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.     

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.     

A bacterial enrichment study and overview of the extractable lipids from paleosols in the Dry Valleys, Antarctica: implications for future Mars reconnaissance

The Dry Valleys of Antarctica are one of the coldest and driest environments on Earth with paleosols in selected areas that date to the emplacement of tills by warm-based ice during the Early Miocene. Cited as an analogue to the martian surface, the ability of the Antarctic environment to support microbial life-forms is a matter of special interest, particularly with the upcoming NASA/ESA 2018 ExoMars mission. Lipid biomarkers were extracted and analyzed by gas chromatography--mass spectrometry to assess sources of organic carbon and evaluate the contribution of microbial species to the organic matter of the paleosols. Paleosol samples from the ice-free Dry Valleys were also subsampled and cultivated in a growth medium from which DNA was extracted with the explicit purpose of the positive identification of bacteria. Several species of bacteria were grown in solution and the genus identified. A similar match of the data to sequenced DNA showed that Alphaproteobacteria, Gammaproteobacteria, Bacteriodetes, and Actinobacteridae species were cultivated. The results confirm the presence of bacteria within some paleosols, but no assumptions have been made with regard to in situ activity at present. These results underscore the need not only to further investigate Dry Valley cryosols but also to develop reconnaissance strategies to determine whether such likely Earth-like environments on the Red Planet also contain life.     

A hydrothermal system associated with the Siljan impact structure, Sweden--implications for the search for fossil life on Mars

The Siljan ring structure (368 +/- 1.1 Ma) is the largest known impact structure in Europe. It isa 65-km-wide, eroded, complex impact structure, displaying several structural units, including a central uplifted region surrounded by a ring-shaped depression. Associated with the impact crater are traces of a post-impact hydrothermal system indicated by precipitated and altered hydrothermal mineral assemblages. Precipitated hydrothermal minerals include quartz veins and breccia fillings associated with granitic rocks at the outer margin of the central uplift, and calcite, fluorite, galena, and sphalerite veins associated with Paleozoic carbonate rocks located outside the central uplift. Two-phase water/gas and oil/gas inclusions in calcite and fluorite display homogenization temperatures between 75 degrees C and 137 degrees C. With an estimated erosional unloading of approximately 1 km, the formation temperatures were probably not more than 10-15 degrees C higher. Fluid inclusion ice-melting temperatures indicate a very low salt content, reducing the probability that the mineralization was precipitated during the Caledonian Orogeny. Our findings suggest that large impacts induce low-temperature hydrothermal systems that may be habitats for thermophilic organisms. Large impact structures on Mars may therefore be suitable targets in the search for fossil thermophilic organisms.     

Ultrastructural and geochemical characterization of Archean-Paleoproterozoic graphite particles: implications for recognizing traces of life in highly metamorphosed rocks

Abundant graphite particles occur in amphibolite-grade quartzite of the Archean-Paleoproterozoic Wutai Metamorphic Complex in the Wutaishan area of North China. Petrographic thin section observations suggest that the graphite particles occur within and between quartzite clasts and are heterogeneous in origin. Using HF maceration techniques, the Wutai graphite particles were extracted for further investigation. Laser Raman spectroscopic analysis of a population of extracted graphite discs indicated that they experienced a maximum metamorphic temperature of 513 +/- 50 degrees C, which is consistent with the metamorphic grade of the host rock and supports their indigenicity. Scanning and transmission electron microscopy revealed that the particles bear morphological features (such as hexagonal sheets of graphite crystals) related to metamorphism and crystal growth, but a small fraction of them (graphite discs) are characterized by a circular morphology, distinct marginal concentric folds, surficial wrinkles, and complex nanostructures. Ion microprobe analysis of individual graphite discs showed that their carbon isotope compositions range from -7.4 per thousand to -35.9 per thousand V-PDB (Vienna Pee Dee Belemnite), with an average of -20.3 per thousand, which is comparable to bulk analysis of extracted carbonaceous material. The range of their size, ultrastructures, and isotopic signatures suggests that the morphology and geochemistry of the Wutai graphite discs were overprinted by metamorphism and their ultimate carbon source probably had diverse origins that included abiotic processes. We considered both biotic and abiotic origins of the carbon source and graphite disc morphologies and cannot falsify the possibility that some circular graphite discs characterized by marginal folds and surficial wrinkles represent deflated, compressed, and subsequently graphitized organic-walled vesicles. Together with reports by other authors of acanthomorphic acritarchs from greenschist-amphibolite-grade metamorphic rocks, this study suggests that it is worthwhile to examine carbonaceous materials preserved in highly metamorphosed rocks for possible evidence of ancient life.     

The Mawrth Vallis region of Mars: A potential landing site for the Mars Science Laboratory (MSL) mission

The primary objective of NASA's Mars Science Laboratory (MSL) mission, which will launch in 2011, is to characterize the habitability of a site on Mars through detailed analyses of the composition and geological context of surface materials. Within the framework of established mission goals, we have evaluated the value of a possible landing site in the Mawrth Vallis region of Mars that is targeted directly on some of the most geologically and astrobiologically enticing materials in the Solar System. The area around Mawrth Vallis contains a vast (>1?×?10? km2) deposit of phyllosilicate-rich, ancient, layered rocks. A thick (>150?m) stratigraphic section that exhibits spectral evidence for nontronite, montmorillonite, amorphous silica, kaolinite, saponite, other smectite clay minerals, ferrous mica, and sulfate minerals indicates a rich geological history that may have included multiple aqueous environments. Because phyllosilicates are strong indicators of ancient aqueous activity, and the preservation potential of biosignatures within sedimentary clay deposits is high, martian phyllosilicate deposits are desirable astrobiological targets. The proposed MSL landing site at Mawrth Vallis is located directly on the largest and most phyllosilicate-rich deposit on Mars and is therefore an excellent place to explore for evidence of life or habitability.     

Survival and germinability of Bacillus subtilis spores exposed to simulated Mars solar radiation: implications for life detection and planetary protection

Bacterial spores have been considered as microbial life that could survive interplanetary transport by natural impact processes or human spaceflight activity. Deposition of terrestrial microbes or their biosignature molecules onto the surface of Mars could negatively impact life detection experiments and planetary protection measures. Simulated Mars solar radiation, particularly the ultraviolet component, has been shown to reduce spore viability, but its effect on spore germination and resulting production of biosignature molecules has not been explored. We examined the survival and germinability of Bacillus subtilis spores exposed to simulated martian conditions that include solar radiation. Spores of B. subtilis that contain luciferase resulting from expression of an sspB-luxAB gene fusion were deposited on aluminum coupons to simulate deposition on spacecraft surfaces and exposed to simulated Mars atmosphere and solar radiation. The equivalent of 42 min of simulated Mars solar radiation exposure reduced spore viability by nearly 3 logs, while germination-induced bioluminescence, a measure of germination metabolism, was reduced by less than 1 log. The data indicate that spores can retain the potential to initiate germination-associated metabolic processes and produce biological signature molecules after being rendered nonviable by exposure to Mars solar radiation.     

Spiders: water-driven erosive structures in the southern hemisphere of Mars

Recent data from space missions reveal that there are ongoing climatic changes and erosive processes that continuously modify surface features of Mars. We have investigated the seasonal dynamics of a number of morphological features located at Inca City, a representative area at high southern latitude that has undergone seasonal processes. By integrating visual information from the Mars Orbiter Camera on board the Mars Global Surveyor and climatic cycles from a Mars' General Circulation Model, and considering the recently reported evidence for the presence of water-ice and aqueous precipitates on Mars, we propose that a number of the erosive features identified in Inca City, among them spiders, result from the seasonal melting of aqueous salty solutions.     

Bacillus subtilis spore survival and expression of germination-induced bioluminescence after prolonged incubation under simulated Mars atmospheric pressure and composition: implications for planetary protection and lithopanspermia

Bacterial endospores in the genus Bacillus are considered good models for studying interplanetary transfer of microbes by natural or human processes. Although spore survival during transfer itself has been the subject of considerable study, the fate of spores in extraterrestrial environments has received less attention. In this report we subjected spores of a strain of Bacillus subtilis, containing luciferase resulting from expression of an sspB-luxAB gene fusion, to simulated martian atmospheric pressure (7-18 mbar) and composition (100% CO(2)) for up to 19 days in a Mars simulation chamber. We report here that survival was similar between spores exposed to Earth conditions and spores exposed up to 19 days to simulated martian conditions. However, germination-induced bioluminescence was lower in spores exposed to simulated martian atmosphere, which suggests sublethal impairment of some endogenous spore germination processes.     

Terraforming Mars: conceptual solutions to the problem of plant growth in low concentrations of oxygen

The widespread growth of higher plants on Mars following ecopoiesis has often been invoked as a method of generating atmospheric oxygen. However, one issue that has been overlooked in this regard is the fact that terrestrial plants do not thrive under conditions of low oxygen tension. A review of the relevant botanical literature reveals that the high oxygen demands of root respiration could limit the introduction of most plants on Mars until after terraforming has raised the atmospheric pO2 to 20-100 mbar. A variety of physiological strategies are discussed which, if it is possible to implement them in a genetically engineered plant specifically designed for life on Mars, might allow this problem to be overcome.     

"Signal" search for intelligence in the galactic nucleus with the array of the Lowlands

In August, 1981, the Westerbork Synthesis Radio Telescope was used for 4 h to search for narrowband pulsing radio beacons in the direction of the Galactic Center. By using both the spatial discrimination and temporal stability available to an interferometric measurement, weak intermittent signals can be detected even in the face of the strong, naturally caused radiation from this region. A radio beacon within our bandwidth, centered on the 21 cm neutral hydrogen line, would be recognizable if it had a repetition period between 40 sec and 1/2 h. The rms sensitivity to point sources was approximately 50 mJy/cycle, and the detection limit was 500 mJy/cycle. The limit degrades for pulse widths < 0.02s. No repetitive signals were found. For a swept, narrow-band radio beacon constrained to the Galactic Disk (beamwidth = 0.02 rad), our detection limit corresponds to a transmitter power of 10(11) MW at the Galactic Center.     

Origin of informational polymers and the search for non-terran life: protection of the polymeric state of DNA by phosphate minerals

An in-depth analysis of the effects exerted on the DNA backbone by 25 crystal phosphate minerals is reported. Degradation of DNA oligomers was performed with two different reactions: Hydrolysis following Nucleophilic Degradation (HND), initiated by the nucleophilic addition of formamide on both purine and pyrimidine nucleobases, and Hydrolysis following Nucleophilic Substitution (HNS) carried on by water and starting with the removal of a nondegraded base. A complete panel of effects on the phosphoester bonds, from protection to enhanced instability to absence of interference, is described. These effects differ in the different degradation pathways and in different physical-chemical conditions. The relationship between the hardness of the mineral and its protective ability is discussed. In addition to its interest per se, this study was prompted by the observed catalytic abilities of soluble and mineral phosphates (Saladino et al., 2006c) on the synthetic reactions by formamide. The relevance of these observations in the search for nonterran life is discussed.     

Discovery of abundant cellulose microfibers encased in 250 Ma Permian halite: a macromolecular target in the search for life on other planets

In this study, we utilized transmission electron microscopy to examine the contents of fluid inclusions in halite (NaCl) and solid halite crystals collected 650 m below the surface from the Late Permian Salado Formation in southeastern New Mexico (USA). The halite has been isolated from contaminating groundwater since deposition approximately 250 Ma ago. We show that abundant cellulose microfibers are present in the halite and appear remarkably intact. The cellulose is in the form of 5 nm microfibers as well as composite ropes and mats, and was identified by resistance to 0.5 N NaOH treatment and susceptibility to cellulase enzyme treatment. These cellulose microfibers represent the oldest native biological macromolecules to have been directly isolated, examined biochemically, and visualized (without growth or replication) to date. This discovery points to cellulose as an ideal macromolecular target in the search for life on other planets in our Solar System.     

Subsurface filamentous fabrics: an evaluation of origins based on morphological and geochemical criteria, with implications for exopaleontology

The fossil record of the subsurface biosphere is sparse. Results obtained on subsurface filamentous fabrics (SFF) from >225 paleosubsurface sites in volcanics, oxidized ores, and paleokarst of subrecent to Proterozoic age are presented. SFF are mineral encrustations on filamentous or fibrous substrates that formed in subsurface environments. SFF occur in association with low-temperature aqueous mineral assemblages and consist of tubular, micron-thick (median 1.6 micron) filaments in high spatial density, which occur as irregular masses, matted fabrics, and vertically draped features that resemble stalactites. Micron-sized filamentous centers rule out a stalactitic origin. Morphometric analysis of SFF filamentous forms demonstrates that their shape more closely resembles microbial filaments than fibrous minerals. Abiogenic filament-like forms are considered unlikely precursors of most SFF, because abiogenic forms differ in the distribution of widths and have a lower degree of curvature and a lower number of direction changes. Elemental analyses of SFF show depletion in immobile elements (e.g., Al, Th) and a systematic enrichment in As and Sb, which demonstrates a relation to environments with high flows of water. Sulfur isotopic analyses are consistent with a biological origin of a SFF sample from a Mississippi Valley-Type deposit, which is consistent with data in the literature. Fe isotopes in SFF and active analogue systems, however, allow no discrimination between biogenic and abiogenic origins. The origin of most SFF is explained as permineralized remains of microbial filaments that possibly record rapid growth during phases of high water flow that released chemical energy. It is possible that some SFF formed due to encrustation of mineral fibers. SFF share similarities with Microcodium from soil environments. SFF are a logical target in the search for past life on Mars. The macroscopic nature of many SFF allows for their relatively easy in situ recognition and targeting for more detailed microstructural and geochemical analysis.