Sieglinde Ott

Microbial rock inhabitants survive hypervelocity impacts on Mars-like host planets: First phase of lithopanspermia experimentally tested

The scenario of lithopanspermia describes the viable transport of microorganisms via meteorites. To test the first step of lithopanspermia, i.e., the impact ejection from a planet, systematic shock recovery experiments within a pressure range observed in martian meteorites (5-50 GPa) were performed with dry layers of microorganisms (spores of Bacillus subtilis, cells of the endolithic cyanobacterium Chroococcidiopsis, and thalli and ascocarps of the lichen Xanthoria elegans) sandwiched between gabbro discs (martian analogue rock). Actual shock pressures were determined by refractive index measurements and Raman spectroscopy, and shock temperature profiles were calculated. Pressure-effect curves were constructed for survival of B. subtilis spores and Chroococcidiopsis cells from the number of colony-forming units, and for vitality of the photobiont and mycobiont of Xanthoria elegans from confocal laser scanning microscopy after live/dead staining (FUN-I). A vital launch window for the transport of rock-colonizing microorganisms from a Mars-like planet was inferred, which encompasses shock pressures in the range of 5 to about 40 GPa for the bacterial endospores and the lichens, and a more limited shock pressure range for the cyanobacterium (from 5-10 GPa). The results support concepts of viable impact ejections from Mars-like planets and the possibility of reseeding early Earth after asteroid cataclysms.

Survival of Rock-Colonizing Organisms After 1.5 Years in Outer Space

Cryptoendolithic microbial communities and epilithic lichens have been considered as appropriate candidates for the scenario of lithopanspermia, which proposes a natural interplanetary exchange of organisms by means of rocks that have been impact ejected from their planet of origin. So far, the hardiness of these terrestrial organisms in the severe and hostile conditions of space has not been tested over extended periods of time. A first long-term (1.5 years) exposure experiment in space was performed with a variety of rock-colonizing eukaryotic organisms at the International Space Station on board the European EXPOSE-E facility. Organisms were selected that are especially adapted to cope with the environmental extremes of their natural habitats. It was found that some-but not all-of those most robust microbial communities from extremely hostile regions on Earth are also partially resistant to the even more hostile environment of outer space, including high vacuum, temperature fluctuation, the full spectrum of extraterrestrial solar electromagnetic radiation, and cosmic ionizing radiation. Although the reported experimental period of 1.5 years in space is not comparable with the time spans of thousands or millions of years believed to be required for lithopanspermia, our data provide first evidence of the differential hardiness of cryptoendolithic communities in space.

Artificial cultures of lichens in the natural environment

The growth and development of several lichens under natural conditions are studied with the SEM. The results permit observations on the influence of the microclimate and of pollution on thalline development and on some principles of lichen systematics.

Survival Potential and Photosynthetic Activity of Lichens Under Mars-Like Conditions: A Laboratory Study

Lichens were repetitively exposed over 22 days to thermophysical Mars-like conditions at low-and mid-latitudes. The simulated parameters and the experimental setup are described. Natural samples of the lichen Xanthoria elegans were used to investigate their ability to survive the applied Mars-like conditions. The effects of atmospheric pressure, CO(2) concentration, low temperature, water availability, and light on Mars were also studied. The results of these experiments indicate that no significant decrease in the vitality of the lichen occurred after exposure to simulated martian conditions, which was demonstrated by confocal laser scanning microscopy analysis, and a 95% CO(2) atmosphere with 100% humidity, low pressure (partial pressure of CO(2) was 600 Pa), and low temperature has a balancing effect on photosynthetic activity as a function of temperature. This means a starting low photosynthetic activity at high CO(2) concentrations with Earth-like pressure has a reduction of 60%. But, if the simulated atmospheric pressure is reduced to Mars-like conditions with the maintenance of the same Mars-like 95% CO(2) concentration, the photosynthetic activity increases and again reaches similar values as those exhibited under terrestrial atmospheric pressure and concentration. Based on these results, we presume that, in any region on Mars where liquid water might be available, even for short periods of time, a eukaryotic symbiotic organism would have the ability to survive, at least over weeks, and to temporarily photosynthesize.

Viability of the lichen Xanthoria elegans and its symbionts after 18 months of space exposure and simulated Mars conditions on the ISS

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.

Extremotolerance and Resistance of Lichens: Comparative Studies on Five Species Used in Astrobiological Research I. Morphological and Anatomical Characteristics

Lichens are symbioses of two organisms, a fungal mycobiont and a photoautotrophic photobiont. In nature, many lichens tolerate extreme environmental conditions and thus became valuable models in astrobiological research to fathom biological resistance towards non-terrestrial conditions; including space exposure, hypervelocity impact simulations as well as space and Martian parameter simulations. All studies demonstrated the high resistance towards non-terrestrial abiotic factors of selected extremotolerant lichens. Besides other adaptations, this study focuses on the morphological and anatomical traits by comparing five lichen species—Circinaria gyrosa, Rhizocarpon geographicum, Xanthoria elegans, Buellia frigida, Pleopsidium chlorophanum—used in present-day astrobiological research. Detailed investigation of thallus organization by microscopy methods allows to study the effect of morphology on lichen resistance and forms a basis for interpreting data of recent and future experiments. All investigated lichens reveal a common heteromerous thallus structure but diverging sets of morphological-anatomical traits, as intra-/extra-thalline mucilage matrices, cortices, algal arrangements, and hyphal strands. In B. frigida, R. geographicum, and X. elegans the combination of pigmented cortex, algal arrangement, and mucilage seems to enhance resistance, while subcortex and algal clustering seem to be crucial in C. gyrosa, as well as pigmented cortices and basal thallus protrusions in P. chlorophanum. Thus, generalizations on morphologically conferred resistance have to be avoided. Such differences might reflect the diverging evolutionary histories and are advantageous by adapting lichens to prevalent abiotic stressors. The peculiar lichen morphology demonstrates its remarkable stake in resisting extreme terrestrial conditions and may explain the high resistance of lichens found in astrobiological research.

Soil properties of an Antarctic inland site: implications for ecosystem development

Inland Antarctic nunataks typically have simple physically weathered soils and limited ecosystem complexity. In this paper we present quantitative measurements of soil physical and chemical properties at one Antarctic nunatak. We measured pH, grain size, field capacity, soil organic carbon, phosphate, nitrate, ammonium and the cations magnesium, calcium and potassium along two transects. The data obtained indicated that very low levels of nutrients were present/available to biota, and that liquid water was absent, at least from the surface depths of soil, except during periods of active snow melt. Consequently, biological activity is severely limited. We conclude that, due to the climatic and microclimatic conditions at this location, the development of biological communities and soils is maintained in an extremely simple but still apparently stable ‘quasi climax’ state. Increased soil development and biological complexity can be expected if the contemporary rapid regional warming in the Antarctic Peninsula region continues.

Mitochondrial and nuclear ribosomal DNA data do not support the separation of the Antarctic lichens Umbilicaria kappenii and Umbilicaria antarctica as distinct species

The Antarctic endemics Umbilicaria kappenii and U. antarctica are morphologically close, but mainly distinguished by their reproductive strategies. Umbilicaria antarctica propagates by means of thalloconidia. Umbilicaria kappenii lacks thalloconidia, but exhibits a variety of asexual propagules: soredia, adventive lobes and thallyles. We have now employed molecular data from three gene regions to examine the phylogenetic relationships of these two morphotypes. The phylogeny of ten samples and four outgroup taxa ( Umbilicaria decussata , U. krascheninnikovii , U. nylanderiana , U. umbilicarioides ) was reconstructed using Bayesian and maximum parsimony analyses of a combined data set of nuclear ITS, nuclear LSU rDNA and mitochondrial LSU rDNA sequences. Forty two new partial sequences of 14 specimens were generated. Our results indicate that all samples morphologically referred to U. antarctica and U. kappenii form a monophyletic group. A topology separating the two morphotypes as phylogenetic species is significantly rejected with the data set. It is proposed to place U. kappenii into synonymy with U. antarctica .

PHOTOSYMBIODEMES AND THEIR DEVELOPMENT IN PELTIGERA VENOSA

A photosymbiodeme is described for Peltigera venosa. One member consists of small black squamules containing cyanobacteria whereas the other, the more familiar thallus of the species, contains green algae. New aspects of morphogenetic influences are discussed. Introduction In several lichens the mycobiont can be associated with either green algae or cyanobacteria, or with both at the same time. Such groups of thalli with differ- ent photobionts have been described by different names including morpho- types, phycotypes, chimeroid associations and phycosymbiodemes. Renner & Galloway (1982) discussed these terms and named such lichens phycosymbio- demes. Principally following their terminology, the term photosymbiodeme is used here. They often occur in the genera Lobaria, Peltigera, Pseudocyphellaria and Sticta and in other cephalodiate lichens. Usually some connection can be observed between the external or internal cephalodia and the development of photosymbiodemes with their green algae or cyanobacteria, respectively. The significance and development of cephalodia and their influence on the green thalli has been investigated many times (Forssell 1883, Bitter 1909, Jordan 1970, 1972, Jordan & Rickson 1971). The most important and detailed dis- cussion of the problems involved has been given by James & Henssen (1976) in a comprehensive review of the relationship of the symbionts in many lichens. Photosymbiodemes in the genus Peltigera have been reported by Brodo & Richardson (1978). They discovered thalli of Peltigera aphthosa containing cyanobacteria in place of green algae, sometimes growing from the cephalodia of normal thalli. Tensberg & Holtan-Hartwig (1983) observed that the green cephalodiate thalli of Peltigera venosa were often associated on the substratum with a brown or black layer developing into small black squamules. Normal thalli, containing green algae, seemed to grow from this dark crust, which contained cyanobacteria and could be interpreted as the second member of a photosymbiodeme. Tonsberg & Holtan-Hartwig (1983) considered a more detailed examination of the early stages to be necessary. The complex system of developmental pathways and morphogenetic interactions expressed in the two morphologies of Peltigera venosa is the subject of this paper.

UV-C tolerance of symbiotic Trebouxia sp. in the space-tested lichen species Rhizocarpon geographicum and Circinaria gyrosa : role of the hydration state and cortex/screening substances

Many experiments were carried out in order to evaluate the survival capacity of extremotolerant lichens when facing harsh conditions, including those of outer space or of simulated Martian environment. For further progress, a deeper study on the physiological mechanisms is needed that confer the unexpected levels of resistance detected on these symbiotic organisms. In this work, the response of the lichenized green algae Trebouxia sp. (a predominant lichen photobiont) to increasing doses of UV-C radiation is studied. UV-C (one of the most lethal factorsto be found in spacetogether with vacuum and cosmic-ionizing radiation with high atomic number and energy (HZE) particles) has been applied in the present experiments up to a maximum dose analogue to 67 days in Low Earth Orbit (LEO). For that purpose we selected two extremotolerant and space-tested lichen species in which Trebouxia sp. is the photosynthetic partner: the crustose lichenRhizocarpon geographicumand the fruticose lichenCircinaria gyrosa.In orderto evaluatethe effect of the physiological state of the lichen thallus (active when wet and dormant when dry) and of protective structures (cortex and photoprotective pigments) on the resistance of the photobiont to UV-C, four different experimental conditions were tested: (1) dry intact samples, (2) wet intact samples, (3) dry samples without cortex/acetone-rinsed and (4) wet samples without cortex/acetone-rinsed. After irradiation and a 72 hours period of recovery, the influence of UV-C on the two lichens photobiont under each experimental approach was assessed by two complimentary methods: (1) By determining the photosystem II (PSII) activity in three successive 24 hours intervals (Mini-PAM fluorometer) to investigate the overall state of the photosynthetic process and the resilience of Trebouxia sp. (2) By performing high performance liquid chromatography (HPLC)-quantification of four essential photosynthetic pigments (chlorophyll a, chlorophyll b, β-carotene and lutein) of one sample of each species and dose. Results indicate that the physiological state of the thallus is the most important factor impairing the tolerance of Trebouxia sp. to UV-C radiation in both lichen species. Desiccated thalli were demonstrated to be more resistant to UV-C. No clear influence of UV-C radiation on the carotenoid content was detected. Comparing the respective doses applied, the individuals of R. geographicum are more sensitive than C. gyrosa.