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The Nostoc sp. strain CCCryo 231-06 is a cyanobacterial strain capable of surviving under extreme conditions and thus is of great interest for the astrobiology community. The knowledge of its complete genome sequence would serve as a guide for further studies. However, a major concern has been placed on the effects of contamination on the quality of sequencing data without a reference genome. Here, we report the use of microfluidic technology combined with single cell sequencing and de novo assembly to minimize the contamination and recover the complete genome of the Nostoc strain CCCryo 231-06 with high quality. 100% of the whole genome was recovered with all contaminants removed and a strongly supported phylogenetic tree. The data reported can be useful for comparative genomics for phylogenetic and taxonomic studies. The method used in this work can be applied to studies that require high-quality assemblies of genomes of unknown microorganisms.

The Moon is characterized by extremely harsh conditions due to ultraviolet irradiation, wide temperature extremes, vacuum resulting from the absence of an atmosphere and high ionizing radiation. Therefore, its surface may provide a unique platform to investigate the effects of such conditions. For lunar exploration with the Lunar Gateway platform, exposure experiments in Low Earth Orbit are useful testbeds to prepare for lunar space experiments and to understand how and if potential biomarkers are influenced by extra-terrestrial conditions. During the BIOMEX (BIOlogy and Mars EXperiment) project, dried colonies of the fungus Cryomyces antarcticus grown on Lunar Regolith Analogue (LRA) were exposed to space conditions for 16 months aboard the EXPOSE-R2 payload outside the International Space Station. In this study, we investigated the stability/degradation of fungal biomarkers in LRA after exposure to (i) simulated space and (ii) real space conditions, using Raman spectroscopy, gas chromatography-mass spectrometry and DNA amplification. The results demonstrated that fungal biomarkers were detectable after 16 months of real space exposure. This work will contribute to the interpretation of data from future biological experiments in the Cislunar orbit with the Lunar Gateway platform and/or on the lunar surface, in preparation for the next step of human exploration.

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.

© Copyright © 2021 Kozyrovska, Reva, Podolich, Kukharenko, Orlovska, Terzova, Zubova, Trovatti Uetanabaro, Góes-Neto, Azevedo, Barh, Verseux, Billi, Kołodziejczyk, Foing, Demets and Vera.Humankind has entered a new era of space exploration: settlements on other planetary bodies are foreseen in the near future. Advanced technologies are being developed to support the adaptation to extraterrestrial environments and, with a view on the longer term, to support the viability of an independent economy. Biological processes will likely play a key role and lead to the production of life-support consumables, and other commodities, in a way that is cheaper and more sustainable than exclusively abiotic processes. Microbial communities could be used to sustain the crews’ health as well as for the production of consumables, for waste recycling, and for biomining. They can self-renew with little resources from Earth, be highly productive on a per-volume basis, and be highly versatile—all of which will be critical in planetary outposts. Well-defined, semi-open, and stress-resistant microecosystems are particularly promising. An instance of it is kombucha, known worldwide as a microbial association that produces an eponymous, widespread soft drink that could be valuable for sustaining crews’ health or as a synbiotic (i.e., probiotic and prebiotic) after a rational assemblage of defined probiotic bacteria and yeasts with endemic or engineered cellulose producers. Bacterial cellulose products offer a wide spectrum of possible functions, from leather-like to innovative smart materials during long-term missions and future activities in extraterrestrial settlements. Cellulose production by kombucha is zero-waste and could be linked to bioregenerative life support system (BLSS) loops. Another advantage of kombucha lies in its ability to mobilize inorganic ions from rocks, which may help feed BLSS from local resources. Besides outlining those applications and others, we discuss needs for knowledge and other obstacles, among which is the biosafety of microbial producers.

As a part of the European Space Agency mission "EXPOSE-R2" on the International Space Station (ISS), the BIOMEX (Biology and Mars Experiment) experiment investigates the habitability of Mars and the limits of life. In preparation for the mission, experimental verification tests and scientific verification tests simulating different combinations of abiotic space- and Mars-like conditions were performed to analyze the resistance of a range of model organisms. The simulated abiotic space- and Mars-stressors were extreme temperatures, vacuum, and Mars-like surface ultraviolet (UV) irradiation in different atmospheres. We present for the first time simulated space exposure data of mosses using plantlets of the bryophyte genus Grimmia, which is adapted to high altitudinal extreme abiotic conditions at the Swiss Alps. Our preflight tests showed that severe UVR200-400nm irradiation with the maximal dose of 5 and 6.8 x 10(5) kJ center dot m(-2), respectively, was the only stressor with a negative impact on the vitality with a 37% (terrestrial atmosphere) or 36% reduction (space- and Mars-like atmospheres) in photosynthetic activity. With every exposure to UVR200-400nm 10(5) kJ center dot m(-2), the vitality of the bryophytes dropped by 6%. No effect was found, however, by any other stressor. As the mosses were still vital after doses of ultraviolet radiation (UVR) expected during the EXPOSE-R2 mission on ISS, we show that this earliest extant lineage of land plants is highly resistant to extreme abiotic conditions.

BIOMEX (BIOlogy and Mars EXperiment) is an ESA/Roscosmos space exposure experiment housed within the exposure facility EXPOSE-R2 outside the Zvezda module on the International Space Station (ISS). The design of the multiuser facility supports—among others—the BIOMEX investigations into the stability and level of degradation of space-exposed biosignatures such as pigments, secondary metabolites, and cell surfaces in contact with a terrestrial and Mars analog mineral environment. In parallel, analysis on the viability of the investigated organisms has provided relevant data for evaluation of the habitability of Mars, for the limits of life, and for the likelihood of an interplanetary transfer of life (theory of lithopanspermia). In this project, lichens, archaea, bacteria, cyanobacteria, snow/permafrost algae, meristematic black fungi, and bryophytes from alpine and polar habitats were embedded, grown, and cultured on a mixture of martian and lunar regolith analogs or other terrestrial minerals. The organisms and regolith analogs and terrestrial mineral mixtures were then exposed to space and to simulated Mars-like conditions by way of the EXPOSE-R2 facility. In this special issue, we present the first set of data obtained in reference to our investigation into the habitability of Mars and limits of life. This project was initiated and implemented by the BIOMEX group, an international and interdisciplinary consortium of 30 institutes in 12 countries on 3 continents. Preflight tests for sample selection, results from ground-based simulation experiments, and the space experiments themselves are presented and include a complete overview of the scientific processes required for this space experiment and postflight analysis. The presented BIOMEX concept could be scaled up to future exposure experiments on the Moon and will serve as a pretest in low Earth orbit., This research was supported by the Italian Space Agency (ASI grant BIOMEX Cyano 051-R.0 to D.B., ASI grant BIOMEX MicroColonial Fungi 063-R.0 to S.O.); the German Aerospace Center (DLR-grants: Department of Infrastructure and Management, Astrobiology Laboratories through a grant DLR-FuW-Project BIOMEX (2474128)/ Department of Radiation Biology supported by the grant DLR-FuE-Projekt ISS LIFE, Programm RF-FuW, Teilprogramm 475); the German Helmholtz Association through the Helmholtz-Alliance ‘‘Planetary Evolution and Life’’; the Spanish Ministry of Economy, Industry and Competitiveness (MINECO, project SUBLIMAS ‘‘SUrvival of Bacteria and LIchens on Mars Analogs and Space,’’ ESP2015- 69810-R, 2015, to R. de la Torre, and project ‘‘CTM2015- 64728-C2-1-R’’ to L.G. Sancho); and the National Academy of Sciences of Ukraine (grant 47/2017).

The next two rover missions to Mars, ESA/Roscosmos’s ExoMars2020 and NASA’s Mars2020, will carry for the first time Raman spectrometers potentially able to dectect traces of past or present life. To support and interpret future spectroscopic data correctly a better understanding of potential habitable environments and putative biosignatures, using analogue environments such as the Atacama Desert, is of paramount importance. In the Atacama Desert, one of the driest places on Earth, life has developed adaptive strategies to decreasing amounts of water: from refuges inside or below rocks as endoliths or hypoliths to inside salts in hygroscopic niches (Davila & Schulze-Makuch, 2016). In the hyperarid core one of the last refuges for life are inside salt crusts using deliquescence as a water source or being in the subsurface waiting for transitory episodes of increased moisture (Schulze-Makuch et al., 2018). These adaptive strategies might also apply to putative Martian life which endured a transition from a water rich past to the very harsh surface conditions of the present giving us clues on where to best look for traces of life on the Red Planet. Salt crusts and salt nodules are particularly interesting targets in this regard because they reside on or very near the surface and are thus easily accessible to future robotic missions. In the Atacama, salt nodules have been shown to host photosynthetic organisms containing easily identifiable pigments by Raman spectroscopy such as carotenoids and scytonemin (Vítek et al., 2014). One of the most damaging factors for life and its remains, both in the Atacama and on Mars, is solar radiation. To investigate the spatial distribution of potential Raman signatures in micro-niches we generated georeferenced 3D-reconstructions of the sampling areas using photogrammetry techniques and plotted the dose received according to the nodules’ orientation. We then analysed salt nodule along dry cut north-south thick sections using Raman mapping to infer any relations between the amount of light received and the presence of detectable signal. Preliminary data show an increased presence of carotenoids, scytonemin, and other biomolecules signals on the nodules oriented towards the south, which are the-more-protected sections of the nodules. References Davila, A.F. & Schulze-Makuch, D. (2016) The Last Possible Outposts for Life on Mars. Astrobiology 16:159–168. Schulze-Makuch, D., Wagner, D., Kounaves, S.P., Mangelsdorf, K., Devine, K.G., Vera, J.-P. de, Schmitt-Kopplin, P., Grossart, H.-P., Parro, V., Kaupenjohann, M., Galy, A., Schneider, B., Airo, A., Frösler, J., Davila, A.F., Arens, F.L., Cáceres, L., Cornejo, F.S., Carrizo, D., Dartnell, L., DiRuggiero, J., Flury, M., Ganzert, L., Gessner, M.O., Grathwohl, P., Guan, L., Heinz, J., Hess, M., Keppler, F., Maus, D., McKay, C.P., Meckenstock, R.U., Montgomery, W., Oberlin, E.A., Probst, A.J., Sáenz, J.S., Sattler, T., Schirmack, J., Sephton, M.A., Schloter, M., Uhl, J., Valenzuela, B., Vestergaard, G., Wörmer, L. & Zamorano, P. (2018) Transitory microbial habitat in the hyperarid Atacama Desert. PNAS 115:2670–2675. Vítek, P., Jehlička, J., Ascaso, C., Mašek, V., Gómez-Silva, B., Olivares, H. & Wierzchos, J. (2014) Distribution of scytonemin in endolithic microbial communities from halite crusts in the hyperarid zone of the Atacama Desert, Chile. FEMS Microbiology Ecology 90:351–366.

Cryptoendolithic microbial communities, discovered in the extremely cold, hyper-arid McMurdo Dry Valleys of Antarctica (Friedmann 1982), the most similar terrestrial environments to Mars surface (Wynn-Williams and Edwards 2000; Onofri et al., 2004), have been considered as a candidate in supporting the search of life in Mars exploration. In such harsh conditions, microorganisms grow in airspaces among mineral grains and show some adaptations, as the accumulation of protective pigments and compatible solutes, assuring their survival. In the frame of the Lichen and Fungi experiment (LIFE, P.I. Silvano Onofri; Onofri et al. 2012, 2015), small samples of these communities, were exposed to space, in the EXPOSE-E facility for 1.5 years, with an exposition to vacuum (10-7 to 10-4 Pa) (Horneck et al., 2010), galactic cosmic radiation (≤190mGy) (Berger et al., 2012), and the full spectrum of solar extraterrestrial electromagnetic radiation to which cryptoendolithic microorganisms demonstrated to survive (Scalzi et al., 2012). The search for trace of extant or extinct life is one of the main goals of the future space mission beyond Earth. The future rover missions ExoMars 2020 (ESA-Roscomos) and Mars 2020 (NASA) are exploring chemical and biological indicators of life, called biomarkers. A good biomarker must have a biogenic origin and thus must unequivocally be identified as possible trace of life. The detection of biomarkers on Mars was the aim of BIOMEX (BIOlogy and Mars Experiment) project, in which has been investigated the alteration of different biomarkers, after exposure to space and Mars-like conditions outside the International Space Station. In this contest, the aim of this work was to characterize fungal biomarkers from these exposed rock samples, with different approaches: i) Raman spectroscopy and InfraRed analyses, which has been considered excellent tools for the detection of inorganic and organic molecules, such as microbial pigments and ii) -omics approaches, as lipidomic and metabolomic techniques, performed to detect biological macromolecules and to determine their stability after space exposure. The focus of lipidomics and metabolomics has been on biomarker discovery, with the aim of identifying metabolites that are correlated with environmental exposures. The results suggest that microbial molecules can be detected through different techniques. In particular, our attention was focused on pigments, such as melanin and carotenoids that maintain their stability also after 1.5 years of space exposure. These results are of importance for the upcoming life-detection missions on Mars finalized for the search for past, extant or extinct life outside the Earth.