Serpentinization is a well-known aqueous alteration process that may have played important roles in the origins and early evolution of life on Earth, and perhaps Mars, but there are still aspects ...related to biomarker distribution, partitioning, and preservation that merit further study. To assess the role that precipitation of carbonate phases in serpentinization settings may have on biomarker preservation, we search for life signs in one of the world's largest outcrops of subcontinental peridotites (Ronda, South Spain). We investigate the organic record of groundwater and associated carbonate deposits (travertines) in seven hyperalkaline springs, and reconstruct the biological activity and metabolic interactions of the serpentinization-hosted ecosystem. We identified lipid biomarkers and isotopic evidences of life, whose concentration and variety were much lower in groundwater than travertine deposits (ppb/ppt versus ppm level). Groundwater carried organics of abiotic (n-alkanes with values of CPI ∼ 1) and/or biotic origin, of fresher (e.g. acids or alcohols) or more diagenetized (mature hopanes and n-alkanes) nature. In contrast, associated travertines held a more prolific record of biomarkers incorporating (molecular and isotopic) fingerprints of surface (mostly phototrophs) and subsurface (chemolithotrophs, methanogens and/or methanotrophs) life. Serpentinization-associated travertines seem to act as biomolecule archives over time fed by autochthonous and allochthonous sources, hence amplifying the dim biological signal of groundwater. These results illustrate the relevance of serpentinization-associated surface mineral deposits in searching for traces of life on analogous environments on Mars. We highlight the diversity of lipids produced in serpentinizing land environments and emphasize the potential of these geostable biomolecules to preserve fingerprints of life.
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•Interest of serpentinization for understanding origins and early evolution of life•Seven serpentinization-associated hyperalkaline springs from Ronda are explored.•The organic record of groundwater and associated carbonates is investigated.•Lipid biomarkers and isotopic evidences of serpentinization-hosted life are found.•The travertines preserve more and more varied biomarkers than groundwaters.
Adequate read filtering is critical when processing high-throughput data in marker-gene-based studies. Sequencing errors can cause the mis-clustering of otherwise similar reads, artificially ...increasing the number of retrieved Operational Taxonomic Units (OTUs) and therefore leading to the overestimation of microbial diversity. Sequencing errors will also result in OTUs that are not accurate reconstructions of the original biological sequences. Herein we present the Poisson binomial filtering algorithm (PBF), which minimizes both problems by calculating the error-probability distribution of a sequence from its quality scores. In order to validate our method, we quality-filtered 37 publicly available datasets obtained by sequencing mock and environmental microbial communities with the Roche 454, Illumina MiSeq and IonTorrent PGM platforms, and compared our results to those obtained with previous approaches such as the ones included in mothur, QIIME and USEARCH. Our algorithm retained substantially more reads than its predecessors, while resulting in fewer and more accurate OTUs. This improved sensitiveness produced more faithful representations, both quantitatively and qualitatively, of the true microbial diversity present in the studied samples. Furthermore, the method introduced in this work is computationally inexpensive and can be readily applied in conjunction with any existent analysis pipeline.
Viruses control natural microbial communities. Identification of virus-host pairs relies either on their cultivation or on metagenomics and tentative assignment based on genomic signatures. Both ...approaches have severe drawbacks when aiming to target such pairs within the uncultured majority. Here we present an unambiguous way to assign viruses to hosts that does not rely on any previous information about either of them nor requires their cultivation. First, genomic contents of individual cells present in an environmental sample are retrieved by means of single-cell genomic technologies. Then, individual cell genomes are hybridized against a set of individual viral genomes from the same sample, previously immobilized on a microarray. Infected cells will yield positive hybridization as they carry viral genomes, which can be then sequenced and characterized. Using this method, we pinpoint viruses infecting the ubiquitous hyperhalophilic Nanohaloarchaeota, included in the so-called 'microbial dark matter' (the uncultured fraction of the microbial world).
Detecting signs of potential extant/extinct life on Mars is challenging because the presence of organics on that planet is expected to be very low and most likely linked to radiation-protected ...refugia and/or preservative strategies (e.g., organo-mineral complexes). With scarcity of organics, accounting for biomineralization and potential relationships between biomarkers, mineralogy, and geochemistry is key in the search for extraterrestrial life. Here we explored microbial fingerprints and their associated mineralogy in Icelandic hydrothermal systems analog to Mars (i.e., high sulfur content, or amorphous silica), to identify potentially habitable locations on that planet. The mineralogical assemblage of four hydrothermal substrates (hot springs biofilms, mud pots, and steaming and inactive fumaroles) was analyzed concerning the distribution of biomarkers. Molecular and isotopic composition of lipids revealed quantitative and compositional differences apparently impacted by surface geothermal alteration and environmental factors. pH and water showed an influence (i.e., greatest biomass in circumneutral settings with highest supply and turnover of water), whereas temperature conditioned the mineralogy that supported specific microbial metabolisms related with sulfur. Raman spectra suggested the possible coexistence of abiotic and biomediated sources of minerals (i.e., sulfur or hematite). These findings may help to interpret future Raman or GC-MS signals in forthcoming Martian missions.
The Late Miocene–Pliocene aged hyperarid evaporitic system of Salar Grande is a unique, halite-rich sedimentary basin in the Cordillera de la Costa of the Central Andes (Chile) whose bio-sedimentary ...record is poorly understood. The persistence of hyperacidity over millions of years, the hypersalinity, and the intense UV radiation make it a terrestrial analogue to assess the potential presence of organic matter in the halite deposits found on Mars. We investigated the occurrence and distribution of biomolecules along a 100-m depth drill down to the ~ 9 Ma old detrital deposits topped by La Soledad Formation (ESF). We have identified two well-defined mineralogical and geochemical units by X-ray diffractometry (XRD) and ion chromatography: a nearly pure halite down to 40 m, and a detrital one down to 100 m depth. One-dimensional GC–MS and two-dimensional GC × GC-TOF–MS gas chromatography–mass spectrometry techniques allowed us to detect a variety of lipidic compounds (n-alkanes, n-alkanols, isoprenoids, steroids, and hopanoids), and a relative abundance of functionalized hydrocarbons (n-fatty acids or n-aldehydes), mostly in the upper halite. We also detected biopolymers and microbial markers by fluorescence sandwich-microarray immunoassays. A dominant prokaryotic origin was associated with halophile bacteria and archaea, with minor contributions of lichens, macrophytes, or higher plants. The lipidic record was also imprinted by oxic (high pristane over phytane ratios) and saline (squalane, and mono-methyl n-alkanes) signatures. The vertical abundance and distribution of biomarkers in the Salar Grande was explained by a generalized effect of xeropreservation, combined with salt encapsulation in the upper halite deposits, or with protective organics-mineral interactions in the deeper detrital unit. The results contribute to the interpretation of terrestrial bio-sedimentary records of halite deposits and their association to environmental conditions. The high potential for preservation of biosignatures at Salar Grande suggests that similar evaporitic deposits in Mars should be priority targets for searching for signs of life.
Ancient peptides are remnants of early biochemistry that continue to play pivotal roles in current proteins. They are simple molecules yet complex enough to exhibit independent functions, being ...products of an evolved biochemistry at the interface of life and nonlife. Their adsorption to minerals may contribute to their stabilization and preservation over time. To investigate the feasibility of conserved peptide sequences and structures as target biomarkers for the search for life on Mars or other planetary bodies, we conducted a bioinformatics selection of well-conserved ancient peptides and produced polyclonal antibodies for their detection using fluorescence microarray immunoassays. Additionally, we explored how adsorbing peptides to Mars-representative minerals to form organomineral complexes could affect their immunological detection. The results demonstrated that the selected peptides exhibited autonomous folding, with some of them regaining their structure, even after denaturation. Furthermore, their cognate antibodies detected their conformational features regardless of amino acid sequences, thereby broadening the spectrum of target peptide sequences. While certain antibodies displayed unspecific binding to bare minerals, we validated that peptide–mineral complexes can be detected using sandwich immunoassays, as confirmed through desorption and competitive assays. Consequently, we conclude that the diversity of peptide sequences and structures suitable for use as target biomarkers in astrobiology can be constrained to a few well conserved sets, and they can be detected even if they are adsorbed in organomineral complexes.
Although phosphate and carbonate are important constituents in ancient and modern environments, it is not yet clear their biogeochemical relationships and their mechanisms of formation. Microbially ...mediated carbonate formation has been widely studied whereas little is known about the formation of phosphate minerals. Here we report that a new bacterial strain, Tessarococcus lapidicaptus, isolated from the subsurface of Rio Tinto basin (Huelva, SW Spain), is capable of precipitating Fe-rich phosphate and carbonate minerals. We observed morphological differences between phosphate and carbonate, which may help us to recognize these minerals in terrestrial and extraterrestrial environments. Finally, considering the scarcity and the unequal distribution and preservation patterns of phosphate and carbonates, respectively, in the geological record and the biomineralization process that produces those minerals, we propose a hypothesis for the lack of Fe-phosphates in natural environments and ancient rocks.
A novel actinobacterium, designated IPBSL-7T, was isolated from a drilling core 297 m deep obtained from the Iberian Pyrite Belt. The strain was isolated anaerobically using nitrate as the electron ...acceptor. 16S rRNA gene sequence analysis revealed that it was related to Tessaracoccus flavescens SST-39T (95.7 % similarity), Tessaracoccus bendigoensis Ben 106T (95.7 %), Tessaracoccus lubricantis KSS-17SeT (95.6 %) and Tessaracoccus oleiagri SL014B-20A1T (95.0 %), while its similarity to any other member of the family Propionibacteriaceae was less than 94 %. Cells were non-motile, non-spore-forming, Gram-positive, oval to rod-shaped, and often appeared in pairs or small groups. The strain was facultatively anaerobic, oxidase-negative, catalase-positive and capable of reducing nitrate. Colonies were circular, convex, smooth and colourless. The organism could grow at between 15 and 40 °C, with an optimal growth at 37 °C. The pH range for growth was from pH 6 to 9, with pH 8 being the optimal value. Strain IPBSL-7T had peptidoglycan type A3-γ′, with ll-diaminopimelic acid as the diagnostic diamino-acid and glycine at position 1 of the peptide subunit. The dominant menaquinone was MK-9(H4) (93.8 %). The major cellular fatty acid was anteiso-C15 : 0 (55.0 %). The DNA G+C content was 70.3 mol%. On the basis of phenotypic and phylogenetic results, strain IPBSL-7T can be differentiated from previously described species of the genus Tessaracoccus and, therefore, represents a novel species, for which the name Tessaracoccus lapidicaptus sp. nov. is proposed. The type strain is IPBSL-7T ( = CECT 8385T = DSM 27266T).
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