Desert Cyanobacterium Research Articles

Deciphering the Role of Trehalose in Chroococcidiopsis sp. 029's High-Desiccation Resistance: Sequence Determination, Structural Modelling and Simulative Analysis of the 30S Ribosomal Subunit.

Desert strains of the genus Chroococcidiopsis are among the most desiccation-resistant cyanobacteria capable of anhydrobiosis. The accumulation of two sugars, sucrose and trehalose, facilitates the entrance of anhydrobiotes into a reversible state of dormancy by stabilizing cellular components upon water removal. This study aimed to evaluate, at the atomistic level, the role of trehalose in desiccation resistance by using as a model system the 30S ribosomal subunit of the desert cyanobacterium Chroococcidiopsis sp. 029. Molecular dynamic simulations provided atomistic evidence regarding its protective role on the 30S molecular structure. Trehalose forms an enveloping shell around the ribosomal subunit and stabilizes the structures through a network of direct interactions. The simulation confirmed that trehalose actively interacts with the 30S ribosomal subunit and that, by replacing water molecules, it ensures ribosomal structural integrity during desiccation, thus enabling protein synthesis to be carried out upon rehydration.

Far-red light photoacclimation in a desert Chroococcidiopsis strain with a reduced FaRLiP gene cluster and expression of its chlorophyll f synthase in space-resistant isolates.

Some cyanobacteria can use far-red light (FRL) to drive oxygenic photosynthesis, a phenomenon known as Far-Red Light Photoacclimation (FaRLiP). It can expand photosynthetically active radiation beyond the visible light (VL) range. Therefore, it holds promise for biotechnological applications and may prove useful for the future human exploration of outer space. Typically, FaRLiP relies on a cluster of ~20 genes, encoding paralogs of the standard photosynthetic machinery. One of them, a highly divergent D1 gene known as chlF (or psbA4), is the synthase responsible for the formation of the FRL-absorbing chlorophyll f (Chl f) that is essential for FaRLiP. The minimum gene set required for this phenotype is unclear. The desert cyanobacterium Chroococcidiopsis sp. CCMEE 010 is unusual in being capable of FaRLiP with a reduced gene cluster (15 genes), and it lacks most of the genes encoding FR-Photosystem I. Here we investigated whether the reduced gene cluster of Chroococcidiopsis sp. CCMEE 010 is transcriptionally regulated by FRL and characterized the spectral changes that occur during the FaRLiP response of Chroococcidiopsis sp. CCMEE 010. In addition, the heterologous expression of the Chl f synthase from CCMEE 010 was attempted in three closely related desert strains of Chroococcidiopsis. All 15 genes of the FaRLiP cluster were preferentially expressed under FRL, accompanied by a progressive red-shift of the photosynthetic absorption spectrum. The Chl f synthase from CCMEE 010 was successfully expressed in two desert strains of Chroococcidiopsis and transformants could be selected in both VL and FRL. In Chroococcidiopsis sp. CCME 010, all the far-red genes of the unusually reduced FaRLiP cluster, are transcriptionally regulated by FRL and two closely related desert strains heterologously expressing the chlF010 gene could grow in FRL. Since the transformation hosts had been reported to survive outer space conditions, such an achievement lays the foundation toward novel cyanobacteria-based technologies to support human space exploration.

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.

Functional specialization of expanded orange carotenoid protein paralogs in subaerial Nostoc species.

Orange carotenoid protein (OCP) is a photoactive protein that participates in the photoprotection of cyanobacteria. There are 2 full-length OCP proteins, 4 N-terminal paralogs (helical carotenoid protein [HCP]), and 1 C-terminal domain-like carotenoid protein (CCP) found in Nostoc flagelliforme, a desert cyanobacterium. All HCPs (HCP1 to 3 and HCP6) from N. flagelliforme demonstrated their excellent singlet oxygen quenching activities, in which HCP2 was the strongest singlet oxygen quencher compared with others. Two OCPs, OCPx1 and OCPx2, were not involved in singlet oxygen scavenging; instead, they functioned as phycobilisome fluorescence quenchers. The fast-acting OCPx1 showed more effective photoactivation and stronger phycobilisome fluorescence quenching compared with OCPx2, which behaved differently from all reported OCP paralogs. The resolved crystal structure and mutant analysis revealed that Trp111 and Met125 play essential roles in OCPx2, which is dominant and long acting. The resolved crystal structure of OCPx2 is maintained in a monomer state and showed more flexible regulation in energy quenching activities compared with the packed oligomer of OCPx1. The recombinant apo-CCP obtained the carotenoid pigment from holo-HCPs and holo-OCPx1 of N. flagelliforme. No such carotenoid transferring processes were observed between apo-CCP and holo-OCPx2. The close phylogenetic relationship of OCP paralogs from subaerial Nostoc species indicates an adaptive evolution toward development of photoprotection: protecting cellular metabolism against singlet oxygen damage using HCPs and against excess energy captured by active phycobilisomes using 2 different working modes of OCPx.

Feasibility as feedstock of the cyanobacterium Chroococcidiopsis sp. 029 cultivated with urine-supplemented moon and mars regolith simulants

Biology-driven in situ resource utilization (bio-ISRU) should provide a breakthrough in supporting long-term human permanence on the Moon and Mars that will rely on the ability to produce consumables on site. Lithotrophic cyanobacteria are gaining interest for linking ISRU to life-support systems: In the “PowerCell” concept, cyanobacteria are grown with raw materials available on the Moon and Mars and used to feed bacteria producing consumables. Here we showed that lunar and martian regolith simulants as well as water-released minerals (supplemented with a nitrogen source) can support planktonic and biofilm growth of the desert cyanobacterium Chroococcidiopsis sp. 029. Such a growth versality is relevant to meet different technological requirements of future cultivation hardware to be used on the Moon (and eventually on Mars). We showed that this cyanobacterium can use urea as a nitrogen source and be cultivated with lunar and martian regolith simulants supplemented with synthetic human urine. Notably, the biomass yielded from each experimental set up could be used as feedstock for bacteria. Results provided a proof-of-concept of the feasibility of Chroococcidiopsis grown with raw materials available on site on the Moon and Mars and human waste, to “power” bio-production by feeding bacteria useful for bioprocesses to support human settlements beyond Earth.

Mass spectrometry and spectroscopic characterization of a tetrameric photosystem I supercomplex from Leptolyngbya ohadii, a desiccation-tolerant cyanobacterium

Cyanobacteria inhabiting desert biological soil crusts face the harsh conditions of the desert. They evolved a suite of strategies toward desiccation-hydration cycles mixed with high light irradiations, etc. In this study we purified and characterized the structure and function of Photosystem I (PSI) from Leptolyngbya ohadii, a desiccation-tolerant desert cyanobacterium. We discovered that PSI forms tetrameric (PSI-Tet) aggregate. We investigated it by using sucrose density gradient centrifugation, clear native PAGE, high performance liquid chromatography, mass spectrometry (MS), time-resolved fluorescence (TRF) and time-resolved transient absorption (TA) spectroscopy. MS analysis identified the presence of two PsaB and two PsaL proteins in PSI-Tet and uniquely revealed that PsaLs are N-terminally acetylated in contrast to non-modified PsaL in the trimeric PSI from Synechocystis sp. PCC 6803. Chlorophyll (Chl) a fluorescence decay profiles of the PSI-Tet performed at 77 K revealed two emission bands at ∼690 nm and 725 nm with the former appearing only at early delay time. The main fluorescence emission peak, associated with emission from the low energy Chls a, decays within a few nanoseconds. TA studies demonstrated that the 725 nm emission band is associated with low energy Chls a with absorption band clearly resolved at ∼710 nm at 77 K. In summary, our work suggests that the heterogenous composition of PsaBs and PsaL in PSI-Tet is related with the adaptation mechanisms needed to cope with stressful conditions under which this bacterium naturally grows.

Protection and Damage Repair Mechanisms Contributed To the Survival of Chroococcidiopsis sp. Exposed To a Mars-Like Near Space Environment.

Chroococcidiopsis spp. can withstand extremely harsh environments, including a Mars-like environment. However, studies are lacking on the molecular mechanisms of Chroococcidiopsis sp. surviving in Mars-like environments. In the HH-21-5 mission, the desert cyanobacterium Chroococcidiopsis sp. was exposed to a Mars-like environment (near space; 35 km altitude) for 4 h, and a single-factor environment of near space was simulated on the ground. We investigated the survival and endurance mechanisms of Chroococcidiopsis sp. ASB-02 after exposing it to near space by studying its physiological and transcriptional properties. After the exposure, Chroococcidiopsis sp. ASB-02 exhibited high cell viability, although photosystem II activity decreased and the levels of reactive oxygen species increased. The single-factor simulation experiments revealed that for the survival of Chroococcidiopsis sp. ASB-02 in near space, UV radiation was the most important limiting factor, and it was followed by temperature. The near space environment triggered multiple metabolic pathway responses in Chroococcidiopsis sp. ASB-02. The upregulation of extracellular polysaccharides as well as carotenoid and scytonemin biosynthesis genes in response to UV radiation attenuated the extent of radiation reaching the cells. At the same time, genes related to protein synthesis were upregulated in response to the low temperature, overcoming the decrease in metabolic activity that was caused by the low temperature. In near space and after rehydration, the genes involved in various DNA and photosystem II repair pathways were upregulated. This reflected the damage to the DNA and photosystem II protein subunits in cells during the flight and suggested that repair mechanisms play an important role in the recovery of Chroococcidiopsis sp. ASB-02. IMPORTANCE This study reported that the protective and repair mechanisms of Chroococcidiopsis sp. ASB-02 contributed to its endurance ability in a Mars-like near space environment. In Chroococcidiopsis sp. ASB-02, a Mars-like near space environment activated the expression of genes involved in extracellular polysaccharides (EPS), carotenoid, scytonemin, and protein syntheses, which provided additional protection. Additionally, the cell damage repair process enhanced the recovery rate of Chroococcidiopsis sp. ASB-02 after the flight. This study will help to enhance the understanding of the tolerance mechanism of Chroococcidiopsis sp. and to provide important guidance as to the survival requirements for microbial life in a Mars-like environment.

Mars-like UV Flux and Ionizing Radiation Differently Affect Biomarker Detectability in the Desert Cyanobacterium Chroococcidiopsis as Revealed by the Life Detector Chip Antibody Microarray.

The effect of a Mars-like UV flux and γ-radiation on the detectability of biomarkers in dried cells of Chroococcidiopsis sp. CCMEE 029 was investigated using a fluorescence sandwich microarray immunoassay. The production of anti-Chroococcidiopsis antibodies allowed the immunoidentification of a reduced, though still detectable, signal in dried cells mixed with phyllosilicatic and sulfatic Mars regolith simulants after exposure to 6.8 × 105 kJ/m2 of a Mars-like UV flux. No signal was detected in dried cells that were not mixed with minerals after 1.4 × 105 kJ/m2. For γ-radiation (60Co), no detectable variations of the fluorescence signal occurred in dried cells exposed to 113 kGy compared to non-irradiated dried cells. Our results suggest that immunoassay-based techniques could be used to detect life tracers eventually present in the martian subsurface in freshly excavated materials only if shielded from solar UV. The high structural integrity of biomarkers irradiated with γ-radiation that mimics a dose accumulated in 13 Myr at 2 m depth from the martian surface has implications for the potential detectability of similar organic molecules/compounds by future life-detection missions such as the ExoMars Rosalind Franklin rover.

Absence of increased genomic variants in the cyanobacterium Chroococcidiopsis exposed to Mars-like conditions outside the space station

Despite the increasing interest in using microbial-based technologies to support human space exploration, many unknowns remain not only on bioprocesses but also on microbial survivability and genetic stability under non-Earth conditions. Here the desert cyanobacterium Chroococcidiopsis sp. CCMEE 029 was investigated for robustness of the repair capability of DNA lesions accumulated under Mars-like conditions (UV radiation and atmosphere) simulated in low Earth orbit using the EXPOSE-R2 facility installed outside the International Space Station. Genomic alterations were determined in a space-derivate of Chroococcidiopsis sp. CCMEE 029 obtained upon reactivation on Earth of the space-exposed cells. Comparative analysis of whole-genome sequences showed no increased variant numbers in the space-derivate compared to triplicates of the reference strain maintained on the ground. This result advanced cyanobacteria-based technologies to support human space exploration.

Expansion of bilin-based red light sensors in the subaerial desert cyanobacterium Nostoc flagelliforme.

Light is the crucial environmental signal for desiccation-tolerant cyanobacteria to activate photosynthesis and prepare for desiccation at dawn. However, the photobiological characteristics of desert cyanobacteria adaptation to one of the harshest habitats on Earth remain unresolved. In this study, we surveyed the genome of a subaerial desert cyanobacterium Nostoc flagelliforme and identified two phytochromes and seven cyanobacteriochromes (CBCRs) with one or more bilin-binding GAF (cGMP phosphodiesterase/adenylyl cyclase/FhlA) domains. Biochemical and spectroscopic analyses of 69 purified GAF-containing proteins from recombinant phycocyanobilin (PCB), biliverdin or phycoerythrobilin-producing Escherichia coli indicated that nine of these proteins bind chromophores. Further investigation revealed that 11 GAFs form covalent adducts responsive to near-UV and visible light: eight GAFs contained PCB chromophores, three GAFs contained biliverdin chromophores and one contained the PCB isomer, phycoviolobilin. Interestingly, COO91_03972 is the first-ever reported GAF-only CBCR capable of sensing five wavelengths of light. Bioinformatics and biochemical analyses revealed that residue P132 of COO91_03972 is essential for chromophore binding to dual-cysteine CBCRs. Furthermore, the complement of N. flagelliforme CBCRs is enriched in red light sensors. We hypothesize that these sensors are critical for the acclimatization of N. flagelliforme to weak light environments at dawn.

Exposure of cyanobacterium Nostoc sp. to the Mars-like stratosphere environment.

During the HH-19-2 flight mission of the Chinese Scientific Experimental System, dried Nostoc sp. cells were exposed to the stratosphere environment (32,508 m altitude) for 3 h and 22 min. The atmospheric pressure, temperature, relative humidity, and ionizing and non-ionizing radiation levels at that altitude are similar to those on the surface of Mars. Although analyses revealed decreased photosynthetic activity, a decline in autofluorescence, and damage to the cellular morphology in the flight-exposed sample, the death rate was low (28%). Physiological changes were not obvious after the exposure to the Mars-like vacuum conditions. The ground-exposed samples showed a similar trend to the flight-exposed samples, but the damage was relatively slight. RNA-sequencing data revealed a number of affected metabolic pathways: photosynthetic system and CO2 fixation function, activation of antioxidant systems, heat shock protein, DNA repair, and protein synthesis. Results suggest that Nostoc sp. has the potential to survive in a Mars-like environment and that it may be a suitable pioneer species to colonize Mars in the future in closed life-support systems (base) or in localities with relatively suitable conditions for life, such as localities with water available.

New types of ATP-grasp ligase are associated with the novel pathway for complicated mycosporine-like amino acid production in desiccation-tolerant cyanobacteria.

Mycosporine-like amino acids (MAAs) were widespread in diverse organisms to attenuate UV radiation. We recently characterized the large, complicated MAA mycosporine-2-(4-deoxygadusolyl-ornithine) in desert cyanobacterium Nostoc flagelliforme. Synthesis of this MAA requires the five-gene cluster mysABDC2C3. Here, bioinformatic analysis indicated that mysC duplication within five-gene mys clusters is strictly limited to drought-tolerant cyanobacteria. Phylogenic analysis distinguished these duplicated MysCs into two clades that separated from canonical MysCs. Heterologous expression of N. flagelliforme mys genes in Escherichia coli showed that MysAB produces 4-deoxygadusol. The ATP-grasp ligase of MysC3 catalyses the linkage of the δ- or ε-amino group of ornithine/lysine to 4-deoxygadusol, yielding mycosporine-ornithine or mycosporine-lysine respectively. The ATP-grasp ligase of MysC2 strictly condenses the α-amino group of mycosporine-ornithine to another 4-deoxygadusol. MysD (D-Ala-D-Ala ligase) functions following MysC2 to catalyse the formation of mycosporine-2-(4-deoxygadusolyl-ornithine). High arginine content likely provides a greater pool of ornithine over other amino acids during rehydration of desiccated N. flagelliforme. Duplication of ATP-grasp ligases is specific for the use of substrates that have two amino groups (such as ornithine) for the production of complicated MAAs with multiple chromophores. This five-enzyme biosynthesis pathway for complicated MAAs is a novel adaptation of cyanobacteria for UV tolerance in drought environments.

Light intensity stimulates the production of extracellular polymeric substances (EPS) in a culture of the desert cyanobacterium Trichormus sp

Light intensity stimulates the production of extracellular polymeric substances (EPS) in a culture of the desert cyanobacterium Trichormus sp

Biomarker Preservation and Survivability Under Extreme Dryness and Mars-Like UV Flux of a Desert Cyanobacterium Capable of Trehalose and Sucrose Accumulation

Unravelling how long life can persist under extreme dryness and what kind of environmental extremes can be faced by dried microorganisms, is relevant to understand Mars habitability and to search for life on planets with transient liquid water availability. Since trehalose and sucrose stabilize dried anhydrobiotes, an in silico survey of the genome of the desert cyanobacterium Chroococcidiopsis sp. CCMEE 029 was performed to identify pathways for trehalose and sucrose biosynthesis. The expression of the identified genes was induced in response to desiccation and trehalose and sucrose accumulation was detected in dried cells. This adaptation strategy enabled viability and biomarker permanence under extreme dryness and Mars-like UV flux. Chroococcidiopsis survivors were scored in 7-year dried biofilms mixed with phyllosilicatic Mars regolith simulant and exposed to 5.5 x 103 kJ/m2 of a Mars-like UV flux. No survivors occurred after exposure to 5.5 x 105 kJ/m2, although in dead cells, photosynthetic pigments and nucleic acids, both DNA and RNA, were still detectable. This suggested that dried biofilms mixed with phyllosilicatic Martian regolith simulant are suitable candidates to identify biosignatures embedded in planetary analogue minerals as planned in the future BioSigN (BioSignatures and habitable Niches) space mission to be performed outside the International Space Station.

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.

Effects of ambient temperature on the redistribution efficiency of nutrients by desert cyanobacteria- Scytonema javanicum

Cultures of Scytonema javanicum obtained from artificial medium are used to control desertification, and through the effective redistribution of nutrients, related environmental problems can be alleviated. Wastewater is considered to be a potential alternative medium for S. javanicum. However, the effect of temperature on the nutrient redistribution ability of S. javanicum cultured in wastewater has rarely been considered. Therefore, this study explores the effect of temperature on S. javanicum in wastewater. The results showed that a sufficient temperature increase (from 25 °C to 30 °C) increased the photosynthetic activity of photosynthetic system II (PSII), accelerated the accumulation rate of S. javanicum biomass, and improved the removal efficiency of nutrients in wastewater. However, an increasing temperature caused a decrease in the final accumulated biomass. When the temperature was above 35 °C, the ratio of the variable to maximal fluorescence (Fv/Fm) of S. javanicum decreased, thus, causing damage to PSII. The average Fv/Fm at 35 °C and 40 °C decreased by 10.49% and 72.37%, respectively, compared to that at 25 °C. By analysing the chlorophyll fluorescence induction kinetics (OJIP) curve after 30 days, the P phase at 30 °C increased by 15.47% relative to that at 25 °C, whereas that at 35 °C and 40 °C decreased by 45.54% and 86.37%, respectively. In particular, at 40 °C, the O-J-I-P phase transformed into the O-J (J = I = P) phase, which caused irreversible damage to the PSII of S. javanicum. Comprehensive scores were determined using the entropy weight method and revealed that 30 °C was the optimal temperature for the wastewater culture of S. javanicum. This temperature improved the biomass accumulation rate and wastewater transfer efficiency. These results provide a scientific basis for improving the efficiency of the coupling technology of wastewater treatment and desert algal cultivation.

Inoculation concentration modulating the secretion and accumulation pattern of exopolysaccharides in desert cyanobacterium Microcoleus vaginatus.

Cyanobacterial exopolysaccharides (EPS) accumulated during microalgal cultivation have significant application potential in antioxidation, pharmaceutical products, and so on. Inoculation concentration strongly affects the cultivation cost, biomass, and EPS accumulation. In this study, a high-EPS-excreted desert cyanobacterium Microcoleus vaginatus was isolated, and the effects of inoculation concentration on biomass, photosynthetic activity, and EPS accumulation were explored. The results showed that the original fluorescence (Fo ) provided a good indication to cyanobacterial biomass, when Chl-a concentration was lower than 10mgL-1 . Inoculation concentration significantly affected cyanobacterial biomass and EPS concentration (P<0.001), whereas did not affect photosynthetic activity (Fv /Fm ; P>0.05). The two fractions of EPS, capsular exopolysaccharides (CPS) and released exopolysaccharides (RPS) were strongly affect by inoculation concentration. Other than forming thick sheath (CPS) surrounded the filaments, M. vaginatus excreted higher proportions of RPS to culture medium, and the ratio of RPS to CPS ranged from 1.08 to 1.58 depending on the inoculation concentration. Additionally, although the biomass and EPS accumulation increased with inoculation concentration, the increasing inoculation concentration did not bring to the proportionate increase of the final biomass and EPS yield. Altogether, comprehensively considering the EPS yield and productivity, inoculation concentration of 0.04mgChl-aL-1 is recommended for M. vaginatus to produce EPS, with an EPS yield of 94.32mgL-1 and EPS productivity of 184.86mg (mgChl-a)-1 L-1 d-1 at the end of experiment.

Promoting desert biocrust formation using aquatic cyanobacteria with the aid of MOF-based nanocomposite

Desertification and eutrophication are two global environmental problems human beings face. Inoculating cyanobacteria to form biocrusts is considered an effective technology to inhibit desertification. The main limitation of biocrust formation is the lack of propagules and nutrients in deserts. A possible low cost source of propagules and nutrients is eutrophic water containing aquatic cyanobacteria (AC), nitrogen and phosphorus. In this study, we fabricated a network-structured nanocomposite (designated as MC) using a metal-organic framework (MOF) and carboxymethyl cellulose (CMC). MC, with a large specific surface area and numerous surface groups, had a high retention capacity for water and nutrients and good biosafety. The combination of AC-containing water (ACW) and MC could provide a suitable microenvironment in the soil, promote the growth of desert cyanobacteria (DC), formation of biocrusts and inhibition of desertification. This study provides a novel approach to simultaneously relieve desertification and eutrophication.

Over-Expression of UV-Damage DNA Repair Genes and Ribonucleic Acid Persistence Contribute to the Resilience of Dried Biofilms of the Desert Cyanobacetrium Chroococcidiopsis Exposed to Mars-Like UV Flux and Long-Term Desiccation.

The survival limits of the desert cyanobacterium Chroococcidiopsis were challenged by rewetting dried biofilms and dried biofilms exposed to 1.5 × 103 kJ/m2 of a Mars-like UV, after 7 years of air-dried storage. PCR-stop assays revealed the presence of DNA lesions in dried biofilms and an increased accumulation in dried-UV-irradiated biofilms. Different types and/or amounts of DNA lesions were highlighted by a different expression of uvrA, uvrB, uvrC, phrA, and uvsE genes in dried-rewetted biofilms and dried-UV-irradiated-rewetted biofilms, after rehydration for 30 and 60 min. The up-regulation in dried-rewetted biofilms of uvsE gene encoding an UV damage endonuclease, suggested that UV-damage DNA repair contributed to the repair of desiccation-induced damage. While the phrA gene encoding a photolyase was up-regulated only in dried-UV-irradiated-rewetted biofilms. Nucleotide excision repair genes were over-expressed in dried-rewetted biofilms and dried-UV-irradiated-rewetted biofilms, with uvrC gene showing the highest increase in dried-UV-irradiated-rewetted biofilms. Dried biofilms preserved intact mRNAs (at least of the investigated genes) and 16S ribosomal RNA that the persistence of the ribosome machinery and mRNAs might have played a key role in the early phase recovery. Results have implications for the search of extra-terrestrial life by contributing to the definition of habitability of astrobiologically relevant targets such as Mars or planets orbiting around other stars.

A Desert Cyanobacterium under Simulated Mars-like Conditions in Low Earth Orbit: Implications for the Habitability of Mars

In the ESA space experiment BIOMEX (BIOlogy and Mars EXperiment), dried Chroococcidiopsis cells were exposed to Mars-like conditions during the EXPOSE-R2 mission on the International Space Station. The samples were exposed to UV radiation for 469 days and to a Mars-like atmosphere for 722 days, approaching the conditions that could be faced on the surface of Mars. Once back on Earth, cell survival was tested by growth-dependent assays, while confocal laser scanning microscopy and PCR-based assay were used to analyze the accumulated damage in photosynthetic pigments (chlorophyll a and phycobiliproteins) and genomic DNA, respectively. Survival occurred only for dried cells (4-5 cell layers thick) mixed with the martian soil simulants P-MRS (phyllosilicatic martian regolith simulant) and S-MRS (sulfatic martian regolith simulant), and viability was only maintained for a few hours after space exposure to a total UV (wavelength from 200 to 400 nm) radiation dose of 492 MJ/m2 (attenuated by 0.1% neutral density filters) and 0.5 Gy of ionizing radiation. These results have implications for the hypothesis that, during Mars's climatic history, desiccation- and radiation-tolerant life-forms could have survived in habitable niches and protected niches while transported.