Measurement of gases entrapped in clean ice from basal portions of the Taylor Glacier, Antarctica, revealed that CO 2 ranged from 229 to 328 ppmv and O 2 was near 20% of the gas volume. In contrast, vertically adjacent sections of the sediment laden basal ice contained much higher concentrations of CO 2 (60,000 to 325,000 ppmv), whereas O 2 represented 4 to 18% of the total gas volume. The deviation in gas composition from atmospheric values occurred concurrently with increased microbial cell concentrations in the basal ice profile, suggesting that in situ microbial processes ( i.e., aerobic respiration) may have altered the entrapped gas composition. Molecular characterization of 16S rRNA genes amplified from samples of the basal ice indicated a low diversity of bacteria, and most of the sequences characterized (87%) were affiliated with the phylum, Firmicutes. The most abundant phylotypes in libraries from ice horizons with elevated CO 2 and depleted O 2 concentrations were related to the genus Paenisporosarcina, and 28 isolates from this genus were obtained by enrichment culturing. Metabolic experiments with Paenisporosarcina sp. TG14 revealed its capacity to conduct macromolecular synthesis when frozen in water derived from melted basal ice samples and incubated at ？15 °C. The results support the hypothesis that the basal ice of glaciers and ice sheets are cryospheric habitats harboring bacteria with the physiological capacity to remain metabolically active and biogeochemically cycle elements within the subglacial environment.
References
[1]
Price, P.B. A habitat for psychrophiles in deep Antarctic ice. Proc. Natl. Acad. Sci. USA 2000, 97, 1247–1251, doi:10.1073/pnas.97.3.1247.
[2]
Doyle, S.M.; Dieser, M.; Broemsen, E.; Christner, B.C. General characteristics of cold-adapted microorganisms. In Polar Microbiology: Life In a Deep Freeze; Whyte, L., Miller, R.V., Eds.; ASM Press: Washington, DC, USA, 2012; pp. 103–125.
[3]
Miteva, V.I.; Sheridan, P.P.; Brenchley, J.E. Phylogenetic and physiological diversity of microorganisms isolated from a deep Greenland glacier ice core. Appl. Environ. Microbiol. 2004, 70, 202–213, doi:10.1128/AEM.70.1.202-213.2004.
[4]
Xiang, S.-R.; Shang, T.-C.; Chen, Y.; Yao, T.-D. Deposition and postdeposition mechanisms as possible drivers of microbial population variability in glacier ice. FEMS Microbiol. Ecol. 2009, 70, 165–176, doi:10.1111/j.1574-6941.2009.00759.x.
[5]
Amato, P.; Doyle, S.M.; Battista, J.R.; Christner, B.C. Implications of subzero metabolic activity on long-term microbial survival in terrestrial and extraterrestrial permafrost. Astrobiology 2010, 10, 789–798, doi:10.1089/ast.2010.0477.
[6]
Abyzov, S.S. Microorganisms in the Antarctic ice. Antarct. Microbiol. 1993, 1, 265–296.
[7]
Abyzov, S.S.; Lipenkov, V.Y.; Bobin, N.E.; Koudryashov, B.B. The microflora of the central Antarctic glacier and the control methods of the sterile isolation of the ice core for microbiological analysis. Akad. Nauk SSSR Izv. Ser. Biol. 1982, 4, 537–548.
[8]
Antony, R.; Krishnan, K.P.; Laluraj, C.M.; Thamban, M.; Dhakephalkar, P.K.; Engineer, A.S.; Shivaji, S. Diversity and physiology of culturable bacteria associated with a coastal Antarctic ice core. Microbiol. Res. 2012, 167, 372–380, doi:10.1016/j.micres.2012.03.003.
[9]
Christner, B.C.; Mosley-Thompson, E.; Thompson, L.G.; Reeve, J.N. Bacterial recovery from ancient ice. Environ. Microbiol. 2003, 5, 433–436, doi:10.1046/j.1462-2920.2003.00422.x.
[10]
Christner, B.C.; Mosley-Thompson, E.; Thompson, L.G.; Zagorodnov, V.; Sandman, K.; Reeve, J.N. Recovery and identification of viable bacteria immured in glacial ice. Icarus 2000, 144, 479–485, doi:10.1006/icar.1999.6288.
[11]
Miteva, V.I.; Brenchley, J.E. Detection and isolation of ultrasmall microorganisms from a 120,000-year-old Greenland glacier ice core. Appl. Environ. Microbiol. 2005, 71, 7806–7818, doi:10.1128/AEM.71.12.7806-7818.2005.
[12]
Skidmore, M.L.; Foght, J.M.; Sharp, M.J. Microbial life beneath a high Arctic glacier. Appl. Environ. Microbiol. 2000, 66, 3214–3220, doi:10.1128/AEM.66.8.3214-3220.2000.
[13]
Ahn, J.; Wahlen, M.; Deck, B.L.; Brook, J.; Mayewski, P.A.; Taylor, K.C.; White, J.W.C. A record of atmospheric CO2 during the last 40,000 years from the siple dome, Antarctica ice core. J. Geophys. Res. 2004, 109, doi:10.1029/2003JD004415.
[14]
Flückiger, J.; Blunier, T.; Stauffer, B.; Chappellaz, J.; Spahni, R.; Kawamura, K.; Schwander, J.; Stocker, T.F.; Dahl-Jensen, D. N2O and CH4 variations during the last glacial epoch: Insight into global processes. Glob. Biogeochem. Cy. 2004, 18, doi:10.1029/2003GB002122.
[15]
Souchez, R.; Janssens, L.; Lemmens, M.; Stauffer, B. Very low oxygen concentration in basal ice from summit, central Greenland. Geophys. Res. Lett. 1995, 22, 2001–2004, doi:10.1029/95GL01995.
[16]
Sowers, T. N2O record spanning the penultimate deglaciation from the vostok ice core. J. Geophys. Res. 2001, 106, 31903–31914, doi:10.1029/2000JD900707.
[17]
Campen, R.K.; Sowers, T.; Alley, R.B. Evidence of microbial consortia metabolizing within a low-latitude mountain glacier. Geology 2003, 31, 231–234, doi:10.1130/0091-7613(2003)031<0231:EOMCMW>2.0.CO;2.
[18]
Souchez, R.; Jouzel, J.; Landais, A.; Chappellaz, J.; Lorrain, R.; Tison, J.-L. Gas isotopes in ice reveal a vegetated central Greenland during ice sheet invasion. Geophys. Res. Lett. 2006, 33, doi:10.1029/2006GL028424.
[19]
Rohde, R.A.; Price, P.B.; Bay, R.C.; Bramall, N.E. In situ microbial metabolism as a cause of gas anomalies in ice. Proc. Natl. Acad. Sci. USA 2008, 105, 8667–8672, doi:10.1073/pnas.0803763105.
[20]
Tung, H.C.; Bramall, N.E.; Price, P.B. Microbial origin of excess methane in glacial ice and implications for life on Mars. Proc. Natl. Acad. Sci. USA 2005, 102, 18292–18296, doi:10.1073/pnas.0507601102.
[21]
Bakermans, C.; Skidmore, M. Microbial respiration in ice at subzero temperatures (？4 °C to ？33 °C). Environ. Microbiol. Rep. 2011, 3, 774–782, doi:10.1111/j.1758-2229.2011.00298.x.
[22]
Panikov, N.; Flanagan, P.; Oechel, W.; Mastepanov, M.; Christensen, T. Microbial activity in soils frozen to below ？39 °C. Soil Biol. Biochem. 2006, 38, 785–794, doi:10.1016/j.soilbio.2005.07.004.
[23]
Christner, B.C. Incorporation of DNA and protein precursors into macromolecules by bacteria at ？15 °C. Appl. Environ. Microbiol. 2002, 68, 6435–6438, doi:10.1128/AEM.68.12.6435-6438.2002.
[24]
Junge, K.; Eicken, H.; Swanson, B.D.; Deming, J.W. Bacterial incorporation of leucine into protein down to ？20 °C with evidence for potential activity in sub-eutectic saline ice formations. Cryobiology 2006, 52, 417–429, doi:10.1016/j.cryobiol.2006.03.002.
[25]
Knight, P.G. The basal ice layer of glaciers and ice sheets. Quat. Sci. Rev. 1997, 16, 975–993, doi:10.1016/S0277-3791(97)00033-4.
[26]
Skidmore, M.; Anderson, S.P.; Sharp, M.; Foght, J.; Lanoil, B.D. Comparison of microbial community compositions of two subglacial environments reveals a possible role for microbes in chemical weathering processes. Appl. Environ. Microbiol. 2005, 71, 6986–6997, doi:10.1128/AEM.71.11.6986-6997.2005.
[27]
Sharp, M.; Parkes, J.; Cragg, B.; Fairchild, I.J.; Lamb, H.; Tranter, M. Widespread bacterial populations at glacier beds and their relationship to rock weathering and carbon cycling. Geology 1999, 27, 107–110, doi:10.1130/0091-7613(1999)027<0107:WBPAGB>2.3.CO;2.
[28]
Tung, H.C.; Price, P.B.; Bramall, N.E.; Vrdoljak, G. Microorganisms metabolizing on clay grains in 3-km-deep Greenland basal ice. Astrobiology 2006, 6, 69–86, doi:10.1089/ast.2006.6.69.
[29]
Hubbard, B.; Cook, S.; Coulson, H. Basal Ice facies: A review and unifying approach. Quat. Sci. Rev. 2009, 28, 1956–1969, doi:10.1016/j.quascirev.2009.03.005.
[30]
Montross, S.N. Biogeochemistry of Basal Ice from Taylor Glacier, Antarctica. Ph.D. Dissertation, Montana State University, Bozeman, MT, USA, 2012.
[31]
Montross, S.M.; Skidmore, M.; Christner, B.; Samyn, D.; Tison, J.L.; Lorrain, R.; Doyle, S.; Fitzsimons, S. Debris-rich basal ice as a microbial habitat, Taylor Glacier, Antarctica. Geomicrobiol. J. 2013, doi:10.1080/01490451.2013.811316.
[32]
Christner, B.C.; Mikucki, J.A.; Foreman, C.M.; Denson, J.; Priscu, J.C. Glacial ice cores: A model system for developing extraterrestrial decontamination protocols. Icarus 2005, 174, 572–584, doi:10.1016/j.icarus.2004.10.027.
[33]
Trevors, J.T.; Cook, S. A Comparison of plating media and diluents for enumeration of aerobic bacteria in a loam soil. J. Microbiol. Meth. 1992, 14, 271–275, doi:10.1016/0167-7012(92)90060-H.
[34]
Samyn, D.; Svensson, A.; Fitzsimons, S. Dynamic implications of discontinuous recrystallization in cold basal ice: Taylor Glacier, Antarctica. J. Geophys. Res. 2008, 113, doi:10.1029/2006JF000600.
[35]
Robe, P.; Nalin, R.; Capellano, C.; Vogel, T.M.; Simonet, P. Extraction of DNA from soil. Eur. J. Soil. Biol. 2003, 39, 183–190, doi:10.1016/S1164-5563(03)00033-5.
[36]
Lane, D.J. 16S/23S rRNA sequencing. In Nucleic Acid Techniques in Bacterial Systematics; Stackebrandt, E., Goodfellow, M., Eds.; Wiley: New York, NY, USA, 1991; pp. 115–175.
Edgar, R.C.; Haas, B.J.; Clemente, J.C.; Quince, C.; Knight, R. UCHIME improves sensitivity and speed of chimera detection. Bioinformatics 2011, 27, 2194–2200, doi:10.1093/bioinformatics/btr381.
[41]
Tamura, K.; Peterson, D.; Peterson, N.; Stecher, G.; Nei, M.; Kumar, S. MEGA5: Molecular evolutionary genetics analysis using maximum likelihood, evolutionary distance, and maximum parsimony methods. Mol. Biol. Evol. 2011, 28, 2731–2739, doi:10.1093/molbev/msr121.
[42]
Kim, O.S.; Cho, Y.J.; Lee, K.; Yoon, S.H.; Kim, M.; Na, H.; Park, S.C.; Jeon, Y.S.; Lee, J.H.; Yi, H.; et al. Introducing EzTaxon-E: A prokaryotic 16S rRNA gene sequence database with phylotypes that represent uncultured species. Int. J. Syst. Evol. Microbiol. 2012, 62, 716–721, doi:10.1099/ijs.0.038075-0.
[43]
Wang, Q.; Garrity, G.M.; Tiedje, J.M.; Cole, J.R. Naive bayesian classifier for rapid assignment of rRNA sequences into the new bacterial taxonomy. Appl. Environ. Microbiol. 2007, 73, 5261–5267, doi:10.1128/AEM.00062-07.
[44]
Felsenstein, J. Confidence limits on phylogenies: An approach using the bootstrap. Evolution 1985, 783–791.
[45]
Bell, R.E.; Ferraccioli, F.; Creyts, T.T.; Braaten, D.; Corr, H.; Das, I.; Damaske, D.; Frearson, N.; Jordan, T.; Rose, K.; et al. Widespread persistent thickening of the east Antarctic ice sheet by freezing from the base. Science 2011, 331, 1592–1595, doi:10.1126/science.1200109.
[46]
Wadham, J.; Arndt, S.; Tulaczyk, S.; Stibal, M.; Tranter, M.; Telling, J.; Lis, G.; Lawson, E.; Ridgwell, A.; Dubnick, A. Potential methane reservoirs beneath Antarctica. Nature 2012, 488, 633–637, doi:10.1038/nature11374.
[47]
Byrne, S.; Dundas, C.M.; Kennedy, M.R.; Mellon, M.T.; McEwen, A.S.; Cull, S.C.; Daubar, I.J.; Shean, D.E.; Seelos, K.D.; Murchie, S.L. Distribution of mid-latitude ground ice on Mars from new impact craters. Science 2009, 325, 1674–1676, doi:10.1126/science.1175307.
[48]
Samyn, D.; Fitzsimons, S.J.; Lorrain, R.D. Strain-induced phase changes within cold basal ice from Taylor Glacier, Antarctica, indicated by textural and gas analyses. J. Glaciol. 2005, 51, 611–619, doi:10.3189/172756505781829098.
[49]
Souchez, R.; Lemmens, M.; Chappellaz, J. Flow-induced mixing in the GRIP basal ice deduced from the CO2 and CH4 records. Geophys. Res. Lett. 1995, 22, 41–44, doi:10.1029/94GL02863.
[50]
Steven, B.; Briggs, G.; McKay, C.P.; Pollard, W.H.; Greer, C.W.; Whyte, L.G. Characterization of the microbial diversity in a permafrost sample from the Canadian High Arctic using culture-dependent and culture-independent methods. FEMS Microbiol. Ecol. 2007, 59, 513–523.
[51]
Brinkmeyer, R.; Knittel, K.; Jürgens, J.; Weyland, H.; Amann, R.; Helmke, E. Diversity and structure of bacterial communities in Arctic versus Antarctic pack ice. Appl. Environ. Microbiol. 2003, 69, 6610–6619, doi:10.1128/AEM.69.11.6610-6619.2003.
[52]
Cheng, S.M.; Foght, J.M. Cultivation-independent and -dependent characterization of bacteria resident beneath John Evans Glacier. FEMS Microbiol. Ecol. 2007, 59, 318–330, doi:10.1111/j.1574-6941.2006.00267.x.
[53]
Yde, J.C.; Finster, K.W.; Raiswell, R.; Steffensen, J.P.; Heinemeier, J.; Olsen, J.; Gunnlaugsson, H.P.; Nielsen, O.B. Basal ice microbiology at the margin of the Greenland ice sheet. Ann. Glaciol. 2010, 51, 71–79.
[54]
Lacelle, D.; Radtke, K.; Clark, I.D.; Fisher, D.; Lauriol, B.; Utting, N.; Whyte, L.G. Geomicrobiology and occluded O2-CO2-Ar gas analyses provide evidence of microbial respiration in ancient terrestrial ground ice. Earth Planet. Sci. Lett. 2011, 306, 46–54, doi:10.1016/j.epsl.2011.03.023.
[55]
Steven, B.; Pollard, W.H.; Greer, C.W.; Whyte, L.G. Microbial diversity and activity through a permafrost/ground ice core profile from the Canadian High Arctic. Environ. Microbiol. 2008, 10, 3388–3403, doi:10.1111/j.1462-2920.2008.01746.x.
[56]
Christner, B.C.; Skidmore, M.L. Montana State University, Bozeman, MT, USA, 2005. Unpublished work.
[57]
Perreault, N.N.; Greer, C.W.; Andersen, D.T.; Tille, S.; Lacrampe-Couloume, G.; Lollar, B.S.; Whyte, L.G. Heterotrophic and autotrophic microbial populations in cold perennial springs of the high Arctic. Appl. Environ. Microbiol. 2008, 74, 6898–6907, doi:10.1128/AEM.00359-08.
[58]
Reddy, G.; Manasa, B.P.; Singh, S.K.; Shivaji, S. Paenisporosarcina indica sp. nov., a psychrophilic bacterium from pindari glacier of the Himalayan Mountain Ranges and reclassification of sporosarcina Antarctica Yu et al., 2008 as Paenisporosarcina antarctica comb. nov. and emended description of the genus Paenisporosarcina. Int. J. Syst. Evol. Microbiol. 2013. in press.
[59]
Reddy, G.; Matsumoto, G.; Shivaji, S. Sporosarcina macmurdoensis sp. nov., from a cyanobacterial mat sample from a pond in the McMurdo Dry Valleys, Antarctica. Int. J. Syst. Evol. Microbiol. 2003, 53, 1363–1367, doi:10.1099/ijs.0.02628-0.
[60]
Price, P.B.; Sowers, T. Temperature dependence of metabolic rates for microbial growth, maintenance, and survival. Proc. Natl. Acad. Sci. USA 2004, 101, 4631–4636, doi:10.1073/pnas.0400522101.
[61]
Barletta, R.E.; Priscu, J.C.; Mader, H.M.; Jones, W.L.; Roe, C.H. Chemical analysis of ice vein microenvironments: II. analysis of glacial samples from Greenland and Antarctica. J. Glaciol. 2012, 58, 1109–1118.
