Raman spectroscopic detection of key biomarkers of cyanobacteria and lichen symbiosis in extreme Antarctic habitats: Evaluation for Mars Lander missions

With proposals that micro-miniaturised Raman spectrometers could soon be part of a suite of analytical instrumentation on the surface of Mars, it is critically important to examine the spectral information that could be forthcoming from attempts to determine key molecular biosignatures under the hostile conditions of extra-terrestrial planetary exploration. Current approaches include the analysis of genuine martian geological material in the form of the SNC class meteorites, the formulation of simulated martian regoliths and the examination of putative martian terrestrial analogues; the latter provide the basis of this paper in the form of Antarctic extremophile habitats. In particular, specimens of epilithic, chasmolithic and endolithic lichen and cyanobacterial colonies sampled along a progressively worsening transect towards a “limits of life” situation, beyond which survival of organisms becomes impossible, provide what is arguably the best terrestrial proving-ground for prototype Raman spectrometers for martian exploration. Here, we report the results of experiments on these extant Antarctic extremophile colonies using a range of Raman excitation wavelengths and experimental conditions and also include a compilation of molecular spectral biosignatures, which may be considered as a suitable database for recognition of bioorganic modification of geological strata.     

Evaluation of carbonate abundance in putative martian paleolake basins

Carbonate deposits have not been found so far on Mars, although there appears to have been sufficient water to have supported their formation. Many hypotheses have been proposed in order to explain this. In the present work we explore the possibility that the missed detection of carbonate deposits on the martian surface could be simply due to the fact that the concentration of carbonates, when mixed with other materials present in the sedimentary deposits, may be below the detection limit of the various instruments used so far in this search. In the present study we consider 21 putative paleolacustrine basins and use a sediment transport model to estimate the abundance of carbonates which could be present in the sediments deposited on the basin floor. In this way we find that for all the selected basins the estimated carbonate abundances are in general less than a few percent, and such values are below (or at best comparable to) the detection limits of the spectrometers flown around Mars during the recent space missions. Furthermore, applying the sediment transport model to the well studied Eberswalde crater, we conclude that the fluvio-lacustrine activity in this basin should have lasted for a period on the order of 10³–10⁴ years, in good agreement with previous work. Our results suggest that a hydrological cycle, able to move large volumes of water and to create relatively stable lakes, could have been active intermittently on Mars in the past, producing carbonate deposits that could escape detection by the instruments that have flown to date.     

Detectability of minor constituents in the martian atmosphere by infrared and submillimeter spectroscopy

Spectroscopic remote sensing in the infrared and (sub)millimeter range is a powerful technique that is well suited for detecting minor species in planetary atmospheres (Planet Space Sci. 43(1995) 1485). Yet, only a handful of molecules in the Mars atmosphere (CO₂, CO and H₂O along with their isotopic species, O_3, and more recently H₂O₂ and CH₄) have been detected so far by this method. New high performance spectroscopic instruments will become available in the future in the infrared and (sub)millimeter range, for observations from the ground (infrared spectrometers on 8 m class telescopes, large millimeter and submillimeter interferometers) and from space, in particular the Planetary Fourier Spectrometer (PFS) aboard Mars Express (MEx), and the Heterodyne Instrument for the Far-Infrared (HIFI) aboard the Herschel Space Observatory (HSO). In this paper we will present results of a study that determines detectability of minor species in the atmosphere of Mars, taking into account the expected performance of the above spectroscopic instruments. In the near future, a new determination of the D/H value is expected with the PFS, especially during times of maximum H₂O abundance in the martian atmosphere. PFS is also expected to place constraints on the abundance of several minor species (H₂O₂,CH₄,CH₂O, SO₂, H₂S, OCS, HCl) above any local outgassing sources, the hot spots. It will be possible to obtain complementary information on some minor species (O₃,H₂O₂, CH₄) from ground-based infrared spectrometers on large telescopes. In the more distant future, HIFI will be ideally suited for measuring the isotopic ratios with unprecedented accuracy. Moreover, it should be able to observe O₂, which has not yet been detected spectroscopically in the IR/submm range, as well as H₂O₂. HIFI should also provide upper limits for several species that have not yet been detected (HCl, NH₃, PH₃) in the atmosphere of Mars. Some species (SO, SO₂,H₂S, OCS, CH₂O) that may be observable from the ground could be searched for with present single-dish antennae and arrays, and in the future with the Atacama Large Millimeter Array (ALMA) submillimeter interferometer.     

Assessing spectral evidence of aqueous activity in two putative martian paleolakes

We evaluate the evidence for the presence of mineral spectral signatures indicative of the past presence of water at two putative paleolakes on Mars using observations by the Mars Reconnaissance Orbiter (MRO) Compact Reconnaissance Image Spectrometer for Mars (CRISM). CRISM spectra of both sites are consistent with laboratory spectra of Mg-rich phyllosilicates. Our analysis represents the first detailed evaluation of these locations. The spatial occurrence and association with topographic features within the craters is distinctly different for the two sites. The occurrence of these minerals supports the conclusion that water was once active in the areas sampled by these craters. The distribution of the phyllosilicates in Luqa does not provide distinctive evidence for the presence of a previous standing body of water and is consistent with either impact emplacement or post-impact alteration. For Cankuzo, the phyllosilicate distribution provides evidence of a layer in the crater wall indicative of aqueous activity, but does not require a paleolake.     

Quantitative geochemical mapping of martian elemental provinces

We present an exploratory approach to the interpretation of the elemental maps produced by the Odyssey Gamma-Ray Spectrometer (GRS). These maps benefit from a direct detection of elemental mass fractions and are used to delineate and characterize elementally homogeneous provinces in the mid-latitudinal martian surface on the basis of chemistry alone. This approach is different from assessing the elemental composition of regions previously defined by their geologic context. Multivariate statistical approaches are discussed and a combination of principal component and clustering analyses is applied on the GRS-based hydrogen, chlorine, potassium, silicon, iron, and calcium maps. At least three principal components must be considered to properly describe the compositional variability seen in the maps. The main component is likely driven by the degree of mantling through a GRS perspective, i.e. by materials enriched in mobile elements (Cl, H) and finer than 10-20 cm, at horizontal and depth scales of hundreds of kilometers and tens of centimeters, respectively. Elemental diversity is found in both mantled and less-mantled provinces, suggesting both local and regional sources for the surficial materials. The less-mantled regions appear to have compositions which include basaltic igneous rocks. Although there is an absence of obvious natural clusters in the data, a solution between five and eight elemental provinces seems optimal and is discussed (Amazonis-Tharsis and Sabaea-Arabia, Tempe and the southern highlands, Chryse and Utopia, Elysium-Tartarus, Acidalia-Arabia). Future investigation of the defined elemental provinces will involve integrating other types of data and geological information.     

Raman spectroscopic and scanning electron microscopic analysis of a novel biological colonisation of volcanic rocks

A novel type of colonisation of a basaltic rock, collected on the Arctic island of Svalbard, Norway, during the AMASE expedition in 2004, was characterised using Raman spectroscopy and Scanning Electron Microscopy (SEM). The sample contains two different types of extremophile communities, one occurring behind a radial white crystallisation and the other occurring inside a dark vacuole. Several types of minerals and microbial colonies have been identified by both Raman spectroscopy and SEM analyses. It is the first time that photosynthetic communities have been documented to colonise the inside of dark basaltic rocks. Our discovery has important implications for planetary exploration because it extends the analytical capability and our understanding of microbial rock colonisations to subaerial volcanic outcrops and has wide implications towards the search for life in extraterrestrial planets. In this work we also demonstrate that the use of different laser wavelengths for Raman spectroscopic studies and complementary microscopic analysis are critical for a comprehensive organic and inorganic compound identification.     

Accretion, loss, and fractionation of martian water

A recently published model for accretion of water by Mars from asteroids and comets beyond 2.5 AU calls for Mars to have accreted 6-27% of Earth's ocean (equivalent to 600-2700 meters depth on Mars' surface) with a D/H ratio 1.2-1.6 times Standard Mean Ocean Water. This model is shown to provide enough outgassed water to account for the subsequent evolution of D/H because of loss of hydrogen and the D/H ratio in water of hydration found in martian meteoric minerals, but enhanced rate loss of hydrogen from early Mars is necessary to permit adequate fractionation to occur during the lifetime of the planet.     

Stability analysis of the martian obliquity during the Noachian era

We performed numerical simulations of the obliquity evolution of Mars during the Noachian era, at which time the giant planets were on drastically different orbits than today. For the preferred primordial configuration of the planets we find that there are two large zones where the martian obliquity is stable and oscillates with an amplitude lower than 20°. These zones occur at obliquities below 30° and above 60°; intermediate values show either resonant or chaotic behaviour depending on the primordial orbits of the terrestrial planets.     

Spatial inhomogeneity of the martian subsurface water distribution: implication from a global water cycle model

In an effort to test and to understand the global hydrogen distribution in the shallow subsurface of Mars retrieved by the Mars Odyssey gamma-ray spectrometer, the present state and movement of water are investigated by a coupled global subsurface-atmosphere water cycle model. It was found that the observed global subsurface hydrogen distribution is largely consistent with the modeled global water cycle, so a large fraction of hydrogen is likely to exist as water, at low and mid latitudes in the form of adsorbed water. Under the present climate the water content in the shallow subsurface becomes higher in the northern hemisphere than in the southern hemisphere as a result of global water cycle, regardless of the initial water distribution in the soil or adsorptive capacity. The higher annual maximum soil temperature in the south, stronger net northward transport of atmospheric water vapor, and the emission of vapor from the northern residual polar cap in northern summer contribute to this hemispheric asymmetry. The generally higher adsorptive capacity of clay minerals in the northern plains may further increase this bias. The longitudinal inhomogeneity is caused by several factors, such as thermal inertia, adsorptive capacity, and atmospheric surface pressure. The water abundance is locally high in low thermal inertia regions (e.g., Arabia Terra) and at deep places where the surface pressure is high (e.g., Hellas); it is low in soil with a low adsorptive capacity (e.g., Tharsis) and high thermal inertia regions (e.g., Solis Planum). Most of the soil humidity near the surface at low and mid latitudes may originate from the atmosphere. The model implies that the upper soil layer should be largely ice-free because otherwise an excessive sublimation and vapor emission into the atmosphere in warm seasons would violate the observational constraints. Moreover, the more uniform latitudinal variation of the observed hydrogen abundance near the surface compared to that of deeper layers is indicative of the presence of adsorbed water instead of ground ice because the adsorbed water content does not as steeply depend on latitude as the ground ice stability. Concerning the regolith mineralogy, montmorillonite can much better account for the observed water cycle than palagonite. While the presence of permanent ground ice appears likely in the polar region below a thin layer, large seasonal cycle of phase change between pore ice and adsorbed water may be possible. Regolith adsorption/desorption is neither negligible nor crucial for the seasonal atmospheric water cycle, but the surface-atmosphere coupling is a major prerequisite for the long-term evolution of subsurface water distribution.     

Metastable methane clathrate particles as a source of methane to the martian atmosphere

The observations of methane made by the PFS instrument onboard Mars Express exhibit a definite correlation between methane mixing ratio, water vapor mixing ratio, and cloud optical depth. The recent data obtained from ground-based telescopes seem to confirm the correlation between methane and water vapor. In order to explain this correlation, we suggest that the source of gaseous methane is atmospheric, rather than at the solid surface of the planet, and that this source may consist of metastable submicronic particles of methane clathrate hydrate continuously released to the atmosphere from one or several clathrate layers at depth, according to the phenomenon of “anomalous preservation” evidenced in the laboratory. These particles, lifted up to middle atmospheric levels due to their small size, and therefore filling the whole atmosphere, serve as condensation nuclei for water vapor. The observed correlation between methane and water vapor mixing ratios could be the signature of the decomposition of the clathrate crystals by condensation-sublimation processes related to cloud activity. Under the effect of water condensation on crystal walls, metastability could be broken and particles be eroded, resulting in a subsequent irreversible release of methane to the gas phase. Using PFS data, and according to our hypothesis, the lifetime of gaseous methane is estimated to be smaller than an upper limit of 6 ± 3 months, much smaller than the lifetime of 300 yr calculated from atmospheric chemical models. The reason why methane has a short lifetime might be the occurrence of heterogeneous chemical decomposition of methane in the subsurface, where it is known since Viking biology experiments that oxidants efficiently decompose organic matter. If true, it is shown by using existing models of H₂O₂ penetration in the regolith that methane could prevent H₂O₂ from penetrating in the subsurface, and further oxidizing the soil, at depths larger than a few millimeters. The present source of methane clathrate, acting over the last few hundred thousand or million years, could have given rise to the thin CO₂-ice layer covering the permanent water ice south polar cap. The hypothesis proposed in this paper requires, to be validated, a number of laboratory experiments studying the stability of methane clathrates in martian atmospheric conditions, and the kinetics and amplitude of clathrate particle erosion in presence of condensing water vapor. Detailed future observations of methane, and associated modeling, will allow to more accurately quantify the production rate of methane clathrate, its temporal variability at seasonal scale, and possibly to locate the source(s) of clathrates at the surface.     

Magnitude of global contraction on Mars from analysis of surface faults: Implications for martian thermal history

Faults provide a record of a planet’s crustal stress state and interior dynamics, including volumetric changes related to long-term cooling. Previous work has suggested that Mars experienced a pulse of large-scale global contraction during Hesperian time. Here we evaluate the evidence for martian global contraction using a recent compilation of thrust faults. Fault-related strains were calculated for wrinkle ridges and lobate scarps to provide lower and upper bounds, respectively, on the magnitude of global contraction from contractional structures observed on the surface of Mars. During the hypothesized pulse of global contraction, contractional strain of −0.007% to −0.13% is indicated by the structures, corresponding to decreases in planetary radius of 112 m to 2.24 km, respectively. By contrast, consideration of all recognized thrust faults regardless of age produces a globally averaged contractional strain of −0.011% to −0.22%, corresponding to a radius decrease of 188 m to 3.77 km since the Early Noachian. The amount of global contraction predicted by thermal models is larger than what is recorded by the faults at the surface, paralleling similar studies for Mercury and the Moon, which suggests that observations of fault populations at the surface may provide tighter bounds on planetary thermal evolution than models alone.     

THEMIS-VIS observations of clouds in the martian mesosphere: Altitudes, wind speeds, and decameter-scale morphology

We present measurements of the altitude and eastward velocity component of mesospheric clouds in 35 imaging sequences acquired by the Mars Odyssey (ODY) spacecraft’s Thermal Emission Imaging System visible imaging subsystem (THEMIS-VIS). We measure altitude by using the parallax drift of high-altitude features, and the velocity by exploiting the time delay in the THEMIS-VIS imaging sequence.We observe two distinct classes of mesospheric clouds: equatorial mesospheric clouds observed between 0° and 180° L_s; and northern mid-latitude clouds observed only in twilight in the 200–300° L_s period. The equatorial mesospheric clouds are quite rare in the THEMIS-VIS data set. We have detected them in only five imaging sequences, out of a total of 2048 multi-band equatorial imaging sequences. All five fall between 20° south and 0° latitude, and between 260° and 295° east longitude. The mid-latitude mesospheric clouds are apparently much more common; for these we find 30 examples out of 210 northern winter mid-latitude twilight imaging sequences. The observed mid-latitude clouds are found, with only one exception, in the Acidalia region, but this is quite likely an artifact of the pattern of THEMIS-VIS image targeting. Comparing our THEMIS-VIS images with daily global maps generated from Mars Orbiter Camera Wide Angle (MOC-WA) images, we find some evidence that some mid-latitude mesospheric cloud features correspond to cloud features commonly observed by MOC-WA. Comparing the velocity of our mesospheric clouds with a GCM, we find good agreement for the northern mid-latitude class, but also find that the GCM fails to match the strong easterly winds measured for the equatorial clouds.Applying a simple radiative transfer model to some of the equatorial mesospheric clouds, we find good model fits in two different imaging sequences. By using the observed radiance contrast between cloud and cloud-free regions at multiple visible-band wavelengths, these fits simultaneously constrain the optical depths and particles sizes of the clouds. The particle sizes are constrained primarily by the relative contrasts at the available wavelengths, and are found to be quite different in the two imaging sequences: r_eff = 0.1 μm and r_eff = 1.5 μm. The optical depths (constrained by the absolute contrasts) are substantial: 0.22 and 0.5, respectively. These optical depths imply a mass density that greatly exceeds the saturated mass density of water vapor at mesospheric temperatures, and so the aerosol particles are probably composed mainly of CO₂ ice. Our simple radiative transfer model is not applicable to twilight, when the mid-latitude mesospheric clouds were observed, and so we leave the properties of these clouds as a question for further work.     

Spatial distribution of putative water related features in Southern Acidalia/Cydonia Mensae, Mars

Many putative water-related features exist in the northern lowlands of Mars. These features may provide clues to the abundance and timing of water or ice that existed there in the past. The Cydonia Mensae and Southern Acidalia area was chosen as the study area owing to the abundance of two of these features: giant polygons and pitted cones. In addition a section of the Deuteronilus shoreline is located there. The abundance and close proximity of the features makes this area an excellent place to study the spatial relationships between these landforms, as well as the morphological characteristics of pitted cones. The features were mapped into a GIS for spatial analyses. The highest densities of pitted cones and giant polygons are adjacent but distinctly separated by a knobby ridge that is surrounded by the Deuteronilus putative shoreline. Pitted cones were measured and examined to determine if a classification by morphology is possible, but the results were inconclusive. Statistical tests on pit-to-cone diameter ratios and tests of surface temperatures of cone material suggest, but do not verify, a single cone origin. The various shapes, sizes, and putative ages of pitted cones may be attributed to temporal variation in emplacement and spatial variation in material properties. Among the possible scenarios put forth for pitted cone genesis on Mars two are likely candidates in Cydonia Mensae: (1) the sublimation of a cold-based glacier, and (2) a buried lens of methane and/or CO₂ clathrates.     

Origin of the martian dichotomy and Tharsis from a giant impact causing massive magmatism

The origin of the ancient martian crustal dichotomy and the massive magmatic province of Tharsis remains an open problem. Here, we explore numerically a hypothesis for the origin of these two features involving both exogenic and endogenic processes. We propose a giant impact event during the late stage of planetary formation as the source of the southern highland crust. In a second stage, the extraction of excess heat by vigorous mantle convection on the impacted hemisphere leads to massive magmatism, forming a distinct Tharsis-like volcanic region. By coupling short-term and long-term numerical simulations, we are able to investigate both the early formation as well as the 4.5 Gyr evolution of the martian crust. We demonstrate numerically that this exogenic-endogenic hypothesis is in agreement with observational data from Mars.     

Photochemistry of the martian atmosphere: Seasonal, latitudinal, and diurnal variations

There is a significant progress in the observational data relevant to Mars photochemistry in the current decade. These data are not covered by and sometimes disagree with the published models. Therefore we consider three types of models for Mars photochemistry. A steady-state model for global-mean conditions is currently the only way to calculate the abundances of long living species (H₂, O₂, and CO). However, our model does not fit the observed CO abundance using gas-phase chemistry and reasonable values of heterogeneous loss of odd hydrogen on the water ice aerosol. The second type of the calculated models is steady-state models for local conditions. The MGS/TES data on temperature profiles, H₂O, and dust are input parameters for these models. The calculations have been made for nine seasonal points spread over the martian year and for twelve latitudes with a step of 10° for each season. The only adopted heterogeneous reaction is a weak loss of H₂O₂ on water ice with probability of 5×¹⁰⁻⁴. The results are in good agreement with the recent observations of the O₂ dayglow at 1.27 μm and the O₃ and H₂O₂ abundances. Global maps of the seasonal and latitudinal behavior of these species have been made. The third type of models is a time-dependent model for local conditions. These models show that odd hydrogen quickly converts to H₂O₂ at the nighttime and the chemistry is switched off while the association of O, the heterogeneous loss of H₂O₂, and eddy diffusion continue. This requires significant changes in the global-mean and local steady-state models discussed above, and these changes have been properly done. The calculated diurnal variations of Mars photochemistry are discussed. The martian photochemistry at low and middle latitudes is significantly different in the aphelion period at LS=10°-130° from that in the remaining part of the year.     

Quantitative compositional analysis of martian mafic regions using the MEx/OMEGA reflectance data 1. Methodology, uncertainties and examples of application

The Mars Express Observatoire pour la Minéralogie, l'Eau, les Glaces et l'Activité (OMEGA) collected an unprecedented visible and near-infrared hyperspectral dataset covering the low albedo regions of Mars. We investigate the ability to infer modal abundance of surfaces of these regions from a radiative transfer model developed by Shkuratov et al. [Shkuratov, Y., Starukhina, L., Hoffmann, H., Arnold, G., 1999. Icarus 137, 235-246] and adapted to basaltic surfaces by Poulet and Erard [Poulet F., Erard, S., 2004. J. Geophys. Res. 109 (E2), doi:10.1029/2003JE002179]. From OMEGA measurements of mafic surfaces, we develop several sensitivity tests to assess the extent to which the model can be applied to predict pyroxene composition (high-calcium phase and low-calcium phase), abundance of almost neutral components (plagioclase) in the near-infrared wavelength as well as grain sizes, by using a library of selected end-members. Results of the sensitivity tests indicate that the scattering model can estimate both abundances and grain sizes of major basaltic materials of low albedo regions within uncertainties (±5 to 15 vol%). The model is then applied to data from dissected cratered terrains located in Terra Meridiani. The derived grain size including uncertainties is in the 50-500 μm range. This is consistent with the thermal inertia and albedo of this region, which indicates a fine sand-sized surface with little dust. The abundances of plagioclase (43-57%) and pyroxenes (35-45±10%, including 11±5% of low-calcium phase) are in good agreement with previous basalt-like compositions of low albedo regions from thermal infrared spectral measurements. The method presented in this paper will provide a valuable tool for evaluating the modal mineralogy of other mafic regions of Mars observed in the near-infrared wavelength range.     

Interactive effects of hypobaria, low temperature, and CO2 atmospheres inhibit the growth of mesophilic Bacillus spp. under simulated martian conditions

Robotic spacecraft are launched with finite levels of terrestrial microorganisms that are similar to the microbial communities within facilities in which spacecraft are assembled. In particular, spores of mesophilic aerobic Bacillus species are common spacecraft contaminants considered most likely to survive interplanetary transfer to Mars. During the cruise phase to Mars, and then again during surface operations, microbial bioloads are exposed to a diversity of biocidal factors that are likely to render the microbial species either dead or significantly inhibited from active metabolic activity and replication. We report here, for the first time, that interactive effects of low pressure, low temperature, and high CO₂ atmospheres approaching conditions likely to be encountered on the martian surface strongly inhibit the growth and replication of seven common Bacillus spp. isolated from spacecraft. Tests were conducted within a small glass bell-jar system maintained in a low-temperature microbial incubator. Atmospheric pressures were controlled at 1013 (Earth-normal), 100, 50, 35, 25, or 15 mb, and temperatures were maintained at 30, 20, 15, 10, or 5 °C. Experiments were carried out for 48 h or 7 days under either Earth-normal O₂/N₂ or pure CO₂ atmospheres. Results indicated that low pressure, low temperature, and high CO₂ atmospheres, applied separately or in combination, were capable of inhibiting the growth and replication of B. pumilus SAFR-032, B. pumilus FO-36B, B. subtilis HA-101, B. subtilis 42HS-1, B. megaterium KL-197, B. licheniformis KL-196, and B. nealsonii FO-092 under simulated martian conditions. Endospores of all seven Bacillus spp. strains failed to germinate and grow at 25 mb at 30 °C. Although, vegetative cells of these strains exhibited a slightly greater ability to replicate at lower pressures than did endospores, vegetative cells of these species failed to grow at pressures below 25 mb. Interactive effects of these environmental parameters acted to generally increase the inhibitory nature of the low-pressure conditions on growth and replication of the seven Bacillus spp. tested.     

On Potential Spectroscopic Detection of Microfossils on Mars

There is much evidence about the ancient presence of water on Mars and it is reasonable to suppose that simple forms of life may have developed during the geological evolution of the planet. In such a case traces of this extinct life could still be present on the planet in form of microfossils included into some geological layer. The rover payloads planned for the next decades will include spectrometers in order to accomplish various scientific tasks. In this respect, we have developed a quantitative model for microfossil inclusions into a crystalline matrix. Such a method foresees some visible effects on measurements obtained via spectroscopic techniques such as infrared reflectance and Raman spectroscopy. In this work we present the quantitative model of the fossilization process and the effects that the microfossil inclusions should have in the real spectra, evaluated by means of computer simulations. Preliminary measurements, in order to provide examples of future model testing, have been performed on samples of homogeneous composition, but with detectable microfossils content, collected at the K–T sequence placed near Gubbio (Italy). The preliminary results are presented and discussed in order to investigate the potential application of such spectroscopic techniques for the detection of extinct life.     

Impact of two DNA repair pathways, homologous recombination and non-homologous end joining, on bacterial spore inactivation under simulated martian environmental conditions

Spores of Bacillus subtilis were used as a model system to study the impact of the two major DNA double-strand break (DSB) repair mechanisms [homologous recombination (HR) and non-homologous end-joining (NHEJ)] on the survivability of air-dried mono- and multilayers of bacterial spores under a simulated martian environment; i.e., an environment with low temperature (−10 °C), pure CO₂ atmosphere (99.99% CO₂), 200-1100 nm UV-VIS-NIR radiation, and 0.69 kPa pressure. Spores in multilayers exhibited low inactivation rates compared to monolayers, mainly due to shadowing effects of overlying spores. Simulated martian UV irradiation reduced dramatically spore viability, whereas when shielded from martian UV radiation, spores deficient in NHEJ- and HR-mediated DNA repair were significantly more sensitive to simulated martian environmental conditions than were wild-type spores. In addition, NHEJ-deficient spores were consistently more sensitive than HR-deficient spores to simulated Mars environmental conditions, suggesting that DSBs were an important type of DNA damage. The results indicated that both HR and NHEJ provide an efficient set of DNA repair pathways ensuring spore survival after exposure to simulated martian environmental conditions.