Environmental analogs from yellowstone hot springs on geochemical and microbial diversity with implications for the search for life on Mars

From Viking landers to Perseverance rover, Mars has been explored by several in situ missions capable of analyzing organic compounds. Results from the SAM and SHERLOC on Curiosity and Perseverance, respectively, support the detection of lean organic matter (at ppb-ppm levels) in the top surface samples, although the source(s) and preservation mechanisms are still ambiguous. Perseverance is currently exploring a fluvio-lacustrine system at Jezero crater and may explore an ancient volcanic terrain after exiting the crater. As Perseverance would collect samples for potential return to Earth, preparation is needed for sample return efforts through various means including i) the detection of trace organic compounds in various matrices, ii) validation of compounds identified by Martian rovers, and iii) better understanding of mechanisms of their production on Mars. On these returned samples, the community may be able to resolve the timing of organic matter formation and refine hypotheses regarding organic preservation in Martian soils despite the presence of numerous oxidants, salts, and pH-temperature intra and inter-site variations that are less conductive to long-term preservation of organic matter. For instance, acidic conditions promote clay catalyzed isomerization, but seem to benefit for the fatty acid preservation producing organic-salts or favoring salt dissolution in the matrix to protect organic compounds from radiations and water alteration. With a similar aim, we selected samples from Yellowstone National Park hot springs and silica sinters as analogs to locations visited by Curiosity and Perseverance or – in the future – Rosalind Franklin rover. The hot springs in this study developed over hundreds to thousands of years, providing optimal conditions (i.e., matrix composition, temperature, pH) of preservation for organic molecules, extremophilic and mesophilic cells. In our study, the most well preserved organic matter and biosignatures were detected in acidic silica sinters with a surface (water) temperature below 50 °C and a minor crystalline phase. The gas chromatography – mass spectrometry molecular analysis revealed a variety of organic compounds we classified as bioindicators (such as amino acids, nucleobases, and sugars), and biosignatures (such as long-chain branched and/or (poly)unsaturated lipids, secondary metabolites involved in the quorum sensing or communication between individuals). We validated with a SAM/MOMA-like benchtop extracting oven the organic matter extraction protocols performed with the SAM experiment. We identified using the different SAM and MOMA extraction protocols (pyrolysis and wet-chemistry derivatizations) eight microbial classes through a unique untargeted environmental metabolomics’ method embracing space flight technology constraints. Additionally, we identified one (and likely two) agnostic biosignature(s): i) the concomitance of some elements and organic compounds in the analogs (correlation of organic matter elements: C, N, S, P and organic molecules co-located with essential biological elements: Fe, Mg, V, Mn and non-essential biological elements concentrated by microorganisms: As, Cs, Ga), and ii) the negative isotope C and N ratio demonstrating organic molecules rich in ¹²C and ¹⁴N: archaeal, bacterial, and eukaryotic lipids for an efficient low energy-consuming metabolism.