Early Evolution Of Life Research Articles

Processive RNA polymerization and promoter recognition in an RNA World.

Early life is thought to have required the self-replication of RNA by RNA replicases. However, how such replicases evolved and subsequently enabled gene expression remains largely unexplored. We engineered and selected a holopolymerase ribozyme that uses a sigma factor-like specificity primer to first recognize an RNA promoter sequence and then, in a second step, rearrange to a processive elongation form. Using its own sequence, the polymerase can also program itself to polymerize from certain RNA promoters and not others. This selective promoter-based polymerization could allow an RNA replicase ribozyme to define "self" from "nonself," an important development for the avoidance of replicative parasites. Moreover, the clamp-like mechanism of this polymerase could eventually enable strand invasion, a critical requirement for replication in the early evolution of life.

MgO surface lattice phonons observation during interstellar ice transition

Relevant information on the origins of the solar system and the early evolution of life itself can be derive from systematic and controlled exploration of water ice here on Earth. Therefore, over the last decades, a huge effort on experimental methodologies has been made to study the multiple crystal ice phases, which are observed outside our home–gravitational–potential. By employing (100)–oriented MgO lattice surface as a microcantilever sensor, we conducted the first ever study on the dynamics of the Structural Phase Transition at 185 K in water ice by means of coherent elastic scattering of electron diffraction. We estimate the amount of phonons caused by this transition applying precise quantum computing key tools, and resulting in a maximum value of 1.23 ± 0.02. Further applications of our microcantilever sensor were assessed using unambiguous mapping of the surface stress induced by the c(4 times 2) → p(3 times 2) Structural Phase Transition of the interstellar ice formulated on the Williamsom–Hall model. This development paves the way and thus establishes an efficient characterization tool of the surface mechanical strains of materials with potential applications arising from interstellar ice inclusive glaciers to the wide spectrum of solid–state physics.

Apatite nanoparticles in 3.46–2.46 Ga iron formations: Evidence for phosphorus-rich hydrothermal plumes on early Earth

AbstractPhosphorus is an essential nutrient that is thought to have regulated primary productivity in global oceans after the advent of oxygenic photosynthesis. The prime source of seawater phosphorus is regarded to be continental weathering of phosphate minerals. Ancient seawater phosphorus concentrations have been constrained using the phosphorus content of iron-rich chemical sediments—banded iron formations (BIFs); however, the removal processes and depositional phases remain unclear. Here we report that nanometer-sized apatite crystals (<500 nm) are ubiquitous in 3.46–2.46 Ga BIFs and cherts from the Kaapvaal (South Africa) and Yilgarn, and Pilbara (Western Australia) cratons. The apatite is uniformly dispersed in a chemical sediment comprising greenalite nanoparticles, which were encased in very early diagenetic silica cement that limited compaction and chemical reactions. The lack of organic carbon (below detection; <0.3 wt%) and absence of primary iron oxides implies that the phosphorus was not derived from the degradation of organic matter or seawater scavenging by oxide particles. Instead, the occurrence of apatite in sediments derived from hydrothermally sourced Fe2+ and SiO2(aq) suggests that phosphorus too was derived from vent plumes. Today, seawater P is rapidly removed from vent fluids due to scavenging by oxidized Fe2+. However, prior to the Great Oxidation Event (2.45–2.32 Ga), dissolved phosphorus released during anoxic alteration of seafloor basalts escaped the iron-oxidation trap. Our results point to the existence of a submarine hydrothermal flux of dissolved phosphorus that supplied nutrients to the early anoxic oceans. High amounts of seawater P may help to explain why phosphorus is ubiquitous in cell biology—it was not limiting during the origin and early evolution of life.

The Semi-Enzymatic Origin of Metabolic Pathways: Inferring a Very Early Stage of the Evolution of Life.

The early evolution of life is a period with many important events with a lot of big and open questions. One of them is the evolution of metabolic pathways, which means the origin and assembly of enzymes that act together. The retrograde hypothesis was the first attempt to explain the origin and evolutionary history of metabolic pathways; Norman Horowitz developed this first significant hypothesis. This idea was followed by relevant proposals developed by Sam Granick, who proposed the "forward direction hypothesis," and then the successful idea of "Patchwork" assembly proposed independently by Martynas Yčas and Roy Jensen. Since then, a few new hypotheses were proposed; one of the most influential was made by Antonio Lazcano and Stanley Miller in the Journal Molecular Evolution, the "semi-enzymatic origin" of metabolic pathways. This article was cited more than 160 times, including in most papers published about the early evolution of metabolism, placing it as influential work in the field. The ideas proposed in this work and their effects on studying the origin and early evolution of life are analyzed.

Unexpectedly high mutation rate of a deep-sea hyperthermophilic anaerobic archaeon.

Deep-sea hydrothermal vents resemble the early Earth, and thus the dominant Thermococcaceae inhabitants, which occupy an evolutionarily basal position of the archaeal tree and take an obligate anaerobic hyperthermophilic free-living lifestyle, are likely excellent models to study the evolution of early life. Here, we determined that unbiased mutation rate of a representative species, Thermococcus eurythermalis, exceeded that of all known free-living prokaryotes by 1-2 orders of magnitude, and thus rejected the long-standing hypothesis that low mutation rates were selectively favored in hyperthermophiles. We further sequenced multiple and diverse isolates of this species and calculated that T. eurythermalis has a lower effective population size than other free-living prokaryotes by 1-2 orders of magnitude. These data collectively indicate that the high mutation rate of this species is not selectively favored but instead driven by random genetic drift. The availability of these unusual data also helps explore mechanisms underlying microbial genome size evolution. We showed that genome size is negatively correlated with mutation rate and positively correlated with effective population size across 30 bacterial and archaeal lineages, suggesting that increased mutation rate and random genetic drift are likely two important mechanisms driving microbial genome reduction. Future determinations of the unbiased mutation rate of more representative lineages with highly reduced genomes such as Prochlorococcus and Pelagibacterales that dominate marine microbial communities are essential to test these hypotheses.

Biogeochemical cycling of sulfur, manganese and iron in ferruginous limnic analog of Archean ocean

The early evolution of life on Earth was intimately coupled with the evolution of ocean chemical composition and redox conditions in Archean ocean. However, the measurements of chemical and isotopic compositions of Archean sedimentary rocks does not provide sufficient information for understanding the biogeochemical processes that characterized oceans during this period. In this research, the cycling of the redox-sensitive elements in the water column and sediments of the seasonally stratified Lake Sihailongwan, China, was studied. It was found that during the stratification, the concentrations of sulfate ions in the epilimnion were <100 μmol L−1. The hypolimnion of the lake becomes anoxic and contains iron, manganese and low concentrations of hydrogen sulfide. These conditions are similar to those suggested to exist in the Archean ocean, and therefore the biogeochemical cycles in Lake Sihailongwan may be analogous to those in the Archean ocean. In Lake Sihailongwan, the chemocline is always deeper than the thermocline. The hypolimnion is supersaturated with respect to iron sulfide in August, while in May and October, this supersaturation is present only in the lower hypolimnion. The concentration profiles of the redox species in the water column and in the sediments show that dissolved iron(II) and manganese(II) diffuse from the sediments into the water column, while hydrogen sulfide and sulfate diffuse into the sediment. At the hypolimnion, the sulfur isotope fractionation values are consistent with microbial sulfate reduction, possibly combined, to a minor extent, with disproportionation of zero-valent sulfur. The absolute sulfur isotope fractionation factor increases between spring and autumn and decreases with water depth, possibly due to precipitation of iron sulfide in the lower hypolimnion. The fraction of highly reactive iron in the sediments is mainly composed of iron(II) carbonates and the most reactive iron(III) (hydr)oxide phases. Iron-based redox proxies are consistent with sediment deposition under ferruginous water column conditions. In the sediment, concentrations of sulfate and hydrogen sulfide are low, and sulfur isotope composition of sedimentary pyrite is equal to that of the epilimnetic sulfate. The obtained results demonstrated that despite a low sulfate concentration in the water column, sulfur isotope fractionation is high. However, due to the complete consumption of the sulfate reservoir, the large isotope fractionation is not preserved in the sedimentary record.

The Universal Evolution and the Origin of Life

The origin of life occupies a very important place in the study of the evolution. Its liminal location between life and non-life poses special challenges to researchers who study this subject. Current approaches in studying the origin and evolution of early life are reductive: they either reduce the domain of non-life to the domain of life or vice versa. This contribution seeks to provide a perspective that would avoid reductionism of any kind. Its goal is to outline a frame that would include both domains and their respective evolutions as its particular cases. The study examines the main theoretical perspectives on the origin and evolution of early life and provides a constructive critique of these perspectives. An objective view requires viewing an object or a phenomenon from all available points of view. The goal of this contribution is not to prove the current perspectives wrong and to deny their achievements. It seeks to provide an angle that would be sufficiently wide and would allow synthesizing current perspectives for a comprehensive and objective interpretation of the origin and evolution of early life. In other words, it seeks to outline a frame for an objective view that will help understand life’s place within the universe.

The proto-Nucleic Acid Builder: a software tool for constructing nucleic acid analogs.

The helical structures of DNA and RNA were originally revealed by experimental data. Likewise, the development of programs for modeling these natural polymers was guided by known structures. These nucleic acid polymers represent only two members of a potentially vast class of polymers with similar structural features, but that differ from DNA and RNA in the backbone or nucleobases. Xeno nucleic acids (XNAs) incorporate alternative backbones that affect the conformational, chemical, and thermodynamic properties of XNAs. Given the vast chemical space of possible XNAs, computational modeling of alternative nucleic acids can accelerate the search for plausible nucleic acid analogs and guide their rational design. Additionally, a tool for the modeling of nucleic acids could help reveal what nucleic acid polymers may have existed before RNA in the early evolution of life. To aid the development of novel XNA polymers and the search for possible pre-RNA candidates, this article presents the proto-Nucleic Acid Builder (https://github.com/GT-NucleicAcids/pnab), an open-source program for modeling nucleic acid analogs with alternative backbones and nucleobases. The torsion-driven conformation search procedure implemented here predicts structures with good accuracy compared to experimental structures, and correctly demonstrates the correlation between the helical structure and the backbone conformation in DNA and RNA.

324 Milk Fat Globule Membrane from Bovine Milk on Brain Development of Early Life

Abstract The milk fat globule membrane (MFGM) is instrumental for the fat delivery system into human and bovine milk, but is typically removed during the manufacture of infant formula. MFGM contains components that may impact neurodevelopment, including sialic acid, gangliosides, sphingomyelin, choline, glycerophospholipids, proteins, and cholesterol. This presentation will review the clinical trial evidence linking MFGM supplementation to beneficial outcomes in infants and will describe potential mechanistic evidence linking MFGM with neurocognitive outcomes arising from preclinical studies in piglets. Infants fed formula supplemented with a MFGM (4% total protein) from 2 to 6 months of age had improved neurocognitive development at 1 year of age compared to infants fed standard formula. Infants fed formula with MFGM (5.0 g/L) and lactoferrin (0.6 g/L) for 1 year had an accelerated neurodevelopmental profile at 1 year and improved language subcategories at 18 months compared to infants fed a standard formula. To investigate potential mechanisms, piglets were fed a CONT formula or a TEST formula with MFGM and lactoferrin at the same concentrations from 2 to 31 days of age. Piglets underwent spatial T-maze testing to assess learning and memory, and magnetic resonance imaging to measure brain micro- and macrostructure. TEST piglets had lower radial and mean diffusivities in the internal capsule, suggesting greater myelination. The internal capsule contains motor and sensory projections from the cortex to corticospinal tract. Piglets on the CONT diet displayed shorter latency to choice in the T-maze compared to TEST piglets, potentially indicating anxiety-like behaviors or greater impulsivity. Aspects of the microbiome-gut-brain-axis (MGBA) were investigated to uncover potential mechanisms. TEST piglets had higher protein abundance of tyrosine hydroxylase and vasoactive intestinal peptide, longer villi and greater disaccharidase activity in the small intestine and differences in microbial abundances in the ascending colon and feces, suggesting potential modulation of the MGBA by MFGM.

A distinct lineage of Caudovirales that encodes a deeply branching multi-subunit RNA polymerase

Bacteriophages play critical roles in the biosphere, but their vast genomic diversity has obscured their evolutionary origins, and phylogenetic analyses have traditionally been hindered by their lack of universal phylogenetic marker genes. In this study we mine metagenomic data and identify a clade of Caudovirales that encodes the β and β′ subunits of multi-subunit RNA polymerase (RNAP), a high-resolution phylogenetic marker which enables detailed evolutionary analyses. Our RNAP phylogeny revealed that the Caudovirales RNAP forms a clade distinct from cellular homologs, suggesting an ancient acquisition of this enzyme. Within these multimeric RNAP-encoding Caudovirales (mReC), we find that the similarity of major capsid proteins and terminase large subunits further suggests they form a distinct clade with common evolutionary origin. Our study characterizes a clade of RNAP-encoding Caudovirales and suggests the ancient origin of this enzyme in this group, underscoring the important role of viruses in the early evolution of life on Earth.

Discovery of an Abundance of Biosynthetic Gene Clusters in Shark Bay Microbial Mats.

Microbial mats are geobiological multilayered ecosystems that have significant evolutionary value in understanding the evolution of early life on Earth. Shark Bay, Australia has some of the best examples of modern microbial mats thriving under harsh conditions of high temperatures, salinity, desiccation, and ultraviolet (UV) radiation. Microorganisms living in extreme ecosystems are thought to potentially encode for secondary metabolites as a survival strategy. Many secondary metabolites are natural products encoded by a grouping of genes known as biosynthetic gene clusters (BGCs). Natural products have diverse chemical structures and functions which provide competitive advantages for microorganisms and can also have biotechnology applications. In the present study, the diversity of BGC were described in detail for the first time from Shark Bay microbial mats. A total of 1477 BGCs were detected in metagenomic data over a 20 mm mat depth horizon, with the surface layer possessing over 200 BGCs and containing the highest relative abundance of BGCs of all mat layers. Terpene and bacteriocin BGCs were highly represented and their natural products are proposed to have important roles in ecosystem function in these mat systems. Interestingly, potentially novel BGCs were detected from Heimdallarchaeota and Lokiarchaeota, two evolutionarily significant archaeal phyla not previously known to possess BGCs. This study provides new insights into how secondary metabolites from BGCs may enable diverse microbial mat communities to adapt to extreme environments.

Geochemical fingerprints of seawater in the Late Mesoproterozoic Midcontinent Rift, North America: Life at the marine-land divide

The 1.1 Ga Midcontinent Rift (MCR) is a thick volcanic-sedimentary succession that forms a curvilinear belt through central North America and crops out along its northern apex around Lake Superior. Sedimentary units of the MCR have been long interpreted as fluvial-lacustrine and invited a number of studies on the early evolution of life in non-marine habitats. One of the key units is the siliciclastic Nonesuch Formation, thought to record deposition in a large lake. However, recent sedimentological observations indicate the presence of marine incursions. To further test this interpretation, we analysed trace element abundances in a broad suite of samples from multiple drill cores through the Nonesuch Formation. We aimed to differentiate geochemical influences of sediment provenance from post-depositional hydrothermal overprint and thereby identify authigenic enrichments in fluid-mobile elements that are indicators of primary environmental conditions. Our results reveal discrete enrichments in Mo and U in organic- and sulphide-rich horizons, which are most parsimoniously interpreted as marine signatures. This conclusion is supported by Sr/Ba ratios, which suggest mixing between freshwater and saltwater, and by rare cm-thick gypsum in the upper Copper Harbor Formation immediately below the Nonesuch rocks. The gypsum displays δ34S values of +25.9 ± 0.6‰, consistent with input of marine sulphate at least during parts of the basin's history. Collectively, our geochemical data support the sedimentological interpretation that this portion of the MCR archives a marine-influenced estuarine system. Although this conclusion rules out that microbial life documented from the MCR was living in exclusively freshwater habitats, the Nonesuch Fm and associated rocks still hold important clues about organisms that were capable of withstanding salinity gradients and bridging the gap between the marine and non-marine environments of the mid-Proterozoic.

Prebiotic Syntheses of Noncanonical Nucleosides and Nucleotides.

The origin of nucleotides is a major question in origins-of-life research. Given the central importance of RNA in biology and the influential RNA World hypothesis, a great deal of this research has focused on finding possible prebiotic syntheses of the four canonical nucleotides of coding RNA. However, the use of nucleotides in other roles across the tree of life might be evidence that nucleotides have been used in noncoding roles for even longer than RNA has been used as a genetic polymer. Likewise, it is possible that early life utilized nucleotides other than the extant nucleotides as the monomers of informational polymers. Therefore, finding plausible prebiotic syntheses of potentially ancestral noncanonical nucleotides may be of great importance for understanding the origins and early evolution of life. Experimental investigations into abiotic noncanonical nucleotide synthesis reveal that many noncanonical nucleotides and related glycosides are formed much more easily than the canonical nucleotides. An analysis of the mechanisms by which nucleosides and nucleotides form in the solution phase or in drying-heating reactions from pre-existing sugars and heterocycles suggests that a wide variety of noncanonical nucleotides and related glycosides would have been present on the prebiotic Earth, if any such molecules were present.

Absence of hexavalent chromium in marine carbonates: implications for chromium isotopes as paleoenvironment proxy.

The oxygenation of Earth's atmosphere is widely regarded to have played an important role in early-life evolution. Chromium (Cr) isotopes recorded in sedimentary rocks have been used to constrain the atmospheric oxygen level (AOL) over geological times based on the fact that a positive Cr isotopic signature is linked to the presence of Cr(VI) as a result of oxidative continental weathering. However, there is no direct evidence of the presence of Cr(VI) in sedimentary rocks yet. Carbonates are most widely distributed over geological times and were thought to have incorporated Cr(VI) directly from seawater. Here, we present results of Cr valence states in carbonates which show Cr(III) is the dominant species in all samples spanning a wide range of geological times. These findings indicate that Cr(VI) in seawater was reduced either before or after carbonate precipitation, which might have caused Cr isotopic fractionation between seawater and carbonates, or marine carbonates preferentially uptake Cr(III) from seawater. As Cr(III) can come from non-redox Cr cycling, which also can cause isotopic fractionation, we suggest that positively fractionated Cr isotopic values do not necessarily correspond to the rise in AOL.

How to build complexity

Summary The ribosome is the most complex molecular machine shared by all cellular life forms. Details of its evolution from an RNA world precursor are still encrypted in its molecular sequences, which are gradually yielding information beyond the reach of phylogenetic trees. Meanwhile, researchers can now study the assembly of the structure in real time, yielding further insights into the origins of complexity. Michael Gross reports.

A Constructive Way to Think about Different Hydrothermal Environments for the Origins of Life

The question of where life originated has been contentious for a very long time. Scientists have invoked many environments to address this question. Often, we find ourselves beholden to a location, especially if we think life originated once and then evolved into the myriad forms we now know today. In this brief commentary, we wish to lay out the following understanding: hydrothermal environments are energetically robust locations for the origins and early evolution of life as we know it. Two environments typify hydrothermal conditions, hydrothermal fields on dry land and submarine hydrothermal vents. If life originated only once, then we must choose between these two environments; however, there is no reason to assume life emerged only once. We conclude with the idea that rather than having an “either or” mind set about the origin of life a “yes and” mind set might be a better paradigm with which to problem solve within this field. Finally, we shall discuss further research with regards to both environments.

Microbial mineralization of botryoidal laminations in the Upper Ediacaran dolostones, Western Yangtze Platform, SW China

Botryoidal dolostones are abundant in the Second Member of the Upper Ediacaran Dengying Formation in the Western Yangtze Platform. Although they have been extensively discussed as important host rocks for giant gas reservoirs, there is no consensus on the microbial impact on mineralization processes in the botryoidal laminations. Typical botryoidal laminations were studied under optical microscopes and scanning electron microscopes (SEMs) to investigate their origin. There are three macroscopic morphological types of botryoidal lamination: stromatoid lamination, cavity-lining lamination, and spherical-encrusting lamination. These three types of lamination are composed of fibrous and bladed crystals growing perpendicular to the botryoidal laminae, which have hitherto been implicitly considered to be from abiogenic chemical precipitation. There are many lines of new evidence that demonstrate microbial activities in the interbedded dark and bright laminae constituting the laminations. In the dark laminae, euhedral dolomite rhombs are observed sticking to linear and tubular filaments, or wrapped in sheets of extracellular polymeric substances (EPS) with dolomite crystal sizes of generally <5 μm. The bright laminae are dominated by coarser dolomites and connected by radial sheaths and trichomes with dolomite crystal sizes of primarily around 10 μm. Different species of bacteria, which are closely related to the macroscopic morphology of the laminations, are present in both the bright and dark laminae. The results are useful in understanding the origin and formation mechanisms of the Neoproterozoic botryoidal dolostones. The new findings may also shed new light on the occurrence and evolution of early life on Earth in the Precambrian Era.

The Evolution of Ecosystem Phenotypes

Evolution by natural selection (ENS) has been extended to several supraorganismic levels, but whether it can apply to ecosystems remains controversial on two main counts. First, local ecosystems are loosely individuated, so that it is unclear how they manifest heredity and fitness. Second, even if they did, the meta-ecosystem formed by this population of local ecosystems will also suffer from a very low degree of cohesion, which will jeopardize any ENS. We suggest a way to overcome both issues, focusing on ecosystem properties (or phenotypes) rather than on ecosystemic individuals. First, we follow recent theoretical developments which deny the centrality of reproduction in ENS. This leads us to merge heredity and fitness in a single process, in which a local ecosystem (the unit of selection) causally influences its neighbors, in a way responsible for their phenotypic similarity. Second, we suggest that the resulting meta-ecosystem (the unit of evolution) need not meet the criteria of individuality (sensu Ghiselin–Hull). Instead, the evolving meta-ecosystem is best understood as a homeostatic property cluster, i.e., a set of phenotypes that together evolve by ENS at the ecosystem level. We illustrate this conceptual framework with a hypothetical example of local ecosystems that vary in terms of stability, productivity, diversity, and complementarity between species. Finally, we stress that our property-based account of ecosystemic ENS can help understand evolution at other biological levels, such as early life evolution and holobionts.

Surface-Assisted Self-Assembly of Fatty Acids to Cell-Like Compartments

In contemporary biological cells, lipid membrane-enclosed, spatially distinct microenvironments confine and protect biochemical reactions and other processes. On the early Earth, the formation of independent compartments is believed to have led to the encapsulation of nucleotides, possibly aiding and facilitating the emergence of life. In this context, solid surfaces have recently been investigated as potential supports for the self-assembly of prebiotic membrane containers, as significantly enhanced compartment formation was reported in the presence of solid interfaces. Particularly, the spontaneous transformation of solid-surface-adhered phospholipid reservoirs to unilamellar vesicular compartments was shown, where the deposits undergo a well-defined progression of topological changes, starting from two-dimensional molecular lipid films, via a network of interconnected lipid nanotubes to unilamellar giant compartments. However fatty acids, having only one hydrophobic tail, are structurally simpler than phospholipids, and could have been synthesized earlier than the more intricate phospholipids, and were therefore available earlier to form protocellular compartments. We show in our latest work that, starting from simple fatty acids on solid supports, the formation of protocells can occur in the same fashion as observed in earlier experiments with phospholipids. The compartments are generated from initially unstructured deposits of fatty acids on high energy surfaces, such as SiO2, where spontaneous surface wetting, film rupturing and giant vesicle formation occurs. The new findings indicate that the earlier observed surface-assisted protocell formation route is not exclusive to phospholipids, but may have been a more general pathway in the development of early life.

Recurrent anoxia recorded in shallow marine facies at Zhangcunping (western Hubei, China) throughout the Ediacaran to earliest Cambrian

In order to reconstruct the redox history of surface seawater for Ediacaran shallow marine facies on the Yangtze Platform, as well as to understand its implications for geological events (oxidation and phosphatization) and early evolution of life, we have conducted a systematic high-resolution investigation on microfacies analyses, pyrite morphology as well as sulfur and carbon contents in a continuous carbonate Ediacaran–Cambrian succession (drilled 407-core) in the Zhangcunping area of western Hubei (South China). The framboidal pyrites and SNAV/TOC (NAV = non-acid volatile sulfur) based redox reconstruction show a dynamic history of recurrent shallow marine anoxia. The early aftermath of the Marinoan Glaciation (cap dolomite to phosphorites) was characterized by either absence of pyrite or dominated by large framboids, low TOC contents and phosphorite deposition as well as well-preserved microfossils, indicating a minor oxygenated (dysoxic to oxic) photic shallow marine ecosystem. Meanwhile, the uppermost Doushantuo and Dengying formations were potentially fully oxygenated due to the lack of pyrites as well as low SNAV and TOC contents. Correlating with macrofossil assemblages (Miaohe biota and Ediacara biota) found in western Hubei, a major oxygenated marine ecosystem can be outlined for the late Ediacaran. In addition, a complicated diagenetic microfabric overprints the fine microstructure of microfossils preserved in phosphorites, leaving remarkable taphonomic puzzles for future study.