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The lichen Buellia frigida was exposed to space and simulated Mars analog conditions in the Biology and Mars Experiment (BIOMEX) project operated outside the International Space Station (ISS) for 1.5 years. To determine the effects of the Low Earth Orbit (LEO) conditions on the lichen symbionts, a LIVE/DEAD staining analysis test was performed. After return from the ISS, the lichen symbionts demonstrated mortality rates of up to 100% for the algal symbiont and up to 97.8% for the fungal symbiont. In contrast, the lichen symbiont controls exhibited mortality rates of 10.3% up to 31.9% for the algal symbiont and 14.5% for the fungal symbiont. The results performed in the BIOMEX Mars simulation experiment on the ISS indicate that the potential for survival and the resistance of the lichen B. frigida to LEO conditions are very low. It is unlikely that Mars could be inhabited by this lichen, even for a limited amount of time, or even not habitable planet for the tested lichen symbionts.

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.