Tanja Bosak

Large Sulfur Isotope Fractionation Does Not Require Disproportionation

In the absence of oxygenation, microbial activity can explain the magnitude of sulfur-isotope traces in sediments. The composition of sulfur isotopes in sedimentary sulfides and sulfates traces the sulfur cycle throughout Earth’s history. In particular, depletions of sulfur-34 (34S) in sulfide relative to sulfate exceeding 47 per mil (‰) often serve as a proxy for the disproportionation of intermediate sulfur species in addition to sulfate reduction. Here, we demonstrate that a pure, actively growing culture of a marine sulfate-reducing bacterium can deplete 34S by up to 66‰ during sulfate reduction alone and in the absence of an extracellular oxidative sulfur cycle. Therefore, similar magnitudes of sulfur isotope fractionation in sedimentary rocks do not unambiguously record the presence of other sulfur-based metabolisms or the stepwise oxygenation of Earth’s surface environment during the Proterozoic.

Microbial nucleation of calcium carbonate in the Precambrian

Microbial sulfate reduction is thought to stimulate carbonate precipitation in modern stromatolites, yet whether this metabolism was important in shaping Precambrian stromatolites is unknown. Here we use geochemical modeling to suggest that the influence of sulfate reduction on the saturation index of calcite (SI) is negligible when seawater is in equilibrium with high pCO_2, as is thought for the Precambrian. Our laboratory experiments with heterotrophic bacteria in a medium mimicking Precambrian seawater chemistry show that even if sulfate reduction does not significantly change the SI, the presence of bacteria stimulates calcite precipitation over sterile controls by effectively increasing the SI over a pH range from 7.3 to 7.8. Under our experimental conditions, dead cells stimulate in situ carbonate precipitation equally, if not more, than active sulfate-reducing bacteria. Heterogeneous nucleation of calcite by microbial cell material appears to be the driving mechanism that explains this phenomenon.

Morphological record of oxygenic photosynthesis in conical stromatolites

Conical stromatolites are thought to be robust indicators of the presence of photosynthetic and phototactic microbes in aquatic environments as early as 3.5 billion years ago. However, phototaxis alone cannot explain the ubiquity of disrupted, curled, and contorted laminae in the crests of many Mesoproterozoic, Paleoproterozoic, and some Archean conical stromatolites. Here, we demonstrate that cyanobacterial production of oxygen in the tips of modern conical aggregates creates contorted laminae and submillimeter-to-millimeter-scale enmeshed bubbles. Similarly sized fossil bubbles and contorted laminae may be present only in the crestal zones of some conical stromatolites 2.7 billion years old or younger. This implies not only that cyanobacteria built Proterozoic conical stromatolites but also that fossil bubbles may constrain the timing of the evolution of oxygenic photosynthesis.

Formation and stability of oxygen-rich bubbles that shape photosynthetic mats

Gas release in photic-zone microbialites can lead to preservable morphological biosignatures. Here, we investigate the formation and stability of oxygen-rich bubbles enmeshed by filamentous cyanobacteria. Sub-millimetric and millimetric bubbles can be stable for weeks and even months. During this time, lithifying organic-rich laminae surrounding the bubbles can preserve the shape of bubbles. Cm-scale unstable bubbles support the growth of centimetric tubular towers with distinctly laminated mineralized walls. In environments that enable high photosynthetic rates, only small stable bubbles will be enclosed by a dense microbial mesh, while in deep waters extensive microbial mesh will cover even larger photosynthetic bubbles, increasing their preservation potential. Stable photosynthetic bubbles may be preserved as sub-millimeter and millimeter-diameter features with nearly circular cross-sections in the crests of some Proterozoic conical stromatolites, while centrimetric tubes formed around unstable bubbles provide a model for the formation of tubular carbonate microbialites that are not markedly depleted in (13)C.

A polycyclic terpenoid that alleviates oxidative stress

Polycyclic terpenoid lipids such as hopanes and steranes have been widely used to understand ancient biology, Earth history, and the oxygenation of the ocean–atmosphere system. Some of these lipids are believed to be produced only by aerobic organisms, whereas others actually require molecular oxygen for their biosynthesis. A persistent question remains: Did some polycyclic lipids initially evolve in response to certain environmental or metabolic stresses, including the presence of oxygen? Here, we identify tetracyclic isoprenoids in spores of the bacterium Bacillus subtilis. We call them sporulenes. They are produced by cyclization of regular polyprenes, a reaction that is more favorable chemically than the formation of terpenoids such as hopanoids and steroids from squalene. The simplicity of the reaction suggests that the B. subtilis cyclase may be analogous to evolutionarily ancient cyclases. We show that these molecules increase the resistance of spores to a reactive oxygen species, demonstrating a specific physiological role for a nonpigment bacterial lipid biomarker. Geostable derivatives of these compounds in sediments could thus be used as direct indicators of oxidative stress and aerobic environments.

Biophysical basis for the geometry of conical stromatolites

Stromatolites may be Earth’s oldest macroscopic fossils; however, it remains controversial what, if any, biological processes are recorded in their morphology. Although the biological interpretation of many stromatolite morphologies is confounded by the influence of sedimentation, conical stromatolites form in the absence of sedimentation and are, therefore, considered to be the most robust records of biophysical processes. A qualitative similarity between conical stromatolites and some modern microbial mats suggests a photosynthetic origin for ancient stromatolites. To better understand and interpret ancient fossils, we seek a quantitative relationship between the geometry of conical stromatolites and the biophysical processes that control their growth. We note that all modern conical stromatolites and many that formed in the last 2.8 billion years display a characteristic centimeter-scale spacing between neighboring structures. To understand this prominent—but hitherto uninterpreted—organization, we consider the role of diffusion in mediating competition between stromatolites. Having confirmed this model through laboratory experiments and field observation, we find that organization of a field of stromatolites is set by a diffusive time scale over which individual structures compete for nutrients, thus linking form to physiology. The centimeter-scale spacing between modern and ancient stromatolites corresponds to a rhythmically fluctuating metabolism with a period of approximately 20 hr. The correspondence between the observed spacing and the day length provides quantitative support for the photosynthetic origin of conical stromatolites throughout geologic time.

Possible early foraminiferans in post-Sturtian (716–635 Ma) cap carbonates

Foraminifera are an ecologically important group of modern heterotrophic amoeboid eukaryotes whose naked and testate ancestors are thought to have evolved ∼1 Ga ago. However, the single-chambered agglutinated tests of these protists appear in the fossil record only after ca. 580 Ma, coinciding with the appearance of macroscopic and mineralized animals. Here we report the discovery of small, slender tubular microfossils in the Sturtian (ca. 716–635 Ma) cap carbonate of the Rasthof Formation in Namibia. The tubes are 200–1300 μm long and 20–70 μm wide, and preserve apertures and variably wide lumens, folds, constrictions, and ridges. Their sometimes flexible walls are composed of carbonaceous material and detrital minerals. This combination of morphologic and compositional characters is also present in some species of modern single-chambered agglutinated tubular foraminiferans, and is not found in other agglutinated eukaryotes. The preservation of possible early Foraminifera in the carbonate rocks deposited in the immediate aftermath of Sturtian low-latitude glaciation indicates that various morphologically modern protists thrived in microbially dominated ecosystems, and contributed to the cycling of carbon in Neoproterozoic oceans much before the rise of complex animals.

Putative Cryogenian ciliates from Mongolia

Major lineages of modern eukaryotes, represented primarily by microscopic taxa, are thought to have originated during the Neoproterozoic, but microfossils older than 635 Ma rarely have unambiguous relationships to modern microscopic eukaryotes. Here we report exceptionally preserved 715–635 m.y. old eukaryotic tests in limestone strata of Mongolia. The ∼100-μm-long organic-rich three-dimensional tests have flask-like shapes, constricted necks, distinct and often thickened collars, and flexible walls composed of densely packed alveolar structures. The combined morphological and ultrastructural characters of these Cryogenian tests are remarkably similar to the tests of tintinnids, modern planktonic ciliates. Eukaryotes forming recalcitrant organic or mineral-rich tests before 635 Ma may have increased export and burial fraction of organic carbon, driving an increase in atmospheric oxygen and the subsequent radiation of metazoans.

Cyanobacterial diversity and activity in modern conical microbialites

Modern conical microbialites are similar to some ancient conical stromatolites, but growth, behavior and diversity of cyanobacteria in modern conical microbialites remain poorly characterized. Here, we analyze the diversity of cyanobacterial 16S rRNA gene sequences in conical microbialites from 14 ponds fed by four thermal sources in Yellowstone National Park and compare cyanobacterial activity in the tips of cones and in the surrounding topographic lows (mats), respectively, by high-resolution mapping of labeled carbon. Cones and adjacent mats contain similar 16S rRNA gene sequences from genetically distinct clusters of filamentous, non-heterocystous cyanobacteria from Subsection III and unicellular cyanobacteria from Subsection I. These sequences vary among different ponds and between two sampling years, suggesting that coniform mats through time and space contain a number of cyanobacteria capable of vertical aggregation, filamentous cyanobacteria incapable of initiating cone formation and unicellular cyanobacteria. Unicellular cyanobacteria are more diverse in topographic lows, where some of these organisms respond to nutrient pulses more rapidly than thin filamentous cyanobacteria. The densest active cyanobacteria are found below the upper 50 μm of the cone tip, whereas cyanobacterial cells in mats are less dense, and are more commonly degraded or encrusted by silica. These spatial differences in cellular activity and density within macroscopic coniform mats imply a strong role for diffusion limitation in the development and the persistence of the conical shape. Similar mechanisms may have controlled the growth, morphology and persistence of small coniform stromatolites in shallow, quiet environments throughout geologic history.

Micrometer-scale porosity as a biosignature in carbonate crusts

We formed calcite crusts in the presence and absence of the heterotrophic bacterium Desulfovibrio desulfuricans strain G20 to investigate microbial morphological signatures in fast-accreting carbonate precipitates. Submicrometer- to micrometer-sized pores (micropores) were present and ubiquitous in the G20 crusts but absent in abiotically precipitated crusts. Bacterial micropores resemble inclusions under transmitted light, but have distinct size, biological shapes and patterns (swirling or dendritic) and are distributed differently from common fluid inclusions. We observed similar porosity in both modern and ancient carbonate crusts of putative biotic origin. Our experiments support the microbial origin of micropores and help define specific criteria whereby to recognize these features as biosignatures in the rock record.