Crucial crises in biology: life in the deep biosphere.

The origin and evolution of life on Earth are the result of a series of crises that have taken place on the planet over about 4500 millions of years since it originated. Biopoiesis (origin of life), ecopoiesis (origin of ecosystems) and the first ecosystems (stromatolites and microbial mats), as well as eukaryopoiesis (origin of nucleated cells) are revised. The paper then focuses on the study of the deep biosphere, describing ecosystems never found before, which are independent of solar radiation and have changed previous assumptions about the requirements of life; even the concept of biosphere, as Vernadsky defined it, has increased its scope. Since the discovery, in 1987, of bacteria growing in the crevices of rocks at 500 m deep, in boreholes drilled near the Savanna River, Aiken, South Carolina, other bacteria have been found in the deep subsurface reaching depths of about 3 km (e.g., in the Columbia River Basalt Group, near Richland, Washington state), in an anaerobic, hot, high-pressure environment. Some kinds of microorganisms can thrive at such depths, living in many cases a geochemical existence, by using very specialized metabolisms, which depend on the local environments. The existence of organisms independent from photosynthetic production is the most outstanding, novel feature of the deep biosphere. Living beings might not need other energy and chemical sources than those which occur in the development of all planetary bodies. Life, therefore, could even be an ineluctable outcome of planetary evolution and, as a corollary, a natural continuation of the usual development of physical phenomena in the universe.     

Are microorganisms everywhere they can be?

Baas-Becking is famously attributed with the conjecture that ‘everything is everywhere, but the environment selects’. Although this aphorism is largely challenged by microbial biogeographical data, even weak versions of the claim leave unanswered the question about whether all environments that could theoretically support life contain life. In the last decade, the discovery of thermally sterilized habitable environments disconnected from inhabited regions, and habitats within organisms such as the sterile, but habitable human fetal gut, suggest the existence of a diversity of macroscopic habitable environments apparently devoid of actively metabolizing or reproducing life. Less clear is the status of such environments at the micron scale, for example, between colonies of organisms within rock interstices or on and within other substrates. I discuss recent evidence for these types of environments. These environments have practical uses in: (i) being negative controls for understanding the role of microbial processes in geochemical cycles and geological processes, (ii) yielding insights into the extent to which the biosphere extends into all spaces it theoretically can, (iii) suggesting caution in interpreting the results of life detection instrumentation, and (iv) being useful for establishing the conditions for the origin of life and its prevalence on other planetary bodies.     

Nitric Oxide Cycle in Mammals and the Cyclicity Principle

This paper continues a series of reports considering nitric oxide (NO) and its cyclic conversions in mammals. Numerous facts are summarized with the goal of developing a general concept that would allow the statement of the multiple effects of NO on various systems of living organisms in the form of a short and comprehensive law. The current state of biological aspects of NO research is analyzed in term of elucidation of possible role of these studies in the system of biological sciences. The general concept is based on a notion on cyclic conversions of NO and its metabolites. NO cycles in living organisms and nitrogen turnover in the biosphere and also the Bethe nitrogen–carbon cycle in star matter are considered. A hypothesis that the cyclic organization of processes in living organisms and the biosphere reflects the evolution of life is proposed: the development of physiological functions and metabolism are suggested to be closely related to space and evolution of the Earth as a planet of the Solar System.     

Giant viruses coexisted with the cellular ancestors and represent a distinct supergroup along with superkingdoms Archaea, Bacteria and Eukarya

ABSTRACT: BACKGROUND: The discovery of giant viruses with genome and physical size comparable to cellular organisms, remnants of protein translation machinery and virus-specific parasites (virophages) have raised intriguing questions about their origin. Evidence advocates for their inclusion into global phylogenomic studies and their consideration as a distinct and ancient form of life. RESULTS: Here we reconstruct phylogenies describing the evolution of proteomes and protein domain structures of cellular organisms and double-stranded DNA viruses with medium-to-very-large proteomes (giant viruses). Trees of proteomes define viruses as a 'fourth supergroup' along with superkingdoms Archaea, Bacteria, and Eukarya. Trees of domains indicate they have evolved via massive and primordial reductive evolutionary processes. The distribution of domain structures suggests giant viruses harbor a significant number of protein domains including those with no cellular representation. The genomic and structural diversity embedded in the viral proteomes is comparable to the cellular proteomes of organisms with parasitic lifestyles. Since viral domains are widespread among cellular species, we propose that viruses mediate gene transfer between cells and crucially enhance biodiversity. CONCLUSIONS: Results call for a change in the way viruses are perceived. They likely represent a distinct form of life that either predated or coexisted with the last universal common ancestor (LUCA) and constitute a very crucial part of our planet's biosphere.     

Search for life in deep biospheres]

The life in deep biospheres bridges conventional biology and future exobiology. This review focuses the microbiological studies from the selected deep biospheres, i.e., deep-sea hydrothermal vents, sub-hydrothermal vents, terrestrial subsurface and a sub-glacier lake. The dark biospheres facilitate the emergence of organisms and communities dependent on chemolithoautotrophy, which are overwhelmed by photoautotrophy (photosynthesis) in the surface biospheres. The life at deep-sea hydrothermal vents owes much to chemolithoautotrophy based on the oxidation of sulfide emitted from the vents. It is likely that similarly active bodies such as the Jovian satellite Europa may have hydrothermal vents and associated biological communities. Anoxic or anaerobic condition is characteristic of deep subsurface biospheres. Subsurface microorganisms exploit available oxidants, or terminal electron acceptors (TEA), for anaerobic respiration. Sulfate, nitrate, iron (III) and CO2 are the representative TEAs in the deep subsurface. Below the 3000-4000 m-thick glacier on Antarctica, there have been >70 lakes with liquid water located. One of such sub-glacial lakes, Lake Vostok, is about to be drill-penetrated for microbiological studies. These deep biosphere "platforms" provide new knowledge about the diversity and potential of the Earth's life. The expertise obtained from the deep biosphere expeditions will facilitate the capability of exobiologial exploration.     

Review: from chemosphere to biosphere

An understanding of how the Earth's chemosphere was transformed to a biosphere is central to our understanding of the origin of life and the search for extraterrestrial life or life signatures. Once early prokaryotic life originated and colonized the Earth, the biosphere was well on its way to being formed. In this paper, information and knowledge is integrated to examine the possibility how life first self-assembled and transformed a lifeless chemosphere into a complex biosphere that we still do not understand today.     

A New Frontier for Palaeobiology: Earth's Vast Deep Biosphere

Diverse micro‐organisms populate a global deep biosphere hosted by rocks and sediments beneath land and sea, containing more biomass than any other biome except forests. This paper reviews an emerging palaeobiological archive of these dark habitats: microfossils preserved in ancient pores and fractures in the crust. This archive, seemingly dominated by mineralized filaments (although rods and coccoids are also reported), is presently far too sparsely sampled and poorly understood to reveal trends in the abundance, distribution, or diversity of deep life through time. New research is called for to establish the nature and extent of the fossil record of Earth's deep biosphere by combining systematic exploration, rigorous microanalysis, and experimental studies of both microbial preservation and the formation of abiotic pseudofossils within the crust. It is concluded that the fossil record of Earth's largest microbial habitat may still have much to tell us about the history of life, the evolution of biogeochemical cycles, and the search for life on Mars.     

The origin of life—did it occur at high temperatures?

A high-temperature origin of life has been proposed, largely for the reason that the hyperthermophiles are claimed to be the last common ancestor of modern organisms. Even if they are the oldest extant organisms, which is in dispute, their existence can say nothing about the temperatures of the origin of life, the RNA world, and organisms preceding the hyperthermophiles. There is no geological evidence for the physical setting of the origin of life because there are no unmetamorphosed rocks from that period. Prebiotic chemistry points to a low-temperature origin because most biochemicals decompose rather rapidly at temperatures of 100°C (e.g., half-lives are 73 min for ribose, 21 days for cytosine, and 204 days for adenine). Hyperthermophiles may appear at the base of some phylogenetic trees because they outcompeted the mesophiles when they adapted to lower temperatures, possibly due to enhanced production of heat-shock proteins. Correspondence to: S.L. Miller     

Superbugs viewed from cosmobiology]

Superbugs (microorganisms living in unfamiliar and very harsh environments) are located in the center of scientific interests in the sense that 1) most of their habitats belong to marginal regions of the biosphere, 2) clues for the elucidation of the origin of life can be deduced from them, and 3) they are deeply correlated to the extraterrestrial life. Not only for the basic scientific interests, but also for the applied fields, the spot light is shed to them. We, human beings, have been deeply dependent on other organisms through the global material flow they make. Microorganisms together with plants are the leading characters in this process. Also implicit, our future is surely correlated to microorganisms including superbugs. In this special issue, superbugs; thermophilic, psychrophilic barophilic etc. are summarized, and future development on the study on the superbugs are introduced.     

First ecosystems on Earth

Theories attempting to explain the origin of life on Earth should be based on the assumption that habitability precedes habitation. The hypotheses of the first organism should be based on the evaluation of its possible life-supporting ecosystem. The ecosystem should necessarily include primary autotrophic producers, and hydrogenotrophy appears to be an adequate physiological type for primitive ecosystems. Consideration of life on Earth should differentiate between the origin of organisms in situ and the transportation of organisms from outside with cosmic bodies in the framework of life as a widespread phenomenon of the Universe. In the case of transport of life with cosmic bodies, there are no limitations on the transfer of a community rather than an individual cell. In the case of the transport of the community with a large piece of “dirty ice,” the problem lies in the correspondence between the community and its ecosystem on the parent body and the conditions on the primeval Earth rather than functional divergence from a primary ancestor. Subsequent events are within the framework of paleontologically observed evolution and can be described as biogeochemical succession without any additional speculations.     

On the early evolutionary stage of the geosphere and biosphere and the problem of early glaciations

The early evolutionary stages of the geosphere and biosphere are determined by three interrelated factors: (1) continuous cooling of the surface and interior (mantle) of the Earth (the mean temperatures of the mantle and surface decreased by a factor of 1.5–2 and 3–4, respectively; the mean heat flow was reduced by approximately one order of magnitude, and viscosity, by three orders); (2) continuous stepwise oxidation of the surface, which was particularly well pronounced from 3.8 to 1.8 Ga; and (3) periodic and correlated fluctuations of conditions in the geosphere and biosphere of varying extent and nature. The major boundaries of this evolution were about 4 Ga (the origin of rather thick and heterogeneous earth’s crust, the origin of life); about 3 Ga (appearance of a strong magnetic field, an increase in photosynthetic activity); about 1.8–1.9 Ga (appearance of an oxidized atmosphere, the first supercontinent, possibly, the first superplumes from the nucleus); and about 0.75 Ga (acceleration of subduction, “watering” of the upper mantle, elevation of continents with vast land masses, shelves, large rivers, and the first great glaciations). The significance and correlations of the earliest events (before and about 4 Ga) and events about 750 Ma are widely debated. In the Late Archean and Early Proterozoic (before 1.8 Ga), the biosphere was dominated by cyanobacteria, the dynamics and developmental peaks of which are marked by the presence of widespread stromatolite buildups in carbonaceous rocks (initially, mostly dolomitic matter). About 700–750 Ma, intense and frequent glaciations developed, marking the cooling of the Earth. The greatest glaciation apparently occurred about 640 Ma, which gave rise to the discussion of the model of the Snowball Earth. The emergence and evolution of skeletons in animals is sometimes thought to be connected with glaciations. These events are correlated and accounted for by great endogenous changes. One of the major events in endogenous history is the onset about 750 Ma of periodic manifestation of mantle flows (superplumes), which explain further periodicity of the biosphere evolution. In conclusion, extrapolation of future evolution and successive collapse of biosphere segments in the course of transformation of the Sun into a red star and warming of the Earth surface are proposed.     

Geochemical ecology and problems of health.

The state of health or disease is determined by the nature of the organism, the properties of the biosphere, the heterogeneity of its natural geochemical composition and changes brought about by technology (technogenic changes). For a systematic study of the conditions of health and endemic diseases we have suggested a system of biogeochemical regionalizing of the biosphere with the aid of biospheric taxa: regions of the biosphere, subregions of the biosphere, biogeochemical provinces. The main criteria of the regionalizing are biogenous cycles of chemical elements (links of the biogeochemical food chain from soil-forming rocks to man). An important criterion of the biogeochemical regionalizing is threshold concentrations of chemical elements. The organism regulates its metabolism within the ranges of chemical element concentration between the upper and lower thresholds (necessity range). When chemical elements are present in concentrations above the upper threshold and below the lower threshold, dysfunctions and endemic diseases are observed. Hence, the biogeochemical food chain allows us to establish critical links responsible for the state of health or endemic disease. Principles of optimizing the conditions of the environment and life have been worked out. The creation by us in the U.S.S.R. of biogeochemical maps relating conditions of the environment to biological reactions of organisms has proved a useful method of studying the ecological structure of the biosphere.     

Microbes on the edge of global biosphere]

The search for life on the edge of global biosphere is a frontier to bridge conventional bio/ecology and exo/astrobiology. This communication reviews the foci of microbiological studies on the inhabitants of the selected "edges", i.e., deep-sea, deep subsurface and Antarctic habitats. The deep-sea is characterized as the no-light (non-photosynthetic) habitat, and the primary production is mostly due to the chemosynthetic autotrophy at the hydrothermal vents and methane-rich seeps. Formation of the chemosynthesis-dependent animal communities in the deep leads to the idea that such communities may be found in "ocean" of the Jovian satellite, Europa. The oxygen minimal layer (OML) in mid-water provides another field of deep-sea research. Modern OML is a relatively thin layer, found between the water depth of 200 and 1000 m, but was much thicker during the periods of oceanic anoxia events (OAEs) in the past. The history of oceanic biosphere is regarded as the cycle of OAE and non-OAE periods, and the remnants of the past OAEs may be seen in the modem OML. Anoxic (no-O2) condition is also characteristic of deep subsurface biosphere. Microorganisms in deep subsurface biosphere exploit every available oxidant, or terminal electron acceptor (TEA), for anaerobic respiration. Sulfate, nitrate, iron (III) and CO2 are the representative TEAs in the deep subsurface. Subsurface of hydrothermal vents, or sub-vent biosphere, may house brine (high salt) habitats and halophilic microorganisms. Some sub-vent halophiles were phylogenetically closely similar to the ones found in the Antarctic habitats which are extremely dry by the liophilizing climate. Below the 3000-4000 m-thick glacier on Antarctica, there have been >70 lakes with liquid water located. One of such sub-glacial lakes, Lake Vostok, has been a target of "life in extreme environments" and is about to be drill-penetrated for microbiological studies. These 'microbiological platforms' will provide new knowledge about the diversity and potential of the Earth's life and facilitate the capability of astrobiologial exploration.     

Abiogenic synthesis of prebiotic matter for the Earth’s biosphere as a stage of self-organization on an astrophysical and paleontological time scale

Existing data suggest that an early circumstellar preplanetary disk was the most likely location for primary abiogenic synthesis of prebiotic organic matter from simple molecules along with the “RNA world” and the origin of life. This paper discusses the stages of self-organization that have resulted in the Earth’s modern biosphere, and the relationships between astrophysical and paleontological events in evolution.     

Application of alkaline elution, Fast Micromethod and flow cytometry in detection of marine contamination.

DNA damage is an inescapable aspect of life in the biosphere. The presented investigations were an attempt to examine the response of a DNA damage as a biomarker of environmental quality in the mussels Mytilus galloprovincialis sampled at differently contaminated areas of Istrian coast, Northern Adriatic. The investigations were performed in order to get information about the genotoxic risk for marine organisms exposed to mixed environmental pollution, as well as the information about the presence of unknown mixture of genotoxic contaminants in the marine environment. Types of DNA damage detected are alkali-labile sites and single-strand breaks measured by Fast Micromethod, interstrand cross-links and DNA protein cross-links by alkaline filter elution and cell cycle disturbation by flow cytometry. The applicability of all three methods for marine quality control is discussed.     

The origin and evolution of model organisms

The phylogeny and timescale of life are becoming better understood as the analysis of genomic data from model organisms continues to grow. As a result, discoveries are being made about the early history of life and the origin and development of complex multicellular life. This emerging comparative framework and the emphasis on historical patterns is helping to bridge barriers among organism-based research communities.     

Biomass recycling and the origin of phenotype in fungal mycelia

Fungi are one of the most important and widespread components of the biosphere, and are essential for the growth of over 90% of all vascular plants. Although they are a separate kingdom of life, we know relatively little about the origins of their ubiquitous existence. This reflects a wider ignorance arising from their status as indeterminate organisms epitomized by extreme phenotypic plasticity that is essential for survival in complex environments. Here we show that the fungal phenotype may have its origins in the defining characteristic of indeterminate organisms, namely their ability to recycle locally immobilized internal resources into a mobilized form capable of being directed to new internal sinks. We show that phenotype can be modelled as an emergent phenomenon resulting from the interplay between simple local processes governing uptake and remobilization of internal resources, and macroscopic processes associated with their transport. Observed complex growth forms are reproduced and the sensitive dependence of phenotype on environmental context may be understood in terms of nonlinearities associated with regulation of the recycling apparatus.     

Origins of highly mosaic mycobacteriophage genomes

Bacteriophages are the most abundant organisms in the biosphere and play major roles in the ecological balance of microbial life. The genomic sequences of ten newly isolated mycobacteriophages suggest that the bacteriophage population as a whole is amazingly diverse and may represent the largest unexplored reservoir of sequence information in the biosphere. Genomic comparison of these mycobacteriophages contributes to our understanding of the mechanisms of viral evolution and provides compelling evidence for the role of illegitimate recombination in horizontal genetic exchange. The promiscuity of these recombination events results in the inclusion of many unexpected genes including those implicated in mycobacterial latency, the cellular and immune responses to mycobacterial infections, and autoimmune diseases such as human lupus. While the role of phages as vehicles of toxin genes is well established, these observations suggest a much broader involvement of phages in bacterial virulence and the host response to bacterial infections.     

Deep‐biosphere consortium of fungi and prokaryotes in Eocene subseafloor basalts

The deep biosphere of the subseafloor crust is believed to contain a significant part of Earth's biomass, but because of the difficulties of directly observing the living organisms, its composition and ecology are poorly known. We report here a consortium of fossilized prokaryotic and eukaryotic micro‐organisms, occupying cavities in deep‐drilled vesicular basalt from the Emperor Seamounts, Pacific Ocean, 67.5 m below seafloor (mbsf). Fungal hyphae provide the framework on which prokaryote‐like organisms are suspended like cobwebs and iron‐oxidizing bacteria form microstromatolites (Frutexites). The spatial inter‐relationships show that the organisms were living at the same time in an integrated fashion, suggesting symbiotic interdependence. The community is contemporaneous with secondary mineralizations of calcite partly filling the cavities. The fungal hyphae frequently extend into the calcite, indicating that they were able to bore into the substrate through mineral dissolution. A symbiotic relationship with chemoautotrophs, as inferred for the observed consortium, may be a pre‐requisite for the eukaryotic colonization of crustal rocks. Fossils thus open a window to the extant as well as the ancient deep biosphere.     

Viruses in extreme environments

The tolerance limits of extremophiles in term of temperature, pH, salinity, desiccation, hydrostatic pressure, radiation, anaerobiosis far exceed what can support non-extremophilic organisms. Like all other organisms, extremophiles serve as hosts for viral replication. Many lines of evidence suggest that viruses could no more be regarded as simple infectious “fragments of life” but on the contrary as one of the major components of the biosphere. The exploration of niches with seemingly harsh life conditions as hypersaline and soda lakes, Sahara desert, polar environments or hot acid springs and deep sea hydrothermal vents, permitted to track successfully the presence of viruses. Substantial populations of double-stranded DNA virus that can reach 10⁹ particles per milliliter were recorded. All these viral communities, with genome size ranging from 14 kb to 80 kb, seem to be genetically distinct, suggesting specific niche adaptation. Nevertheless, at this stage of the knowledge, very little is known of their origin, activity, or importance to the in situ microbial dynamics. The continuous attempts to isolate and to study viruses that thrive in extreme environments will be needed to address such questions. However, this topic appears to open a new window on an unexplored part of the viral world. Marc Le Romancer and Mélusine Gaillard contributed equally to this work.