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.

Two rover missions to Mars aim to detect biomolecules as a sign of extinct or extant life with, among other instruments, Raman spectrometers. However, there are many unknowns about the stability of Raman-detectable biomolecules in the martian environment, clouding the interpretation of the results. To quantify Raman-detectable biomolecule stability, we exposed seven biomolecules for 469 days to a simulated martian environment outside the International Space Station. Ultraviolet radiation (UVR) strongly changed the Raman spectra signals, but only minor change was observed when samples were shielded from UVR. These findings provide support for Mars mission operations searching for biosignatures in the subsurface. This experiment demonstrates the detectability of biomolecules by Raman spectroscopy in Mars regolith analogs after space exposure and lays the groundwork for a consolidated space-proven database of spectroscopy biosignatures in targeted environments.

The concept of a biosignature is widely used in astrobiology to suggest a link between some observation and a biological cause, given some context. The term itself has been defined and used in several ways in different parts of the scientific community involved in the search for past or present life on Earth and beyond. With the ongoing acceleration in the search for life in distant time and/or deep space, there is a need for clarity and accuracy in the formulation and reporting of claims. Here, we critically review the biosignature concept(s) and the associated nomenclature in light of several problems and ambiguities emphasized by recent works. One worry is that these terms and concepts may imply greater certainty than is usually justified by a rational interpretation of the data. A related worry is that terms such as “biosignature” may be inherently misleading, for example, because the divide between life and non-life—and their observable effects—is fuzzy. Another worry is that different parts of the multidisciplinary community may use non-equivalent or conflicting definitions and conceptions, leading to avoidable confusion. This review leads us to identify a number of pitfalls and to suggest how they can be circumvented. In general, we conclude that astrobiologists should exercise particular caution in deciding whether and how to use the concept of biosignature when thinking and communicating about habitability or life. Concepts and terms should be selected carefully and defined explicitly where appropriate. This would improve clarity and accuracy in the formulation of claims and subsequent technical and public communication about some of the most profound and important questions in science and society. With this objective in mind, we provide a checklist of questions that scientists and other interested parties should ask when assessing any reported detection of a “biosignature” to better understand exactly what is being claimed.

The future robotic exploration missions to Mars—European Space Agency/Roscosmos's ExoMars2020 and National Aeronautics and Space Administration's Mars2020 rovers—will search for signs of extant or extinct life using, among other instruments, Raman spectrometers for the first time. The question remains whether organic biosignatures—such as pigments and cellular components—may be detected by this method. Evaluating their detection limit under simulated extraterrestrial conditions is therefore crucial for the success of future life detection missions. Ionizing radiation can be considered as the most deleterious factor for the long‐term preservation of potential biomarkers on Mars. Here, we report on the preservation potential of Raman signatures in the Antarctic strain CCCryo 231‐06 of the cyanobacterium Nostoc sp. after high doses of gamma irradiation. The carotenoids' signals, a well‐established biosignature model, dominate the Raman spectra at 532‐nm excitation wavelength due to resonance effects. But comparing their distribution and quantifying their preservation are still problematic in natural samples. To standardize the analyses, we successfully applied Raman mapping and signal‐to‐noise ratio masks to quantify the effects of irradiation. The typical in vivo Raman signatures of carotenoids could be detected after exposure to up to 56 kGy with significant deterioration in terms of signal coverage and signal‐to‐noise ratio. But they remained stable even after the highest dose of γ rays (117 kGy) tested in this study for colonies embedded in two different Martian mineral analogues. Data gathered during these ground‐based irradiation experiments contribute to interpret results from space experiments and will guide our search for life on Mars and other bodies of interest.

Single-cell sequencing is a powerful technology that provides the capability of analyzing a single cell within a population. This technology is mostly coupled with microfluidic systems for controlled cell manipulation and precise fluid handling to shed light on the genomes of a wide range of cells. So far, single-cell sequencing has been focused mostly on human cells due to the ease of lysing the cells for genome amplification. The major challenges that bacterial species pose to genome amplification from single cells include the rigid bacterial cell walls and the need for an effective lysis protocol compatible with microfluidic platforms. In this work, we present a lysis protocol that can be used to extract genomic DNA from both gram-positive and gram-negative species without interfering with the amplification chemistry. Corynebacterium glutamicum was chosen as a typical gram-positive model and Nostoc sp. as a gram-negative model due to major challenges reported in previous studies. Our protocol is based on thermal and chemical lysis. We consider 80% of single-cell replicates that lead to >5 ng DNA after amplification as successful attempts. The protocol was directly applied to Gloeocapsa sp. and the single cells of the eukaryotic Sphaerocystis sp. and achieved a 100% success rate.

Mars is a primary target of astrobiological interest: its past environmental conditions may have been favourable to the emergence of a prebiotic chemistry and, potentially, biological activity. In situ exploration is currently underway at the Mars surface, and the subsurface (2 m depth) will be explored in the future ESA ExoMars mission. In this context, BIOlogy and Mars EXperiment was performed to evaluate the stability and detectability of organic biomarkers under space and Mars-like conditions. Our data suggested that some target molecules, namely melanin, azelaic acid and nucleic acids, can be detected even after 16 months exposure to Low Earth Orbit conditions by multidisciplinary approaches. We used the same techniques as onboard the ExoMars rover, as Raman and infrared spectroscopies and gas chromatograph-mass spectrometer, and polymerase chain reaction even if this is not planned for the imminent mission to Mars. These results should be taken into account for future Mars exploration.