Sulfate reducing bacteria (SRB) have existed throughout much of Earth's history and remain major contributors to carbon cycling in modern systems. Despite their importance, misconceptions about SRB ...are prevalent. In particular, SRB are commonly thought to lack oxygen tolerance and to exist only in anoxic environments. Through the last two decades, researchers have discovered that SRB can, in fact, tolerate and even respire oxygen. Investigations of microbial mat systems have demonstrated that SRB are both abundant and active in the oxic zones of mats. Additionally, SRB have been found to be highly active in the lithified zones of microbial mats, suggesting a connection between sulfate reduction and mat lithification. In the present paper, we review recent research on SRB distribution and present new preliminary findings on both the diversity and distribution of δ-proteobacterial SRB in lithifying and non-lithifying microbial mat systems. These preliminary findings indicate the unexplored diversity of SRB in a microbial mat system and demonstrate the close microspatial association of SRB and cyanobacteria in the oxic zone of the mat. Possible mechanisms and further studies to elucidate mechanisms for carbonate precipitation via sulfate reduction are also discussed.
Microbialites (benthic microbial carbonate deposits) were discovered in a hypersaline alkaline lake on Eleuthera Island (Bahamas). From the edge towards the centre of the lake, four main zones of ...precipitation could be distinguished: (1) millimetre‐sized clumps of Mg‐calcite on a thin microbial mat; (2) thicker and continuous carbonate crusts with columnar morphologies; (3) isolated patches of carbonate crust separated by a dark non‐calcified gelatinous mat; and (4) a dark microbial mat without precipitation. In thin section, the precipitate displayed a micropeloidal structure characterized by micritic micropeloids (strong autofluorescence) surrounded by microspar and spar cement (no fluorescence). Observations using scanning electron microscopy (SEM) equipped with a cryotransfer system indicate that micrite nucleation is initiated within a polymer biofilm that embeds microbial communities. These extracellular polymeric substances (EPS) are progressively replaced with high‐Mg calcite. Discontinuous EPS calcification generates a micropeloidal structure of the micrite, possibly resulting from the presence of clusters of coccoid or remnants of filamentous bacteria. At high magnification, the microstructure of the initial precipitate consists of 200–500 nm spheres. No precipitation is observed in or on the sheaths of cyanobacteria, and only a negligible amount of precipitation is directly associated with the well‐organized and active filamentous cyanobacteria (in deeper layers of the mat), indicating that carbonate precipitation is not associated with CO2 uptake during photosynthesis. Instead, the precipitation occurs at the uppermost layer of the mat, which is composed of EPS, empty filamentous bacteria and coccoids (Gloeocapsa spp.). Two‐dimensional mapping of sulphate reduction shows high activity in close association with the carbonate precipitate at the top of the microbial mat. In combination, these findings suggest that net precipitation of calcium carbonate results from a temporal and spatial decoupling of the various microbial metabolic processes responsible for CaCO3 precipitation and dissolution. Theoretically, partial degradation of EPS by aerobic heterotrophs or UV fuels sulphate‐reducing activity, which increases alkalinity in microdomains, inducing CaCO3 precipitation. This degradation could also be responsible for EPS decarboxylation, which eliminates Ca2+‐binding capacity of the EPS and releases Ca2+ ions that were originally bound by carboxyl groups. At the end of these processes, the EPS biofilm is calcified and exhibits a micritic micropeloidal structure. The EPS‐free precipitate subsequently serves as a substrate for physico‐chemical precipitation of spar cement from the alkaline water of the lake. The micropeloidal structure has an intimate mixture of micrite and microspar comparable to microstructures of some fossil microbialites.
In modern stromatolites, mineralization results from a complex interplay between microbial metabolisms, the organic matrix, and environmental parameters. Here, we combined biogeochemical, ...mineralogical, and microscopic analyses with measurements of metabolic activity to characterize the mineralization processes and products in an emergent (<18 months) hypersaline microbial mat. While the nucleation of Mg silicates is ubiquitous in the mat, the initial formation of a Ca‐Mg carbonate lamina depends on (i) the creation of a high‐pH interface combined with a major change in properties of the exopolymeric substances at the interface of the oxygenic and anoxygenic photoautotrophic layers and (ii) the synergy between two major players of sulfur cycle, purple sulfur bacteria, and sulfate‐reducing bacteria. The repetition of this process over time combined with upward growth of the mat is a possible pathway leading to the formation of a stromatolite.
Mineral precipitation in microbial mats may have been the key to their preservation as fossil stromatolites, potentially documenting evidence of the earliest life on Earth. Two factors that ...contribute to carbonate mineral precipitation are the saturation index (SI) and the presence of nucleation sites. Both of these can be influenced by micro‐organisms, which can either alter SI through their metabolisms, or produce and consume organic substances such as extracellular polymeric substances (EPS) that can affect nucleation. It is the balance of individual metabolisms within the mat community that determines the pH and the dissolved inorganic carbon concentration, thereby potentially increasing the alkalinity and consequently the SI. Sulfate‐reducing bacteria (SRB) are an important component of this ‘alkalinity engine.’ The activity of SRB often peaks in layers where CaCO3 precipitates, and mineral precipitation has been demonstrated in SRB cultures; however, the effect of their metabolism on the alkalinity engine and actual contribution to mineral precipitation is the subject of controversy. Here, we show through culture experiments, theoretical calculations, and geochemical modeling studies that the pH, alkalinity, and organomineralization potential will vary depending on the type of electron donor. Specifically, hydrogen and formate can increase the pH, but electron donors like lactate and ethanol, and to a lesser extent glycolate, decrease the pH. The implication of this for the lithification of mats is that the combination of processes supplying electron donors and the utilization of these compounds by SRB may be critical to promoting mineral precipitation.
Microbial mats are dynamic and complex ecosystems exhibiting spatial and temporal heterogeneity. The physical/chemical environment is typified by steep gradients and distinct microenvironments. These ...microenvironments support a great diversity of species with a wide range of metabolic processes. These processes often result in coupled reactions and biogeochemical cycles, and produce important end products such as trace gases and mineral precipitates. The latter can impact the composition and character of the sediment, imparting a “biosignature.” These biosignatures can be preserved in the rock record and are useful in the interpretation of fossil record on Earth and possibly as an indication of life on other planetary bodies. The modern marine stromatolites of the Exuma Cays, Bahamas, provide an ideal system for studying the populations, processes, and products in a microbial ecosystem using a multidisciplinary approach. In order to acquire redox energy, microbial populations need to carry out metabolic reactions at rates faster than the equivalent chemical (abiotic) reactions. As such, microbes can be viewed as bioreactors that preferably oxidize carbon to CO
2 to maximize the energy yield. The study of the microbial role in carbonate sedimentation and lithification in these stromatolites provides a picture of microbial mats as bioreactors producing a biosignature.
The use of metals as biosignatures in the fossil stromatolite record requires understanding of the processes controlling the initial metal(loid) incorporation and diagenetic preservation in living ...microbialites. Here, we report the distribution of metals and the organic fraction within the lithifying microbialite of the hypersaline Big Pond Lake (Bahamas). Using synchrotron‐based X‐ray microfluorescence, confocal, and biphoton microscopies at different scales (cm–μm) in combination with traditional geochemical analyses, we show that the initial cation sorption at the surface of an active microbialite is governed by passive binding to the organic matrix, resulting in a homogeneous metal distribution. During early diagenesis, the metabolic activity in deeper microbialite layers slows down and the distribution of the metals becomes progressively heterogeneous, resulting from remobilization and concentration as metal(loid)‐enriched sulfides, which are aligned with the lamination of the microbialite. In addition, we were able to identify globules containing significant Mn, Cu, Zn, and As enrichments potentially produced through microbial activity. The similarity of the metal(loid) distributions observed in the Big Pond microbialite to those observed in the Archean stromatolites of Tumbiana provides the foundation for a conceptual model of the evolution of the metal distribution through initial growth, early diagenesis, and fossilization of a microbialite, with a potential application to the fossil record.
The initial lamination in young, metabolically active Scytonema knobs developing in Storr's Lake (Bahamas) results from the iterative succession of two different stages of microbial growth at the top ...of this microbialite. Stage 1 is dominated by vertically oriented cyanobacterial filaments and is characterized by a high porosity of the fabric. Stage 2 shows a higher microbial density with the filaments oriented horizontally and with higher carbonate content. The more developed, dense microbial community associated with Stage 2 of the Scytonema knobs rapidly degrades extracellular organic matter (EOM) and coupled to this, precipitates carbonate. The initial nucleation forms high‐Mg calcite nanospheroids that progressively replace the EOM. No precipitation is observed within the thick sheath of the Scytonema filaments, possibly because of strong cross‐linking of calcium and EOM (forming EOM‐Ca‐EOM complexes), which renders Ca unavailable for carbonate nucleation (inhibition process). Eventually, organominerals precipitate and form an initial lamina through physicochemical and microbial processes, including high rates of photosynthetic activity that lead to 13C‐enriched DIC available for initial nucleation. As this lamina moves downward by the iterative production of new laminae at the top of the microbialite, increased heterotrophic activity further alters the initial mineral product at depth. Although some rare relic preservation of ‘Stage 1–Stage 2’ laminae in subfossil knobs exists, the very fine primary lamination is considerably altered and almost completely lost when the knobs develop into larger and more complex morphologies due to the increased accommodation space and related physicochemical and/or biological alteration. Despite considerable differences in microstructure, the emerging ecological model of community succession leading to laminae formation described here for the Scytonema knobs can be applied to the formation of coarse‐grained, open marine stromatolites. Therefore, both fine‐ and coarse‐grained extant stromatolites can be used as model systems to understand the formation of microbialites in the fossil record.
Extracellular polymeric secretions (EPS) that are produced by cyanobacteria represent potential structuring agents in the formation of marine stromatolites. The abundance, production, and degradation ...of EPS in the upper layers of a microbial mat forming shallow subtidal stromatolites at Highborne Cay, Bahamas, were determined using
14C tracer experiments and were integrated with measurements of other microbial community parameters. The upper regions of a Type 2 Reid, R.P., Visscher, P.T., Decho, A.W., Stolz, J., Bebout, B., MacIntyre, I.G., Dupraz, C., Pinckney, J., Paerl, H., Prufert-Bebout, L., Steppe, T., Des Marais, D., 2000. The role of microbes in accretion, lamination and early lithification of modern marine stromatolites. Nature (London) 406, 989–992 stromatolite mat exhibited a distinct layering of alternating “green” cyanobacteria-rich layers (Layers 1 and 3) and “white” layers (Layers 2 and 4), and the natural abundance of EPS varied significantly depending on the mat layer. The highest EPS abundance occurred in Layer 2. The production of new EPS, as estimated by the incorporation of
14C-bicarbonate into EPS, occurred in all layers examined, with the highest production in Layer 1 and during periods of photosynthesis (i.e., daylight hours). A large pool (i.e., up to 49%) of the total
14C-bicarbonate uptake was released as low molecular-weight (MW) dissolved organic carbon (DOC). This DOC was rapidly mineralized to CO
2 by heterotrophic bacteria. EPS degradation, as determined by the conversion of
14C-EPS to
14CO
2, was slowest in Layer 2. Results of slurry experiments, examining O
2 uptake following additions of organic substrates, including EPS, supported this degradation trend and further demonstrated selective utilization by heterotrophs of specific monomers, such as acetate, ethanol, and uronic acids. Results indicated that natural EPS may be rapidly transformed post-secretion by heterotrophic degradation, specifically by sulfate-reducing bacteria, to a more-refractory remnant polymer that is relatively slow to accumulate. A mass balance analysis suggested that a layer-specific pattern in EPS and low-MW DOC turnover may contribute to major carbonate precipitation events within stromatolites. Our findings represent the first estimate of EPS turnover in stromatolites and support an emerging idea that stromatolite formation is limited by a delicate balance between evolving microbial activities and environmental factors.
ABSTRACT
Sulfate‐reducing bacteria (SRB) have been recognized as key players in the precipitation of calcium carbonate in lithifying microbial communities. These bacteria increase the alkalinity by ...reducing sulfate ions, and consuming organic acids. SRB also produce copious amounts of exopolymeric substances (EPS). All of these processes influence the morphology and mineralogy of the carbonate minerals. Interactions of EPS with metals, calcium in particular, are believed to be the main processes through which the extracellular matrix controls the precipitation of the carbonate minerals. SRB exopolymers were purified from lithifying mat and type cultures, and their potential role in CaCO3 precipitation was determined from acid‐base titrations and calcium‐binding experiments. Major EPS characteristics were established using infrared spectroscopy and gas chromatography to characterize the chemical functional groups and the sugar monomers composition. Our results demonstrate that all of the three SRB strains tested were able to produce large amounts of EPS. This EPS exhibited three main buffering capacities, which correspond to carboxylic acids (pKa = 3.0), sulfur‐containing groups (thiols, sulfonic and sulfinic acids – pKa = 7.0–7.1) and amino groups (pKa = 8.4–9.2). The calcium‐binding capacity of these exopolymers in solution at pH 9.0 ranged from 0.12gCa gEPS−1–0.15 gCa gEPS−1. These results suggest that SRB could play a critical role in the formation of CaCO3 in lithifying microbial mats. The unusually high sulfur content, which has not been reported for EPS before, indicates a possible strong interaction with iron. In addition to changing the saturation index through metabolic activity, our results imply that SRB affect the rock record through EPS production and its effect on the CaCO3 precipitation. Furthermore, EPS produced by SRB may account for the incorporation of metals (e.g. Sr, Fe, Mg) associated with carbonate minerals in the rock record.
Microbialites are sedimentary deposits formed by the metabolic interactions of microbes and their environment. These lithifying microbial communities represent one of the oldest ecosystems on Earth, ...yet the molecular mechanisms underlying the function of these communities are poorly understood. In this study, we used comparative metagenomic and metatranscriptomic analyses to characterize the spatial organization of the thrombolites of Highborne Cay, The Bahamas, an actively forming microbialite system. At midday, there were differences in gene expression throughout the spatial profile of the thrombolitic mat with a high abundance of transcripts encoding genes required for photosynthesis, nitrogen fixation and exopolymeric substance production in the upper three mm of the mat. Transcripts associated with denitrification and sulfate reduction were in low abundance throughout the depth profile, suggesting these metabolisms were less active during midday. Comparative metagenomics of the Bahamian thrombolites with other known microbialite ecosystems from across the globe revealed that, despite many shared core pathways, the thrombolites represented genetically distinct communities. This study represents the first time the metatranscriptome of living microbialite has been characterized and offers a new molecular perspective on those microbial metabolisms, and their underlying genetic pathways, that influence the mechanisms of carbonate precipitation in lithifying microbial mat ecosystems.
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