Microbialites from the Freshwater System of Cuatro Ciénegas, Mexico: Genomic, Molecular Organic, and Stable Isotopic Perspectives

Modern microbialites are carbonate-precipitating microbial mats and represent the closest living analogues to ancient stromatolites. These ancient carbonate formations are the oldest fossil evidence of life on Earth; however, our comprehension of their relationship to early earth ecosystems relies heavily on understanding the formation of modern microbialites. Research regarding these formation processes has suggested that chemical constraints of CaCO 3 precipitation vary on sub-millimeter spatial scales within the living microbial community. In an attempt to shed light on the importance of these chemical microenvironments, this study focused on understanding the spatial distribution of the organisms and processes involved in the formation of modern microbialites. This was accomplished by isolating five visually distinct layers from the upper 2 – 3 cm of an actively forming microbialite found in the freshwater system of Cuatro Ciénegas, Mexico. Each layer was analyzed using genomic, molecular organic, and stable isotopic techniques. Bacterial diversity was determined by 16S rRNA gene analyses, lipid biomarker content was detected by GC-MS, and carbon isotope composition of organic matter and CaCO 3 were used as indicators of specific microbial processes. Results of the 16S rRNA gene analysis showed that there is little overlap in the community composition of individual layers. Approximately 90% of the ribotypes identified in the microbialite were unique to a single layer. Furthermore, the relative accretion of CaCO 3 at each layer was used to connect the distribution of organisms and processes with two specific zones of CaCO 3 precipitation. The first zone of CaCO3 accretion, which accounted for approximately 55% of total CaCO 3 accumulation, is found in the surface two layers of the microbialites and dominated by photoautotrophic cyanobacteria and algae. The second zone of CaCO 3 precipitation, found at the interior (layers 4 and 5), is composed primarily of heterotrophic proteobacteria and dominated by sulfate-reducing !-proteobacteria. The lipid content of the microbialite reflected the community structure as determined by genomics. Numerous photosynthetic biomarkers were detected and decreased in abundance with depth, indicating the important function of heterotrophic degradation. Additionally, the detection of sulfurized phytol compounds in layer 5 highlighted an important mechanism for the preservation of biogenic signatures, and reflected both the abundance of phototrophic organisms and sulfatereducing bacteria. In combination, these interdisciplinary analyses provided an understanding of microbial community composition and metabolism while indicating the spatial relationship to CaCO 3 formation and the preservation of distinct biochemical signatures.