Metatranscriptomic and Metagenomic Analysis of Biological Diversity in Subglacial Lake Vostok (Antarctica).

A combined metatranscriptomic and metagenomic study of Vostok (Antarctica) ice core sections from glacial, basal, and lake water accretion ice yielded sequences that indicated a wide variety of species and possible conditions at the base of the glacier and in subglacial Lake Vostok. Few organisms were in common among the basal ice and accretion ice samples, suggesting little transmission of viable organisms from the basal ice meltwater into the lake water. Additionally, samples of accretion ice, each of which originated from water in several locations of the shallow embayment, exhibit only small amounts of mixing of species. The western-most portion of the embayment had very low numbers of organisms, likely due to biologically challenging conditions. Increasing numbers of organisms were found progressing from west to east, up to approximately 7 km into the embayment. At that point, the numbers of unique sequences and sequence reads from thermophilic, thermotolerant, psychrophilic, and psychrotolerant organisms increased dramatically, as did sequences from alkaliphilic, alkalitolerant, acidophilic, and acidotolerant sequences. The number of unique and total sequences were positively associated with increases in concentrations of Na+, Ca2+, Mg2+, SO42-, Cl-, total amino acids, and non-purgeable organic carbon. The numbers of unique sequences from organisms reported from soil, sediment, ice, aquatic, marine, animal, and plant (probably pollen) sources also peaked in this region, suggesting that this was the most biologically active region. The confluence of the high numbers of organisms, physiologies, and metabolic capabilities suggests the presence of energy and nutrient sources in the eastern half of the embayment. Data from the main basin suggested a cold oligotrophic environment containing fewer organisms. In addition to bacteria, both the basal ice and accretion ice contained sequences from a diverse assemblage of eukaryotes, as well as from bacteria that are known to be associated with multicellular eukaryotes.

Physiological Ecology of Microorganisms in Subglacial Lake Whillans.

Subglacial microbial habitats are widespread in glaciated regions of our planet. Some of these environments have been isolated from the atmosphere and from sunlight for many thousands of years. Consequently, ecosystem processes must rely on energy gained from the oxidation of inorganic substrates or detrital organic matter. Subglacial Lake Whillans (SLW) is one of more than 400 subglacial lakes known to exist under the Antarctic ice sheet; however, little is known about microbial physiology and energetics in these systems. When it was sampled through its 800 m thick ice cover in 2013, the SLW water column was shallow (~2 m deep), oxygenated, and possessed sufficient concentrations of C, N, and P substrates to support microbial growth. Here, we use a combination of physiological assays and models to assess the energetics of microbial life in SLW. In general, SLW microorganisms grew slowly in this energy-limited environment. Heterotrophic cellular carbon turnover times, calculated from 3H-thymidine and 3H-leucine incorporation rates, were long (60 to 500 days) while cellular doubling times averaged 196 days. Inferred growth rates (average ~0.006 d-1) obtained from the same incubations were at least an order of magnitude lower than those measured in Antarctic surface lakes and oligotrophic areas of the ocean. Low growth efficiency (8%) indicated that heterotrophic populations in SLW partition a majority of their carbon demand to cellular maintenance rather than growth. Chemoautotrophic CO2-fixation exceeded heterotrophic organic C-demand by a factor of ~1.5. Aerobic respiratory activity associated with heterotrophic and chemoautotrophic metabolism surpassed the estimated supply of oxygen to SLW, implying that microbial activity could deplete the oxygenated waters, resulting in anoxia. We used thermodynamic calculations to examine the biogeochemical and energetic consequences of environmentally imposed switching between aerobic and anaerobic metabolisms in the SLW water column. Heterotrophic metabolisms utilizing acetate and formate as electron donors yielded less energy than chemolithotrophic metabolisms when calculated in terms of energy density, which supports experimental results that showed chemoautotrophic activity in excess of heterotrophic activity. The microbial communities of subglacial lake ecosystems provide important natural laboratories to study the physiological and biogeochemical behavior of microorganisms inhabiting cold, dark environments.

Microbial Community Structure of Subglacial Lake Whillans, West Antarctica.

Subglacial Lake Whillans (SLW) is located beneath ∼800 m of ice on the Whillans Ice Stream in West Antarctica and was sampled in January of 2013, providing the first opportunity to directly examine water and sediments from an Antarctic subglacial lake. To minimize the introduction of surface contaminants to SLW during its exploration, an access borehole was created using a microbiologically clean hot water drill designed to reduce the number and viability of microorganisms in the drilling water. Analysis of 16S rRNA genes (rDNA) amplified from samples of the drilling and borehole water allowed an evaluation of the efficacy of this approach and enabled a confident assessment of the SLW ecosystem inhabitants. Based on an analysis of 16S rDNA and rRNA (i.e., reverse-transcribed rRNA molecules) data, the SLW community was found to be bacterially dominated and compositionally distinct from the assemblages identified in the drill system. The abundance of bacteria (e.g., Candidatus Nitrotoga, Sideroxydans, Thiobacillus, and Albidiferax) and archaea (Candidatus Nitrosoarchaeum) related to chemolithoautotrophs was consistent with the oxidation of reduced iron, sulfur, and nitrogen compounds having important roles as pathways for primary production in this permanently dark ecosystem. Further, the prevalence of Methylobacter in surficial lake sediments combined with the detection of methanogenic taxa in the deepest sediment horizons analyzed (34-36 cm) supported the hypothesis that methane cycling occurs beneath the West Antarctic Ice Sheet. Large ratios of rRNA to rDNA were observed for several operational taxonomic units abundant in the water column and sediments (e.g., Albidiferax, Methylobacter, Candidatus Nitrotoga, Sideroxydans, and Smithella), suggesting a potentially active role for these taxa in the SLW ecosystem. Our findings are consistent with chemosynthetic microorganisms serving as the ecological foundation in this dark subsurface environment, providing new organic matter that sustains a microbial ecosystem beneath the West Antarctic Ice Sheet.

Stable water isotopic composition of the Antarctic subglacial Lake Vostok: implications for understanding the lake's hydrology.

We estimated the stable isotopic composition of water from the subglacial Lake Vostok using two different sets of samples: (1) water frozen on the drill bit immediately after the first lake unsealing and (2) water frozen in the borehole after the unsealing and re-drilled one year later. The most reliable values of the water isotopic composition are: -59.0 ± 0.3 ‰ for oxygen-18, -455 ± 1 ‰ for deuterium and 17 ± 1 ‰ for d-excess. This result is also confirmed by the modelling of isotopic transformations in the water which froze in the borehole, and by a laboratory experiment simulating this process. A comparison of the newly obtained water isotopic composition with that of the lake ice (-56.2 ‰ for oxygen-18, -442.4 ‰ for deuterium and 7.2 ‰ for d-excess) leads to the conclusion that the lake ice is very likely formed in isotopic equilibrium with water. In turn, this means that ice is formed by a slow freezing without formation of frazil ice crystals and/or water pockets. This conclusion agrees well with the observed physical and chemical properties of the lake's accreted ice. However, our estimate of the water's isotopic composition is only valid for the upper water layer and may not be representative for the deeper layers of the lake, so further investigations are required.

Microbiology of the subglacial Lake Vostok: first results of borehole-frozen lake water analysis and prospects for searching for lake inhabitants.

This article examines the question of the possible existence of microbial life inhabiting the subglacial Lake Vostok buried beneath a 4 km thick Antarctic ice sheet. It represents the results of analysis of the only available frozen lake water samples obtained upon the first lake entry and subsequent re-coring the water frozen within the borehole. For comparison, results obtained by earlier molecular microbiological studies of accretion ice are included in this study, with the focus on thermophiles and an unknown bacterial phylotype. A description of two Lake Vostok penetrations is presented for the first time from the point of view of possible clean water sampling. Finally, the results of current studies of Lake Vostok frozen water samples are presented, with the focus on the discovery of another unknown bacterial phylotype w123-10 distantly related to the above-mentioned unknown phylotype AF532061 detected in Vostok accretion ice, both successfully passing all possible controls for contamination. The use of clean-room facilities and the establishment of a contaminant library are considered to be prerequisites for research on microorganisms from Lake Vostok. It seems that not yet recorded microbial life could exist within the Lake Vostok water body. In conclusion, the prospects for searching for lake inhabitants are expressed with the intention to sample the lake water as cleanly as possible in order to make sure that further results will be robust.

Geology and environments of subglacial Lake Vostok.

The reconstruction of the geological (tectonic) structure and environments of subglacial Lake Vostok is based on geophysical surveys and the study of mineral particles found in cores of accreted ice and frozen lake water (sampled after the lake was unsealed). Seismic reflection and refraction investigations conducted in the southern part of Lake Vostok show very thin (200-300 m) sedimentary cover overlying a crystalline basement. Most of this thin veneer is thought to have been deposited during temperate-glacial conditions in Oligocene to Middle Miocene time (ca 34-14 Ma). The composition of the lake-bottom sediments can be deduced from mineral inclusions found in cores of accreted ice. Inclusions are represented by soft aggregates consisting mainly of clay-mica minerals and micrometre-sized quartz grains. Some of these inclusions contain subangular to semi-rounded rock clasts (siltstones and sandstones) ranging from 0.3 to 8 mm in size. In total, 31 zircon grains have been identified in two rock clasts and dated using SHRIMP-II. The ages of the studied zircons range from 0.6 to 2.0 Ga with two distinct clusters between 0.8 and 1.15 Ga and between 1.6 and 1.8 Ga. Rock clasts obviously came from the western lake shore, which is thus composed of terrigenous strata with an age of not older than 600 Ma. The sedimentary nature of the western lake shore is also confirmed by seismic refraction data showing seismic velocities there of 5.4-5.5 km s(-1) at the bedrock surface. After Lake Vostok was unsealed, its water (frozen and sampled next season) was also studied with scanning electron microscopy and X-ray microprobe analysis. This study showed the existence of calcium carbonate and silica microparticles (10-20 µm across) in frozen water.

High geothermal heat flux measured below the West Antarctic Ice Sheet.

The geothermal heat flux is a critical thermal boundary condition that influences the melting, flow, and mass balance of ice sheets, but measurements of this parameter are difficult to make in ice-covered regions. We report the first direct measurement of geothermal heat flux into the base of the West Antarctic Ice Sheet (WAIS), below Subglacial Lake Whillans, determined from the thermal gradient and the thermal conductivity of sediment under the lake. The heat flux at this site is 285 ± 80 mW/m(2), significantly higher than the continental and regional averages estimated for this site using regional geophysical and glaciological models. Independent temperature measurements in the ice indicate an upward heat flux through the WAIS of 105 ± 13 mW/m(2). The difference between these heat flux values could contribute to basal melting and/or be advected from Subglacial Lake Whillans by flowing water. The high geothermal heat flux may help to explain why ice streams and subglacial lakes are so abundant and dynamic in this region.