Identification of far-red light acclimation in an endolithic Chroococcidiopsis strain and associated genomic features: Implications for oxygenic photosynthesis on exoplanets

Deserts represent extreme habitats where photosynthetic life is restricted to the lithic niche. The ability of rock-inhabiting cyanobacteria to modify their photosynthetic apparatus and harvest far-red light (near-infrared) was investigated in 10 strains of the genus Chroococcidiopsis, previously isolated from diverse endolithic and hypolithic desert communities. The analysis of their growth capacity, photosynthetic pigments, and apcE2-gene presence revealed that only Chroococcidiopsis sp. CCMEE 010 was capable of far-red light photoacclimation (FaRLiP). A total of 15 FaRLiP genes were identified, encoding paralogous subunits of photosystem I, photosystem II, and the phycobilisome, along with three regulatory elements. CCMEE 010 is unique among known FaRLiP strains by undergoing this acclimation process with a significantly reduced cluster, which lacks major photosystem I paralogs psaA and psaB. The identification of an endolithic, extremotolerant cyanobacterium capable of FaRLiP not only contributes to our appreciation of this phenotype’s distribution in nature but also has implications for the possibility of oxygenic photosynthesis on exoplanets

Carotenoid Raman Signatures Are Better Preserved in Dried Cells of the Desert Cyanobacterium Chroococcidiopsis than in Hydrated Counterparts after High-Dose Gamma Irradiation

Carotenoids are promising targets in our quest to search for life on Mars due to their biogenic origin and easy detection by Raman spectroscopy, especially with a 532 nm excitation thanks to resonance effects. Ionizing radiations reaching the surface and subsurface of Mars are however detrimental for the long-term preservation of biomolecules. We show here that desiccation can protect carotenoid Raman signatures in the desert cyanobacterium Chroococcidiopsis sp. CCMEE 029 even after high-dose gamma irradiation. Indeed, while the height of the carotenoids Raman peaks was considerably reduced in hydrated cells exposed to gamma irradiation, it remained stable in dried cells irradiated with the highest tested dose of 113 kGy of gamma rays, losing only 15-20% of its non-irradiated intensity. Interestingly, even though the carotenoid Raman signal of hydrated cells lost 90% of its non-irradiated intensity, it was still detectable after exposure to 113 kGy of gamma rays. These results add insights into the preservation potential and detectability limit of carotenoid-like molecules on Mars over a prolonged period of time and are crucial in supporting future missions carrying Raman spectrometers to Mars’ surface

Insights into the chaotropic tolerance of the desert cyanobacterium Chroococcidiopsis sp. 029 (Chroococcidiopsales, cyanobacteria)

The mechanism of perchlorate resistance of the desert cyanobacterium Chroococcidiopsis sp. CCMEE 029 was investigated by assessing whether the pathways associated with its desiccation tolerance might play a role against the destabilizing effects of this chaotropic agent. During 3 weeks of growth in the presence of 2.4 mM perchlorate, an upregulation of trehalose and sucrose biosynthetic pathways was detected. This suggested that in response to the water stress triggered by perchlorate salts, these two compatible solutes play a role in the stabilization of macromolecules and membranes as they do in response to dehydration. During the perchlorate exposure, the production of oxidizing species was observed by using an oxidant-sensing fluorochrome and determining the expression of the antioxidant defense genes, namely superoxide dismutases and catalases, while the presence of oxidative DNA damage was highlighted by the over-expression of genes of the base excision repair. The involvement of desiccation-tolerance mechanisms in the perchlorate resistance of this desert cyanobacterium is interesting since, so far, chaotropic-tolerant bacteria have been identified among halophiles. Hence, it is anticipated that desert microorganisms might possess an unrevealed capability of adapting to perchlorate concentrations exceeding those naturally occurring in dry environments. Furthermore, in the endeavor of supporting future human outposts on Mars, the identified mechanisms might contribute to enhance the perchlorate resistance of microorganisms relevant for biologically driven utilization of the perchlorate-rich soil of the red planet

Revival of Anhydrobiotic Cyanobacterium Biofilms Exposed to Space Vacuum and Prolonged Dryness: Implications for Future Missions beyond Low Earth Orbit

Dried biofilms of Chroococcidiopsis sp. CCMEE 029 were revived after a 672-day exposure to space vacuum outside the International Space Station during the EXPOSE-R2 space mission. After retrieval, they were airdried stored for 3.5 years. Space vacuum reduced cell viability and increased DNA damage compared to airdried storage for 6 years under laboratory conditions. Long exposure times to space vacuum and extreme dryness decrease the changes of survival that ultimately depend on DNA damage repair upon rehydration, and hence, an in silico analysis of Chroococcidiopsis sp. CCMEE 029’s genome was performed with a focus on DNA repair pathways. The analysis identified a high number of genes that encode proteins of the homologous recombination RecF pathway and base excision repair that were over-expressed during 1 and 6 h rehydration of space-vacuum exposed biofilms. This suggests that Chroococcidiopsis developed a survival strategy against desiccation, with DNA repair playing a key role, which allowed the revival of biofilms exposed to space vacuum. Unravelling how long anhydrobiotic cyanobacteria can persist under space vacuum followed by prolonged air-dried storage is relevant to future astrobiological experiments that use space platforms and might require prolonged air-dried storage of the exposed samples before retrieval to Earth

Unravelling the secret of the resistance of desert strains of Chroococcidiopsis to desiccation and radiation

Chroococcidiopsis is a unicellular cyanobacterial genus that is growing in extreme dry conditions, either in low or high temperatures. At the lower end of the spectrum, they live as cryptoendoliths in rocks of the Mc Murdo Dry Valleys in Antarctica where they were discovered by Imre Friedmann, while at the higher end, they grow as hypoliths/endoliths in hot deserts, e.g. Negev, Gobi, Atacama. The capacity of desert strains of Chroococcidiopsis to stabilize their sub-cellular organization is so efficient that, when dried, they can cope with simulated space and Martian conditionsas well as with high doses of ionizing and UV radiations .Chroococcidiopsi

Avoidance of protein oxidation correlates with the desiccation and radiation resistance of hot and cold desert strains of the cyanobacterium Chroococcidiopsis

In order to investigate the relationship between desiccation and the extent of protein oxidation in desert strains of Chroococcidiopsis a selection of 10 isolates from hot and cold deserts and the terrestrial cyanobacterium Chroococcidiopsis thermalis sp. PCC 7203, were exposed to desiccation (air-drying) and analysed for survival. Strain CCMEE 029 from the Negev desert and the aquatic cyanobacterium Synechocystis sp. PCC 6803 were further investigated for protein oxidation after desiccation (drying over silica gel), treatment with H2O2 up to 1M and exposure to γ-rays up to 25 kGy. Then a selection of desert strains of Chroococcidiopsis with different survival rates after prolonged desiccation, as well as Synechocystis sp. PCC 6803 and Chroococcidiopsis thermalis sp. PCC 7203, were analysed for protein oxidation after treatment with 10 mM and 100 mM of H2O2. Results suggest that in the investigated strains a tight correlation occurs between desiccation and radiation tolerance and avoidance of protein oxidatio

Dried Biofilms of Desert Strains of Chroococcidiopsis Survived Prolonged Exposure to Space and Mars-like Conditions in Low Earth Orbit

Chroococcidiopsis were exposed to low Earth conditions by using the EXPOSE-R2 facility outside the International Space Station. During the space mission, samples in Tray 1 (space vacuum and solar radiation, from λ ≈ 110 nm) and Tray 2 (Mars-like UV flux, λ > 200 nm and Mars-like atmosphere) received total UV (200–400 nm) fluences of about 4.58 × 102 kJ/m² and 4.92 × 102 kJ/m², respectively, and 0.5 Gy of cosmic ionizing radiation. Postflight analyses were performed on 2.5-year-old samples due to the space mission duration, from launch to sample return to the lab. The occurrence of survivors was determined by evaluating cell division upon rehydration and damage to the genome and photosynthetic apparatus by polymerase chain reaction–stop assays and confocal laser scanning microscopy. Biofilms recovered better than their planktonic counterparts, accumulating less damage not only when exposed to UV radiation under space and Mars-like conditions but also when exposed in dark conditions to low Earth conditions and laboratory control conditions. This suggests that, despite the shielding provided by top-cell layers being sufficient for a certain degree of survival of the multilayered planktonic samples, the enhanced survival of biofilms was due to the presence of abundant extracellular polymeric substances and to additional features acquired upon drying

Evaluation of the Resistance of Chroococcidiopsis spp. to Sparsely and Densely Ionizing Irradiation

Studying the resistance of cyanobacteria to ionizing radiation provides relevant information regarding astrobiology-related topics including the search for life on Mars, lithopanspermia, and biological life-support systems. Here, we report on the resistance of desert cyanobacteria of the genus Chroococcidiopsis, which were exposed (as part of the STARLIFE series of experiments) in both hydrated and dried states to ionizing radiation with different linear energy transfer values (0.2 to 200 keV/μm). Irradiation with up to 1 kGy of He or Si ions, 2 kGy of Fe ions, 5 kGy of X-rays, or 11.59 kGy of γ rays (60Co) did not eradicate Chroococcidiopsis populations, nor did it induce detectable damage to DNA or plasma membranes. The relevance of these results for astrobiology is briefly discussed