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: UV-resistance
The data from the R3DE instrument of ESA’s EXPOSE-E mission outside the ISS at the European Technological Expose Facility (EuTEF) on the ESA Columbus module shows that the docking of the Space Shuttle with the International Space Station (ISS) decreased the South-Atlantic Anomaly (SAA) maxima dose rates from about 1500 Gy h-1 down to 600-700 Gy h-1 or by factor of 2. The dose rate data at the same time from another Bulgarian built instrument (R3DR) of the EXPOSE-R mission outside the Russian “Zvezda” module showed that: 1) before the Space Shuttle docking, the SAA dose rates measured with R3DR were higher (2500 Gy h-1) than the R3DE data; 2) The relative decrease of the SAA dose rates after the shuttle docking was only by a factor of 1.25. These differences are explained by the smaller shielding of R3DR from the body of ISS and by the larger distance of it from the body of Space Shuttle. Very similar data, but with smaller dose rates were obtained with a third Bulgarian built instrument (Liulin-5) inside Russian “Pirs” module. The analysis of the ascending/descending SAA dose rate maxima of the three instruments shows that the effect can be simply explained by the additional shielding against the 30 to 150 MeV protons of the SAA, provided by the 78 tons Shuttle to the instruments and by changing of the ISS 3D mass distribution when the ISS rotates.
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.