Mobile genetic elements, including plasmids, drive the evolution of prokaryotic genomes through the horizontal transfer of genes allowing genetic exchange between bacteria. Moreover, plasmids carry accessory genes, which encode functions that may offer an advantage to the host. Thus, it is expected that in a certain ecological niche, plasmids are enriched in accessory functions, which are important for their hosts to proliferate in that niche. Puquio de Campo Naranja is a high-altitude lake from the Andean Puna exposed to multiple extreme conditions, including high UV radiation, alkalinity, high concentrations of arsenic, heavy metals, dissolved salts, high thermal amplitude and low O 2 pressure. Microorganisms living in this lake need to develop efficient mechanisms and strategies to cope under these conditions. The aim of this study was to characterize the plasmidome of microbialites from Puquio de Campo Naranja, and identify potential hosts and encoded functions using a deep-sequencing approach. The potential ecological impact of the plasmidome, including plasmids from cultivable and non-cultivable microorganisms, is described for the first time in a lake representing an extreme environment of the Puna. This study showed that the recovered genetic information for the plasmidome was novel in comparison to the metagenome derived from the same environment. The study of the total plasmid population allowed the identification of genetic features typically encoded by plasmids, such as resistance and virulence factors. The resistance genes comprised resistances to heavy metals, antibiotics and stress factors. These results highlight the key role of plasmids for their hosts and impact of extrachromosomal elements to thrive in a certain ecological niche.

The acrosome reaction (AR) is a special exocytotic process promoted by signal transduction pathways studied in many laboratories. Progesterone (P4) is one of the trigger molecules proposed. Upon the binding of P4 to its receptor, several molecules could be activated, including G-proteins, phospholipase A(2) (PLA(2)), and phospholipase C (PLC). The role of these molecules was analyzed in this study using the Chlortetracycline (CTC) protocol to detect and quantify the AR. Incubation of capacitated sperm cells with GTPgammas (GTPgammas, a mimetic of G-protein activation), arachidonic acid (AA, product of PLA(2) action), or phorbol ester (PMA, an activator of PLC) for 15 min increased the AR to a similar percentage as P4. Conversely, a decrease in the AR was detected when sperm cells were incubated with P4 after preincubation with: GDPbetaS (GDP, an inhibitor of G-protein activation), ONO RS-82 (ONO, an inhibitor of PLA(2)), or neomycin (Neo, an inhibitor of PLC) for 15 min. To analyze the activation sequence of G proteins, PLA(2), and PLC combinations of these mimetic/inhibitors were used during successive incubation periods. Inhibition promoted by GDP, ONO, and Neo were overcome by 15-min incubation with GTPgammas, AA, or PMA, respectively. But GTPgammas or P4 did not reverse the inhibition due to incubation with Neo and ONO. Interestingly, this dual inhibition was reverted by another 15-min incubation with AA or PMA. Results presented here could indicate that the AR triggered by P4 is driven by activation of G-proteins, that in turn activate PLA(2) and PLC simultaneously, that finally promote acrosomal exocytosis.

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Biofilms, microbial mats, and microbialites dwell under highly limiting conditions (high salinity, extreme aridity, pH, and elevated arsenic concentration) in the Andean Puna. Only recent pioneering studies have described the microbial diversity of different Altiplano lakes and revealed their unexpectedly diverse microbial communities. Arsenic metabolism is proposed to be an ancient mechanism to obtain energy by microorganisms. Members of Bacteria and Archaea are able to exploit arsenic as a bioenergetic substrate in either anaerobic arsenate respiration or chemolithotrophic growth on arsenite. Only six aioAB sequences coding for arsenite oxidase and three arrA sequences coding for arsenate reductase from haloarchaea were previously deposited in the NCBI database. However, no experimental data on their expression and function has been reported. Recently, our working group revealed the prevalence of haloarchaea in a red biofilm from Diamante Lake and microbial mat from Tebenquiche Lake using a metagenomics approach. Also, a surprisingly high abundance of genes used for anaerobic arsenate respiration (arr) and arsenite oxidation (aio) was detected in the Diamante's metagenome. In order to study in depth the role of arsenic in these haloarchaeal communities, in this work, we obtained 18 haloarchaea belonging to the Halorubrum genus, tolerant to arsenic. Furthermore, the identification and expression analysis of genes involved in obtaining energy from arsenic compounds (aio and arr) showed that aio and arr partial genes were detected in 11 isolates, and their expression was verified in two selected strains. Better growth of two isolates was obtained in presence of arsenic compared to control. Moreover, one of the isolates was able to oxidize As[III]. The confirmation of the oxidation of arsenic and the transcriptional expression of these genes by RT-PCR strongly support the hypothesis that the arsenic can be used in bioenergetics processes by the microorganisms flourishing in these environments.

The Salar de Antofalla (salt flat) is located in the Puna region of Catamarca, in northern Argentina. In this paper we report and provide the first descriptive data of Las Quínoas, a modern system of oncoids located in the western margin of the salt flat. Oncoids were studied by insitu logging, polished and thin sections analysis. In addition, the 16s rRNA genes of microbial mats associated with these oncoids were amplified and sequenced to characterize the microbial biodiversity. Oncoids present discoidal to subrounded morphologies and sizes up to 15 cm in diameter. They are scattered along channels, which originate from the groundwater springs of a wetland and enter the salt flat. Its macrostructure is concentric and composed by three zones: 1. A nucleus zone built by clastic material in a carbonate matrix. 2. A well-laminated zone around the nucleus that shows two types of mesostructures: concentrically stacked spheroids (SS-C) and randomly stacked hemispheroids (SS-R), both showing an alternation of dense and dark micritic laminae with light micritic to microsparitic laminae. 3. A poorly-laminated zone, in the outermost sector of oncoids, with two types of mesostructures too: a laminated mesostructure composed also of an alternation of dense and dark micritic laminae with light micritic to microsparitic laminae, and a non-laminated mesostructure composed of agglomerated and cemented clastic material within a calcareous matrix (wackestones-packstones). Regarding the microbial diversity, the analyzed oncoids in this work are mainly inhabited by Proteobacteria (ca. 37.5%), Bacteroidetes (ca. 25.0%), and in less proportion Planctomycetes, Actinobacteria and Cyanobacteria.

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Pozo Bravo is a high-altitude Andean lake that harbors modern microbialites thriving in hypersaline conditions in the Salar de Antofalla, one of the driest sites on Earth and located in the Puna region of Catamarca, northwest Argentine. Due to the lake physiography, microbialites are restricted to a narrow belt following Pozo Bravo lake variations. Microbialites exhibit a wide range of external morphologies including domal, discoidal, tabular, and horseshoe-like bioherms which vary considerably in size, as well as large biostromal terraces. As documented by other studies on modern microbialites, external morphology appears to be mainly the product of the environmental setting. In Pozo Bravo lake, high evaporation rates and hypersalinity (driven by high temperature and strong winds), water-level fluctuations, and lake-bottom topography are major controlling factors. The distinctive feature of Pozo Bravo microbialites is their internal structure, showing a gradual transition from a thrombolitic core to dendrolitic structures and to a sharply overlying stromatolitic layer within a single microbialite. We suggest that these various microbialite textures represent a gradual change within an environmental gradient based on lake-level variations, and the influence of these environmental factors on biological activity, mainly by cyanobacteria and diatoms. The study of this site is particularly relevant given that it represents an active system where progressive changes in microbialite type (from thrombolites to dendrolites and stromatolites) are recorded, providing an excellent natural laboratory to study these textural changes from a mechanistic perspective, and it may provide insights for better understanding of the microbialite geological record. In addition, given that these systems are threatened by human activities (mining of lithium-rich brines), its study and preservation are necessary.