Pieter T. Visscher

The role of microbes in accretion, lamination and early lithification of modern marine stromatolites.

For three billion years, before the Cambrian diversification of life, laminated carbonate build-ups called stromatolites were widespread in shallow marine seas. These ancient structures are generally thought to be microbial in origin and potentially preserve evidence of the Earths earliest biosphere. Despite their evolutionary significance, little is known about stromatolite formation, especially the relative roles of microbial and environmental factors in stromatolite accretion. Here we show that growth of modern marine stromatolites represents a dynamic balance between sedimentation and intermittent lithification of cyanobacterial mats. Periods of rapid sediment accretion, during which stromatolite surfaces are dominated by pioneer communities of gliding filamentous cyanobacteria, alternate with hiatal intervals. These discontinuities in sedimentation are characterized by development of surface films of exopolymer and subsequent heterotrophic bacterial decomposition, forming thin crusts of microcrystalline carbonate. During prolonged hiatal periods, climax communities develop, which include endolithic coccoid cyanobacteria. These coccoids modify the sediment, forming thicker lithified laminae. Preservation of lithified layers at depth creates millimetre-scale lamination. This simple model of modern marine stromatolite growth may be applicable to ancient stromatolites.

Microscale observations of sulfate reduction: Correlation of microbial activity with lithified micritic laminae in modern marine stromatolites

We report for the first time micrometer-scale correlation of geologic and microbial processes in modern marine stromatolites. Precipitation of micritic laminae in these stromatolites was studied by comparing microstructure, as observed in petrographic thin sections, with microbial sulfatereduction activity. Two-dimensional mapping of sulfate-reduction rates was implemented by incubating a vertical section of a stromatolite face on silver foil coated with 35 SO 4 2– . Our results show that sulfate-reduction activity is high in zones of CaCO 3 precipitation and indicate that microbial activity produces lithified micritic laminae near the surface of the stromatolites. Similarities with micritic laminae in ancient stromatolites suggest that sulfate reduction may also have been an important mechanism of carbonate precipitation in these fossilized structures.

Biological and environmental chemistry of DMSP and related sulfonium compounds

Proceedings of the June 1995 symposium, covering topics related to dimethylsulfoniopropionate (DMSP) and providing background and the latest research on the subject. DMSP and related sulfonium compounds are of interest to biological chemists because they are used by organisms to combat osmotic stres

Quorum sensing in natural environments: emerging views from microbial mats.

Much laboratory-based information exists on quorum sensing, a type of bacterial cell-to-cell communication that depends upon exchanges of molecular signals between neighboring cells. However, little is known about how this and other microbial sensing systems operate in nature. Geochemical and biological modifications of signals probably occur in extracellular environments, and these could disrupt intended communication if signals are no longer recognized. However, as we discuss here, signal alterations might result in other outcomes: if a modified signal is able to interact with a different receptor then further environmental information can be gained by the receiving cells. We also postulate that quorum sensing occurs within cell clusters, where signal dispersion might be significantly influenced by extracellular polymers. As a model system to discuss these points we use microbial mats - highly-structured biofilm communities living under sharply-defined, fluctuating geochemical gradients.

Exopolymeric substances (EPS) from Bacillus subtilis: polymers and genes encoding their synthesis.

Bacterial exopolymeric substances (EPS) are molecules released in response to the physiological stress encountered in the natural environment. EPS are structural components of the extracellular matrix in which cells are embedded during biofilm development. The chemical nature and functions of these EPS are dependent on the genetic expression of the cells within each biofilm. Although some bacterial matrices have been characterized, understanding of the function of the EPS is relatively limited, particularly within the Bacillus genus. Similar gaps of knowledge exist with respect to the chemical composition and specific roles of the macromolecules secreted by Bacillus subtilis in its natural environment. In this review, the different EPS from B. subtilis were classified into four main functional categories: structural (neutral polymers), sorptive (charged polymers), surface-active and active polymers. In addition, current information regarding the genetic expression, production and function of the main polymers secreted by B. subtilis strains, particularly those related to biofilm formation and its architecture, has been compiled. Further characterization of these EPS from B. subtilis remains a challenge.

Characteristics and turnover of exopolymeric substances in a hypersaline microbial mat

The properties and microbial turnover of exopolymeric substances (EPS) were measured in a hypersaline nonlithifying microbial mat (Eleuthera, Bahamas) to investigate their potential role in calcium carbonate (CaCO(3)) precipitation. Depth profiles of EPS abundance and enzyme activities indicated that c. 80% of the EPS were turned over in the upper 15-20 mm. Oxic and anoxic mat homogenates amended with low-molecular-weight (LMW) organic carbon, sugar monomers, and different types of EPS revealed rapid consumption of all substrates. When comparing the consumption of EPS with that of other substrates, only marginally longer lag times and lower rates were observed. EPS (5-8%) were readily consumed during the conversion of labile to refractory EPS. This coincided with a decrease in glucosidase activity and a decrease in the number of acidic functional groups on the EPS. Approximately half of the calcium bound to the EPS remained after 10 dialyses steps. This tightly bound calcium was readily available to precipitate as CaCO(3). We present a conceptual model in which LMW organic carbon complexed with the tightly bound calcium is released upon enzyme activity. This increases alkalinity and creates binding sites for carbonate and allows CaCO(3) to precipitate. Therefore, this model explains interactions between EPS and CaCO(3) precipitation, and underscores the critical role of aerobic and anaerobic microorganisms in early diagenesis and lithification processes.

Autoinducers extracted from microbial mats reveal a surprising diversity of N-acylhomoserine lactones (AHLs) and abundance changes that may relate to diel pH.

Microbial mats are highly structured and diverse communities, and one of the earliest-known life assemblages. Mat bacteria interact within an environment marked by strong geochemical gradients and fluctuations. We examined natural mat systems for the presence of autoinducers involved in quorum sensing, a form of cell-cell communication. Our results revealed that a diverse array of N-acylhomoserine lactones (AHLs) including C(4)- to C(14)-AHLs, were identified from mat extracts using mass spectrometry (MS), with further confirmation by MS/MS-collision-induced dissociation (CID), and additions of external standards. Microelectrode measurements showed that mats exhibited diel pH fluctuations, ranging from alkaline (pH 9.4) during daytime (net photosynthesis) to acidic (pH 6.8) during darkness (net respiration/fermentation). Under laboratory conditions, AHLs having shorter acyl-chains were degraded within the time frame that daily alkaline pH (> 8.2) conditions exist in mats. Intensive sampling of mats after full day- or night-time incubations revealed that accumulations of extractable shorter-chain AHLs (e.g. C(8)- and C(10)-AHLs) were significantly (P < 0.001) diminished during daytime. Our study offers evidence that stabilities of AHLs under natural conditions may be influenced by the proximal extracellular environment. We further propose that the ancient periodicity of photosynthesis/respiration in mats may potentially drive a mechanism for diel differences in activities of certain autoinducers, and hence bacterial activities mediated through quorum sensing.

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.

POLYSULFIDE UTILIZATION BY THIOCAPSA-ROSEOPERSICINA

The purple sulfur bacterium Thiocapsa roseopersicina, being the dominant anoxygenic phototroph in microbial mats, was tested for growth on polysulfide as the electron donor for carbon dioxide fixation. Data collected in continuous cultures revealed μmax to be 0.065 h-1 and the saturation affinity constant Ks to be 6.7 μM. The value of the inhibition constant Ki was estimated in batch cultures and was found to be approximately 1100 μM. When grown on monosulfide, the organism was capable of trisulfide utilization without lag. Monosulfide-limited growth was established to have a μmax of 0.091 h-1 and Ks of 8.0 μM. Field observations revealed polysulfide, present at supra-optimal concentrations, as a major pool of reduced sulfur in a laminated marine sediment ecosystem.

Formation and diagenesis of modern marine calcified cyanobacteria.

Calcified cyanobacterial microfossils are common in carbonate environments through most of the Phanerozoic, but are absent from the marine rock record over the past 65 Myr. There has been long-standing debate on the factors controlling the formation and temporal distribution of these fossils, fostered by the lack of a suitable modern analog. We describe calcified cyanobacteria filaments in a modern marine reef setting at Highborne Cay, Bahamas. Our observations and stable isotope data suggest that initial calcification occurs in living cyanobacteria and is photosynthetically induced. A single variety of cyanobacteria, Dichothrix sp., produces calcified filaments. Adjacent cyanobacterial mats form well-laminated stromatolites, rather than calcified filaments, indicating there can be a strong taxonomic control over the mechanism of microbial calcification. Petrographic analyses indicate that the calcified filaments are degraded during early diagenesis and are not present in well-lithified microbialites. The early diagenetic destruction of calcified filaments at Highborne Cay indicates that the absence of calcified cyanobacteria from periods of the Phanerozoic is likely to be caused by low preservation potential as well as inhibited formation.