SignificanceIt has remained an unresolved question whether microorganisms recovered from the most arid environments on Earth are thriving under such extreme conditions or are just dead or dying vestiges of viable cells fortuitously deposited by atmospheric processes. Based on multiple lines of evidence, we show that indigenous microbial communities are present and temporally active even in the hyperarid soils of the Atacama Desert (Chile). Following extremely rare precipitation events in the driest parts of this desert, where rainfall often occurs only once per decade, we were able to detect episodic incidences of biological activity. Our findings expand the range of hyperarid environments temporarily habitable for terrestrial life, which by extension also applies to other planetary bodies like Mars.

Extraterrestrial environments encompass physicochemical conditions and habitats that are unknown on Earth, such as perchlorate-rich brines that can be at least temporarily stable on the martian surface. To better understand the potential for life in these cold briny environments, we determined the maximum salt concentrations suitable for growth (MSCg) of six different chloride and perchlorate salts at 25°C and 4°C for the extremotolerant cold-and saltadapted bacterial strain Planococcus halocryophilus. Growth was measured through colony-forming unit (CFU) counts, while cellular and colonial phenotypic stress responses were observed through visible light, fluorescence, and scanning electron microscopy. Our data show the following: (1) The tolerance to high salt concentrations can be increased through a stepwise inoculation toward higher concentrations. (2) Ion-specific factors are more relevant for the growth limitation of P. halocryophilus in saline solutions than single physicochemical parameters like ionic strength or water activity. (3) P. halocryophilus shows the highest microbial sodium perchlorate tolerance described so far. However, (4) MSCg values are higher for all chlorides compared to perchlorates. (5) The MSCg for calcium chloride was increased by lowering the temperature from 25°C to 4°C, while sodium-and magnesium-containing salts can be tolerated at 25°C to higher concentrations than at 4°C. (6) Depending on salt type and concentration, P. halocryophilus cells show distinct phenotypic stress responses such as novel types of colony morphology on agar plates and biofilm-like cell clustering, encrustation, and development of intercellular nanofilaments. This study, taken in context with previous work on the survival of extremophiles in Mars-like environments, suggests that highconcentrated perchlorate brines on Mars might not be habitable to any present organism on Earth, but extremophilic microorganisms might be able to evolve thriving in such environments.

It is well known that dissolved salts can significantly lower the freezing point of water and thus extend habitability to subzero conditions. However, most investigations thus far have focused on sodium chloride as a solute. In this study, we report on the survivability of the bacterial strain Planococcus halocryophilus in sodium, magnesium, and calcium chloride or perchlorate solutions at temperatures ranging from +25°C to −30°C. In addition, we determined the survival rates of P. halocryophilus when subjected to multiple freeze/thaw cycles. We found that cells suspended in chloride-containing samples have markedly increased survival rates compared with those in perchlorate-containing samples. In both cases, the survival rates increase with lower temperatures; however, this effect is more pronounced in chloride-containing samples. Furthermore, we found that higher salt concentrations increase survival rates when cells are subjected to freeze/thaw cycles. Our findings have important implications not only for the habitability of cold environments on Earth but also for extraterrestrial environments such as that of Mars, where cold brines might exist in the subsurface and perhaps even appear temporarily at the surface such as at recurring slope lineae.

Recurring slope lineae (RSL) are flow‐like features on Mars characterized by a local darkening of the soil thought to be generated by the formation and flow of liquid brines. One possible mechanism responsible for forming these brines could be the deliquescence of salts present in the Martian soil. We show that the JSC Mars‐1a analogue soil undergoes a darkening process when salts dispersed in the soil deliquesce, but forming continuous liquid films and larger droplets takes much longer than previously assumed. Thus, RSL may not necessarily require concurrent flowing liquid water/brine or a salt‐recharge mechanism, and their association with gullies may be the result of previously flowing water and deposited salts during an earlier warmer and wetter period. In addition, our results show that electrical conductivity measurements correlate well with the deliquescence rates and provide better overall characterization than either Raman spectroscopy or estimates based on deliquescence relative humidity.

The habitability of Mars is strongly dependent on the availability of liquid water, which is essential for life as we know it. One of the few places where liquid water might be found on Mars is in liquid perchlorate brines that could form via deliquescence. As these concentrated perchlorate salt solutions do not occur on Earth as natural environments, it is necessary to investigate in lab experiments the potential of these brines to serve as a microbial habitat. Here, we report on the sodium perchlorate (NaClO4) tolerances for the halotolerant yeast Debaryomyces hansenii and the filamentous fungus Purpureocillium lilacinum. Microbial growth was determined visually, microscopically and via counting colony forming units (CFU). With the observed growth of D. hansenii in liquid growth medium containing 2.4 M NaClO4, we found by far the highest microbial perchlorate tolerance reported to date, more than twice as high as the record reported prior (for the bacterium Planococcus halocryophilus). It is plausible to assume that putative Martian microbes could adapt to even higher perchlorate concentrations due to their long exposure to these environments occurring naturally on Mars, which also increases the likelihood of microbial life thriving in the Martian brines.

The question whether organic compounds occur on Mars remained unanswered for decades. However, the recent discovery of various classes of organic matter in martian sediments by the Curiosity rover seems to strongly suggest that indigenous organic compounds exist on Mars. One intriguing group of detected organic compounds were thiophenes, which typically occur on Earth in kerogen, coal, and crude oil as well as in stromatolites and microfossils. Here we provide a brief synopsis of conceivable pathways for the generation and degradation of thiophenes on Mars. We show that the origin of thiophene derivatives can either be biotic or abiotic, for example, through sulfur incorporation in organic matter during early diagenesis. The potential of thiophenes to represent martian biomarkers is discussed as well as a correlation between abundances of thiophenes and sulfate-bearing minerals. Finally, this study provides suggestions for future investigations on Mars and in Earth-based laboratories to answer the question whether the martian thiophenes are of biological origin.

the current understanding of the Martian surface indicates that briny environments at the near-surface are temporarily possible, e.g. in the case of the presumably deliquescence-driven Recurring Slope Lineae (RSL). However, whether such dynamic environments are habitable for terrestrial organisms remains poorly understood. this hypothesis was tested by developing a closed Deliquescence System (CDS) consisting of a mixture of desiccated Martian Regolith Analog (MRA) substrate, salts, and microbial cells, which over the course of days became wetted through deliquescence. The methane produced via metabolic activity for three methanogenic archaea: Methanosarcina mazei, M. barkeri and M. soligelidi, was measured after exposing them to three different MRA substrates using either NaCl or naclo 4 as a hygroscopic salt. Our experiments showed that (1) M. soligelidi rapidly produced methane at 4 °C, (2) M. barkeri produced methane at 28 °C though not at 4 °C, (3) M. mazei was not metabolically reactivated through deliquescence, (4) none of the species produced methane in the presence of perchlorate, and (5) all species were metabolically most active in the phyllosilicate-containing MRA. These results emphasize the importance of the substrate, microbial species, salt, and temperature used in the experiments. Furthermore, we show here for the first time that water provided by deliquescence alone is sufficient to rehydrate methanogenic archaea and to reactivate their metabolism under conditions roughly analogous to the near-subsurface Martian environment. Methane in the atmosphere of Mars was first detected by Formisano, et al. 1 with the Planetary Fourier Spectrometer onboard the Mars Express orbiter. Recent measurements by the Tunable Laser Spectrometer onboard the Curiosity rover show that methane concentrations are currently in the parts per billion range and fluctuate repeatedly throughout the seasons 2. However, the long-term stability and presence of methane in the Martian atmosphere is considered unlikely over geological time periods due to degradation by UV radiation and/ or oxidizing compounds at the surface 3. Hence, it has been proposed that subsurface reservoirs might sporadically release methane and/or that it is actively produced through abiotic, or potentially, as it is on Earth, from microbial processes 4. On Mars, however, the environmental conditions within the surficial regolith generally do not permit a lasting presence of liquid water necessary for microbial metabolism. Nevertheless, besides surficial monolayers of liquid-like water 5 and morning frost 6 , a possible exception are near-surface environments laden with hygroscopic salts which can undergo deliquescence, a process occurring when the relative humidity (RH) exceeds the deliquescence relative humidity (DRH) of a given salt above its eutectic temperature. Such a mechanism was first hypothesized by McEwen, et al. 7 to play a role in the seasonal surface features on Mars known as Recurring Slope Lineae (RSL), dark streaks that appear on steep crat...

The existence of microbial activity hotspots in temperate regions of Earth is driven by soil heterogeneities, especially the temporal and spatial availability of nutrients. Here we investigate whether microbial activity hotspots also exist in lithic microhabitats in one of the most arid regions of the world, the Atacama Desert in Chile. While previous studies evaluated the total DNA fraction to elucidate the microbial communities, we here for the first time use a DNA separation approach on lithic microhabitats, together with metagenomics and other analysis methods (i.e., ATP, PLFA, and metabolite analysis) to specifically gain insights on the living and potentially active microbial community. Our results show that hypolith colonized rocks are microbial hotspots in the desert environment. In contrast, our data do not support such a conclusion for gypsum crust and salt rock environments, because only limited microbial activity could be observed. The hypolith community is dominated by phototrophs, mostly Cyanobacteria and Chloroflexi, at both study sites. The gypsum crusts are dominated by methylotrophs and heterotrophic phototrophs, mostly Chloroflexi, and the salt rocks (halite nodules) by phototrophic and halotolerant endoliths, mostly Cyanobacteria and Archaea. The major environmental constraints in the organic-poor arid and hyperarid Atacama Desert are water availability and UV irradiation, allowing phototrophs and other extremophiles to play a key role in desert ecology.