Transformation of carbonate minerals in a cyano-bacterial mat in the course of laboratory modeling

A laboratory model of a cyano-bacterial mat with mineral layers of carbonates was used to examine the dynamics of the transformation of calcium-magnesium carbonate under the conditions of a soda lake. The activity of various organisms of the cyanobacterial community results in conditions under which the Ca-Mg carbonate precipitate undergoes changes. The crystal lattice of the initial carbonate is restructured; its mineralogical composition changes depending on the conditions of the mat. In magnesium calcites, which are formed under such low-temperature conditions, a rudimentary cation adjustment can occur with the formation of dolomite domains. These experiments confirm the hypothesis that the dolomite found in stromatolites is of a secondary origin and can be formed in the course of transformation of Ca-Mg carbonates under alkaline conditions in an alkaliphilic cyanobacterial community.     

Cyanobacterial life at low O(2): community genomics and function reveal metabolic versatility and extremely low diversity in a Great Lakes sinkhole mat

Cyanobacteria are renowned as the mediators of Earth's oxygenation. However, little is known about the cyanobacterial communities that flourished under the low-O(2) conditions that characterized most of their evolutionary history. Microbial mats in the submerged Middle Island Sinkhole of Lake Huron provide opportunities to investigate cyanobacteria under such persistent low-O(2) conditions. Here, venting groundwater rich in sulfate and low in O(2) supports a unique benthic ecosystem of purple-colored cyanobacterial mats. Beneath the mat is a layer of carbonate that is enriched in calcite and to a lesser extent dolomite. In situ benthic metabolism chambers revealed that the mats are net sinks for O(2), suggesting primary production mechanisms other than oxygenic photosynthesis. Indeed, (14)C-bicarbonate uptake studies of autotrophic production show variable contributions from oxygenic and anoxygenic photosynthesis and chemosynthesis, presumably because of supply of sulfide. These results suggest the presence of either facultatively anoxygenic cyanobacteria or a mix of oxygenic/anoxygenic types of cyanobacteria. Shotgun metagenomic sequencing revealed a remarkably low-diversity mat community dominated by just one genotype most closely related to the cyanobacterium Phormidium autumnale, for which an essentially complete genome was reconstructed. Also recovered were partial genomes from a second genotype of Phormidium and several Oscillatoria. Despite the taxonomic simplicity, diverse cyanobacterial genes putatively involved in sulfur oxidation were identified, suggesting a diversity of sulfide physiologies. The dominant Phormidium genome reflects versatile metabolism and physiology that is specialized for a communal lifestyle under fluctuating redox conditions and light availability. Overall, this study provides genomic and physiologic insights into low-O(2) cyanobacterial mat ecosystems that played crucial geobiological roles over long stretches of Earth history.     

Cyanobacterial formation of intracellular Ca‐carbonates in undersaturated solutions

Cyanobacteria have long been thought to induce the formation of Ca‐carbonates as secondary by‐products of their metabolic activity, by shifting the chemical composition of their extracellular environment to conditions favoring mineral precipitation. Some cyanobacterial species forming Ca‐carbonates intracellularly were recently discovered. However, the environmental conditions under which this intracellular biomineralization process can occur and the impact of cyanobacterial species forming Ca‐carbonates intracellularly on extracellular carbonatogenesis are not known. Here, we show that these cyanobacteria can form Ca‐carbonates intracellularly while growing in extracellular solutions undersaturated with respect to all Ca‐carbonate phases, that is, conditions thermodynamically unfavorable to mineral precipitation. This shows that intracellular Ca‐carbonate biomineralization is an active process; that is, it costs energy provided by the cells. The cost of energy may be due to the active accumulation of Ca intracellularly. Moreover, unlike cyanobacterial strains that have been usually considered before by studies on Ca‐carbonate biomineralization, cyanobacteria forming intracellular carbonates may slow down or hamper extracellular carbonatogenesis, by decreasing the saturation index of their extracellular solution following the buffering of the concentration of extracellular calcium to low levels.     

Microbial geochemical calcium cycle

The participation of microorganisms in the geochemical calcium cycle is the most important factor maintaining neutral conditions on the Earth. This cycle has profound influence on the fate of inorganic carbon, and, thereby, on the removal of CO2 from the atmosphere. The major part of calcium deposits was formed in the Precambrian, when prokaryotic biosphere predominated. After that, calcium recycling based on biogenic deposition by skeletal organisms became the main process. Among prokaryotes, only a few representatives, e.g., cyanobacteria, exhibit a special calcium function. The geochemical calcium cycle is made possible by the universal features of bacteria involved in biologically mediated reactions and is determined by the activities of microbial communities. In the prokaryotic system, the calcium cycle begins with the leaching of igneous rock predominantly through the action of the community of organotrophic organisms. The release of carbon dioxide to the soil air by organotrophic aerobes leads to leaching with carbonic acid and soda salinization. Under anoxic conditions, of major importance is the organic acid production by primary anaerobes (fermentative microorganisms). Calcium carbonate is precipitated by secondary anaerobes (sulfate reducers) and to a smaller degree by methanogens. The role of the cyanobacterial community in carbonate deposition is exposed by stromatolites, which are the most common organo-sedimentary Precambrian structures. Deposition of carbonates in cyanobacterial mats as a consequence of photoassimilation of CO2 does not appear to be a significant process. It is argued that carbonates were deposited at the boundary between the "soda continent", which emerged as a result of subaerial leaching with carbonic acid, and the ocean containing Ca2+. Such ecotones provided favorable conditions for the development of the benthic cyanobacterial community, which was a precursor of stromatolites.     

Laboratory analogs of cyanobacterial mats of the alkaline geochemical barrier

The goal of this work was to illustrate a possible interaction between the "soda continent" and the ocean. A laboratory simulation was undertaken of the development of alkaliphilic mat with calcium carbonate and calcium phosphate interlayers in the zone where ocean waters, containing calcium and manganese, come into contact with carbonate- and phosphate-rich alkaline waters. The macrostructure of the layered cyanobacterial mat turned out to little dependent on the chemical conditions causing sediment formation. The chemical composition of freshly formed mineral interlayers of the mat was found to vary with the medium composition. The mineralogical composition of the sediment is determined by diagenesis conditions in its depth, which can cause mineral phase conversions.     

Metabolic potential of lithifying cyanobacteria-dominated thrombolitic mats

Thrombolites are unlaminated carbonate deposits formed by the metabolic activities of microbial mats and can serve as potential models for understanding the molecular mechanisms underlying the formation of lithifying communities. To assess the metabolic complexity of these ecosystems, high throughput DNA sequencing of a thrombolitic mat metagenome was coupled with phenotypic microarray analysis. Functional protein analysis of the thrombolite community metagenome delineated several of the major metabolic pathways that influence carbonate mineralization including cyanobacterial photosynthesis, sulfate reduction, sulfide oxidation, and aerobic heterotrophy. Spatial profiling of metabolite utilization within the thrombolite-forming microbial mats suggested that the top 5 mm contained a more metabolically diverse and active community than the deeper within the mat. This study provides evidence that despite the lack of mineral layering within the clotted thrombolite structure there is a vertical gradient of metabolic activity within the thrombolitic mat community. This metagenomic profiling also serves as a foundation for examining the active role individual functional groups of microbes play in coordinating metabolisms that lead to mineralization.     

Microbial consumption of dimethyl sulfide and methanethiol in coastal marine sediments

Abstract: Samples were taken from oxic and anoxic zones of three ecosystems: a cyanobacterial mat, a diatom film and a carbonate sediment. Dimethylsulfide (DMS) concentrations were determined by headspace analysis of sediment slurries; maximal amounts were in the upper 5–10 mm of the sediments of 20 μM (cyanobacterial mat), 8 μM (diatom film) and < 1 μM in the carbonate sediment. Dissolved DMS in the cyanobacterial mat, determined by centrifugation and cryogenic trapping, was about two orders of magnitude lower than from slurry estimations but its variation with depth was similar. CH₃SH concentrations in slurried samples, determined after treatment with tributylphosphine, ranged from 2 to 7 μM in the diatom mat and was below the limit of detection (< 0.1 μM) in the carbonate sediment. MPN counts of bacteria that grew on DMS under oxic and anoxic (nitrate added) conditions were determined at all three sites. Aerobic DMS utilizers peaked in the surface and decreased with depth, while the population of anaerobic DMS utilizers was relatively constant in the top 20 mm. Populations of DMS utilizers were highest in the cyanobacterial mat and lowest in the carbonate sediment. MPN's of thiosulfate utilizers, aerobic and anaerobic (nitrate added) were determined in the cyanobacterial mat. Populations of aerobic and anaerobic S₂O₃²⁻ utilizers were similar throughout the top 20 mm and comparable to those of DMS utilizers in the top 5 mm, but higher by about 100-fold below that zone. DMS and CH₃SH consumption rates were measured in slurries of sediments and aerobic rates were similar or only slightly higher than anaerobic rates; the latter were stimulated by nitrate.     

Laboratory Simulations of Cyanobacterial Mats of the Alkaline Geochemical Barrier

The goal of this work was to illustrate a possible interaction between the soda continent and the ocean. A laboratory simulation was undertaken of the development of alkaliphilic mat with calcium carbonate and calcium phosphate interlayers in the zone where ocean waters, containing calcium and manganese, come into contact with carbonate- and phosphate-rich alkaline waters. The macrostructure of the layered cyanobacterial mat turned out to be little dependent on the chemical conditions causing sediment formation. The chemical composition of freshly formed mineral interlayers of the mat was found to vary with the medium composition. The mineralogical composition of the sediment is determined by diagenesis conditions in its depth, which can cause mineral phase conversions.     

Genetic variance in the composition of two functional groups (diazotrophs and cyanobacteria) from a hypersaline microbial mat

Examination of variation in ecological communities can lead to an understanding of the forces that structure communities, the consequences of change at the ecosystem level, and the relevant scales involved. This study details spatial and seasonal variability in the composition of nitrogen-fixing and cyanobacterial (i.e., oxygenic photosynthetic) functional groups of a benthic, hypersaline microbial mat from Salt Pond, San Salvador Island, Bahamas. This system shows extreme annual variability in the salinity of the overlying water and the extent of water coverage. Analysis of molecular variance and F(ST) tests of genetic differentiation of nifH and cyanobacterial 16S rRNA gene clone libraries allowed for changes at multiple taxonomic levels (i.e., above, below, and at the species level) to inform the conclusions regarding these functional groups. Composition of the nitrogen-fixing community showed significant seasonal changes related to salinity, while cyanobacterial composition showed no consistent seasonal pattern. Both functional groups exhibited significant spatial variation, changing with depth in the mat and horizontally with distance from the shoreline. The patterns of change suggest that cyanobacterial composition was more insensitive to water stress, and consequently, cyanobacteria dominated the nitrogen-fixing community during dry months but gave way to a more diverse community of diazotrophs in wet months. This seasonal pattern may allow the mat community to respond quickly to water-freshening events after prolonged dry conditions (system recovery) and maintain ecosystem function in the face of disturbance during the wet season (system resilience).     

Comparative microbial diversity analyses of modern marine thrombolitic mats by barcoded pyrosequencing

Thrombolites are unlaminated carbonate structures that form as a result of the metabolic interactions of complex microbial mat communities. Thrombolites have a long geological history; however, little is known regarding the microbes associated with modern structures. In this study, we use a barcoded 16S rRNA gene-pyrosequencing approach coupled with morphological analysis to assess the bacterial, cyanobacterial and archaeal diversity associated with actively forming thrombolites found in Highborne Cay, Bahamas. Analyses revealed four distinct microbial mat communities referred to as black, beige, pink and button mats on the surfaces of the thrombolites. At a coarse phylogenetic resolution, the domain bacterial sequence libraries from the four mats were similar, with Proteobacteria and Cyanobacteria being the most abundant. At the finer resolution of the rRNA gene sequences, significant differences in community structure were observed, with dramatically different cyanobacterial communities. Of the four mat types, the button mats contained the highest diversity of Cyanobacteria, and were dominated by two sequence clusters with high similarity to the genus Dichothrix, an organism associated with the deposition of carbonate. Archaeal diversity was low, but varied in all mat types, and the archaeal community was predominately composed of members of the Thaumarchaeota and Euryarchaeota. The morphological and genetic data support the hypothesis that the four mat types are distinctive thrombolitic mat communities.     

Genetic Variance in the Composition of Two Functional Groups (Diazotrophs and Cyanobacteria) from a Hypersaline Microbial Mat

Examination of variation in ecological communities can lead to an understanding of the forces that structure communities, the consequences of change at the ecosystem level, and the relevant scales involved. This study details spatial and seasonal variability in the composition of nitrogen-fixing and cyanobacterial (i.e., oxygenic photosynthetic) functional groups of a benthic, hypersaline microbial mat from Salt Pond, San Salvador Island, Bahamas. This system shows extreme annual variability in the salinity of the overlying water and the extent of water coverage. Analysis of molecular variance and F_ST tests of genetic differentiation of nifH and cyanobacterial 16S rRNA gene clone libraries allowed for changes at multiple taxonomic levels (i.e., above, below, and at the species level) to inform the conclusions regarding these functional groups. Composition of the nitrogen-fixing community showed significant seasonal changes related to salinity, while cyanobacterial composition showed no consistent seasonal pattern. Both functional groups exhibited significant spatial variation, changing with depth in the mat and horizontally with distance from the shoreline. The patterns of change suggest that cyanobacterial composition was more insensitive to water stress, and consequently, cyanobacteria dominated the nitrogen-fixing community during dry months but gave way to a more diverse community of diazotrophs in wet months. This seasonal pattern may allow the mat community to respond quickly to water-freshening events after prolonged dry conditions (system recovery) and maintain ecosystem function in the face of disturbance during the wet season (system resilience).     

Sulphate-reducing bacteria induce low-temperature Ca-dolomite and high Mg-calcite formation

In this study, we demonstrate that sulphate‐reducing bacteria induce anoxic low‐temperature Ca‐dolomite formation both in situ in Lagoa Vermelha and Brejo do Espinho, two neighbouring, dolomite‐precipitating hypersaline lagoons in Brazil, and in laboratory culture experiments. The metabolic activity of sulphate‐reducing bacteria facilitates dolomite formation under anoxic conditions, as demonstrated with experiments using dialysis bags. Overall changes in the chemical conditions of the medium exclusively, without the presence of bacteria, did not result in carbonate precipitation. Only pure cultures of metabolizing sulphate‐reducing bacteria induced Ca‐dolomite and high Mg‐calcite precipitates, indicating that the carbonate nucleation takes place in the locally changed microenvironment around the sulphate‐reducing bacterial cells. Not all pure strains, however, produced Ca‐dolomite under similar conditions, suggesting that the bacterial metabolism, activity and the rate of mineral precipitation have an influence on the type of carbonate formed.     

Microbial Geochemical Calcium Cycle

The participation of microorganisms in the geochemical calcium cycle is the most important factor maintaining neutral conditions on the Earth. This cycle has profound influence on the fate of inorganic carbon, and, thereby, on the removal of CO₂ from the primitive atmosphere. Most calcium deposits were formed in the Precambrian, when the prokaryotic biosphere predominated. After that, calcium recycling based on biogenic deposition by skeletal organisms became the main process. Among prokaryotes, only a few representatives, e.g. cyanobacteria, exhibit a special calcium function. The geochemical calcium cycle is made possible by the universal features of bacteria involved in biologically mediated reactions and is determined by the activities of microbial communities. In the prokaryotic system, the calcium cycle begins with the leaching of igneous rocks, predominantly through the action of the community of organotrophic organisms. The release of carbon dioxide to the soil air by organotrophic aerobes leads to leaching with carbonic acid and soda salinization. Under anoxic conditions, of major importance is the organic acid production by primary anaerobes (fermentative microorganisms). Calcium carbonate is precipitated by secondary anaerobes (sulfate reducers) and to a smaller degree by methanogens. The role of the cyanobacterial community in carbonate deposition is recorded by stromatolites, which are the most common organo–sedimentary Precambrian structures. Deposition of carbonates in cyanobacterial mats as a consequence of photoassimilation of CO₂ does not appear to be a significant process. It is argued that carbonates were deposited at the boundary between the soda continent, which emerged as a result of subaerial leaching with carbonic acid, and the ocean containing Ca²⁺. Such ecotones provided favorable conditions for the development of the benthic cyanobacterial communities, which were the precursors of stromatolites.     

Diversity of phototrophic bacteria in microbial mats from Arctic hot springs (Greenland)

We investigated the genotypic diversity of oxygenic and anoxygenic phototrophic microorganisms in microbial mat samples collected from three hot spring localities on the east coast of Greenland. These hot springs harbour unique Arctic microbial ecosystems that have never been studied in detail before. Specific oligonucleotide primers for cyanobacteria, purple sulfur bacteria, green sulfur bacteria and Choroflexus/Roseiflexus-like green non-sulfur bacteria were used for the selective amplification of 16S rRNA gene fragments. Amplification products were separated by denaturing gradient gel electrophoresis (DGGE) and sequenced. In addition, several cyanobacteria were isolated from the mat samples, and classified morphologically and by 16S rRNA-based methods. The cyanobacterial 16S rRNA sequences obtained from DGGE represented a diverse, polyphyletic collection of cyanobacteria. The microbial mat communities were dominated by heterocystous and non-heterocystous filamentous cyanobacteria. Our results indicate that the cyanobacterial community composition in the samples were different for each sampling site. Different layers of the same heterogeneous mat often contained distinct and different communities of cyanobacteria. We observed a relationship between the cyanobacterial community composition and the in situ temperatures of different mat parts. The Greenland mats exhibited a low diversity of anoxygenic phototrophs as compared with other hot spring mats which is possibly related to the photochemical conditions within the mats resulting from the Arctic light regime.     

Transformation of clay minerals caused by an alkaliphilic cyanobacterial community

Transformation of clay minerals (smectite-zeolite, illite, kaolinite, and bentonite) and admixtures of iron oxides (hydroxides) under the action of an alkaline cyanobacterial community was studied. The results demonstrate that the processes of transformation of clay minerals such as intensification of removal of exchange bases and dissolution of silicates and iron oxides occurred in the presence of the alkaliphilic cyanobacterial community. The main factor that determines resistance of a mineral to biochemical weathering is its composition. Transformations of clay minerals in the course of active cyanobacterial photosynthesis (up to 14 days) and at decomposition of organic matter (OM) (28–60 days) are different. For smectite-zeolite and illite, these processes are dissolution of silicates and oxides (removal of Si and Fe) and removal of exchange bases (K), which were observed at both the of biomass production and OM destruction stages. For two other clays, the processes of neosynthesis are more typical: formation of carbonates (most probably siderite for bentonite clay and Mg-calcite for kaolin clay) and transformation of ferrihydrite into the more thermodynamically stable goethite.     

Diurnal Cycles of Sulfate Reduction under Oxic Conditions in Cyanobacterial Mats

Diurnal cycles of sulfate reduction were examined in a well-developed cyanobacterial mat which grew in an outdoor experimental hypersaline pond system at a constant salinity of 75 ± 5% for 3 years. Vertical profiles of sulfate reduction were determined for the upper 12 mm of the microbial mat. Sulfate reduction activities were compared with diurnal variations of oxygen and sulfide concentrations measured by microelectrodes. Significant activity of sulfate-reducing bacteria was detected under aerobic conditions during the daytime, with maximal activity at 2 p.m. When comparing sulfate reduction activities in sediment cores taken at 6 a.m. and 12 a.m. and incubated at a constant temperature in the light and in the dark, a distinct stimulation of the activity in the vertical profile of sulfate reduction by light was evident. It is therefore concluded that the maximal in situ activities, measured at 2 p.m. in the chemocline of the cyanobacterial mat, cannot be attributed to diurnal changes of temperature alone. The response of sulfate-reducing bacteria to the addition of specific carbon sources was significantly different in the cyanobacterial layer, the anoxygenic phototrophic bacterial layer, and the permanently reduced layer of the microbial mat. Sulfate reduction in the mat layer exposed to high oxygen concentrations as a result of cyanobacterial oxygenic photosynthesis was enhanced only by glycolate; in the microzone where the chemocline is found during the daytime, ethanol was the only carbon source to enhance sulfate reduction, while both ethanol and lactate enhanced this activity in the permanently reduced zone.     

Microbial diversity in modern marine stromatolites, Highborne Cay, Bahamas

Living marine stromatolites at Highborne Cay, Bahamas, are formed by microbial mat communities that facilitate precipitation of calcium carbonate and bind and trap small carbonate sand grains. This process results in a laminated structure similar to the layering observed in ancient stromatolites. In the modern marine system at Highborne Cay, lamination, lithification and stromatolite formation are associated with cycling between three types of microbial communities at the stromatolite surface (Types 1, 2 and 3, which range from a leathery microbial mat to microbially fused sediment). Examination of 923 universal small-subunit rRNA gene sequences from these communities reveals that taxonomic richness increases during transition from Type 1 to Type 3 communities, supporting a previous model that proposed that the three communities represent different stages of mat development. The phylogenetic composition also changes significantly between these community types and these community changes occur in concert with variation in biogeochemical rates. The dominant bacterial groups detected in the stromatolites include Alphaproteobacteria , Planctomycetes , Cyanobacteria and Bacteroidetes . In addition, the stromatolite communities were found to contain novel cyanobacteria that may be uniquely associated with modern marine stromatolites. The implications of these findings are discussed in the context of current models for stromatolite formation.     

Long-term compositional changes after transplant in a microbial mat cyanobacterial community revealed using a polyphasic approach

Using a polyphasic approach that included microscopy, cultivation and 16S rRNA-based cultivation-independent molecular fingerprinting, we compared the cyanobacterial composition of Solar Lake microbial mats and samples thereof transplanted and maintained in new settings for extended periods of time. Significant changes in community composition, with clear replacement of the dominant cyanobacterium, Microcoleus chthonoplastes, were detected in all cases. The most dramatic shifts occurred in a sample kept in the laboratory for 3 years, which resulted in dominance by an Oscillatoria -like cyanobacterium whose 16S rRNA closely matched that of a morphologically similar isolate from mats in Mexico. Transfer of Solar Lake mat to an artificial experimental pond with incubation under seminatural conditions resulted in an increase in cyanobacterial diversity. Judging from the molecular signatures, two novel, previously unrecognized and phylogenetically well-delimited cyanobacterial populations became dominant. Through cultivation, one population was shown to correspond to a filamentous, non-heterocystous group of Cyanobacteria with very narrow trichomes (≈ 0.75–1.5 μm). The most dominant novel molecular signature, however, could not be identified by cultivation efforts or correlation with microscopy and, upon phylogenetic analyses, its 16S rRNA genes showed no particular close association to known cyanobacterial groups.     

Microfacies of Cambrian Limestones in Jordan

Summary The limestones of the Wadi Nasb Formation of the uppermost Lower Cambrian of Jordan are under- and overlain by massive sandstones of a near-shore facies. Facies analysis is based on samples from an outcrop at the northeastern shore of the Dead Sea and two oil test wells in the Wadi Sirhan Depression in eastern Jordan. Limestones were deposited in the shallow sea and within the coastal tidal area. Cyanobacteria, algae, echinoderms, trilobites and hyoliths have contributed the bulk of the carbonate and phosphatic material composing the Wadi Nasb limestone. Fine-grained facies types are composed of peloidal carbonate muds with laminar and nodular algal and cyanobacterial mats. They formed within a quiet tidallagoonal environment. The coarse grained facies types consist of carbonate sands with layers of sheell debris deposited in crossbeds in an environment with a rich endobenthic fauna. Here most particles were coated by cyanobacterial crusts. Ooids, oncoids and various coated grains are present. Consolidated sediments were commonly eroded within or near to this environment and their remains were integrated within the sands. Diagenesis is reconstructed step by step with deposition, first cementation, aragonite dissolution, compaction, pore filling, formation of pressure solution, growth of dolomite and anhydrite within the calcitic limestone and final fissure formation and filling.