environ Microbiol 70:6542–6550
Priscu JC, Downes MT, McKay CP (1996) Extreme super-saturation of nitrous oxide in a permanently ice-covered Antarctic lake. Limnol Oceanogr 41:1544–1551
Priscu JC (1997) The biogeochemistry of nitrous oxide in permanently ice-covered lakes of the McMurdo Dry Valleys, Antarctica. Glob Chang Biol 3:301–305
Rappe MS, Connon SA, Vergin KL, Giovannoni SJ (2002) Cultivation of the ubiquitous SAR11 marine bacterioplankton clade. Nature 418:630–633
Sassen R, Roberts HH, Carney R, Milkov AV, DeFreitas DA, Lanoil B, Zhang CL (2004) Free hydrocarbon gas, gas hydrate, and authigenic minerals in chemosynthetic communities of the northern Gulf of Mexico continental slope: relation to microbial processes. Chem Geol 205:195–217
Sattley WM, Madigan MT (2006) Isolation, characterization, and ecology of cold-active, chemolithotrophic, sulfur-oxidizing bacteria from perennially ice-covered Lake Fryxell, Antartica. Appl Environ Microbiol 72:5562–5568
Shivaji S, Reddy GSN, Suresh K, Gupta P, Chintalapati S, Schumann P, Stackebrandt E, Matsumoto GI (2005) Psychrobacter vallis sp nov and Psychrobacter aquaticus sp nov, from Antarctica. Int J Syst Evol Microbiol 55:757–762
Skidmore M, Anderson SP, Sharp M, Foght J, Lanoil BD (2005) Comparison of microbial community compositions of two subglacial environments reveals a possible role for microbes in chemical weathering processes. Appl Environ Microbiol 71:6986–6997
Spigel RH, Priscu JC (1996) Evolution of temperature and salt structure of Lake Bonney, a chemically stratified Antarctic lake. Hydrobiologia 321(3):177–190
Spigel RH, Priscu JC (1998) Physical limnology of the McMurdo Dry Valley lakes. In: Priscu JC (ed) Ecosystem dynamics in a polar desert: the McMurdo Dry Valleys, Antarctica, vol 72. American Geophysical Union, Washington, DC, pp 153–188
Takacs CD, Priscu JC (1998) Bacterioplankton dynamics in the McMurdo Dry Valley lakes, Antarctica: Production and biomass loss over four seasons. Microb Ecol 36:239–250
Voytek MA, Priscu JC, Ward BB (1999) The distribution and relative abundance of ammonia-oxidizing bacteria in lakes of the McMurdo Dry Valley, Antarctica. Hydrobiologia 401:113–130
Wharton Jr RA, Lyons WB, Des Marais D (1993) Stable isotope biogeochemistry of carbon and nitrogen in a perennially ice-covered Antarctic lake. Chem Geol 107:159–172
Acknowledgments
This work was supported by NSF 0522990 to BDL and SJG. Samples were collected by MT Madigan, DO Jung, and WM Sattley. We would like to thank MT Madigan for constructive comments on this manuscript. Data in Table 1 were provided by JC Priscu of the McMurdo Long Term Ecological Research Project, NSF-OPP 432595.
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Stingl, U., Cho, JC., Foo, W. et al. Dilution-to-Extinction Culturing of Psychrotolerant Planktonic Bacteria from Permanently Ice-covered Lakes in the McMurdo Dry Valleys, Antarctica. Microb Ecol 55, 395–405 (2008). https://doi.org/10.1007/s00248-007-9284-4
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DOI: https://doi.org/10.1007/s00248-007-9284-4