An extensive phase space for the potential martian biosphere

We present a comprehensive model of martian pressure-temperature (P-T) phase space and compare it with that of Earth. Martian P-T conditions compatible with liquid water extend to a depth of ～310?km. We use our phase space model of Mars and of terrestrial life to estimate the depths and extent of the water on Mars that is habitable for terrestrial life. We find an extensive overlap between inhabited terrestrial phase space and martian phase space. The lower martian surface temperatures and shallower martian geotherm suggest that, if there is a hot deep biosphere on Mars, it could extend 7 times deeper than the ～5?km depth of the hot deep terrestrial biosphere in the crust inhabited by hyperthermophilic chemolithotrophs. This corresponds to ～3.2% of the volume of present-day Mars being potentially habitable for terrestrial-like life.     

Subfreezing activity of microorganisms and the potential habitability of Mars' polar regions

The availability of water-ice at the surface in the Mars polar cap and within the top meter of the high-latitude regolith raises the question of whether liquid water can exist there under some circumstances and possibly support the existence of biota. We examine the minimum temperatures at which liquid water can exist at ice grain-dust grain and ice grain-ice grain contacts, the minimum subfreezing temperatures at which terrestrial organisms can grow or multiply, and the maximum temperatures that can occur in martian high-latitude and polar regions, to see if there is overlap. Liquid water can exist at grain contacts above about -20 degrees C. Measurements of growth in organisms isolated from Siberian permafrost indicate growth at -10 degrees C and metabolism at -20 degrees C. Mars polar and high-latitude temperatures rise above -20 degrees C at obliquities greater than ~40 degrees, and under some conditions rise above 0 degrees C. Thus, the environment in the Mars polar regions has overlapped habitable conditions within relatively recent epochs, and Mars appears to be on the edge of being habitable at present. The easy accessibility of the polar surface layer relative to the deep subsurface make these viable locations to search for evidence of life.     

Adsorption water-related potential chemical and biological processes in the upper martian surface

Mars Odyssey has given strong evidence for the existence of water in the upper martian surface at equatorial latitudes. The water content, which corresponds to the hydrogen in the soil, can regionally reach values up to about 15%. This water is mainly in the form of structurally and partially irreversibly bound "crystal" water, and of reversibly bound and partially unfrozen adsorption water. This adsorption water, which has "liquid-like" properties as a two dimensional fluid or film, can trigger-in the presence of ultraviolet light and in concentrations similar to what has been measured on Mars-photocatalytic processes that are important for martian surface chemistry. The consequences of the diurnally variable presence of adsorption water on the chemistry and hypothetical biological processes at and in the upper martian surface at equatorial and mid-latitudes are discussed in terms of water-related environmental aspects for chemical and hypothetical life processes on Mars.     

Special regions in Mars exploration: Problems and potential

“Special regions” on Mars are areas designated in the COSPAR planetary protection policy as areas that may support Earth microbes inadvertently introduced to Mars, or that may have a high probability of supporting indigenous martian life. Since absolutely nothing is known about martian life, the operational definition of a special region is a place that may allow the formation and maintenance of liquid water, on or under the surface of Mars. This paper will review the special-regions concept, the implications of recent recommendations on avoiding them, and the work of the Mars science community in providing an operational definition of those areas on Mars that are “non-special.”     

Survival of methanogens during desiccation: implications for life on Mars

The relatively recent discoveries that liquid water likely existed on the surface of past Mars and that methane currently exists in the martian atmosphere have fueled the possibility of extant or extinct life on Mars. One possible explanation for the existence of the methane would be the presence of methanogens in the subsurface. Methanogens are microorganisms in the domain Archaea that can metabolize molecular hydrogen as an energy source and carbon dioxide as a carbon source and produce methane. One factor of importance is the arid nature of Mars, at least at the surface. If one is to assume that life exists below the surface, then based on the only example of life that we know, liquid water must be present. Realistically, however, that liquid water may be seasonal just as it is at some locations on our home planet. Here we report on research designed to determine how long certain species of methanogens can survive desiccation on a Mars soil simulant, JSC Mars-1. Methanogenic cells were grown on JSC Mars-1, transferred to a desiccator within a Coy anaerobic environmental chamber, and maintained there for varying time periods. Following removal from the desiccator and rehydration, gas chromatographic measurements of methane indicated survival for varying time periods. Methanosarcina barkeri survived desiccation for 10 days, while Methanobacterium formicicum and Methanothermobacter wolfeii were able to survive for 25 days.     

The physics,biology, and environmental ethics of making mars habitable

The considerable evidence that Mars once had a wetter, more clement, environment motivates the search for past or present life on that planet. This evidence also suggests the possibility of restoring habitable conditions on Mars. While the total amounts of the key molecules--carbon dioxide, water, and nitrogen--needed for creating a biosphere on Mars are unknown, estimates suggest that there may be enough in the subsurface. Super greenhouse gases, in particular, perfluorocarbons, are currently the most effective and practical way to warm Mars and thicken its atmosphere so that liquid water is stable on the surface. This process could take approximately 100 years. If enough carbon dioxide is frozen in the South Polar Cap and absorbed in the regolith, the resulting thick and warm carbon dioxide atmosphere could support many types of microorganisms, plants, and invertebrates. If a planet-wide martian biosphere converted carbon dioxide into oxygen with an average efficiency equal to that for Earth's biosphere, it would take > 100,000 years to create Earth-like oxygen levels. Ethical issues associated with bringing life to Mars center on the possibility of indigenous martian life and the relative value of a planet with or without a global biosphere.     

The PROCESS experiment: amino and carboxylic acids under Mars-like surface UV radiation conditions in low-earth orbit

The search for organic molecules at the surface of Mars is a top priority of the next Mars exploration space missions: Mars Science Laboratory (NASA) and ExoMars (ESA). The detection of organic matter could provide information about the presence of a prebiotic chemistry or even biological activity on this planet. Therefore, a key step in interpretation of future data collected by these missions is to understand the preservation of organic matter in the martian environment. Several laboratory experiments have been devoted to quantifying and qualifying the evolution of organic molecules under simulated environmental conditions of Mars. However, these laboratory simulations are limited, and one major constraint is the reproduction of the UV spectrum that reaches the surface of Mars. As part of the PROCESS experiment of the European EXPOSE-E mission on board the International Space Station, a study was performed on the photodegradation of organics under filtered extraterrestrial solar electromagnetic radiation that mimics Mars-like surface UV radiation conditions. Glycine, serine, phthalic acid, phthalic acid in the presence of a mineral phase, and mellitic acid were exposed to these conditions for 1.5 years, and their evolution was determined by Fourier transform infrared spectroscopy after their retrieval. The results were compared with data from laboratory experiments. A 1.5-year exposure to Mars-like surface UV radiation conditions in space resulted in complete degradation of the organic compounds. Half-lives between 50 and 150?h for martian surface conditions were calculated from both laboratory and low-Earth orbit experiments. The results highlight that none of those organics are stable under low-Earth orbit solar UV radiation conditions.     

Survival of Bacillus subtilis endospores on ultraviolet-irradiated rover wheels and Mars regolith under simulated Martian conditions

Endospores of Bacillus subtilis HA101 were applied to a simulated Mars Exploration Rover (MER) wheel and exposed to Mars-normal UV irradiation for 1, 3, or 6 h. The experiment was designed to simulate a contaminated rover wheel sitting on its landing platform before rolling off onto the martian terrain, as was encountered during the Spirit and Opportunity missions. When exposed to 1 h of Mars UV, a reduction of 81% of viable endospores was observed compared to the non-UV irradiated controls. When exposed for 3 or 6 h, reductions of 94.6% and 96.6%, respectively, were observed compared to controls. In a second experiment, the contaminated rover wheel was rolled over a bed of heat-sterilized Mars analog soil; then the analog soil was exposed to full martian conditions of UV irradiation, low pressure (6.9 mbar), low temperature (-10°C), and an anaerobic CO(2) martian atmosphere for 24 h to determine whether endospores of B. subtilis on the contaminated rover wheel could be transferred to the surface of the analog soil and survive martian conditions. The experiment simulated conditions in which a rover wheel might come into contact with martian regolith immediately after landing, such as is designed for the upcoming Mars Science Laboratory (MSL) rover. The contaminated rover wheel transferred viable endospores of B. subtilis to the Mars analog soil, as demonstrated by 31.7% of samples showing positive growth. However, when contaminated soil samples were exposed to full martian conditions for 24 h, only 16.7% of samples exhibited positive growth-a 50% reduction in the number of soil samples positive for the transferred viable endospores.     

Bacterial growth at the high concentrations of magnesium sulfate found in martian soils

The martian surface environment exhibits extremes of salinity, temperature, desiccation, and radiation that would make it difficult for terrestrial microbes to survive. Recent evidence suggests that martian soils contain high concentrations of MgSO? minerals. Through warming of the soils, meltwater derived from subterranean ice-rich regolith may exist for an extended period of time and thus allow the propagation of terrestrial microbes and create significant bioburden at the near surface of Mars. The current report demonstrates that halotolerant bacteria from the Great Salt Plains (GSP) of Oklahoma are capable of growing at high concentrations of MgSO? in the form of 2 M solutions of epsomite. The epsotolerance of isolates in the GSP bacterial collection was determined, with 35% growing at 2 M MgSO?. There was a complex physiological response to mixtures of MgSO? and NaCl coupled with other environmental stressors. Growth also was measured at 1 M concentrations of other magnesium and sulfate salts. The complex responses may be partially explained by the pattern of chaotropicity observed for high-salt solutions as measured by agar gelation temperature. Select isolates could grow at the high salt concentrations and low temperatures found on Mars. Survival during repetitive freeze-thaw or drying-rewetting cycles was used as other measures of potential success on the martian surface. Our results indicate that terrestrial microbes might survive under the high-salt, low-temperature, anaerobic conditions on Mars and present significant potential for forward contamination. Stringent planetary protection requirements are needed for future life-detection missions to Mars.     

Glycine identification in natural jarosites using laser desorption Fourier transform mass spectrometry: implications for the search for life on Mars

The jarosite group minerals have received increasing attention since the discovery of jarosite on the martian surface by the Mars Exploration Rover Opportunity. Given that jarosite can incorporate foreign ions within its structure, we have investigated the use of jarosite as an indicator of aqueous and biological processes on Earth and Mars. The use of laser desorption Fourier transform mass spectrometry has revealed the presence of organic matter in several jarosite samples from various locations worldwide. One of the ions from the natural jarosites has been attributed to glycine because it was systematically observed in combinations of glycine with synthetic ammonium and potassium jarosites, Na(2)SO(4) and K(2)SO(4). The ability to observe these organic signatures in jarosite samples with an in situ instrumental technique, such as the one employed in this study, furthers the goals of planetary geologists to determine whether signs of life (e.g., the presence of biomolecules or biomolecule precursors) can be detected in the rock record of terrestrial and extraterrestrial samples.     

Effects of a simulated martian UV flux on the cyanobacterium, Chroococcidiopsis sp. 029

Dried monolayers of Chroococcidiopsis sp. 029, a desiccation-tolerant, endolithic cyanobacterium, were exposed to a simulated martian-surface UV and visible light flux, which may also approximate to the worst-case scenario for the Archean Earth. After 5 min, there was a 99% loss of cell viability, and there were no survivors after 30 min. However, this survival was approximately 10 times higher than that previously reported for Bacillus subtilis. We show that under 1 mm of rock, Chroococcidiopsis sp. could survive (and potentially grow) under the high martian UV flux if water and nutrient requirements for growth were met. In isolated cells, phycobilisomes and esterases remained intact hours after viability was lost. Esterase activity was reduced by 99% after a 1-h exposure, while 99% loss of autofluorescence required a 4-h exposure. However, cell morphology was not changed, and DNA was still detectable by 4',6-diamidino-2-phenylindole staining after an 8-h exposure (equivalent to approximately 1 day on Mars at the equator). Under 1 mm of simulant martian soil or gneiss, the effect of UV radiation could not be detected on esterase activity or autofluorescence after 4 h. These results show that under the intense martian UV flux the morphological signatures of life can persist even after viability, enzymatic activity, and pigmentation have been destroyed. Finally, the global dispersal of viable, isolated cells of even this desiccation-tolerant, ionizing-radiation-resistant microorganism on Mars is unlikely as they are killed quickly by unattenuated UV radiation when in a desiccated state. These findings have implications for the survival of diverse microbial contaminants dispersed during the course of human exploratory class missions on the surface of Mars.     

Magnetism, iron minerals, and life on Mars

A short critical review is provided on two questions linking magnetism and possible early life on Mars: (1) Did Mars have an Earth-like internal magnetic field, and, if so, during which period and was it a requisite for life? (2) Is there a connection between iron minerals in the martian regolith and life? We also discuss the possible astrobiological implications of magnetic measurements at the surface of Mars using two proposed instruments. A magnetic remanence device based on magnetic field measurements can be used to identify Noachian age rocks and lightning impacts. A contact magnetic susceptibility probe can be used to investigate weathering rinds on martian rocks and identify meteorites among the small regolith rocks. Both materials are considered possible specific niches for microorganisms and, thus, potential astrobiological targets. Experimental results on analogues are presented to support the suitability of such in situ measurements.     

Astrobiology through the ages of Mars: the study of terrestrial analogues to understand the habitability of Mars

Mars has undergone three main climatic stages throughout its geological history, beginning with a water-rich epoch, followed by a cold and semi-arid era, and transitioning into present-day arid and very cold desert conditions. These global climatic eras also represent three different stages of planetary habitability: an early, potentially habitable stage when the basic requisites for life as we know it were present (liquid water and energy); an intermediate extreme stage, when liquid solutions became scarce or very challenging for life; and the most recent stage during which conditions on the surface have been largely uninhabitable, except perhaps in some isolated niches. Our understanding of the evolution of Mars is now sufficient to assign specific terrestrial environments to each of these periods. Through the study of Mars terrestrial analogues, we have assessed and constrained the habitability conditions for each of these stages, the geochemistry of the surface, and the likelihood for the preservation of organic and inorganic biosignatures. The study of these analog environments provides important information to better understand past and current mission results as well as to support the design and selection of instruments and the planning for future exploratory missions to Mars.     

Immunological detection of small organic molecules in the presence of perchlorates: relevance to the life marker chip and life detection on Mars

The proposed ExoMars mission, due to launch in 2018, aims to look for evidence of extant and extinct life in martian rocks and regolith. Previous attempts to detect organic molecules of biological or abiotic origin on Mars have been unsuccessful, which may be attributable to destruction of these molecules by perchlorate salts during pyrolysis sample extraction techniques. Organic molecules can also be extracted and measured with solvent-based systems. The ExoMars payload includes the Life Marker Chip (LMC) instrument, capable of detecting biomarker molecules of extant and extinct Earth-like life in liquid extracts of martian samples with an antibody microarray assay. The aim of the work reported here was to investigate whether the presence of perchlorate salts, at levels similar to those at the NASA Phoenix landing site, would compromise the LMC extraction and detection method. To test this, we implemented an LMC-representative sample extraction process with an LMC-representative antibody assay and used these to extract and analyze a model sample that consisted of a Mars analog sample matrix (JSC Mars-1) spiked with a representative organic molecular target (pyrene, an example of abiotic meteoritic infall targets) in the presence of perchlorate salts. We found no significant change in immunoassay function when using pyrene standards with added perchlorate salts. When model samples spiked with perchlorate salts were subjected to an LMC-representative liquid extraction, immunoassays functioned in a liquid extract and detected extracted pyrene. For the same model sample matrix without perchlorate salts, we observed anomalous assay signals that coincided with yellow coloration of the extracts. This unexpected observation is being studied further. This initial study indicates that the presence of perchlorate salts, at levels similar to those detected at the NASA Phoenix landing site, is unlikely to prevent the LMC from extracting and detecting organic molecules from martian samples.     

Low-temperature ionizing radiation resistance of Deinococcus radiodurans and Antarctic Dry Valley bacteria

The high flux of cosmic rays onto the unshielded surface of Mars poses a significant hazard to the survival of martian microbial life. Here, we determined the survival responses of several bacterial strains to ionizing radiation exposure while frozen at a low temperature characteristic of the martian near-subsurface. Novel psychrotolerant bacterial strains were isolated from the Antarctic Dry Valleys, an environmental analogue of the martian surface, and identified by 16S rRNA gene phylogeny as representatives of Brevundimonas, Rhodococcus, and Pseudomonas genera. These isolates, in addition to the known radioresistant extremophile Deinococcus radiodurans, were exposed to gamma rays while frozen on dry ice (-79°C). We found D. radiodurans to exhibit far greater radiation resistance when irradiated at -79°C than was observed in similar studies performed at higher temperatures. This greater radiation resistance has important implications for the estimation of potential survival times of microorganisms near the martian surface. Furthermore, the most radiation resistant of these Dry Valley isolates, Brevundimonas sp. MV.7, was found to show 99% 16S rRNA gene similarity to contaminant bacteria discovered in clean rooms at both Kennedy and Johnson Space Centers and so is of prime concern to efforts in the planetary protection of Mars from our lander probes. Results from this experimental irradiation, combined with previous radiation modeling, indicate that Brevundimonas sp. MV.7 emplaced only 30?cm deep in martian dust could survive the cosmic radiation for up to 100,000 years before suffering 10? population reduction.     

The photochemical stability of carbonates on Mars

Carbonates, predominately MgCO3, have been spectroscopically identified at a level of 2-5% in martian dust. However, in spite of this observation, and a large number of climate studies that suggest 1 to several bars of CO2 should be sequestered in carbonate rocks, no outcrop-scale exposures of carbonate have been detected anywhere on Mars to date. To address one hypothesis for this long-standing puzzle, the effect of ultraviolet (UV) light on the stability of calcium carbonate in a simulated martian atmosphere was experimentally investigated. Using 13C-labeled calcite, we found no experimental evidence of the UV photodecomposition of calcium carbonate in a simulated martian atmosphere. Extrapolating the lower limit of detection of our experimental system to an upper limit of carbonate decomposition on Mars yields a quantum efficiency of 3.5 x 10(-8) molecules/photon over the wavelength interval of 190-390 nm and a maximum UV photodecomposition rate of 1.2 x 10(-13) kg m(-2) s(-1) from a calcite surface. The maximum loss of bulk calcite due to this process would be 2.5 nm year(-1) (Mars year). However, calcite is expected to be thermodynamically stable on the surface of Mars, and potential UV photodecomposition reaction mechanisms indicate that, though calcium carbonate may decompose under vacuum, it would be stable in a CO2 atmosphere. Given the expected stability of carbonate on Mars and our inability to detect carbonate decomposition, we conclude that it is unlikely that the apparent absence of extensive carbonate deposits on the martian surface is due to UV photodecomposition in the current environment.     

Key science questions from the second conference on early Mars: geologic, hydrologic, and climatic evolution and the implications for life

In October 2004, more than 130 terrestrial and planetary scientists met in Jackson Hole, WY, to discuss early Mars. The first billion years of martian geologic history is of particular interest because it is a period during which the planet was most active, after which a less dynamic period ensued that extends to the present day. The early activity left a fascinating geological record, which we are only beginning to unravel through direct observation and modeling. In considering this time period, questions outnumber answers, and one of the purposes of the meeting was to gather some of the best experts in the field to consider the current state of knowledge, ascertain which questions remain to be addressed, and identify the most promising approaches to addressing those questions. The purpose of this report is to document that discussion. Throughout the planet's first billion years, planetary-scale processes-including differentiation, hydrodynamic escape, volcanism, large impacts, erosion, and sedimentation-rapidly modified the atmosphere and crust. How did these processes operate, and what were their rates and interdependencies? The early environment was also characterized by both abundant liquid water and plentiful sources of energy, two of the most important conditions considered necessary for the origin of life. Where and when did the most habitable environments occur? Did life actually occupy them, and if so, has life persisted on Mars to the present? Our understanding of early Mars is critical to understanding how the planet we see today came to be.     

Discrimination of aqueous and aeolian paleoenvironments by atomic force microscopy-- a database for the characterization of martian sediments

In the search for aqueous habitats on Mars direct proof of (ancient) flowing water is still lacking, although remote sensing has provided indications of young fluvial systems. To demonstrate that such proof can be given, we examined surface marks on recent terrestrial sand grains by atomic force microscopy (AFM) and applied a quantitative three-dimensional analysis that can numerically distinguish between aeolian and aquatic transport mechanisms in sedimentary deposits on Earth. The surfaces of natural quartz grains as well as olivine, feldspar pyroxene, and monazite sands of known origin were imaged, each image yielding a three-dimensional map of the mineral surface. A fully automated analysis of distribution patterns of the structural elements that constitute the grain surfaces shows that wind-transported quartz grains have short linear elements irregularly distributed on the surface. Linear elements on water-transported grains, however, are longer with orientations that reflect the mineral symmetry. Because the surface patterns found on aqueous grains are due to preferential etching, they can be used as diagnostic fingerprints for the existence of past or present aqueous transport systems. We used a cluster analysis of the cross-correlation distance of distribution patterns in the structures of aeolian and aquatic sand grains to build a phenogram that provides a map for the relationship of the various sediments found on earth. The analysis shows that the method is highly significant and that water and wind transport can clearly be differentiated. In particular, feldspar and olivine sands contributed even more to the discrimination than quartz grains, which indicated that the method is promising for its application on future missions to Mars. Assuming that martian aqueous sand grains exhibit similar erosional patterns to mineral grains on Earth, simple AFM experiments on a Mars lander would be capable of proving the activity of flowing water in modern runoff systems and of analyzing the paleoenvironments of Mars.     

A microbial oasis in the hypersaline Atacama subsurface discovered by a life detector chip: implications for the search for life on Mars

The Atacama Desert has long been considered a good Mars analogue for testing instrumentation for planetary exploration, but very few data (if any) have been reported about the geomicrobiology of its salt-rich subsurface. We performed a Mars analogue drilling campaign next to the Salar Grande (Atacama, Chile) in July 2009, and several cores and powder samples from up to 5?m deep were analyzed in situ with LDChip300 (a Life Detector Chip containing 300 antibodies). Here, we show the discovery of a hypersaline subsurface microbial habitat associated with halite-, nitrate-, and perchlorate-containing salts at 2?m deep. LDChip300 detected bacteria, archaea, and other biological material (DNA, exopolysaccharides, some peptides) from the analysis of less than 0.5?g of ground core sample. The results were supported by oligonucleotide microarray hybridization in the field and finally confirmed by molecular phylogenetic analysis and direct visualization of microbial cells bound to halite crystals in the laboratory. Geochemical analyses revealed a habitat with abundant hygroscopic salts like halite (up to 260?g kg(-1)) and perchlorate (41.13?μg g(-1) maximum), which allow deliquescence events at low relative humidity. Thin liquid water films would permit microbes to proliferate by using detected organic acids like acetate (19.14?μg g(-1)) or formate (76.06?μg g(-1)) as electron donors, and sulfate (15875?μg g(-1)), nitrate (13490?μg g(-1)), or perchlorate as acceptors. Our results correlate with the discovery of similar hygroscopic salts and possible deliquescence processes on Mars, and open new search strategies for subsurface martian biota. The performance demonstrated by our LDChip300 validates this technology for planetary exploration, particularly for the search for life on Mars.