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Here, we report the draft genome sequence for a new putative genus and species in the family M1A02 within the order Phycisphaerales . Isolated from the metagenome of a benthic pinnacle-shaped mat in the Antarctic Lake Untersee, the members of this family have been found in biofilms and freshwater environments.

Lake Untersee located in Eastern Antarctica, is a perennially ice-covered lake. At the bottom of its southern basin lies 20 m of anoxic, methane rich, stratified water, making it a good analog for Enceladus, a moon of Saturn. Here we present the first metagenomic study of this basin and detail the community composition and functional potential of the microbial communities at 92 m, 99 m depths and within the anoxic sediment. A diverse and well-populated microbial community was found, presenting the potential for Enceladus to have a diverse and abundant community. We also explored methanogenesis, sulfur metabolism, and nitrogen metabolism, given the potential presence of these compounds on Enceladus. We found an abundance of these pathways offering a variety of metabolic strategies. Additionally, the extreme conditions of the anoxic basin make it optimal for testing spaceflight technology and life detection methods for future Enceladus exploration.

Lake Untersee is one of the largest perennially ice-covered lakes in Dronning Maud Land. We investigated the energy and water mass balance of Lake Untersee to understand its state of equilibrium. The thickness of the ice cover is strongly correlated with sublimation rates; variations in sublimation rates across the ice cover are largely determined by wind-driven turbulent heat fluxes and the number of snow-covered days. Lake extent and water level have remained stable for the past 20 years, indicating that the water mass balance is in equilibrium. The lake is damned by the Anuchin Glacier and mass balance calculation suggest that subaqueous melting of terminus ice contributes 40–45% of the annual water budget; since there is no evidence of streams flowing into the lake, the lake must be connected to a groundwater system that contributes 55–60% in order to maintain the lake budget in balance. The groundwater likely flows at a rate of ~8.8 × 10−2m3s−1, a reasonable estimate given the range of subglacial water flux in the region. The fate of its well-sealed ice cover is likely tied to changes in wind regime, whereas changes in water budget are more closely linked to the response of surrounding glaciers to climate change.

Antarctic perennially ice-covered lakes provide a stable low-disturbance environment where complex microbially mediated structures can grow. Lake Untersee, an ultra-oligotrophic lake in East Antarctica, has the lake floor covered in benthic microbial mat communities, where laminated organo-sedimentary structures form with three distinct, sympatric morphologies: small, elongated cuspate pinnacles, large complex cones and flat mats. We examined the diversity of prokaryotes and eukaryotes in pinnacles, cones and flat microbial mats using high-throughput sequencing of 16S and 18S rRNA genes and assessed how microbial composition may underpin the formation of these distinct macroscopic mat morphologies under the same environmental conditions. Our analysis identified distinct clustering of microbial communities according to mat morphology. The prokaryotic communities were dominated by Cyanobacteria, Proteobacteria, Verrucomicrobia, Planctomycetes, and Actinobacteria. While filamentous Tychonema cyanobacteria were common in all mat types, Leptolyngbya showed an increased relative abundance in the pinnacle structures only. Our study provides the first report of the eukaryotic community structure of Lake Untersee benthic mats, which was dominated by Ciliophora, Chlorophyta, Fungi, Cercozoa, and Discicristata. The eukaryote richness was lower than for prokaryote assemblages and no distinct clustering was observed between mat morphologies. These findings suggest that cyanobacterial assemblages and potentially other bacteria and eukaryotes may influence structure morphogenesis, allowing distinct structures to form across a small spatial scale.

Perennially ice-covered lakes that host benthic microbial ecosystems are present in many regions of Antarctica. Lake Untersee is an ultra-oligotrophic lake that is substantially different from any other lakes on the continent as it does not develop a seasonal moat and therefore shares similarities to sub-glacial lakes where they are sealed to the atmosphere. Here, we determine the source of major solutes and carbon to Lake Untersee, evaluate the carbon cycling and assess the metabolic functioning of microbial mats using an isotope geochemistry approach. The findings suggest that the glacial meltwater recharging the closed-basin and well-sealed Lake Untersee largely determines the major solute chemistry of the oxic water column with plagioclase and alumino-silicate weathering contributing 2+–Na+ solutes to the lake. The TIC concentration in the lake is very low and is sourced from melting of glacial ice and direct release of occluded CO2 gases into the water column. The comparison of δ13CTIC of the oxic lake waters with the δ13C in the top microbial mat layer show no fractionation due to non-discriminating photosynthetic fixation of HCO3– in the high pH and carbon-starved water. The 14C results indicate that phototrophs are also fixing respired CO2 from heterotrophic metabolism of the underlying microbial mats layers. The findings provide insights into the development of collaboration in carbon partitioning within the microbial mats to support their growth in a carbon-starved ecosystem.

Lake Untersee is a perennially ice-covered Antarctic lake that consists of two basins. The deepest basin, next to the Anuchin Glacier is aerobic to its maximum depth of 160 m. The shallower basin has a maximum depth of 100 m, is anoxic below 80 m, and is shielded from convective currents. The thermal profile in the anoxic basin is unusual in that the water temperature below 50 m is constant at 4°C but rises to 5°C between 70 m and 80 m depth, then drops to 3.7°C at the bottom. Field measurements were used to conduct a thermal and stability analysis of the anoxic basin. The shape of the thermal maximum implies two discrete locations of energy input, one of 0.11 W m-2 at 71 m depth and one of 0.06 W m-2 at 80 m depth. Heat from microbial activity cannot account for the required amount of energy at either depth. Instead, absorption of solar radiation due to an increase in water opacity at these depths can account for the required energy input. Hence, while microbial metabolism is not an important source of heat, biomass increases opacity in the water column resulting in greater absorption of sunlight.

Calcite-rich columnar stromatolites grew in perennially ice-covered Lake Joyce in the McMurdo Dry Valleys, Antarctica, during a period of environmental change associated with rising lake level. Stromatolite calcite contains carbon and oxygen isotope records of changes to microbial activity in response to variable light environments and water chemistry through time. The stromatolites grew synchronously with correlative calcite zones. The innermost (oldest) calcite zone has a wide range of δ13Ccalcite values consistent with variable photosynthetic effects on local DIC 13C/12C. Subsequent calcite zones preserve a progressive enrichment in δ13Ccalcite values of approximately + 2.6‰ through time, with δ13Ccalcite values becoming less variable. This enrichment likely records the removal of 12C by photosynthesis from the DIC reservoir over decades, with photosynthetic effects decreasing as light levels became lower and more consistent through time. Mean δ18Ocalcite values of the innermost calcified zone were at least 1‰ lower than those of the other calcified zones (t test p-level

Soil formation on the territory of ice-free oasis of the continental Antarctica was studied on example of Untersee Valley, Queen Maud Land (QML), Eastern Antarctica. Soils of the Lake Untersee surroundings were characterized in terms of morphology and main chemical and physical properties for the first time. Dominating soils are basically characterized by a poorly differentiated soil profile, with a horizon Cryic type located directly on the parent material. The predominant parent material types of the Untersee Valley are the glacial, fluvioglacial sandy loams and gravels with a large content of differently sized fragments, as well as glacial-lacustrine sandy-loams and loamy-clays. A characteristic feature of glacial and fluvioglacial deposits is their sorting as a consequence of the cryoturbation process. The main meso-relief forms of the Valley include terminal type complexes of moraine hills and ridges, eskers, drumlins, sandurs, patterned grounds, and pingo. The investigated soils are characterized by a low content of organic matter and low intensity of weathering processes in parent material as a function of low biogenic and abiogenic interactions in severe bioclimatic conditions of the Antarctic intercontinental oases.

Here, we report the draft genome sequence for a new putative genus and species in the Methanoregulaceae family, whose members are generally slow-growing rod-shaped or coccoid methanogenic archaea. The information on this sediment-dwelling organism sheds light on the prokaryotes inhabiting isolated, deep, and extremely cold methane-rich environments.

Microbial pinnacles in ice-covered Lake Vanda, McMurdo Dry Valleys, Antarctica, extend from the base of the ice to more than 50 m water depth. The distribution of microbial communities, their photosynthetic potential, and pinnacle morphology affects the local accumulation of biomass, which in turn shapes pinnacle morphology. This feedback, plus environmental stability, promotes the growth of elaborate microbial structures. In Lake Vanda, all mats sampled from greater than 10 m water depth contained pinnacles with a gradation in size from

Comparative analysis of the presence of bacteriophages was carried out for the water column of a permanently ice-covered, extremely oligotrophic Lake Untersee (East Antarctica) and the ancient ice wedge of the Mamontova Gora outcrop (Aldan River, Central Yakutia). Microscopy revealed bacteriophages in the Mamontova Gora ice samples and in the lysates of the pure cultures of phage-sensitive bacteria isolated from the same samples. Bacteriophages isolated from these cultures were filamentous and interacted with bacteria as moderate (lysogenic) phages. A similar filamentous bacteriophage was isolated from the Lake Untersee water column. The highest morphological diversity of bacteriophages was revealed by microscopy in the oxic Lake Untersee water column in the chemocline zone (70–76 m) and in the sulfide layer (85 m). Detection of similar filamentous bacteriophages in a relic ice sample and in the samples from Antarctic Lake Untersee indicate wide occurrence of bacteriophages and lysogeny in microbial communities of low-temperature ecosystems.

This study describes microbial community compositions, and various cold-responsive stress genes, encompassing cold-induced proteins (CIPs) and cold-associated general stress-responsive proteins (CASPs) in selected Antarctic lake water, sediment, and soil metagenomes. Overall, Proteobacteria and Bacteroidetes were the major taxa in all metagenomes. Prochlorococcus and Thiomicrospira were highly abundant in waters, while Myxococcus, Anaeromyxobacter, Haliangium, and Gloeobacter were dominant in the soil and lake sediment metagenomes. Among CIPs, genes necessary for DNA replication, translation initiation, and transcription termination were highly abundant in all metagenomes. However, genes for fatty acid desaturase (FAD) and trehalose synthase (TS) were common in the soil and lake sediment metagenomes. Interestingly, the Lake Untersee water and sediment metagenome samples contained histone-like nucleoid structuring protein (H-NS) and all genes for CIPs. As for the CASPs, high abundances of a wide range of genes for cryo- and osmo-protectants (glutamate, glycine, choline, and betaine) were identified in all metagenomes. However, genes for exopolysaccharide biosynthesis were dominant in Lake Untersee water, sediment, and other soil metagenomes. The results from this study indicate that although diverse microbial communities are present in various metagenomes, they share common cold-responsive stress genes necessary for their survival and sustenance in the extreme Antarctic conditions.

In November 2008 an automated meteorological station was established at Lake Untersee in East Antarctica, producing a 5-yr data record of meteorological conditions at the lake. This dataset includes five austral summer seasons composed of December, January, and February (DJF). The average solar flux at Lake Untersee for the four years with complete solar flux data is 99.2 ± 0.6 W m−2. The mean annual temperature at Lake Untersee was determined to be −10.6° ± 0.6°C. The annual degree-days above freezing for the five years were 9.7, 37.7, 22.4, 7.0, and 48.8, respectively, with summer (DJF) accounting for virtually all of this. For these five summers the average DJF temperatures were −3.5°, −1.9°, −2.2°, −2.6°, and −2.5°C. The maximum (minimum) temperatures were +5.3°, +7.6°, +5.7°, +4.4°, and +9.0°C (−13.8°, −12.8°, −12.9°, −13.5°, and −12.1°C). The average of the wind speed recorded was 5.4 m s−1, the maximum was 35.7 m s−1, and the average daily maximum was 15 m s−1. The wind speed was higher in the winter, averaging 6.4 m s−1. Summer winds averaged 4.7 m s−1. The dominant wind direction for strong winds is from the south for all seasons, with a secondary source of strong winds in the summer from the east-northeast. Relative humidity averages 37%; however, high values will occur with an average period of ~10 days, providing a strong indicator of the quasi-periodic passage of storms across the site. Low summer temperatures and high wind speeds create conditions at the surface of the lake ice resulting in sublimation rather than melting as the main mass-loss process.

Perennially ice-covered, meromictic lakes in the McMurdo Dry Valleys, Antarctica, are useful models to study the relationship between cyanobacterial and environmental variables. They have rich benthic cyanobacterial mat accumulations and stable stratification of physical and chemical conditions. Here, we evaluated cyanobacteria from benthic mats from multiple depths in three geographically separated ice-covered lakes, Lakes Vanda, Hoare and Joyce, using 16S rRNA gene clone libraries. We identified 19 ribotypes, mostly Oscillatoriales and several Chroococcales, as well as potentially novel cyanobacterial ribotypes. The majority of ribotype diversity was shared between lakes, and only a weak relationship between ribotype community structure and environmental variables was evident. Multivariate analysis of all lake–depth combinations implied that photosynthetically active radiation, dissolved reactive phosphorus and conductivity were potentially important for shaping benthic communities in McMurdo Dry Valley lakes. Cyanobacterial-specific pigment signature analysis by high-performance liquid chromatography showed that the cyanobacterial communities responded to light conditions similarly, irrespective of community composition. The results imply a capability within a suite of cyanobacteria to colonise, adapt and grow across broad environmental ranges and geographic space, and such adaptability may provide a high degree of community resistance and resilience to future climate-driven environmental change in Antarctic terrestrial aquatic ecosystems.

This chapter focuses on the rationale and implementation of a suite of software in the data management, processing, and statistical analysis used towards establishing reliable microbial community compositions, predictive functions, and other useful downstream bioinformatics analyses, using NGS data generated by the Illumina Miseq targeted metagenomics approach. This chapter also emphasizes the importance of using multiple taxonomic databases and software to obtain a comprehensive microbial community composition and associated functional profiles. We have used previously unpublished 16S rRNA amplicon-based next-generation sequencing (NGS) dataset of lithobiont communities (rock-dwelling microorganisms) of the Untersee Oasis, Antarctica, as an example of microbial communities in an atypical environment. The bioinformatics workflow includes “Module A”, which describes obtaining the NGS files from purified lithobiont metacommunity DNA; “Module B” elaborates the steps involved in processing and assessing the sequence quality, and subsequent sequence alignment against publicly available microbial taxonomic databases (i.e. Greengenes and SILVA), to designate identities to a list of operational taxonomic units representing the microbial communities. Lastly, “Module C” presents the microbial community compositions at the highest taxonomic levels, diversity within and between samples, predictive functional attributes, and Co-Occurrence Network (CoNet) analysis of microbial communities. The results generated from this example demonstrated that the lithobiont communities from Lake Untersee had high abundances of Aliihoeflea and Sphingomonas, whereas Acinetobacter and Paenibacillus were dominant in Lake Obersee. Predictive functional profiles of both communities showed high abundances of genes designated for membrane transport and bacterial secretion systems, amino acid metabolism, and UV tolerance.

In this study, we report the distribution of microbial taxa and their predicted metabolic functions observed in the top (U1), middle (U2) and inner (U3) decadal growth laminae of a unique large conical microbial mat from a perennially ice-covered freshwater Lake Untersee of East Antarctica, using NextGen sequencing of the 16S rRNA gene and bioinformatics tools. The results showed that the U1 lamina was dominated by cyanobacteria, specifically Phormidium sp., Leptolyngbya sp. and Pseudanabaena sp. The U2 and U3 laminae had high abundances of Actinobacteria, Verrucomicrobia, Proteobacteria, and Bacteroidetes. Closely related taxa within each abundant bacterial taxon found in each lamina were further differentiated at the highest taxonomic resolution using the oligotyping method. PICRUSt analysis, which determines predicted KEGG functional categories from the gene contents and abundances among microbial communities, revealed a high number of sequences belonging to carbon fixation, energy metabolism, cyanophycin, chlorophyll and photosynthesis proteins in the U1 lamina. The functional predictions of the microbial communities in U2 and U3 represented signal transduction, membrane transport, zinc transport and amino acid-, carbohydrate-, and arsenic- metabolisms. The Nearest Sequenced Taxon Index (NSTI) values processed through PICRUSt were 0.10, 0.13 and 0.11 for U1, U2 and U3 laminae, respectively. These values indicated a close correspondence with the reference microbial genome database, implying high confidence in the predicted metabolic functions of the microbial communities in each lamina. Distribution of microbial taxa observed in each lamina and their predicted metabolic functions reveled in this study contributes to an insight into the Lake Untersee mat microbial ecosystem and lays the foundation for future studies for our understanding of detailed mechanisms that led to the formation of these unique mat structures and their evolutionary significance.

This paper describes the complete genome sequences and methylome analysis of six psychrotrophic strains isolated from perennially ice-covered Lake Untersee in Antarctica.

The Schimacher Oasis, an ice-free plateau in East Antarctic Dronning Maud Land, consists of over 120 freshwater lakes. These lakes are connected largely through four major surface channels. The bacterial diversity in these lake ecosystems remains largely unexplored. In this study, we compared the bacterial diversity in five freshwater lakes (L42, L46, L47, L50, and L51) interconnected by two surface channels using bacterial tag-encoded FLX amplicon pyrosequencing (bTEFAP) method. We further compared the resultant bacterial composition from these five lakes with another freshwater lake in the Schirmacher Oasis, Lake Tawani(P), which is not connected through the same surface channels. Using bTEFAP, we differentiated nine different phyla with the phyla Proteobacteria (especially the class Alphaproteobacteria) and Bacteroidetes (the class Sphingobacteria) dominating in lakes interconnected by surface channel 1, while the phyla Chloroflexi and Firmicutes were highly abundant in lakes interconnected by surface channel 2. The operational taxonomic unit (OTU) network and Principle Coordinate Analysis (PCoA) plot based on unweighted UNIFRAC determined that the bacterial assemblages found in these five lakes are different than the bacterial composition residing in Lake Tawani(P). The distribution and the diversity of the bacterial communities in Schirmacher Oasis freshwater lakes that are connected through surface channels may provide an insight into the role of the extreme physico-chemical parameters that help shape microbially driven functional ecosystems in other oases on this icy continent.

The heterotrophic mesophilic microbial component was studied in microbial communities of the samples of frozen regolith collected from the glacier near Lake Untersee collected in 2011 during the joint Russian-American expedition to central Dronning Maud Land (Eastern Antarctica). Cultural techniques revealed high bacterial numbers in the samples. For enumeration of viable cells, the most probable numbers (MPN) method proved more efficient than plating on agar media. Fluorescent in situ hybridization with the relevant oligonucleotide probes revealed members of the groups Eubacteria (Actinobacteria, Firmicutes) and Archaea. The application of the methods of cell resuscitation, such as the use of diluted media and prevention of oxidative stress, did not result in a significant increase in the numbers of viable cells retrieved from subglacial sediment samples. Our previous investigations demonstrated the necessity for special procedures for efficient reactivation of the cells from microbial communities of replace with buried soil and permafrost samples collected in the Arctic zone. The differing responses to the special resuscitation procedures may reflect the differences in the physiological and morphological state of bacterial cells in microbial communities subject to continuous or periodic low temperatures and dehydration.

Contrary to earlier beliefs, crustaceans are present in ice-covered lakes of Antarctica. Interpretation of the significance of this has been hampered by the absence of robust identification of taxa present. We examine cyclopoid copepods from three widely separated lakes. All belong to the michaelseni group of the genus Diacyclops, which is widespread across Continental Antarctica, but do not fit into any existing species. Two new species were identified from eastern Antarctica, D. walkeri from Pineapple Lake (Vestfold Hills) and D. kaupi from Transkriptsii Gulf (Bunger Hills). Most significant was a dense population of a new epibenthic species (D. joycei) associated with microbial mats in Lake Joyce, one of the smaller McMurdo Dry Valleys lakes. This represents the first record of adult cyclopoid copepods from the ice-covered lakes of the Transantarctic Mountains. Continental Antarctica is the centre of diversity for this group of crustaceans and we argue that this is better explained by persistence through past glacial advances rather than by recent post-glacial colonization. The existence of a species endemic to Lake Joyce but apparently absent from other Dry Valleys lakes is discussed in relation to our understanding of the history of the McMurdo Dry Valleys lakes and their faunas.

The high elevation valleys of the McMurdo Dry Valleys of Antarctica are the only locations on Earth known to contain dry permafrost. The Dry Valleys are a hyper-arid polar desert environment and above 1500 m elevation, air temperatures do not exceed 0 °C and thus, similarly to Mars, liquid water is largely absent and instead the hydrologic cycle is dominated by frozen ice and vapor phase processes such as sublimation. These conditions make the high elevation Dry Valleys a key Mars analog location where periglacial processes and geomorphic features, and their use as a diagnostic for subsurface ice, can be studied in situ. Two valleys in the upper Dry Valleys show a diversity of subsurface ice; University Valley is dominated by dry permafrost overlying ice-cemented to ice-bonded ground and nearby middle Beacon Valley is dominated by massive ground ice. In both cases the ice is 10–60 cm below the surface. Here we compare the surface features in these two valleys to assess any correlation with the nature of the subsurface ice and compare these features to similar features seen at the Phoenix landing site on Mars. We conclude that while surface features may be indicative of ground ice, no specific correlations are possible and more direct methods are required to determine the nature of subsurface ice on Mars.

ISSN 0026!2617, Microbiology, 2013, Vol. 82, No. 3, pp. 383–386. © Pleiades Publishing, Ltd., 2013.Original Russian Text © S.N. Filippova, N.A. Surgucheva, E.E. Kulikov, V.V. Sorokin, V.N. Akimov, A.K. Bej, C. McKay, D. Andersen, V.F. Galchenko, 2013, published in Mikro!biologiya, 2013, Vol. 82, No. 3, pp. 366–369.

The search for evidence of life on Mars is the primary motivation for the exploration of that planet. The results from previous missions, and the Phoenix mission in particular, indicate that the ice-cemented ground in the north polar plains is likely to be the most recently habitable place that is currently known on Mars. The near-surface ice likely provided adequate water activity during periods of high obliquity, ≈ 5 Myr ago. Carbon dioxide and nitrogen are present in the atmosphere, and nitrates may be present in the soil. Perchlorate in the soil together with iron in basaltic rock provides a possible energy source for life. Furthermore, the presence of organics must once again be considered, as the results of the Viking GCMS are now suspect given the discovery of the thermally reactive perchlorate. Ground ice may provide a way to preserve organic molecules for extended periods of time, especially organic biomarkers. The Mars Icebreaker Life mission focuses on the following science goals: (1) Search for specific biomolecules that would be conclusive evidence of life. (2) Perform a general search for organic molecules in the ground ice. (3) Determine the processes of ground ice formation and the role of liquid water. (4) Understand the mechanical properties of the martian polar ice-cemented soil. (5) Assess the recent habitability of the environment with respect to required elements to support life, energy sources, and possible toxic elements. (6) Compare the elemental composition of the northern plains with midlatitude sites. The Icebreaker Life payload has been designed around the Phoenix spacecraft and is targeted to a site near the Phoenix landing site. However, the Icebreaker payload could be supported on other Mars landing systems. Preliminary studies of the SpaceX Dragon lander show that it could support the Icebreaker payload for a landing either at the Phoenix site or at midlatitudes. Duplicate samples could be cached as a target for possible return by a Mars Sample Return mission. If the samples were shown to contain organic biomarkers, interest in returning them to Earth would be high.

In this paper, we describe the UV and cold tolerance of a purple violet pigment (PVP)-producing Antarctic bacterium, Janthinobacterium sp. Ant5-2 (PVP(+)) and compared its physiological adaptations with a pigmentless mutant strain (PVP(-)). A spontaneous deletion of vioA that codes for tryptophan monooxygenase, the first gene involved in the biosynthesis of PVP was found in PVP(-) strain. The PVP(-) culture exhibited significantly reduced survival during exponential and stationary growth phase following exposure to UVB (320 nm) and UVC (254 nm) (dose range: 0-300 J/m²) when compared to wild-type (PVP(+)) cultures. In addition, upon biochemical inhibition of pigment synthesis by 2(5H)-furanone, wild-type PVP(+) cultures exhibited approximately 50-fold growth reduction at a higher dose (300 J/m²) of UV. Increased resistance to UV was observed upon inducing starvation state in both PVP(+) and PVP(-) cultures. There was 80% (SD = ±8) reduction in extrapolymeric substance (EPS) production in the PVP(-) cultures along with a compromised survival to freeze-thaw cycles when compared to the PVP(+) cultures. Perhaps synthesis of PVP and EPS are among the key adaptive features that define the survival of this bacterium in Antarctic extreme conditions, especially during austral summer months.

We report on 475 measurements of depth to ice-cemented ground in four high-elevation valleys of the Quartermain Mountains, McMurdo Dry Valleys, Antarctica. These valleys have pervasive ice-cemented ground, and the depth to ice-cemented ground and the ice composition may be indicators of climate change. In University Valley, the measured depth to ice-cemented ground ranges from 0–98 cm. There is an overall trend of increasing depth to ice-cemented ground with distance from a small glacier at the head of the valley, with a slope of 32 cm depth per kilometre along the valley floor. For Farnell Valley, the depth to ice-cemented ground is roughly constant (c. 30 cm) in the upper and central parts of the valley, but increases sharply as the valley descends into Beacon Valley. The two valleys north of University Valley also have extensive ice-cemented ground, with depths of 20–40 cm, but exhibit no clear patterns of ice depth with location. For all valleys there is a tendency for the variability in depth to ice-cemented ground at a site to increase with increasing depth to ice. Snow recurrence, solar insolation, and surface albedo may all be factors that cause site to site variations in these valleys.

The next step in the exploration of Mars should include a strong and comprehensive life detection component. We propose a mission called BOLD: Biological Oxidant and Life Detection mission. The scientific objectives of the BOLD mission are to characterize habitability of the martian surface and to search for evidence of extinct or extant life. In contrast to the Viking mission, which was designed to detect heterotrophic life on Mars, the BOLD mission incorporates a more comprehensive search for autotrophic microorganisms, as well as detecting a variety of biomarkers and understanding their environment. Six miniature landers are envisioned for BOLD that utilize either an orbital (e.g. Viking) or direct entry (e.g., MER, Phoenix) mission architecture. The number of landers will provide mission redundancy, and each will incorporate a Mars Soil Analyzer, a Multispectral Microscopic Imager, a Nanopore-ARROW that detects biopolymers with single molecule resolution, an Atmospheric Structure and Surface Environment Instrument, a Fluorescent Stain experiment, and a Chirality experiment. A terrain navigation system, coupled with robust propulsion, permits a landing accuracy on the order of meters if required to meet the science objectives. The probes will use existing orbiters for communication relay if the orbiter architecture proves too ambitious.

Buried snowpack deposits are found within the McMurdo Dry Valleys of Antarctica, which offers the opportunity to study these layered structures of sand and ice within a polar desert environment. Four discrete buried snowpacks are studied within Pearse Valley, Antarctica, through in situ observations, sample analyses, O-H isotope measurements and numerical modelling of snowpack stability and evolution. The buried snowpack deposits evolve throughout the year and undergo deposition, melt, refreeze, and sublimation. We demonstrate how the deposition and subsequent burial of snow can preserve the snowpacks in the Dry Valleys. The modelled lifetimes of the buried snowpacks are dependent upon subsurface stratigraphy but are typically less than one year if the lag thickness is less than c. 7 cm and snow thickness is less than c. 10 cm, indicating that some of the Antarctic buried snowpacks form annually. Buried snowpacks in the Antarctic polar desert may serve as analogues for similar deposits on Mars and may be applicable to observations of the north polar erg, buried ice at the Mars Phoenix landing site, and observations of buried ice throughout the martian Arctic. Numerical modelling suggests that seasonal snows and subsequent burial are not required to preserve the snow and ice on Mars.

Many Antarctic lakes provide habitat for extensive microbial mats that respond on various timescales to environmental change. Lake Joyce contains calcifying microbialites and provides a natural laboratory to constrain how environmental changes influence microbialite development. In Lake Joyce, depth-specific distributions of calcitic microbialites, organic carbon, photosynthetic pigments and photosynthetic potential cannot be explained by current growth conditions, but are a legacy of a 7-m lake level rise between 1973 and 2009. In the well-illuminated margins of the lake, photosynthetically active benthic communities colonised surfaces submerged for just a few years. However, observed increases in accumulated organic material with depth from 5 to 20 m (2-40 mg ash-free dry weight cm(-2)) and the presence of decimetre-scale calcite microbialites at 20-22 m depth, apparently related to in situ photosynthetic growth, are inconsistent with the current distributions of irradiance, photosynthetic pigments and mat photosynthetic potential (as revealed by pulse-amplitude-modulated fluorometry). The microbialites appeared photosynthetically active in 1986 and 1997, but were outside the depth zone where significant phototrophic growth was possible and were weakly photosynthetically competent in 2009. These complex microbial structures have persisted after growth has ceased, demonstrating how fluctuating environmental conditions and the hysteresis between environmental change, biological response and microbialite development can be important factors to consider when interpreting modern, and by inference ancient, microbially mediated structures.

Background In this study, we have investigated the chemotherapeutic potential of a purple violet pigment (PVP), which was isolated from a previously undescribed Antarctic Janthinobacterium sp. (Ant5-2), against murine UV-induced 2237 fibrosarcoma and B16F10 melanoma cells. Methods The 2237, B16F10, C50, and NIH3T3 cells were treated with PVP at different doses and for different times, and their proliferation and viability were detected by 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide (MTT) assay. Cell cycle arrest induced by PVP in 2237 fibrosarcoma cells was assessed by flow cytometry and expression analysis of cell cycle regulatory proteins were done by Western blot. Apoptosis induced by PVP in 2237 cells was observed by annexin-V/propidium iodide double staining flow cytometry assay and fluorescence microscopy. To further determine the molecular mechanism of apoptosis induced by PVP, the changes in expression of Bcl-2, Bax and cytochrome c were detected by Western blot. The loss of mitochondrial membrane potential in PVP treated 2237 cells was assessed by staining with JC-1 dye following flow cytometry. Caspase-3, Caspase-9 and PARP cleavage were analyzed by Western blot and Caspase-3 and -9 activities were measured by colorimetric assays. Results In vitro treatment of murine 2237 cells with the PVP resulted in decreased cell viability (13–79%) in a time (24–72 h) and dose (0.1–1 μm)-dependent manner. The PVP-induced growth inhibition in 2237 cells was associated with both G0/G1 and G2/M phase arrest accompanied with decrease in the expression of cyclin dependent kinases (Cdks) and simultaneous increase in the expression of cyclin dependent kinase inhibitors (Cdki) – Cip1/p21 and Kip1/p27. Further, we observed a significant increase in the apoptosis of the 2237 fibrosarcoma cells which was associated with an increased expression of pro-apoptotic protein Bax, decreased expression of anti-apoptotic proteins Bcl-2, disruption of mitochondrial membrane potential, cytochrome c release, activation of caspase-3, caspase-9 and poly-ADP-ribose-polymerase (PARP) cleavage. Conclusions We describe the anti-cancer mechanism of the PVP for the first time from an Antarctic bacterium and suggest that the PVP could be used as a potent chemotherapeutic agent against nonmelanoma skin cancers.

Lake Untersee is one of the largest (11.4 km2) and deepest (>160 m) freshwater lakes in East Antarctica. Located at 71°S the lake has a perennial ice cover, a water column that, with the exception of a small anoxic basin in the southwest of the lake, is well mixed, supersaturated with dissolved oxygen, alkaline (pH 10.4) and exceedingly clear. The floor of the lake is covered with photosynthetic microbial mats to depths of at least 100 m. These mats are primarily composed of filamentous cyanophytes and form two distinct macroscopic structures, one of which – cm-scale cuspate pinnacles dominated by Leptolyngbya spp. – is common in Antarctica, but the second – laminated, conical stromatolites that rise up to 0.5 m above the lake floor, dominated by Phormidium spp. – has not previously been reported in any modern environment. The laminae that form the conical stromatolites are 0.2–0.8 mm in thickness consisting of fine clays and organic material; carbon dating implies that laminations may occur on near decadal timescales. The uniformly steep sides (59.6 ± 2.5°) and the regular laminar structure of the cones suggest that they may provide a modern analog for growth of some of the oldest well-described Archean stromatolites. Mechanisms underlying the formation of these stromatolites are as yet unclear, but their growth is distinct from that of the cuspate pinnacles. The sympatric occurrence of pinnacles and cones related to microbial communities with distinct cyanobacterial compositions suggest that specific microbial behaviors underpin the morphological differences in the structures.

A cold shock domain (CSD)-containing protein, CspD, of molecular mass ~7.28 kDa in a psychrotolerant Antarctic Janthinobacterium sp. Ant5-2 (ATCC BAA-2154) exhibited constitutive expression at 37, 22, 15, 4 and -1°C. The cspD gene encoding the CspD protein of Ant5-2 was cloned, sequenced and analyzed. The deduced protein sequence was highly similar to the conserved domains of the cold shock proteins (Csps) from bacteria belonging to the class Betaproteobacteria. Its expression was both time- and growth phase-dependent and increased when exposed to 37°C and UV radiation (UVC, dose: 1.8 and 2.8 mJ cm(-2)). The results from the electrophoretic mobility shift and subcellular localization study confirmed its single-stranded DNA-binding property. In silico analysis of the deduced tertiary structure of CspD from Ant5-2 showed a highly stable domain-swapped dimer, forming two similar monomeric Csp folds. This study established an overall framework of the structure, function and phylogenetic analysis of CspD from an Antarctic Janthinobacterium sp. Ant5-2, which may facilitate and stimulate the study of CSD fold proteins in the class Betaproteobacteria.

To date, studies of the stability of subsurface ice in the McMurdo Dry Valleys of Antarctica have been mainly based on climate-based vapor diffusion models. In University Valley (1800 m), a small glacier is found at the base of the head of the valley, and adjacent to the glacier, a buried body of massive ice was uncovered beneath 20–40 cm of loose cryotic sediments and sandstone boulders. This study assesses the origin and stability of the buried body of massive ice by measuring the geochemistry and stable O–H isotope composition of the ice and applies a sublimation and molecular diffusion model that accounts for the observed trends. The results indicate that the buried massive ice body represents an extension of the adjacent glacier that was buried by a rock avalanche during a cold climate period. The contrasting δ 18 O profiles and regression slope values between the uppermost 6 cm of the buried massive ice (upward convex δ 18 O profile and S D-18O = 5.1) and that below it (progressive increase in δ 18 O and S D-18O = 6.4) suggest independent post-depositional processes affected the isotope composition of the ice. The upward convex δ 18 O profile in the uppermost 6 cm is consistent with the ice undergoing sublimation. Using a sublimation and molecular diffusion model, and assuming that diffusion occurred through solid ice, the sublimation rate needed to fit the measured δ 18 O profile is 0.2 ⋅ 10 − 3 mm yr − 1 , a value that is more similar to net ice removal rates derived from 3 He data from cobbles in Beacon Valley till (7.0 ⋅ 10 − 3 mm yr − 1 ) than sublimation rates computed based on current climate (0.1–0.2 mm yr −1 ). We suggest that the climate-based sublimation rates are offset due to potential ice recharge mechanisms or to missing parameters, particularly the nature and thermo-physical properties of the overlying sediments (i.e., temperature, humidity, pore structure and ice content, grain size).

The influence of microbial activity on carbonate precipitation was investigated within micro-stromatolitic nodules associated with modern, freshwater microbialites located in Pavilion Lake, B.C. Observed carbonate δ13C values enriched by up to + 3.6‰ as compared to predicted abiotic carbonate δ13C values from measured dissolved inorganic carbon (mean − 1.2‰, n = 13) were consistent with microbial photosynthetic influence on in situ precipitation within the nodule microenvironment. Estimated carbonate precipitation temperatures within the nodules based on δ18O were consistent with recorded summertime temperatures, indicative of precipitation during the period of highest levels of photosynthetic activity. Low δ13C values of organic matter within the nodules (− 30.6 to − 21.1‰) and an average inorganic to organic carbon Δδ13C value of 26.8‰ reflected the preferential uptake of 12C during non-CO2 limited photosynthesis, supporting the generation of 13C-enriched DIC. Microelectrode profiles through the nodules showed oxygen supersaturation of up to ∼ 275%, elevated pH compared to ambient water and a lack of any observable dissolved sulphide, Mn or Fe further indicated that photosynthetic activity was the predominant metabolic process within the nodule during light exposure. Microbial phospholipid fatty acid profiles of the nodule communities were indicative of bacteria rather than eukaryotes and PLFA δ13C values were depleted relative to the bulk cell by 2.6–6.6‰, consistent with a predominance of photosynthetic microbes. Scanning electron microscopy images of the relationship between carbonate minerals and filaments indicated that carbonate precipitation had occurred in situ due to microbial influences on the geochemistry within the nodule microenvironment rather than due to cell surface effects or trapping and binding. The observation of photosynthetically induced 13C-enrichment of in situ precipitated carbonate within the nodule microenvironment is thus a biosignature of the activity of these surface communities and is consistent with the hypothesized role of biology in the formation of microbialites.

We report the first microbiological characterization of a terrestrial methane seep in a cryo-environment in the form of an Arctic hypersaline (∼24% salinity), subzero (-5 °C), perennial spring, arising through thick permafrost in an area with an average annual air temperature of -15 °C. Bacterial and archaeal 16S rRNA gene clone libraries indicated a relatively low diversity of phylotypes within the spring sediment (Shannon index values of 1.65 and 1.39, respectively). Bacterial phylotypes were related to microorganisms such as Loktanella, Gillisia, Halomonas and Marinobacter spp. previously recovered from cold, saline habitats. A proportion of the bacterial phylotypes were cultured, including Marinobacter and Halomonas, with all isolates capable of growth at the in situ temperature (-5 °C). Archaeal phylotypes were related to signatures from hypersaline deep-sea methane-seep sediments and were dominated by the anaerobic methane group 1a (ANME-1a) clade of anaerobic methane oxidizing archaea. CARD-FISH analyses indicated that cells within the spring sediment consisted of ∼84.0% bacterial and 3.8% archaeal cells with ANME-1 cells accounting for most of the archaeal cells. The major gas discharging from the spring was methane (∼50%) with the low CH(4)/C(2+) ratio and hydrogen and carbon isotope signatures consistent with a thermogenic origin of the methane. Overall, this hypersaline, subzero environment supports a viable microbial community capable of activity at in situ temperature and where methane may behave as an energy and carbon source for sustaining anaerobic oxidation of methane-based microbial metabolism. This site also provides a model of how a methane seep can form in a cryo-environment as well as a mechanism for the hypothesized Martian methane plumes.