The lichen Xanthoria elegans has been exposed to space conditions and simulated Mars-analogue conditions in the lichen and fungi experiment (LIFE) on the International Space Station (ISS). After several simulations and short space exposure experiments such as BIOPAN, this was the first long-term exposure of eukaryotic organisms to the hostile space conditions of the low Earth orbit (LEO). The biological samples were integrated in the EXPOSE-E facility and exposed for 1.5 years outside the ISS to the combined impact of insolation, ultraviolet (UV)-irradiation, cosmic radiation, temperatures and vacuum conditions of LEO space. Additionally, a subset of X. elegans samples was exposed to simulated Martian environmental conditions by applying Mars-analogue atmosphere and suitable solar radiation filters. After their return to Earth the viability of the lichen samples was ascertained by viability analysis of LIVE/DEAD staining and confocal laser-scanning microscopy, but also by analyses of chlorophyll a fluorescence. According to the LIVE/DEAD staining results, the lichen photobiont showed an average viability rate of 71%, whereas the even more resistant lichen mycobiont showed a rate of 84%. Post-exposure viability rates did not significantly vary among the applied exposure conditions. This remarkable viability is discussed in the context of particular protective mechanisms of lichens such as anhydrobiosis and UV-screening pigments.
Research Containing: Astrobiology
Characterisation of Growth and Ultrastructural Effects of the Xanthoria elegans Photobiont After 1.5 Years of Space Exposure on the International Space Station
The lichen Xanthoria elegans has been exposed to space and simulated Mars-analogue environment in the Lichen and Fungi Experiment (LIFE) on the EXPOSE-E facility at the International Space Station (ISS). This long-term exposure of 559 days tested the ability of various organisms to cope with either low earth orbit (LEO) or Mars-analogue conditions, such as vacuum, Mars-analogue atmosphere, rapid temperature cycling, cosmic radiation of up to 215 +/- 16 mGy, and insolation of accumulated doses up to 4.87 GJm(-2), including up to 0.314 GJm(-2) of UV irradiation. In a previous study, X. elegans demonstrated considerable resistance towards these conditions by means of photosynthetic activity as well as by post-exposure metabolic activity of 50-80 % in the algal and 60-90 % in the fungal symbiont (Brandt et al. Int J Astrobiol 14(3):411-425, 2015). The two objectives of the present study were complementary: First, to verify the high post-exposure viability by using a qualitative cultivation assay. Second, to characterise the cellular damages by transmission electron microscopy (TEM) which were caused by the space and Mars-analogue exposure conditions of LIFE. Since the algal symbiont of lichens is considered as the more susceptible partner (de Vera and Ott 2010), the analyses focused on the photobiont. The study demonstrated growth and proliferation of the isolated photobiont after all exposure conditions of LIFE. The ultrastructural analysis of the algal cells provided an insight to cellular damages caused by long-term exposure and highlighted that desiccation-induced breakdown of cellular integrity is more pronounced under the more severe space vacuum than under Mars-analogue atmospheric conditions. In conclusion, desiccation-induced damages were identified as a major threat to the photobiont of X. elegans. Nonetheless, a fraction of the photobiont cells remained cultivable after all exposure conditions tested in LIFE.
Preservation of Biomarkers from Cyanobacteria Mixed with Mars-Like Regolith Under Simulated Martian Atmosphere and UV Flux
The space mission EXPOSE-R2 launched on the 24th of July 2014 to the International Space Station is carrying the BIOMEX (BIOlogy and Mars EXperiment) experiment aimed at investigating the endurance of extremophiles and stability of biomolecules under space and Mars-like conditions. In order to prepare the analyses of the returned samples, ground-based simulations were carried out in Planetary and Space Simulation facilities. During the ground-based simulations, Chroococcidiopsis cells mixed with two Martian mineral analogues (phyllosilicatic and sulfatic Mars regolith simulants) were exposed to a Martian simulated atmosphere combined or not with UV irradiation corresponding to the dose received during a 1-year-exposure in low Earth orbit (or half a Martian year on Mars). Cell survival and preservation of potential biomarkers such as photosynthetic and photoprotective pigments or DNA were assessed by colony forming ability assays, confocal laser scanning microscopy, Raman spectroscopy and PCR-based assays. DNA and photoprotective pigments (carotenoids) were detectable after simulations of the space mission (570 MJ/m(2) of UV 200-400 nm irradiation and Martian simulated atmosphere), even though signals were attenuated by the treatment. The fluorescence signal from photosynthetic pigments was differently preserved after UV irradiation, depending on the thickness of the samples. UV irradiation caused a high background fluorescence of the Martian mineral analogues, as revealed by Raman spectroscopy. Further investigation will be needed to ensure unambiguous identification and operations of future Mars missions. However, a 3-month exposure to a Martian simulated atmosphere showed no significant damaging effect on the tested cyanobacterial biosignatures, pointing out the relevance of the latter for future investigations after the EXPOSE-R2 mission. Data gathered during the ground-based simulations will contribute to interpret results from space experiments and guide our search for life on Mars.