S. Ott

The potential of the lichen symbiosis to cope with extreme conditions of outer space – I. Influence of UV radiation and space vacuum on the vitality of lichen symbiosis and germination capacity

The lichen symbiotic organisms Fulgensia bracteata and Xanthoria elegans as well as their isolated photobionts and mycobionts were exposed to conditions simulating the extreme parameters of outer space in order to assess their limits of survival, e.g. during a potential interplanetary transfer or on the surface of an extraterrestrial body. Using the space simulation facilities at DLR, the test parameters under investigation were vacuum (10 −3 Pa) and ultraviolet (UV) radiation at wavelength ranges from vacuum-UV to UV-A, applied separately or in combination. An analysis of vitality was made using confocal laser scanning microscopy and LIVE/DEAD staining using FUN I or SYTOX green. Intact lichens were extremely resistant to vacuum exposure as well as to UV radiation up to doses of about 160 kJ m −2 (200 −2 reduced the vitality rate to nearly 50%; however, the dose effect curves levelled off and continued treatment did not reduce the vitality further. Similar survival curves levelling off at higher UV doses were observed for the isolated photobionts; however, in the latter cases, the saturation occurred at five times lower doses (full UV spectrum). Also spores of the mycobionts showed a remarkable UV (254 nm) resistance up to doses of about 3 kJ m −2 . The data suggest that the symbiotic features peculiar to lichens allow them to cope with the extreme conditions of outer space or even with Martian surface conditions provided suitable niche habitats are available to serve as refuges and complementary endogenous or exogenous protection mechanisms are established.

Life at the Limits: Capacities of Isolated and Cultured Lichen Symbionts to Resist Extreme Environmental Stresses

Lichens are described as a symbiosis formed by a myco- and photobiont, capable of colonizing habitats where their separate symbionts would not be able to survive. Space simulation studies on the separated symbionts of the lichen Xanthoria elegans have been performed to test their capacity to resist the most extreme conditions. The isolated cultured symbiont cells were exposed to different doses of the UV spectrum, and to vacuum. Cultures of both symbionts were analysed by specific vitality tests (LIVE/DEAD-staining detected by Confocal Laser Scanning Microscopy). Growth capacity of symbiont cultures on different media was analysed after exposure to extreme environmental stresses. The data obtained support the hypothesis that the symbiotic state considerably enhances the ability of the respective symbionts to survive exposure to extreme conditions, including the conditions of space simulation. Species such as X. elegans may, therefore, be suitable for use as model organisms in exobiological studies.

Extremotolerance and Resistance of Lichens: Comparative Studies on Five Species Used in Astrobiological Research II. Secondary Lichen Compounds

Lichens, which are symbioses of a fungus and one or two photoautotrophs, frequently tolerate extreme environmental conditions. This makes them valuable model systems in astrobiological research to fathom the limits and limitations of eukaryotic symbioses. Various studies demonstrated the high resistance of selected extremotolerant lichens towards extreme, non-terrestrial abiotic factors including space exposure, hypervelocity impact simulations as well as space and Martian parameter simulations. This study focusses on the diverse set of secondary lichen compounds (SLCs) that act as photo- and UVR-protective substances. Five lichen species used in present-day astrobiological research were compared: Buellia frigida, Circinaria gyrosa, Rhizocarpon geographicum, Xanthoria elegans, and Pleopsidium chlorophanum. Detailed investigation of secondary substances including photosynthetic pigments was performed for whole lichen thalli but also for axenically cultivated mycobionts and photobionts by methods of UV/VIS-spectrophotometry and two types of high performance liquid chromatography (HPLC). Additionally, a set of chemical tests is presented to confirm the formation of melanic compounds in lichen and mycobiont samples. All investigated lichens reveal various sets of SLCs, except C. gyrosa where only melanin was putatively identified. Such studies will help to assess the contribution of SLCs on lichen extremotolerance, to understand the adaptation of lichens to prevalent abiotic stressors of the respective habitat, and to form a basis for interpreting recent and future astrobiological experiments. As most of the identified SLCs demonstrated a high capacity in absorbing UVR, they may also explain the high resistance of lichens towards non-terrestrial UVR.

Resistance of the Lichen Buellia frigida to Simulated Space Conditions during the Preflight Tests for BIOMEX—Viability Assay and Morphological Stability

Samples of the extremotolerant Antarctic endemite lichen Buellia frigida are currently exposed to low-Earth orbit-space and simulated Mars conditions at the Biology and Mars Experiment (BIOMEX), which is part of the ESA mission EXPOSE-R2 on the International Space Station and was launched on 23 July 2014. In preparation for the mission, several preflight tests (Experimental and Scientific Verification Tests, EVT and SVT) assessed the sample preparation and hardware integration procedures as well as the resistance of the candidate organism toward the abiotic stressors experienced under space and Mars conditions. Therefore, we quantified the post-exposure viability with a live/dead staining technique utilizing FUN-1 and confocal laser scanning microscopy (CLSM). In addition, we used scanning electron microscopy (SEM) to investigate putative patterns of morphological-anatomical damage that lichens may suffer under the extreme exposure conditions. The present results demonstrate that Buellia frigida is capable of surviving the conditions tested in EVT and SVT. The mycobiont showed lower average impairment of its viability than the photobiont (viability rates of >83% and >69%, respectively), and the lichen thallus suffered no significant damage in terms of thalline integrity and symbiotic contact. These results will become essential to substantiate and validate the results prospectively obtained from the returning space mission. Moreover, they will help assess the limits and limitations of terrestrial organisms under space and Mars conditions as well as characterize the adaptive traits that confer lichen extremotolerance.

Effects of UVC 254 nm on the photosynthetic activity of photobionts from the astrobiologically relevant lichens Buellia frigida and Circinaria gyrosa

In the past decade, various astrobiological studies on different lichen species investigated the impairment of viability and photosynthetic activity by exposure to simulated or real space parameters (as vacuum, polychromatic ultraviolet (UV)-radiation and monochromatic UVC) and consistently found high post-exposure viability as well as low rates of photosynthetic impairment (de Vera et al. 2003, 2004a; 2004b; de la Torre et al. 2010; Onofri et al. 2012; Sanchez et al. 2012, 2014; Brandt et al. 2014). To achieve a better understanding of the basic mechanisms of resistance, the present study subdued isolated and metabolically active photobionts of two astrobiologically relevant lichens to UVC254 nm, examined its effect on photosynthetic activity by chlorophyll a fluorescence and characterized the UVC-induced damages by quantum yield reduction and measurements of non-photochemical quenching. The results indicate a strong impairment of photosynthetic activity, photoprotective mechanisms and overall photobiont vitality when being irradiated in the isolated and metabolically active state. In conclusion, the present study stresses the higher susceptibility of photobionts towards extreme environmental conditions as UVC-exposure, a stressor that does not occur on the Earth. By comparison with previous studies, the present results highlight the importance of protective mechanisms in lichens, such as morphological-anatomical traits (Meesen et al. 2013), secondary lichen compounds (Meesen et al. 2014) and the symbionts pivotal ability to pass into anhydrobiosis when desiccating. Received 7 May 2014, accepted 16 July 2014