ALBERT

Description: We are currently investigating the biological population present in the highest and least explored perennial lakes on earth in the Bolivian and Chilean Andes, including several volcanic crater lakes of more than 6000 m elevation, in combination of microbiological and molecular biological methods. Our samples were collected in saline lakes of the Laguna Blanca Laguna Verde area in the Bolivian Altiplano and in the Licancabur volcano crater (27 deg. 47 min S/67 deg. 47 min. W) in the ongoing project studying high altitude lakes. The main goal of the project is to look for analogies with Martian paleolakes. These Bolivian lakes can be described as Andean lakes following the classification of Chong. We have attempted to isolate pure cultures and phylogenetically characterize prokaryotes that grew under laboratory conditions. Sediment samples taken from the Licancabur crater lake (LC), Laguna Verde (LV), and Laguna Blanca (LB) were analyzed and cultured using enriched liquid media under both aerobic and anaerobic conditions. All cultures were incubated at room temperature (15 to 20 C) and under light exposure. For the reported isolates, 36 hours incubation were necessary for reaching optimal optical densities to consider them viable cultures. Ten serial dilutions starting from 1% inoculum were required to obtain a suitable enriched cell culture to transfer into solid media. Cultures on solid medium were necessary to verify the formation of colonies in order to isolate pure cultures. Different solid media were prepared using several combinations of both trace minerals and carbohydrates sources in order to fit their nutrient requirements. The microorganisms formed individual colonies on solid media enriched with tryptone, yeast extract and sodium chloride. Cells morphology was studied by optical and electronic microscopy. Rodshape morphologies were observed in most cases. Total bacterial genomic DNA was isolated from 50 ml late-exponential phase culture by using the CTAB miniprep protocol. The 16S rRNA genes were amplified by PCR using both Bacteria- and Archaeauniversal primer sets: 27f and 1492r, 21f and 1492r respectively. Sequences of 16S rRNA gene were determined and initially compared with reference sequences contained in the EMBL nucleotide sequence database by using the BLAST program and were subsequently aligned with 16S rRNA reference sequences in the ARB package (http://www.mikro.biologie.tu-muenchen.de). Aligned sequences were inserted within a stable phylogenetic tree by using the ARB parsimony tool. In this work we report the morphology and phylogenetic characterization of two isolates belonged to Laguna Blanca sediments.

Description: We present data and results from an ongoing project of astrobiological high-altitude expeditions investigating the highest and least explored perennial lakes on Earth in the Bolivian and Chilean Andes, including several volcanic crater lakes nearing and beyond 6,000 m in elevation. In the next five years, they will provide the first integrated long-term astrobiological characterization and monitoring of lacustrine environments and their biology for such altitude. These extreme lakes are natural laboratories. They provide the field data missing beyond 4,000 m to complete our understanding of terrestrial lakes and biota. Research on the effects of UV has been performed in lower altitude lakes and models of UV flux over time are being developed. Lakes showing a high content of dissolved organic material (DOM) shield organisms from UV. DOM acts as a natural sunscreen as it influences the water transparency, therefore is a determinant of photic zone depth. In sparsely vegetated alpine areas, lakes are clearer and offer less protection from UV to organisms living in the water. Transparent water and high UV irradiance may maximize the penetration and effect of UV radiation. Shallow-water communities in these lakes are particularly sensitive to UV radiation. The periphyton can live on various susbtrates. While on rocks, it includes immobile species that cannot seek low UV refuges unlike sediment-dwelling periphyton or alpine phytoflagellates which undergo vertical migration. Inhibition of algal photosynthesis by UV radiation has been documented in laboratory and showed that phytoplankton production is reduced by formation of nucleic acid lesions or production of peroxides and free oxygen radicals. of peroxides and free oxygen radicals. Our project is providing the field data that is missing from natural laboratories beyond 4,000 m and will complement the vision of the effects of UV on life and its adaptation modes (or lack thereof).