In a precursory study for the space experiment ADAPT ("Molecular adaptation strategies of microorganisms to different space and planetary UV climate conditions"), cells of Bacillus subtilis 168 were continuously cultured for 700 generations under periodic polychromatic UV irradiation (200-400 nm) to model the suggested UV radiation environment on early Earth at the origin of the first microbial ecosystem during the Archean eon when Earth lacked a significant ozone layer. Populations that evolved under UV stress were about 3-fold more resistant than the ancestral and non-UV-evolved populations. UV-evolved cells were 7-fold more resistant to ionizing radiation than their non-UV-exposed evolved relatives and ancestor. In addition to the acquired increased UV resistance, further changes in microbial stress response to hydrogen peroxide, increased salinity, and desiccation were observed in UV-evolved cells. This indicates that UV-sensitive ancestral cells are capable of adapting to periodically applied UV stress via the evolution of cells with an increased UV resistance level and further enhanced responses to other environmental stressors, which thereby allows them to survive and reproduce under extreme UV radiation as a selection pressure.

In the space experiment "Molecular adaptation strategies of microorganisms to different space and planetary UV climate conditions" (ADAPT), bacterial endospores of the highly UV-resistant Bacillus subtilis strain MW01 were exposed to low-Earth orbit (LEO) and simulated martian surface conditions for 559 days on board the European Space Agency's exposure facility EXPOSE-E, mounted outside the International Space Station. The survival of B. subtilis MW01 spores from both assays (LEO and simulated martian conditions) was determined by a colony-formation assay after retrieval. It was clearly shown that solar extraterrestrial UV radiation (λ≥110 nm) as well as the martian UV spectrum (λ≥200 nm) was the most deleterious factor applied; in some samples only a few spore survivors were recovered from B. subtilis MW01 spores exposed in monolayers. However, if shielded from solar irradiation, about 8% of MW01 spores survived in LEO conditions, and 100% survived in simulated martian conditions, compared to the laboratory controls. The results demonstrate the effect of shielding against the high inactivation potential of extraterrestrial solar UV radiation, which limits the chances of survival of even the highly UV-resistant strain of B. subtilis MW01 in the harsh environments of outer space and the martian surface.

After 700 generations of a short-term evolution experiment with Bacillus subtilis 168, two strains were isolated, the UV-adapted strain MW01 and the UV-unexposed control strain DE69, and chosen for UV-C radiation resistance studies with respect to growth phase. The ancestral strain from the evolution experiment was used as reference for comparative purposes. Cells of the UV-adapted strain showed signiWcant diVerences in their physiology (growth behavior, doubling time, cell density, and sporulation capacity) and were more resistant to UV in all monitored stages. These Wndings implicate the evolution to an increased UV radioresistance was not limited to a speciWc growth phase and led to reduced growth dynamics, compared with those obtained from the ancestral and the control strain.

Aims:  To determine the effects of cysteine, cystine, proline and thioproline as sporulation medium supplements on Bacillus subtilis spore resistance to hydrogen peroxide (H2O2), wet heat, and germicidal 254 nm and simulated environmental UV radiation. Methods and Results:  Bacillus subtilis spores were prepared in a chemically defined liquid medium, with and without supplementation of cysteine, cystine, proline or thioproline. Spores produced with thioproline, cysteine or cystine were more resistant to environmentally relevant UV radiation at 280–400 and 320–400 nm, while proline supplementation had no effect. Spores prepared with cysteine, cystine or thioproline were also more resistant to H2O2 but not to wet heat or 254‐nm UV radiation. The increases in spore resistance attributed to the sporulation supplements were eliminated if spores were chemically decoated. Conclusions:  Supplementation of sporulation medium with cysteine, cystine or thioproline increases spore resistance to solar UV radiation reaching the Earth’s surface and to H2O2. These effects were eliminated if the spores were decoated, indicating that alterations in coat proteins by different sporulation conditions can affect spore resistance to some agents. Significance and Impact of the Study:  This study provides further evidence that the composition of the sporulation medium can have significant effects on B. subtilis spore resistance to UV radiation and H2O2. This knowledge provides further insight into factors influencing spore resistance and inactivation.

The Planetary Society's Phobos Living Interplanetary Flight Experiment (Phobos LIFE) flew in the sample return capsule of the Russian Federal Space Agency's Phobos Grunt mission and was to have been a test of one aspect of the hypothesis that life can move between nearby planets within ejected rocks. Although the Phobos Grunt mission failed, we present here the scientific and engineering design and motivation of the Phobos LIFE experiment to assist with the scientific and engineering design of similar future experiments. Phobos LIFE flew selected organisms in a simulated meteoroid. The 34-month voyage would have been the first such test to occur in the high-radiation environment outside the protection of Earth's magnetosphere for more than a few days. The patented Phobos LIFE “biomodule” is an 88 g cylinder consisting of a titanium outer shell, several types of redundant seals, and 31 individual Delrin sample containers. Phobos LIFE contained 10 different organisms, representing all three domains of life, and one soil sample. The organisms are all very well characterized, most with sequenced genomes. Most are extremophiles, and most have flown in low Earth orbit. Upon return from space, the health and characteristics of organisms were to have been compared with controls that remained on Earth and have not yet been opened.