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Molecular and microscopic studies were performed to identify Chroococcidiopsis sp., an endolithic cyanobacterium, isolated from gypsum rocks of Atacama Desert (Chile). It was adapted to grow in mineral liquid medium, with 9 mM nitrate, bubbled with CO-enriched air (2.5 % v/v), and continuously illuminated with a white light of 70 μmol photons m s. The obtained biomass (productivity of 0.21 g L d) had a C/N ratio of 6.67, and it contained carbohydrates (45.40 % of dry weight), proteins (36.72 %), lipids (5.60 %) nucleic acids (3.90 %) and ashes (8.28 %). The lipid fraction was particularly rich in palmitic (29.86 % of total fatty acids), linoleic (18.20 %), palmitoleic (12.75 %), linolenic (10.92 %), stearic (9.64 %) and capric acid (6.29 %). Chroococcidiopsis sp. accumulated phycobiliproteins in a light-dependent process and produced 204 mg g, under incident light of 10 μmol photons·m·s, with a relative abundance of 40.9 % for phycocyanin, 23.3 % for phycoerythrin, and 35.8 % for allophycocyanin. The biomass from this cyanobacterium can be a good source of these pigments, especially APC (maximum of 95 mg g dw), which are of interest for pharmacological, cosmetic, and food industries., This work was funded by MCIU/AEI (Spain) and FEDER (UE) (grantreference PGC2018–094076-B-I00) and by a PhD-Grant from the R&DPlan of the University of Huelva, Spain, for Zaida Montero.

Raman imaging allows one to obtain spatially resolved chemical information in a nondestructive manner. Herein, we present analytical aspects of effective in situ and in vivo Raman imaging of algae and cyanobacteria from within their native rock habitats. Specifically, gypsum and halite inhabited by endolithic communities from the hyperarid Atacama Desert were analyzed. Raman imaging of these phototrophic colonization reveals a pigment composition within the aggregates that helps in understanding some of their adaptation strategies to survive in this harsh polyextreme environment. The study is focused on methodical aspects of Raman imaging acquisition and subsequent data processing. Point imaging is compared with line imaging in terms of their image quality, spatial resolution, spectral signal-to-noise ratio, time requirements, and risk of laser-induced sample alteration. The roles of excitation wavelength, exposure time, and step size of the imaging grid on successful Raman imaging results are also discussed. [Figure not available: see fulltext.], This study was supported by the Czech Republic Ministry of Education, Youth and Sports under the Funding Programme INTER-COST, grant number LTC18036 (COST Action NEUBIAS,CA15124), and by grant PGC2018-094076-B-I00 from MCIU/AEI (Spain) and FEDER (UE).

Este artículo está sujeto a una licencia CC BY-NC-ND 4.0, Biological colonization and later biodeterioration damage are well-known phenomena affecting cultural heritage. Glasses, as those from historical stained glass windows, are frequently biocolonized under certain conditions. There is still limited knowledge about biocolonization growth patterns on glass and safe removing procedures to act before irreversible biodegradation damage occurs. This paper presents the results of an experimental study in which biological colonization of three different model historical glasses, namely sodalime, lead crystal, and potash-lime silicate glasses, has been naturally induced. Glass samples were exposed for 13-15 months to natural conditions at the mountain region near Madrid (Spain) and monitored over time. After exposition, samples were first observed through fluorescence microscopy (FM), scanning electron microscopy (SEM), and UV-Vis-NIR spectrophotometry. Next, the Acticide® CF biocide was applied on glass samples exposed and were then again observed through the same techniques to evaluate its effect. Biocolonizers detected were algae, fungi, cyanobacteria and heterotrophic bacteria in different combinations and amounts and sometimes forming a biofilm. Soda-lime silicate glasses were the ones that showed the highest biocolonization growth, while lead crystal silicate glass appeared more biocolonized than the potash-lime one. The biofilm formed appeared intensely damaged and was practically no longer visible after the biocide treatment which proves its efficiency for removing glass biocolonization., This study was funded under research programmes TOP Heritage: Technologies in Heritage Sciences from the Regional Government of Madrid (Ref. S2018/NMT-4372), and Challenges of Society from the Spanish Ministry of Science and Innovation (Ref. PID2019- 104220RB-I00). Microscopy services from MNCN-CSIC are gratefully acknowledged, as well as to Thor Especialidades SA, Barcelona, Spain, for providing with the Acticide® CF biocide. Professional support from the TechnoHeritage network of Science and Technology for the Conservation of Cultural Heritage is also acknowledged. In addition, some of the authors wish to acknowledge professional support of the CSIC Interdisciplinary Thematic Platform Open Heritage: Research and Society (PTI-PAIS)

The efcacy of low-frequency ultrasound pulses in cell disaggregation of Chroococcidiopsis sp. aggregates has been studied as a possible strategy to improve the productivity and operation of the cultures. The modulation of the ultrasound pulses allowed to almost disaggregate most of the cyanobacterial aggregates completely while retaining cellular viability in terms of photosynthetic efciency. In this study we used a strain isolated from the endolithic habitat of the Atacama Desert, the driest desert in the world due to the extremely scarce rainfall, low level of relative humidity and extremely high incident solar radiation. To survive these conditions and reduce the cell exposure to the incident UV radiation, Chroococcidiopsis sp. grows in the form of aggregates, diminishing the associated photo-oxidative damage. However, this adaptation strategy can reduce the availability of both light and nutrients to the growing cells. This study showed that the low-frequency ultrasound pulses were efcient in disaggregating Chroococcidiopsis sp. aggregates, improving light and nutrient availability to the cells. Our results revealed also that the modulated use of ultrasound pulses resulted in a decreased cell sedimentation velocity which becomes advantageous at large scale. The length of the ultrasound pulses can be optimized to achieve complete disaggregation of the aggregates without afecting cell viability. The preservation of cell viability is considered an advantage for eventual large-scale production as disaggregating of the aggregates can result in more homogeneous cultures with less energy needed to perform mechanical agitation. Additionally, our results indicated an improved growth of cyanobacterium in disaggregated cultures., Open Access funding provided thanks to the CRUE-CSIC agreement with Springer Nature.

Cyanobacteria exposed to high solar radiation make use of a series of defense mechanisms, including avoidance, antioxidant systems, and the production of photoprotective compounds such as scytonemin. Two cyanobacterial strains of the genus Chroococcidiopsis from the Atacama Desert – which has one of the highest solar radiation levels on Earth- were examined to determine their capacity to protect themselves from direct photosynthetically active (PAR) and ultraviolet radiation (UVR): the UAM813 strain, originally isolated from a cryptoendolithic microhabitat within halite (NaCl), and UAM816 strain originally isolated from a chasmoendolithic microhabitat within calcite (CaCO3). The oxidative stress induced by exposure to PAR or UVR C PAR was determined to observe their short-term response, as were the long-term scytonemin production, changes in metabolic activity and ultrastructural damage induced. Both strains showed oxidative stress to both types of light radiation. The UAM813 strain showed a lower acclimation capacity than the UAM816 strain, showing an ever-increasing accumulation of reactive oxygen species (ROS) and a smaller accumulation of scytonemin. This would appear to reflect differences in the adaptation strategies followed to meet the demands of their different microhabitats., This study was supported by grant PGC2018-094076-B-I00 from MCIU/AEI (Spain) and FEDER (UE). MC was supported by grant BES 2014-069106 from the Spanish Ministry of Science and Innovation (MCINN).

Endolithic microhabitats have been described as the last refuge for life in arid and hyper-arid deserts where life has to deal with harsh environmental conditions. A number of rock substrates from the hyper-arid Atacama Desert, colonized by endolithic microbial communities such as halite, gypsum crusts, gypcrete, calcite, granite and ignimbrite, have been characterized and compared using different approaches. In this work, three different endolithic microhabitats are described, each one with a particular origin and architecture, found within a lithic substrate known as gypcrete. Gypcrete, an evaporitic rock mainly composed of gypsum (CaSO4 ⋅ 2H2O) and collected in the Cordón de Lila area of the desert (Preandean Atacama Desert), was found to harbour cryptoendolithic (within pore spaces in the rock), chasmoendolithic (within cracks and fissures) and hypoendolithic (within microcave-like pores in the bottom layer of rock) microhabitats. A combination of microscopy investigation and high-throughput sequencing approaches were used to characterize the endolithic communities and their habitats at the microscale within the same piece of gypcrete. Microscopy techniques revealed differences in the architecture of the endolithic microhabitats and the distribution of the microorganisms within those microhabitats. Cyanobacteria and Proteobacteria were dominant in the endolithic communities, of which the hypoendolithic community was the least diverse and hosted unique taxa, as a result of less access to sun radiation. These results show, for the first time, that the differences in the architecture of a microhabitat, even within the same piece of a lithic substrate, play an essential role in shaping the diversity and composition of endolithic microbial communities., The work of MCC was supported by grant BES 2014-069106 from the Spanish Ministry of Science and Innovation (MCINN). We also acknowledge institutional support from the Unit of Information Resources for Research at the Unit of Information Resources for Research at the "Consejo Superior de Investigaciones Científicas" (CSIC) for the article-processing charges contribution

The modern Martian surface is unlikely to be habitable due to its extreme aridity among other environmental factors. This is the reason why the hyperarid core of the Atacama Desert has been studied as an analog for the habitability of Mars for more than 50 years. Here we report a layer enriched in smectites located just 30 cm below the surface of the hyperarid core of the Atacama. We discovered the clay-rich layer to be wet (a phenomenon never observed before in this region), keeping a high and constant relative humidity of 78% (a 0.780), and completely isolated from the changing and extremely dry subaerial conditions characteristic of the Atacama. The smectite-rich layer is inhabited by at least 30 halophilic species of metabolically active bacteria and archaea, unveiling a previously unreported habitat for microbial life under the surface of the driest place on Earth. The discovery of a diverse microbial community in smectite-rich subsurface layers in the hyperarid core of the Atacama, and the collection of biosignatures we have identified within the clays, suggest that similar shallow clay deposits on Mars may contain biosignatures easily reachable by current rovers and landers., The reported research is a contribution from the Project “MarsFirstWater”, funded by the European Research Council, ERC Consolidator Grant No. 818602 to AGF and by the Human Frontiers Science Program grant nº RGY0066/2018 to AAB. This work was also supported by the grant PGC2018-094076-B-I00 to JW and CA from the Ministry of Science, Innovation and Universities of Spain.

Huang et al. (1) describe a supposed mechanism of water extraction from gypsum by cyanobacteria sampled from endoliths inhabiting Ca sulfates in the Atacama Desert, and cultivated in the laboratory. The authors claim that the phase transformation from gypsum (CaSO4·2H2O) to anhydrite (CaSO4) (G→A) occurred under “dry conditions” in the contact zone between a “dry biofilm” and the gypsum, where only {011} planes of gypsum are transformed to anhydrite, supposedly providing water for cyanobacteria. This work (1) has a number of major conceptual problems, as follows. First, the authors show the presence of gypsum and/or anhydrite in the inoculated Ca sulfate samples using X-ray diffraction and, incompletely, by Fourier transform infrared spectroscopy techniques (2), and not by Raman (3) or transmission electron microscopy. Selected area electron diffraction (SAED) data in ref. 1 do not allow any mineral identification, due to their differences with … [↵][1]1To whom correspondence may be addressed. Email: j.wierzchos{at}mncn.csic.es or oartieda{at}unex.es. [1]: #xref-corresp-1-1

Cyanobacteria exposed to high solar radiation make use of a series of defense mechanisms, including avoidance, antioxidant systems, and the production of photoprotective compounds such as scytonemin. Two cyanobacterial strains of the genus Chroococcidiopsis from the Atacama Desert – which has one of the highest solar radiation levels on Earth- were examined to determine their capacity to protect themselves from direct photosynthetically active (PAR) and ultraviolet radiation (UVR): the UAM813 strain, originally isolated from a cryptoendolithic microhabitat within halite (NaCl), and UAM816 strain originally isolated from a chasmoendolithic microhabitat within calcite (CaCO3). The oxidative stress induced by exposure to PAR or UVR + PAR was determined to observe their short-term response, as were the long-term scytonemin production, changes in metabolic activity and ultrastructural damage induced. Both strains showed oxidative stress to both types of light radiation. The UAM813 strain showed a lower acclimation capacity than the UAM816 strain, showing an ever-increasing accumulation of reactive oxygen species (ROS) and a smaller accumulation of scytonemin. This would appear to reflect differences in the adaptation strategies followed to meet the demands of their different microhabitats.

The Atacama Desert is known to be the place on Earth with one of the highest solar radiation limiting the presence of life to endolithic microhabitats and soil microbial ecosystems. Endolithic microbial communities are supported by photosynthetic primary producers, mainly cyanobacteria, which can be injured by UVR. Nevertheless, cyanobacteria exposed to high solar radiation and its harmful effects have developed a series of defense mechanisms: avoidance, antioxidant systems or production of photoprotective compounds such as scytonemin among others. Scytonemin is a liposoluble pigment whose absorption maxima are located in UVA and UVC range and highly absorbing in the UVB range. In order to elucidate the protection capacity of endolithic cyanobacteria against harmful radiation, two cyanobacterial strains from Chroococcidiopsis genus were isolated from different endolithic microhabitats in the Atacama Desert: UAM813 strain, originally from the cryptoendolithic microhabitat of halite (NaCl), and UAM816 strain from chasmoendolithic microhabitat of calcite (CaCO3). Both were exposed to PAR and UVR+PAR conditions studying their short-term response, as oxidative stress and long-term response, as scytonemin production, metabolic activity and ultrastructural damage. The observed response of both strains reveals a high sensitivity to direct light exposure, even to PAR. The differences in their acclimation suggest specific adaptation strategies related to their original microhabitat, revealing their protective potential and the strain specific environmental pressure selection to inhabit different microhabitats and exposed to different light conditions.ImportanceCyanobacteria are photosynthetic prokaryotes that inhabit most types of illuminated environments, even the endolithic microhabitats in cold and hot deserts. The environmental pressure caused by the extreme solar irradiation in the Atacama Desert involve that only those cyanobacterial strains able to cope with it can be found in these endolithic communities, usually dominated by members belonging to the extremotolerant Chroococcidiopsis genus. Here, a comprehensive analysis of multiple lines of defense against harmful sun radiation was conducted to diagnose the response of two Chroococcidiopsis strains isolated from different endolithic microhabitats and lithic substrates, and identify its relation with the original microenvironmental conditions of each strain. Our results contribute to a better understanding of the acclimation strategies developed by these cyanobacterial strains and its potential protective role for the whole endolithic microbial community.

Fibers are found in all environments. However, the impact of their presence on ecosystems and human health is not yet well understood, especially in the case of the atmosphere. In this work, we presented evidence that fibers traveling through the atmosphere act as vectors to spread microorganisms. Here, we investigated the presence of viable microorganisms on fibers collected within and beyond the planetary boundary layer during flights of C-212 aircraft over Central Spain. In total, seven fibers, six of which transported viable microorganisms, were isolated in two flights. The viability of the microorganisms was determined by confocal microscopy by means of the fluorescent probes SYBR-Green to detect microorganisms and CTC redox dye to assess their cellular respiration activity. The fibers that transported viable microorganisms were spectroscopically analyzed by micro-FTIR and identified as wool-silk and cellulose-cotton. Taken together, the results demonstrated that fibers host viable microorganisms when traveling through the lower free troposphere. The authors acknowledge the financial support provided by the Spanish Government (CTM2016-74927-C2-1-R/2-R, CGL2017-92086-EXP, PGC2018-094076-B-100, and RTI2018-094867-B-I00). C.E. thanks the Spanish Ministry of Science, Innovation and Universities for the award of a predoctoral grant (FPI). C-212 airborne access was generously provided by INTA, coordinated by the Aerial Platform for Research team and with the logistic and operational support of Group 47 of the Spanish Air Force; With funding from the Spanish government through the "María de Maeztu Unit of Excellence" accreditation (MDM-2017-0737). Peer review

An extreme halophilic archaeon, strain SGH1, is a novel microorganism isolated from endolithic microbial communities colonizing halites at Salar Grande, Atacama Desert, in northern Chile. Our study provides structural, biochemical, genomic, and physiological information on this new isolate living at the edge of the physical and chemical extremes at the Atacama Desert. SGH1 is a Gram-negative, red-pigmented, non-motile unicellular coccoid organism. Under the transmission electron microscope, strain SGH1 showed an abundant electro-dense material surrounding electron-lucent globular structures resembling gas vacuoles. Strain SGH1 showed a 16S rRNA gene sequence with a close phylogenetic relationship to the extreme halophilic archaea Haloterrigena turkmenica and Haloterrigena salina and has been denominated Haloterrigena sp. strain SGH1. Strain SGH1 grew at 20–40°C (optimum 37°C), at salinities between 15 and 30% (w/v) NaCl (optimum 25%) and growth was improved by addition of 50 mM KCl and 0.5% w/v casamino acids. Growth was severely restricted at salinities below 15% NaCl and cell lysis is avoided at a minimal 10% NaCl. Maximal concentrations of magnesium chloride and sodium or magnesium perchlorates that supported SGH1 growth were 0.5 and 0.15M, respectively. Haloterrigena sp. strain SGH1 accumulates bacterioruberin (BR), a C xanthophyll, as the major carotenoid. Total carotenoids in strain SGH1 amounted to nearly 400 μg BR per gram of dry biomass. Nearly 80% of total carotenoids accumulated as geometric isomers of BR: all-trans-BR (50%), 5-cis-BR (15%), 9-cis-BR (10%), 13-cis-BR (4%); other carotenoids were dehydrated derivatives of BR. Carotenogenesis in SGH1 was a reversible and salt-dependent process; transferring BR-rich cells grown in 25% (w/v) NaCl to 15% (w/v) NaCl medium resulted in depigmentation, and BR content was recovered after transference and growth of unpigmented cells to high salinity medium. Methanol extracts and purified BR isomers showed an 8–9-fold higher antioxidant activity than Trolox or β-carotene. Both, plasma membrane integrity and mitochondrial membrane potential measurements under acute 18-h assays showed that purified BR isomers were non-toxic to cultured human THP-1 cells., This work was supported by Grant CeBiB FB-0001, Comisión Nacional de Investigación Científica y Tecnológica (CONICYT), Chile, Semillero de Investigación, Grant SI-5305 by Universidad de Antofagasta, Chile, and Grant PGC2018-094076-B-I00 from Ministry of Science, Innovation and Universities (MCIU/AEI) (Spain) and FEDER (UE) to JW and CA. We acknowledge the support given by grant FONDEQUIP EQM-120137., With the funding support of the ‘Severo Ochoa Centre of Excellence’ accreditation (CEX2019-000928-S), of the Spanish Research Agency (AEI)

Microorganisms that live in extreme environments have recently attracted the attention of scientists worldwide. Among the harshest environments faced by microbial life are hyperarid deserts. The Preandean and Andean “Puna” form part of the Atacama Desert, and these zones are considered the most life-limited places on Earth. As a consequence of two records of extreme environment—the lowest rainfall and highest surface UV radiation/total solar irradiance ever measured on Earth—the Atacama is considered a polyextreme environment. There is evidence today suggesting that successful microbial colonization in this desert, at the limits of microbial life, is the result of a combination of adaptive strategies used by microorganisms to avoid excess solar irradiance and extreme evapotranspiration rates, as they make good use of the rock’s “habitable architecture”. This new concept evokes all the internal structures of a rock that are essential as a habitat for microbial life. This chapter describes the adversity of life conditions and several microbial habitats in the Preandean Depression and Andean region of the Atacama Desert capable of sustaining active microbial ecosystems. All are endolithic habitats (within rocks) providing refuges or islands of life in the desert.

Endolithic ecosystems are considered environmental refuges for life in arid and hyperarid deserts around the world. The microbial communities colonizing these habitats are of special interest due to their ability to live in a polyextreme environment characterized by scarcity of water and extreme solar radiation. The Preandean Depression of the Atacama Desert harbors a number of these endolithic ecosystems, where various substrates are colonized by rich and diverse communities of phototrophic and heterotrophic microorganisms. Recent studies revealed that these communities are dominated by Cyanobacteria, Actinobacteria, Proteobacteria, and Chloroflexi phyla. Studies of the photosynthetic primary producers demonstrated their essential role in supporting the community and uncovered unique adaptation strategies against extreme solar radiation. The diversity found in the endolithic communities of the Preandean Depression, both at the phylogenetic and functional levels, was found to be strongly dependent on water availability and associated with the specific architecture of the substrate. This chapter describes the composition, functioning, and adaptation strategies of microbial communities inhabiting the endolithic substrates found in the Preandean zone of the Atacama Desert. We address the principal biotic and abiotic drivers explaining their diversity from a multidisciplinary perspective.

© The Author(s) 2018., The hyperarid core of the Atacama Desert, the driest and oldest desert on Earth, has experienced a number of highly unusual rain events over the past three years, resulting in the formation of previously unrecorded hypersaline lagoons, which have lasted several months. We have systematically analyzed the evolution of the lagoons to provide quantitative field constraints of large-scale impacts of the rains on the local microbial communities. Here we show that the sudden and massive input of water in regions that have remained hyperarid for millions of years is harmful for most of the surface soil microbial species, which are exquisitely adapted to survive with meager amounts of liquid water, and quickly perish from osmotic shock when water becomes suddenly abundant. We found that only a handful of bacteria, remarkably a newly identified species of Halomonas, remain metabolically active and are still able to reproduce in the lagoons, while no archaea or eukaryotes were identified. Our results show that the already low microbial biodiversity of extreme arid regions greatly diminishes when water is supplied quickly and in great volumes. We conclude placing our findings in the context of the astrobiological exploration of Mars, a hyperarid planet that experienced catastrophic floodings in ancient times., The research leading to these results is a contribution from the Project “icyMARS”, funded by the European Research Council, Starting Grant No. 307496. A.G.F. and V.P. acknowledge funding support from Grant No. ESP2015-69540-R (MINECO/FEDER). We thank V. Souza-Egipsy and BIOPHYM (IEM-CSIC) for TEM observations; C. Arroyo (MNCN-CSIC), and the Geomaterials 2 Program (S2013/MIT_2914), financed by the Comunidad Autónoma de Madrid and the European Social Fund; and Y. Blanco for providing LDChip antibody microarrays.

Microbial communities composed by eukaryotic microorganisms: melanized fungi and algae colonizing the interior of gypsum crusts in hyperarid zone of the Atacama desert were studied by Raman microspectroscopy. Melanin pigments present in fungi cell walls were identified directly in the colonized gypsum crust and on the cultivated culture of the fungi from Neocatenulostroma genus. Two signature broad bands located at wavenumbers 1605–1580 cm −1 and around 1350 cm −1 have been detected and confirmed melanin as a main fungal protective pigment.

Laser irradiation of lichen thalli on heritage stones serves for the control of epilithic and endolithic biolog- ical colonizations. In this work we investigate rock samples from two quarries traditionally used as source of monumental stone, sandstone from Valonsadero (Soria, Spain) and granite from Alpedrete (Madrid, Spain), in order to find conditions for efficient laser removal of lichen thalli that ensure preservation of the lithic substrate. The samples presented superficial areas colonized by different types of crustose lichens, i.e. Candelariella vitellina, Aspicilia viridescens, Rhizocarpon disporum and Protoparmeliopsis muralis in Valonsadero samples and P. cf. bolcana and A. cf. contorta in Alpedrete samples. A comparative laser cleaning study was carried out on the mentioned samples with ns Q-switched Nd:YAG laser pulses of 1064 nm (fundamental radiation), 355 nm (3rd harmonic) and 266 nm (4th harmonic) and sequences of IR-UV pulses. A number of techniques such as UV-Vis absorption spectroscopy, stereomicroscopy, scan- ning electron microscopy (SEM) at low vacuum, SEM with backscattered electron imaging (SEM-BSE), electron dispersive spectroscopy (EDS) and FT-Raman spectroscopy were employed to determine the best laser irradiation conditions and to detect possible structural, morphological and chemical changes on the irradiated surfaces. The results show that the laser treatment does not lead to the complete removal of the studied lichen thalli, although clearly induces substantial damage, in the form of loss of the lichen upper cortex and damage to the algal layer. In the medium term these alterations could result in the destruction of the lichen thalli, thus providing a degree of control of the biodeterioration processes of the lithic substrate and reducing the chances of subsequent lichen recolonization., The present work was supported by programs Geomateriales 2 (S2013/MIT-2914) financed by Comunidad de Madrid and Structural Funds (FSE and FEDER) and IPERION-CH Ref. H2020-INFRAIA-2014-2015, no. 654028. MO thanks CSIC for contract. SPO is supported by the Spanish Ministry of Economy and Competitiveness (MINECO) grant RYC-2014-16784. We are grateful to M. Furio, A. Jorge and L. Tormo from Servicio de Microscopía, MNCN-CSIC.

Since the description of microbial communities colonizing Ca-sulfate crusts in the Atacama Desert, there has been much interest in the mechanisms that could lead to the formation and preservation of biosignatures or microbial fossils of these communities. A key to understanding physico-chemical processes of taphonomy and early diagenesis is to examine microfossils in their natural environment. In this study, we characterize organomineral traces and microbial fossils found around microbial communities present in these Ca-sulfate crusts. Through scanning electron microscopy, microanalytical (EDS) and Raman spectroscopy techniques, calcium carbonate precipitates were detected around remnants of cryptoendolithic algae beneath the crust surface. As what seems to be the final step in the organomineralization of these cryptoendolithic communities, we also observed alga cell remains permineralized by Mg-Si-rich minerals inside gypsum crystals. Additionally, Mg-Si bearing minerals formed a web-like structure within the hypoendolithic cyanobacterial habitat via permineralization of extracellular polymeric substances. Our observations indicate that despite the extremely hyperarid environment, microenvironmental conditions may be appropriate for the formation of biosignatures and microbial fossils of extinct endolithic microbial communities. A model of the possible organomineralization processes involved is presented., This work was supported by grants awarded by MINECO: CGL2013-42509P and CTM2015-64728-C2-2-R.

Raman imaging allows one to obtain spatially resolved chemical information in a nondestructive manner. Herein, we present analytical aspects of effective in situ and in vivo Raman imaging of algae and cyanobacteria from within their native rock habitats. Specifically, gypsum and halite inhabited by endolithic communities from the hyperarid Atacama Desert were analyzed. Raman imaging of these phototrophic colonization reveals a pigment composition within the aggregates that helps in understanding some of their adaptation strategies to survive in this harsh polyextreme environment. The study is focused on methodical aspects of Raman imaging acquisition and subsequent data processing. Point imaging is compared with line imaging in terms of their image quality, spatial resolution, spectral signal-to-noise ratio, time requirements, and risk of laser-induced sample alteration. The roles of excitation wavelength, exposure time, and step size of the imaging grid on successful Raman imaging results are also discussed. Graphical abstract.

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The Raman imaging method was successfully applied for mapping the distribution of biomolecules (e.g., pigments) associated with cryptoendolithic and hypoendolithic microorganisms, as well as the inorganic host mineral matrix that forms the habitat for the biota. To the best of our knowledge, this is the first comprehensive study in the field of geomicrobiology based on this technique. The studied microbial ecosystem was located nearly 3000 m above sea level within the driest desert on Earth, the Atacama in Chile. Enhancement of carotenoid Raman signal intensity close to the surface was registered at different areas of endolithic colonization dominated by algae, with cyanobacteria present as well. This is interpreted as an adaptation mechanism to the excessive solar irradiation. On the other hand, cyanobacteria synthesize scytonemin as a passive UV-screening pigment (found at both the hypoendolithic and cryptoendolithic positions). The distribution of the scytonemin Raman signal was mapped simultaneously with the surrounding mineral matrix. Thus, mapping was done of the phototrophic microorganisms in their original microhabitat together with the host rock environment. Important information which was resolved from the Raman imaging dataset of the host rock is about the hydration state of Ca-sulfate, demonstrated on the presence of gypsum (CaSO·2HO) and the absence of both anhydrite (CaSO) and bassanite (CaSO·1/2HO). Obtaining combined “in situ” simultaneous information from the geological matrix (inorganic) together with the microbial biomolecules (organic) is discussed and concluded as an important advantage of this technique. We discuss how selection of the laser wavelength (785 and 514.5-nm) influences the Raman imaging results., PV wishes to thank the Ministry of Education, Youth and Sport for the NPU I grant (Grant No. LO1415); JW, OA, and CA wish to thank MINECO, Spain, for financial support via Grant No. CGL-2013-42509P.

Halite endoliths in the Atacama Desert represent one of the most extreme ecosystems on Earth. Cultivation-independent methods were used to examine the functional adaptations of the microbial consortia inhabiting halite nodules. The community was dominated by haloarchaea and functional analysis attributed most of the autotrophic CO2 fixation to one unique cyanobacterium. The assembled 1.1 Mbp genome of a novel nanohaloarchaeon, Candidatus Nanopetramus SG9, revealed a photoheterotrophic life style and a low median isoelectric point (pI) for all predicted proteins, suggesting a 'salt-in' strategy for osmotic balance. Predicted proteins of the algae identified in the community also had pI distributions similar to 'salt-in' strategists. The Nanopetramus genome contained a unique CRISPR/Cas system with a spacer that matched a partial viral genome from the metagenome. A combination of reference-independent methods identified over 30 complete or near complete viral or proviral genomes with diverse genome structure, genome size, gene content and hosts. Putative hosts included Halobacteriaceae, Nanohaloarchaea and Cyanobacteria. Despite the dependence of the halite community on deliquescence for liquid water availability, this study exposed an ecosystem spanning three phylogenetic domains, containing a large diversity of viruses and predominance of a 'salt-in' strategy to balance the high osmotic pressure of the environment.

Biodeterioration damage is an important issue in conservation and restoration of built heritage, especially when ceramic materials are used. Biological colonization of ceramic roofing tiles by lichens is a common phenomenon. However, there are no reports to date of lichens removal from unglazed roofing tiles for conservation purposes. This paper for the first time reveals the results of a combined procedure undertaken to assess the removal of lichens on different kinds of unglazed ceramic roofing tiles by treatments based on both dual sequential laser irradiation and treatment using Acticide CF biocide. Three species of lichens were identified: Verrucaria nigrescens, Calogaya decipiens and Pyrenodesmia teicholyta. The chemical and mineralogical composition of roofing tiles were characterized by X-ray fluorescence (XRF) spectrometry, optical polarized petrographic microscopy, and X-ray diffraction (XRD). Laser irradiation was accomplished by applying sequences of nanosecond laser pulses at two wavelengths (1064 and 266 nm). After dual sequential laser irradiation a biocide was applied. To assess the combined effect of both treatments several techniques were used, including stereo and fluorescence (FM) microscopies, scanning (SEM) and transmission (TEM) electron microscopies, and FT-Raman spectroscopy. Chemical composition of the analyzed roofing tiles was shown as a relevant factor regarding the degree of interaction between the biological colonization and the substrate, and hence, the bioweathering effect. The combined procedure has proved to be very effective in ablating cortical layers in all species, or even complete areolae in V. nigrescens, enhancing biocide effect in the thalli of C. decipiens and P. teicholyta, and producing the complete damage of both bionts.

The extremely harsh conditions of hyperarid deserts are a true challenge for microbial life. Microorganisms thriving in such polyextreme environments are fascinating as they can tell us more about life, its strategies and its boundaries than other groups of organisms. The Atacama Desert (North Chile) holds two world records of extreme environmental characteristics: the lowest rainfall and greatest surface ultraviolet radiation and total solar irradiance ever measured on Earth. Despite these limiting conditions for life, we recently identified several remarkable examples of endolithic habitats colonized by phototrophic and heterotrophic microorganisms in the hyperarid core of the Atacama Desert., This work was supported by a grant CGL2013-42509P for all authors from MINECO, Spain.

In hyperarid deserts, endolithic microbial communities colonize the rocks’ interior as a survival strategy. Yet, the composition of these communities and the drivers promoting their assembly are still poorly understood. We analysed the diversity and community composition of endoliths from four different lithic substrates – calcite, gypsum, ignimbrite and granite – collected in the hyperarid zone of the Atacama Desert, Chile. By combining microscopy, mineralogy, spectroscopy and high throughput sequencing, we found these communities to be highly specific to their lithic substrate, although they were all dominated by the same four main phyla, Cyanobacteria, Actinobacteria, Chloroflexi and Proteobacteria. Our finding indicates a fine scale diversification of the microbial reservoir driven by substrate properties. The data suggest that the overall rock chemistry and the light transmission properties of the substrates are not essential drivers of community structure and composition. Instead, we propose that the architecture of the rock, i.e., the space available for colonization and its physical structure, linked to water retention capabilities, is ultimately the driver of community diversity and composition at the dry limit of life., This work was funded by grants NNX15AP18G and NNX15AK57G from NASA, grant DEB1556574 from the NSF to JDR and grant CGL2013‐42509P from the Spanish Ministry of Economy, Industry and Competitiveness to JW, CA, OA, JDR and MCC.

8 páginas y 8 figuras, Laser removal of biodeteriogen layers warrants detailed studies due to the advantages it brings withrespect to mechanical elimination or the use of biocides. We have investigated elimination of biologicalcrusts on dolomite stones from heritage sites in central Spain. The samples were colonized by epilithiccrustose lichens of different species, such as Caloplaca sp. and Verrucaria nigrescens. A comparative studywas carried out by applying infrared (1064 nm) and ultraviolet (355 nm) nanosecond laser pulses andsequences pulses of the two wavelengths using a Q-switched Nd:YAG system. To detect anatomical andultrastructural damage to the lichens, and to assess possible morphological and chemical changes on theunderlying stone induced by laser irradiation, we used stereomicroscopy, scanning electron microscopywith backscattered electron imaging and Fourier transform Raman spectroscopy. The optimal conditionsfor removal of the colonization crust, while ensuring preservation of the lithic substrate, were obtainedfor dual infrared-ultraviolet sequential irradiation., Work funded by Program Geomateriales 2 (S2013/MIT-2914)financed by Comunidad de Madrid and Structural Funds (FSE andFEDER) S2013/MIT-2914. M.O acknowledges contract by CSIC. Weare grateful to M. Furio, A. Jorge and L. Tormo from Servicio de Micro-scopia, MNCN-CSIC and to support by Red de Ciencia y Tecnología para la Conservación del Patrimonio Cultural (Ministerio de Ciencia e Innovación, Spain, Acción Complementaria HAR2010-11432-E).

Miocene continental saltpans are scattered in the Central Valley of the Atacama Desert, one of the driest regions on Earth. These evaporitic deposits are hydrologically inactive, and are detached from groundwater brines or aquifers. The surface of the saltpans, also known as salars, comprises desiccation polygons, commonly with nodular salt structures along their sides. The morphology and bulk mineralogy of salt polygons differs between and within salars, and the shape and internal structure of salt nodules varies between different polygon types. Based on field observation, and mineralogy and crystallography data, we generated a conceptual model for the genesis and evolution of these surface features, whereby rare rainfall events are responsible for the transformation of desiccation salt polygons and the initial formation of salt nodules along polygon borders. In addition, frequent, but less intense, deliquescence events further drive the evolution of salt nodules, resulting in a characteristic internal structure that includes laminations, and changes in porosity and crystal morphologies. As a result, and despite the extreme dryness, the surfaces of fossil salars are dynamic on timescales of several years to decades, in response to daily cycles in atmospheric moisture, and also to rare and meager rainfall events. We propose that fossil salars in the Atacama Desert represent an end stage in the evolution of evaporitic deposits under extreme and prolonged dryness.

© 2015, Springer Science+Business Media New York. The “Great View of Lisbon” is one of the most remarkable blue-and-white Portuguese tile panels, which depicts the city before the tragic earthquake of 1755. This panel presents visible colored alteration in the glaze of tiles both from the exhibition and the depository of the Museu Nacional do Azulejo (Portugal). This work is a contribution to identify the origin of green stains in the glaze by using microbiological techniques, scanning electron microscopy with back-scattered electron imaging (SEM-BSE), Raman spectroscopy, and proton-induced X-ray emission (PIXE). The microbiological assays showed that the filamentous fungi were detected only at a frequency, JW and CA are thankful for the support by Grant CGL2010-16004 from the Spanish Ministry of Science and Innovation.

The Atacama Desert is the driest and one of the most life-limiting places on Earth. Despite the extreme conditions, microbial endolithic communities have been found inside halite rocks. The presence of these microbial communities is possible due to the hygroscopic properties of evaporitic rocks composed of sodium chloride. It is important to elucidate every possible water source in such a hyperarid environment. Therefore, in the present study, an artificial neural network (ANN) based model has been designed to predict the presence of liquid water on the surface of halite pinnacles. The model predicts the moisture formation using two basic meteorological variables, air temperature, and air relative humidity. ANNs have been successfully employed for the first time as a tool for predicting the appearance of liquid water, a key factor for the endolithic microbial communities living in the driest part of the Atacama Desert. The model developed is able to correctly predict the formation of water on the surface of the halite pinnacles 83% of the cases. We anticipate the future application of this model as an important tool for the prediction of the water availability and therefore potential habitability of lithic substrates in extreme environments on Earth and perhaps elsewhere.

Halite deposits from the hyperarid zone of the Atacama Desert reveal the presence of endolithic microbial colonization dominated by cyanobacteria associated with heterotrophic bacteria and archaea. Using the λ-scan confocal laser scanning microscopy (CLSM) option, this study examines the autofluorescence emission spectra produced by single cyanobacterial cells found inside halite rocks and by their photosynthetic pigments. Photosynthetic pigments could be identified according to the shapes of the emission spectra and wavelengths of fluorescence peaks. According to their fluorescence fingerprints, three groups of cyanobacterial cells were identified within this natural extreme microhabitat: (i) cells producing a single fluorescence peak corresponding to the emission range of phycobiliproteins and chlorophyll a, (ii) cells producing two fluorescence peaks within the red and green signal ranges, and (iii) cells emitting only low-intensity fluorescence within the nonspecific green fluorescence signal range. Photosynthetic pigment fingerprints emerged as indicators of the preservation state or viability of the cells. These observations were supported by a cell plasma membrane integrity test based on Sytox Green DNA staining and by transmission electron microscopy ultrastructural observations of cyanobacterial cells. © 2014, American Society for Microbiology., This study was funded by grant CGL2010-16004 from MINECO

Summary The Atacama Desert is one of the oldest and driest deserts in the world, and its hyper-arid core is described as ‘the most barren region imaginable’. We used a combination of high-throughput sequencing and microscopy methods to characterize the endolithic microbial assemblages of halite pinnacles (salt rocks) collected in several hyper-arid areas of the desert. We found communities dominated by archaea that relied on a single phylotype of Halothece cyanobacteria for primary production. A few other phylotypes of salt-adapted bacteria and archaea, including Salinibacter, Halorhabdus, and Halococcus were major components of the halite communities, indicating specific adaptations to the unique halite environments. Multivariate statistical analyses of diversity metrics clearly separated the halite communities from that of the surrounding soil in the Yungay area. These analyses also revealed distribution patterns of halite communities correlated with atmospheric moisture. Microbial endolithic communities from halites exposed to coastal fogs and high relative humidity were more diverse; their archaeal and bacterial assemblages were accompanied by a novel algae related to oceanic picoplankton of the Mamiellales. In contrast, we did not find any algae in the Yungay pinnacles, suggesting that the environmental conditions in this habitat might be too extreme for eukaryotic photosynthetic life.

The hyperarid core of the Atacama Desert is one of the driest and most inhospitable places on Earth, where life is most commonly found in the interior of rocks (i.e., endolithic habitats). Due to the extreme dryness, microbial activity in these habitats is expected to be low; however, the rate of carbon cycling within these microbial communities remains unknown. We address this issue by characterizing the isotopic composition (13C and 14C) of phospholipid fatty acids (PLFA) and glycolipid fatty acids (GLFA) in colonized rocks from four different sites inside the hyperarid core. δ13C results suggest that autotrophy and/or quantitative conversion of organic matter to CO2 are the dominant processes occurring with the rock. Most Δ14C signatures of PLFA and GLFA were consistent with modern atmospheric CO2, indicating that endoliths are using atmospheric carbon as a primary carbon source and are also cycling carbon quickly. However, at one site the PLFA contained 14C from atmospheric nuclear weapons testing that occurred during the 1950s and 1960s, indicating a decadal rate of carbon cycling. At the driest site (Yungay), based on the relative abundance and 14C content of GLFA and PLFA, there was evidence of possible preservation. Hence, in low-moisture conditions, glycolipids may persist while phospholipids are preferentially hydrolyzed. Key Words: Endoliths—Extremophile—Carbon isotopes—Radiocarbon—Lipids. Astrobiology 13, 607–616.

Rhyolitic obsidian was reacted with natural waters to study the effect of water chemistry and biological activity on the composition and formation mechanisms of clay. Two sets of experiments (18 months, 6 years) used fresh, hypersaline water (Mg-Na-SO4-Cl- and NaCl-rich) and seawater. The 6-year experiments produced the transformation of obsidian into quartz, apparently by in situ re-crystallization (Cuadros et al., 2012). The most abundant neoformed clay was dioctahedral (typically montmorillonite), indicating chemical control by the glass (where Al > Mg). Altered glass morphology and chemistry in the 18-months experiments indicated in situ transformation to clay. Magnesium-rich (saponite) clay formed under water-chemistry control in the bulk and within biofilms with elevated Mg concentration (Cuadros et al., 2013). The contact between microbial structures and glass was very intimate. Glass transformation into quartz may be due to some characteristic of the obsidian and/or alteration conditions. Such combination needs not to be uncommon in nature and opens new possibilities of quartz origin.

Summary Endolithic cyanobacteria are found in halite nodules in the hyperarid core of the Atacama Desert. Using Pulse Amplitude Modulated Fluorometry, we show here that photosynthetic systems of these cyanobacteria become active when the relative humidity rises above 70% and the salt becomes wet by way of deliquescence. This is the first evidence of active metabolism in the hyperarid core of the Atacama, and supports the view of a microbial community sustained by deliquescence. Our results expand the water activity envelope of life on Earth.

Biological activity plays a substantial role in the geochemistry of the Earth’s surface. Particularly interesting are effects on clay formation because clays are abundant and have high surface-to-volume ratio, resulting in clays making up a large fraction of the overall mineral-fluid interface and having an effective control of mineral reactions. Thus, biological control on clay composition would affect element budget globally and the mineralogy of subsequent diagenetic processes. Biological acceleration of clay production would result in enhanced clay control of mineral reactions and faster organic C sequestration, by adsorption on clay minerals, with implications for the C and related cycles. We investigated the combined effect of microbial activity and water chemistry on the composition of neoformed clay by reacting volcanic glass with natural waters covering a large composition range (fresh water from a lake and a spring, seawater, and hypersaline water). The microbes (bacteria, fungi, and algae) were totally or partially identified using molecular and microscopy techniques. The solid alteration products were analyzed using cryo-SEM to investigate the mineral-microbe interface and TEM-AEM to study the composition of the neoformed clay. The solution chemistry was also investigated. We found that clay composition was controlled mainly by glass chemistry, rather than biological activity, through a mechanism of in situ transformation. The resulting clay was Al-rich (dioctahedral composition). In one case (inorganic experiment, freshwater lake), the specific inorganic conditions of pH and Mg and Si concentration promoted formation of Mg-rich (trioctahedral clay). Microbes, however, did influence clay composition by confining glass grains in biofilms where water chemistry is significantly different from the bulk solution. Alteration in such conditions generated significant amounts of trioctahedral, Mg-rich clay in the hypersaline water experiment, whereas it favored production of dioctahedral, Al-rich clay in the freshwater lake experiment. It is thus demonstrated that biofilms can exert an effective control on clay mineralogy, We thank T. Wing-Dudek for her contribution in the planning of the experiments, V. Dekov and E. Neykova for providing the volcanic glass, F. Pinto for his expert technical support in the cryo-SEM study, and W. Huff and an anonymous reviewer for their comments that helped to improve the clarity of the manuscript. This work was funded by the Marie Curie Fellowship program, project Bio-Clays (2009–2011).

Under extreme water deficit, endolithic (inside rock) microbial ecosystems are considered environmental refuges for life in cold and hot deserts, yet their diversity and functional adaptations remain vastly unexplored. The metagenomic analyses of the communities from two rock substrates, calcite and ignimbrite, revealed that they were dominated by Cyanobacteria, Actinobacteria, and Chloroflexi. The relative distribution of major phyla was significantly different between the two substrates and biodiversity estimates, from 16S rRNA gene sequences and from the metagenomic data, all pointed to a higher taxonomic diversity in the calcite community. While both endolithic communities showed adaptations to extreme aridity and to the rock habitat, their functional capabilities revealed significant differences. ABC transporters and pathways for osmoregulation were more diverse in the calcite chasmoendolithic community. In contrast, the ignimbrite cryptoendolithic community was enriched in pathways for secondary metabolites, such as non-ribosomal peptides (NRP) and polyketides (PK). Assemblies of the metagenome data produced population genomes for the major phyla found in both communities and revealed a greater diversity of Cyanobacteria population genomes for the calcite substrate. Draft genomes of the dominant Cyanobacteria in each community were constructed with more than 93% estimated completeness. The two annotated proteomes shared 64% amino acid identity and a significantly higher number of genes involved in iron update, and NRPS gene clusters, were found in the draft genomes from the ignimbrite. Both the community-wide and genome-specific differences may be related to higher water availability and the colonization of large fissures and cracks in the calcite in contrast to a harsh competition for colonization space and nutrient resources in the narrow pores of the ignimbrite. Together, these results indicated that the habitable architecture of both lithic substrates- chasmoendolithic versus cryptoendolithic - might be an essential element in determining the colonization and the diversity of the microbial communities in endolithic substrates at the dry limit for life., ThisworkwasfundedbygrantEXOB08-0033fromNASAand grant NSF-0918907fromtheNationalSciencefoundationtoJR and byMINECO(Spain)grantsCGL2013-42509PtoJW,CA,JR, and OA.

In this study, we combined chlorophyll a fluorescence (ChlaF) measurements, using pulseamplitude-modulate (PAM) equipment, with scanning electron microscopy in backscattered electron mode (SEM-BSE) and transmission electron microscopy (TEM) images to evaluate the actions of Koretrel at lower concentrations on Verrucaria nigrescens colonising a dolostone. ChlaF measurements are good indicators of the damaging effects of biocides. However, these indicators only provide an incomplete view of the mechanism of biocides used to control biodeterioration agents. The death of the V. nigrescens photobiont at two biocide concentrations was revealed by PAM, SEM-BSE and TEM. Once Koretrel was applied, the Fv/Fm ratios markedly fell in the first few hours after the 1.5% treatment, and ratios for the 3% dilution remained close to zero throughout the study. The algal zone shows the plasmolysed appearance of the photobiont cells, and important aspects related to the action of the biocide on free and lichenised fungi were also detected using SEM-BSE. Many of the mycobiont cells had only their cell walls preserved; although, some fungal hyphae in lichen thalli and some microorganisms in endolithic clusters maintained lipid storage in their cytoplasm. These results indicated that the combination of physiological and microscopy techniques improves the assessment of biocide action in situ and this will help to optimize protocols in order to reduce the emission of these compounds to the environment., This work was funded by Programa Geomateriales (CAM, S2009/Mat-1629) and CGL 2010-16004 and CTM 2009-12838-CO4-03 from MICINN.

The hyperarid core of the Atacama Desert, Chile, is possibly the driest and most life-limited place on Earth, yet endolithic microorganisms thrive inside halite pinnacles that are part of ancient salt flats. The existence of this microbial community in an environment that excludes any other life forms suggests biological adaptation to high salinity and desiccation stress, and indicates an alternative source of water for life other than rainfall, fog or dew. Here, we show that halite endoliths obtain liquid water through spontaneous capillary condensation at relative humidity (RH) much lower than the deliquescence RH of NaCl. We describe how this condensation could occur inside nano-pores smaller than 100 nm, in a newly characterized halite phase that is intimately associated with the endolithic aggregates. This nano-porous phase helps retain liquid water for long periods of time by preventing its evaporation even in conditions of utmost dryness. Our results explain how life has colonized and adapted to one of the most extreme environments on our planet, expanding the water activity envelope for life on Earth, and broadening the spectrum of possible habitats for life beyond our planet.

This paper reports on the differential elimination of surface lichen thalli and their penetrating hyphae and epilithic and endolithic cyanobacteria by two biocide treatments applied on a small scale to the cloister of the Segovia cathedral. Both chemical treatments were followed by mechanical cleaning, and their effects were determined by scanning electron microscopy with backscattered electron imaging (SEM–BSE). Both treatments were effective at killing epilithic lichens and cyanobacteria, although they showed different effects on endolithic microbiota. The cleaning treatment involving Biotin T, dissolved in water, applied to areas extensively colonized by crustose lichens, resulted in a cleaner stone surface that was practically free of surface epilithic lichen remains, but the presence of living fungal hyphae was detected in fissures of the stone under these remnants in samples examined two months after treatment. The treatment using Biotin R, dissolved in White Spirit, left more superficial lichen remains but was better at eliminating mycobiont hyphae penetrating the stone. Endolithic cyanobacteria from zones showing green and black colourations were also more affected by Biotin R treatment. Our SEM–BSE observations highlight the need for a complete evaluation of treatment effects, including those produced on both epilithic lichen thalli and their penetrating components and on epilithic and endolithic microbiota. The causes and consequences of the different efficacy of treatments observed, as well as the potential risks of recolonization by viable cells left behind after treatment, are discussed., This work was supported by TRYCSA and grants GEOMATERIALES (S2009/Mat-1629/GEOMATERIALES) from the CAMand CTM2009-122838-C04-03 from the Spanish Ministry of Science and Innovation.

The Lithopanspermia space experiment was launched in 2007 with the European Biopan facility for a 10-day spaceflight on board a Russian Foton retrievable satellite. Lithopanspermia included for the first time the vagrant lichen species Aspicilia fruticulosa from Guadalajara steppic highlands (Central Spain), as well as other lichen species. During spaceflight, the samples were exposed to selected space conditions, that is, the space vacuum, cosmic radiation, and different spectral ranges of solar radiation (λ ≥ 110, ≥200, ≥290, or ≥400 nm, respectively). After retrieval, the algal and fungal metabolic integrity of the samples were evaluated in terms of chlorophyll a fluorescence, ultrastructure, and CO(2) exchange rates. Whereas the space vacuum and cosmic radiation did not impair the metabolic activity of the lichens, solar electromagnetic radiation, especially in the wavelength range between 100 and 200 nm, caused reduced chlorophyll a yield fluorescence; however, there was a complete recovery after 72 h of reactivation. All samples showed positive rates of net photosynthesis and dark respiration in the gas exchange experiment. Although the ultrastructure of all flight samples showed some probable stress-induced changes (such as the presence of electron-dense bodies in cytoplasmic vacuoles and between the chloroplast thylakoids in photobiont cells as well as in cytoplasmic vacuoles of the mycobiont cells), we concluded that A. fruticulosa was capable of repairing all space-induced damage. Due to size limitations within the Lithopanspermia hardware, the possibility for replication on the sun-exposed samples was limited, and these first results on the resistance of the lichen symbiosis A. fruticulosa to space conditions and, in particular, on the spectral effectiveness of solar extraterrestrial radiation must be considered preliminary. Further testing in space and under space-simulated conditions will be required. Results of this study indicate that the quest to discern the limits of lichen symbiosis resistance to extreme environmental conditions remains open.

The scarcity of liquid water in the hyperarid core of the Atacama Desert makes this region one of the most challenging environments for life on Earth. The low numbers of microbial cells in the soils suggest that within the Atacama Desert lies the dry limit for life on our planet. Here, we show that the Ca-sulfate crusts of this hyperarid core are the habitats of lithobiontic micro-organisms. This microporous, translucent substrate is colonized by epilithic lichens, as well as endolithic free-living algae, fungal hyphae, cyanobacteria and non photosynthetic bacteria. We also report a novel type of endolithic community, "hypoendoliths", colonizing the undermost layer of the crusts. The colonization of gypsum crusts within the hyperarid core appears to be controlled by the moisture regime. Our data shows that the threshold for colonization is crossed within the dry core, with abundant colonization in gypsum crusts at one study site, while crusts at a drier site are virtually devoid of life. We show that the cumulative time in 1 year of relative humidity (RH) above 60% is the best parameter to explain the difference in colonization between both sites. This is supported by controlled humidity experiments, where we show that colonies of endolithic cyanobacteria in the Ca-sulfate crust undergo imbibition process at RH >60%. Assuming that life once arose on Mars, it is conceivable that Martian micro-organisms sought refuge in similar isolated evaporite microenvironments during their last struggle for life as their planet turned arid.