The cyanobacterium Chroococcidiopsis, overlain by 3 mm of Antarctic sandstone, was exposed as dried multilayers to simulated space and martian conditions. Ground-based experiments were conducted in the context of Lichens and Fungi Experiments (EXPOSE-E mission, European Space Agency), which were performed to evaluate, after 1.5 years on the International Space Station, the survival of cyanobacteria (Chroococcidiopsis), lichens, and fungi colonized on Antarctic rock. The survival potential and the role played by protection and repair mechanisms in the response of dried Chroococcidiopsis cells to ground-based experiments were both investigated. Different methods were employed, including evaluation of the colony-forming ability, single-cell analysis of subcellular integrities based on membrane integrity molecular and redox probes, evaluation of the photosynthetic pigment autofluorescence, and assessment of the genomic DNA integrity with a PCR-based assay. Desiccation survivors of strain CCMEE 123 (coastal desert, Chile) were better suited than CCMEE 134 (Beacon Valley, Antarctica) to withstand cellular damage imposed by simulated space and martian conditions. Exposed dried cells of strain CCMEE 123 formed colonies, maintained subcellular integrities, and, depending on the exposure conditions, also escaped DNA damage or repaired the induced damage upon rewetting. Key Words: Astrobiology—Endolithic cyanobacteria—Chroococcidiopsis—Ground-based experiments. Astrobiology 11, 65–73.

Dried biofilms and dried multilayered planktonic counterparts obtained from three desert strains of Chroococcidiopsis were exposed to low Earth conditions by using the EXPOSE-R2 facility outside the International Space Station. During the space mission, samples in Tray 1 (space vacuum and solar radiation, from λ ≈ 110 nm) and Tray 2 (Mars-like UV flux, λ > 200 nm and Mars-like atmosphere) received total UV (200–400 nm) fluences of about 4.58 × 10 ² kJ/m ² and 4.92 × 10 ² kJ/m ² , respectively, and 0.5 Gy of cosmic ionizing radiation. Postflight analyses were performed on 2.5-year-old samples due to the space mission duration, from launch to sample return to the lab. The occurrence of survivors was determined by evaluating cell division upon rehydration and damage to the genome and photosynthetic apparatus by polymerase chain reaction–stop assays and confocal laser scanning microscopy. Biofilms recovered better than their planktonic counterparts, accumulating less damage not only when exposed to UV radiation under space and Mars-like conditions but also when exposed in dark conditions to low Earth conditions and laboratory control conditions. This suggests that, despite the shielding provided by top-cell layers being sufficient for a certain degree of survival of the multilayered planktonic samples, the enhanced survival of biofilms was due to the presence of abundant extracellular polymeric substances and to additional features acquired upon drying.

In the ESA space experiment BIOMEX (BIOlogy and Mars EXperiment), dried Chroococcidiopsis cells were exposed to Mars-like conditions during the EXPOSE-R2 mission on the International Space Station. The samples were exposed to UV radiation for 469 days and to a Mars-like atmosphere for 722 days, approaching the conditions that could be faced on the surface of Mars. Once back on Earth, cell survival was tested by growth-dependent assays, while confocal laser scanning microscopy and PCR-based assay were used to analyze the accumulated damage in photosynthetic pigments (chlorophyll a and phycobiliproteins) and genomic DNA, respectively. Survival occurred only for dried cells (4–5 cell layers thick) mixed with the martian soil simulants P-MRS (phyllosilicatic martian regolith simulant) and S-MRS (sulfatic martian regolith simulant), and viability was only maintained for a few hours after space exposure to a total UV (wavelength from 200 to 400 nm) radiation dose of 492 MJ/m ² (attenuated by 0.1% neutral density filters) and 0.5 Gy of ionizing radiation. These results have implications for the hypothesis that, during Mars's climatic history, desiccation- and radiation-tolerant life-forms could have survived in habitable niches and protected niches while transported.

Studying the resistance of cyanobacteria to ionizing radiation provides relevant information regarding astrobiology-related topics including the search for life on Mars, lithopanspermia, and biological life-support systems. Here, we report on the resistance of desert cyanobacteria of the genus Chroococcidiopsis, which were exposed (as part of the STARLIFE series of experiments) in both hydrated and dried states to ionizing radiation with different linear energy transfer values (0.2 to 200 keV/μm). Irradiation with up to 1 kGy of He or Si ions, 2 kGy of Fe ions, 5 kGy of X-rays, or 11.59 kGy of γ rays (⁶⁰Co) did not eradicate Chroococcidiopsis populations, nor did it induce detectable damage to DNA or plasma membranes. The relevance of these results for astrobiology is briefly discussed. Key Words: Ionizing radiation—Linear energy transfer—Lithopanspermia—Cyanobacterial radioresistance—Chroococcidiopsis—Mars. Astrobiology 17, 118–125.