Xanthoria parietina is a widespread foliose lichen growing on barks and rocks showing a broad spectrum of tolerance to air pollutants such as NOX and heavy metals, and resistance to UV-radiation because of the screening properties provided by the secondary metabolite parietin. The aim of this study was to evaluate the ability of this lichen species to survive in the following simulated space conditions, UV-radiation in N2 atmosphere and UV-radiation in vacuum. The efficiency of the photosynthetic apparatus was used as an indicator of vitality, and was expressed in terms of chlorophyll a fluorescence (FV/FM) and Normalized Difference Vegetation Index (NDVI), which were measured within 72 h from the exposure. Additionally, during the irradiation, the IR reflectance spectrum of the lichen was monitored in situ to assess changes in spectral bands. The results showed significant differences in physiological recovery trends between the treatments, highlighting that UV-radiation in vacuum causes stronger effects on FV/FM values. The IR analysis revealed several spectral band changes in the fingerprint region. The most visible variation was the 5200 cm−1 water band that disappeared in the overtone region. Nevertheless, X. parietina was able to survive UV-radiation in N2 atmosphere and in vacuum, and for this reason it may be considered a candidate for further evaluations on its survival capacity in extreme conditions.