Stable carbon isotope fractionation in the search for life on early Mars.

Isotopic measurements and, more specifically, ratios of 13C to 12C in organic relative to inorganic deposits, are useful in reconstructing past biological activity on Earth. Organic matter has a lower ratio of 13C to 12C due largely to the preferential fixation of 12C over the heavier isotope by the major carbon-fixation enzyme, ribulose-1,5-bisphosphate carboxylase/oxygenase, although other factors (e.g., availability of source carbon, fixation by other carboxylating enzymes and diagenesis of organic material) also contribute to fractionation. Would carbon isotope discrepancies between inorganic and organic carbon indicate past biological activity on Mars? In order to answer this question, we analyse what is known about terrestrial biologic and abiologic carbon fixation and its preservation in the fossil record, and suggest what the isotope discrimination during possible biologic and abiologic carbon fixation on Mars might have been like. Primarily because isotopic signatures of abiotically fixed carbon overlap with those of biotic fixation, but also because heterotrophy does not significantly alter the isotopic signature of ingested carbon, fractionation alone would not be definitive evidence for life. However, a narrow range of fractionation, including no fractionation, would suggest biotic processes. Never-the-less, isotopic ratios in organic deposits on Mars would be extremely useful in analysing prebiotic, if not biotic, carbon transformations on Mars.     

The initiation of biological processes on Earth: summary of empirical evidence.

With the currently available geological record at band, the existence of life on this planet as from at least 3.8 Gyr ago seems so firmly established as to be virtually unassailable. Specifically, various disparate lines of evidence have merged to indicate (1) that the surface of the Archaean Earth had hosted prolific microbial ecosystems as is testified by a quasi-continuous record of microbialites ("stromatolites") and associated microfossils of prokaryotic affinity over 3.5, if not 3.8 Gyr of geological history, and (2) that the sedimentary carbon record has preserved the isotopic signature of autotrophic (notably photosynthetic) carbon fixation over the same time span. With the observed enrichment of isotopically light carbon in sedimentary organic matter largely consonant with the bias in favor of 12C during photosynthesis, the mainstream of the carbon isotope record can be best explained as geochemical manifestation of the isotope discriminating properties of the ribulose-1,5-bisphosphate (RuBP) carboxylase reaction of the Calvin cycle suggesting an extreme degree of evolutionary conservatism in the biochemistry of autotrophic carbon fixation. As a consequence, partial biological control of the geochemical carbon cycle was established already during Early Archaean times and fully operative by the time of formation of the Earth's earliest sediments.     

Stable carbon isotopes: possible clues to early life on Mars.

Organic and inorganic carbon in terrestrial near-surface environments are characterized by a marked difference in their 13C/12C ratios which can be traced back in the Earth's sedimentary record over almost 4 billion years. There is no doubt that the bias in favour of 12C displayed by biogenic matter derives, for the most part, from the isotope-selecting properties of the carbon-fixing enzyme (ribulose-1,5-bisphosphate carboxylase) that is operative in the principal photosynthetic pathway and promotes most of the carbon transfer from the non-living to the living realm. Postulating a universality of biological principles in analogy to the proven universality of the laws of physics and chemistry, we may expect enzymatic reactions in exobiological systems to be beset with B similar kinetic fractionation effects. Hence, the retrieval from the oldest Martian sediments of isotopic fractionations between reduced and oxidized (carbonate) carbon may substantially constrain current conjectures on the possible existence of former life on Mars.     

Biological modulation of the terrestrial carbon cycle: Isotope clues to early organic evolution

The characteristic difference of some 20 to 30 ‰ in the ¹³C/¹²C rations of reduced (organic) and oxidized (carbonate) carbon as observed in the contemporary environment can be traced back over 3.5 (if not 3.8) Ga of recorded Earth history. There is little doubt that the enrichment of isotopically light carbon in fossil organic matter ultimately derives from the bias in favor of ¹²C during photosynthetic carbon fixation (notably the RuBP carboxylase reaction of the Calvin cycle). Accordingly, the mainstream of the sedimentary δ ¹³C_org record can be most reasonably explained as geochemical evidence of the isotope-discriminating properties of the principal CO₂-fixing enzymatic reaction of the assimilatory pathway, thus giving eloquent testimony to an extreme degree of evolutionary conservatism in the biochemistry of autotrophic carbon fixation.     

Stable isotope analysis at the molecular level: a new approach for determining the origins of amino acids in the Murchison meteorite.

A combined gas chromatography/isotope ratio mass spectrometry (GC/IRMS) method has been developed that permits the direct stable carbon isotope analysis of N(O)-trifluoroacetyl-isopropyl esters of individual amino acids and their respective enantiomers at nanomole abundances. Calculation of the original delta 13C values of the amino acids is accomplished via a correction for the carbon introduced during the derivatization process. Previous GC/IRMS analyses of individual amino acids in the non-hydrolyzed water extract of an interior sample of a Murchison meteorite stone revealed an enrichment in 13C relative to terrestrial organic matter, in agreement with previous findings for bulk extracts. The range of amino acid delta 13C values (+5 to +30%, PDB) suggests possible kinetic effects during synthesis. In this study, an apparent kinetic isotope effect was also observed for the amino acid products of a spark discharge experiment. These preliminary results are supportive of a similar mechanism for the abiotic synthesis of amino acids in the Murchison meteorite.     

From Titan’s tholins to Titan’s aerosols: Isotopic study and chemical evolution at Titan’s surface

In the present work, we focused on the possible isotopic fractionation of carbon during the processes involved in the formation of Titan’s tholins. We present the first results obtained on the ¹²C/¹³C isotopic ratios measured on Titan’s tholins synthesized in laboratory with cold plasma discharges. Measurements of isotopic ratio ¹²C/¹³C, done both on tholins and on the initial gas mixture (N₂:CH₄ (98:2)) used to produce them, do not show any evident deficit or enrichment in ¹³C relatively to ¹²C in the synthesized tholins, compared to the initial gas mixture. This observation allows to go further in the analyses of the ACP experiment data, including part of the Cassini–Huygens mission.     

The origin of organic matter in the Martian meteorite ALH84001.

Stable carbon isotope measurements of the organic matter associated with the carbonate globules and the bulk matrix material in the ALH84001 Martian meteorite indicate that two distinct sources are present in the sample. The delta 13C values for the organic matter associated with the carbonate globules averaged -26% and is attributed to terrestrial contamination. In contrast, the delta 13C values for the organic matter associated with the bulk matrix material yielded a value of -15%. The only common carbon sources on the Earth that yield similar delta 13C values, other then some diagenetically altered marine carbonates, are C4 plants. A delta 13C value of -15%, on the other hand, is consistent with a kerogen-like component, the most ubiquitous form of organic matter found in carbonaceous chondrites such as the Murchison meteorite. Examination of the carbonate globules and bulk matrix material using laser desorption mass spectrometry (LDMS) indicates the presence of a high molecular weight organic component which appears to be extraterrestrial in origin, possibly derived from the exogenous delivery of meteoritic or cometary debris to the surface of Mars.     

Functional, regulatory and indicator features of microorganisms in man-made ecosystems.

Functional, regulatory and indicator features of microorganisms in development and functioning of the systems and sustaining stability of three man-made ecosystem types has been studied. 1) The functional (metabolic) feature was studied in aquatic ecosystems of biological treatment of sewage waters for the reducer component. 2) The regulatory feature of bacteria for plants (producer component) was studied in simple terrestrial systems "wheat plants-rhizospheric microorganisms-artificial soil" where the behavior of the system varied with activity of the microbial component. For example with atmospheric carbon dioxide content elevated microbes promote intensification of photosynthesis processes, without binding the carbon in the plant biomass. 3) The indicator feature for the humans (consumer component) was studied in Life Support Systems (LSS). High sensitivity of human microflora to system conditions allowed its use as an indicator of the state of both system components and the entire ecosystem. Grant numbers: N99-04-96017, N15.     

The use of mineral crystals as bio-markers in the search for life on Mars.

Photographs that depict presumed fluvial features on the martian surface have led geologists to hypothesize that water flowed across the early martian terrain. From this, it has been further hypothesized that the surface and atmospheric conditions on early Mars were similar to those on early Earth. Because the oldest fossil evidence of life on Earth dates back to this early period, at least 3.5 billion years ago, the possibility exists that the early Martian environment could have also been conducive to the origin of life. To investigate this possibility, universal signatures or bio-markers indicative of past (or present) biological activity must be identified for use in the search for life on Mars. Several potentially applicable biomarkers have been identified and include: organics (e.g., specific classes of lipids and hopanes), suites of specific inorganic and organic compounds, as well as the isotopic ratios of C, N, and S. Unfortunately, all of these bio-markers may be of biologic or abiotic origin; these origins are often difficult to distinguish. Thus, the discovery of any one of these compounds alone is not a bio-marker. Because minerals produced under biologic control have distinctive crystallographies, morphologies, and isotopic ratios that distinguishable from abiotically produced minerals with the same chemical composition, and are stable through geologic time, we propose the use of minerals resulting from biologically controlled mineralization processes as bio-markers.     

Utilization of white potatoes in CELSS.

Potatoes (Solanum tuberosum) have a strong potential as a useful crop species in a functioning CELSS. The cultivar Denali has produced 37.5 g m-2 d-1 when grown for 132 days with the first 40 days under a 12-h photoperiod and a light:dark temperature cycle of 20 degrees C:16 degrees C, and then 92 days under continuous irradiance and a temperature of 16 degrees C. Irradiance was at 725 micromoles m-2 s-1 PPF and carbon dioxide at 1000 micromoles mol-1. The dried tubers had 82% carbohydrates, 9% protein and 0.6% fat. Other studies have shown that carbon dioxide supplementation (1000 micromoles mol-1) is of significant benefit under 12-h irradiance but less benefit under 24 h irradiance. Irradiance cycles of 60 minutes light and 30 minutes dark caused a reduction of more than 50% in tuber weight compared to cycles of 16 h light and 8 h dark. A diurnal temperature change of 22 degrees C for the 12-h light period to 14 degrees C during the 12-h dark period gave increased yields of 30% and 10% for two separate cultivars, compared with plants grown under a constant 18 degrees C temperature. Cultivar screening under continuous irradiance and elevated temperatures (28 degrees C) for 8 weeks of growth indicated that the cvs Haig, Denali, Atlantic, Desiree and Rutt had the best potential for tolerance to these conditions. Harvesting of tubers from plants at weekly intervals, beginning at 8 weeks after planting, did not increase yield over a single final harvest. Spacing of plants on 0.055 centers produced greater yield per m2 than spacing at 0.11 or 0.22 m2. Plants maintained 0.33 meters apart (0.111 m2 per plant) in beds produced the same yields when separated by dividers in the root matrix as when no separation was made.     

Soil carbon distribution and site characteristics in hyper-arid soils of the Atacama Desert: A site with Mars-like soils

The soil carbon content and its relation to site characteristics are important in evaluating current local, regional, and global soil C storage and projecting future variations in response to climate change. In this study we analyzed the concentration of organic and inorganic carbon and their relationship with in situ climatic and geological characteristics in 485 samples of surface soil and 17 pits from the hyper-arid area and 51 samples with 2 pits from the arid–semiarid region from the Atacama Desert located in Peru and Chile. The soil organic carbon (SOC) in hyperarid soils ranged from 1.8 to 50.9 μg C per g of soil for the 0–0.1 m profile and from 1.8 to 125.2 μg C per g of soil for the 0–1 m profile. The analysis of climatic (temperature and precipitation), elevation, and some geologic characteristics (landforms) associated with hyper-arid soils explained partially the SOC variability. On the other hand, soil inorganic carbon (SIC) contents, in the form of carbonates, ranged from 200 to 1500 μg C per g of soil for the 0–0.1 m profile and from 200 to 3000 μg C per g of soil for the 0–1.0 m profile in the driest area. The largest accumulations of organic and inorganic carbon were found near to arid–semiarid areas. In addition, the elemental carbon concentrations show that the presence of other forms of inorganic carbon (e.g. graphite, etc.) was negligible in these hyperarid soils. Overall, the top 1 m soil layer of hyperarid lands contains ∼11.6 Tg of organic carbon and 344.6 Tg of carbonate carbon. The total stored carbon was 30.8-fold the organic carbon alone. To our knowledge, this is the first study evaluating the total budget carbon on the surface and shallow subsurface on ∼160,000 km² of hyperarid soils.     

A "cryptic" microbial mat: a new model ecosystem for extant life on Mars.

If life were present on Mars to day, it would face potentially lethal environmental conditions such as a lack of water, frigid temperatures, ultraviolet radiation, and soil oxidants. In addition, the Viking missions did not detect near-surface organic carbon available for assimilation. Autotrophic organisms that lived under a protective layer of sand or gravel would be able to circumvent the ultraviolet radiation and lack of fixed carbon. Two terrestrial photosynthetic near-surface microbial communities have been identified, one in the inter- and supertidal of Laguna Ojo de Liebre (Baja California Sur, Mexico) and one in the acidic gravel near several small geysers in Yellowstone National Park (Wyoming, U.S.A.). Both communities have been studied with respect to their ability to fix carbon under different conditions, including elevated levels of inorganic carbon. Although these sand communities have not been exposed to the entire suite of Martian environmental conditions simultaneously, such communities can provide a useful model ecosystem for a potential extant Martian biota.     

Mass exchange in an experimental new-generation life support system model based on biological regeneration of environment.

An experimental model of a biological life support system was used to evaluate qualitative and quantitative parameters of the internal mass exchange. The photosynthesizing unit included the higher plant component (wheat and radish), and the heterotrophic unit consisted of a soil-like substrate, California worms, mushrooms and microbial microflora. The gas mass exchange involved evolution of oxygen by the photosynthesizing component and its uptake by the heterotroph component along with the formation and maintaining of the SLS structure, growth of mushrooms and California worms, human respiration, and some other processes. Human presence in the system in the form of "virtual human" that at regular intervals took part in the respirative gas exchange during the experiment. Experimental data demonstrated good oxygen/carbon dioxide balance, and the closure of the cycles of these gases was almost complete. The water cycle was nearly 100% closed. The main components in the water mass exchange were transpiration water and the watering solution with mineral elements. Human consumption of the edible plant biomass (grains and roots) was simulated by processing these products by a unique physicochemical method of oxidizing them to inorganic mineral compounds, which were then returned into the system and fully assimilated by the plants. The oxidation was achieved by "wet combustion" of organic biomass, using hydrogen peroxide following a special procedure, which does not require high temperature and pressure. Hydrogen peroxide is produced from the water inside the system. The closure of the cycle was estimated for individual elements and compounds. Stoichiometric proportions are given for the main components included in the experimental model of the system. Approaches to the mathematical modeling of the cycling processes are discussed, using the data of the experimental model. Nitrogen, as a representative of biogenic elements, shows an almost 100% closure of the cycle inside the system. The proposed experimental model of a biological system is discussed as a candidate for potential application in the investigations aimed at creating ecosystems with largely closed cycles of the internal mass exchange. The formation and maintenance of sustainable cycling of vitally important chemical elements and compounds in biological life support systems (BLSS) is an extremely pressing problem. To attain the stable functioning of biological life support systems (BLSS) and to maintain a high degree of closure of material cycles in than, it is essential to understand the character of mass exchange processes and stoichiometnc proportions of the initial and synthesized components of the system.     

ECOSIMP2 model: prediction of CO2 concentration changes and carbon status in closed ecosystems.

The ECOSIMP2 model, simulating the Plant-Soil-Atmosphere interactions, was developed as a tool for the management of an experimental artificial ecosystem. It consists in three main carbon compartments for production, consumption and decomposition of the biomass. The main biological parameters concern photosynthesis (apparent Km, CO2 compensation point), the harvest index, the rate of consumption, and the kinetics of litter decomposition. From realistic assumptions of kinetics of soil compartments, a steady-state case was obtained, simulating a terrestrial ecosystem. The stability of the atmospheric CO2 concentration was studied after a virtual enclosure of the system in a 20-m high greenhouse. In natural lighting the conditions of stability are severe because of the small size of the atmospheric compartment which amplifies any imbalance between carbon fluxes. The positive consequence of that amplification for research on artificial ecosystems was emphasized.     

Biogeochemical evidence of microbial activity on Mars.

We suggest a new interpretation of the data on so-called SNC meteorites and delta 13C values of the calcium carbonate minerals and organic matter discovered in them. The delta 13C value of calcite (up to 15 ppt) is accounted for by the microbial reaction CO2 + H2 ---> CH4 + H2O. Methane-forming bacteria also synthesize organic carbon (in the form of biomass) from CO2, and this process is accompanied by 12C fractionation. Therefore, the organic carbon of SNC meteorites is enriched with 12C (delta 13C as low as -35 ppt). The environmental conditions under which the calcite of SNC meteorites was formed were favorable for the activity of methanogens.     

Life in the solar system.

Life, defined as a chemical system capable of transferring its molecular information via self-replication and also capable of evolving, must develop within a liquid to take advantage of the diffusion of complex molecules. On Earth, life probably originated from the evolution of reduced organic molecules in liquid water. Organic matter might have been formed in the primitive Earth's atmosphere or near hydrothermal vents. A large fraction of prebiotic organic molecules might have been brought by extraterrestrial-meteoritic and cometary dust grains decelerated by the atmosphere. Any celestial body harboring permanent liquid water may therefore accumulate the ingredients that generated life on the primitive Earth. The possibility that life might have evolved on early Mars when water existed on the surface marks it as a prime candidate in a search for bacterial life beyond the Earth. Europa has an icy carapace. However, cryovolcanic flows at the surface point to a possible water subsurface region which might harbor a basic life form. The atmosphere and surface components of Titan are also of interest to exobiology for insight into a hydrocarbon-rich chemically evolving world. One-handed complex molecules and preferential isotopic fractionation of carbon, common to all terrestrial life forms, can be used as basic indicators when searching for life beyond the Earth.     

Amino acids in meteorites.

Carbonaceous chondrites carry a record of chemical evolution that is unparalleled among presently accessible natural materials. Within the complex suite of organic compounds that characterize these meteorites, amino acids occur at a total concentration that may reach 0.6 micromole g-1 meteorite (approximately 60 ppm). Both free amino acids and acid-labile amino acid derivatives have been found in hot-water extracts of a CI1 and seven CM2 chondrites. Although the amino acid composition of all CM2 chondrites is not the same, differences may be largely explicable on the basis of spontaneous and biologically-caused decomposition occurring during their terrestrial residence. The amino acids of the Murchison meteorite (CM2) have been extensively analyzed and 52 amino acids have been positively identified. Thirty three of these amino acids are unknown in natural materials other than carbonaceous chondrites. Thus the Murchison meteorite has recently been the major source of new naturally-occurring amino acids. The Murchison amino acids comprise a mixture of C2 through C8 cyclic and acyclic monoamino alkanoic and alkandioic acids of nearly complete structural diversity. Within the acyclic monoamino alkanoic acid series, primary alpha-amino alpha-branched amino acids are predominant. The concentrations of individual amino acids decline exponentially with increasing carbon number within homologous series. Amino acid enantiomers are found in approximately equal amounts. Eight of the terrestrial protein amino acids have been found.     

Solar radiation variability in the last 1400 years recorded in the carbon isotope ratio of a mediterranean sea core

We present measurements and data analysis of the carbon stable isotopes (δ¹³C) in the planktonic Globigerinoides ruber extracted from the GT90/3 shallow water Ionian sea core, dated with high precision. It is commonly accepted that δ¹³C variations in symbiontic foraminifera mainly record the effects of productivity and of photosynthetic activity, varying with the ambient light level. Therefore from this time series we can deduce information on the sea surface illumination at the time of the planktonic foraminifera growth. The profile (359 points) covers the period 590–1979 AD, with a resolution of 3.87 years and it is an extension of the time series (215 points) previously published in this journal. The spectral analysis of the longer time series confirms the presence of the 11 y signal, with amplitude 0.08‰ (peak-to-trough), found in the shorter time series in phase with the sunspot solar cycle; furthermore it shows the presence of two centennial cycles of 100 and 200 years, with amplitude 0.08‰ and 0.02‰ respectively. These components are identified at high significance level by Monte Carlo singular spectrum analysis (MC-SSA). A comparison between the δ¹³C profile and the historical aurorae series (600–1500 AD) shows that the long-term δ¹³C variations are at least partially generated by the solar activity modulation and in phase with the solar output, as represented by the solar wind interaction with the magnetosphere.     

Cometary constraints on the planet forming environment.

Molecular elemental and isotopic abundances of comets provide sensitive diagnostics for models of the primitive solar nebula. New measurements of the N2, NH and NH2 abundances in comets together with the in situ Giotto mass spectrometer and dust analyzer data provide new constraints for models of the comet forming environment in the solar nebula. An inventory of nitrogen-containing species in comet Halley indicates that NH3 and CN are the dominant N carriers observed in the coma gas. The elemental nitrogen abundance in the gas component of the coma is found to be depleted by a factor approximately 75 relative to the solar photosphere. Combined with the Giotto dust analyzer results for the coma dust component, we find for comet Halley Ngas + dust approximately 1/6 the solar value. The measurement of the CN carbon isotope ratio from the bulk coma gas and dust in comet Halley indicates a significantly lower value, 12C/13C = 65 +/- 9 than the solar system value of 89 +/- 2. Because the dominant CN carrier species in comets remains unidentified, it is not yet possible to attribute the low isotope ratio predominantly to the bulk gas or dust components. The large chemical and isotopic inhomogeneities discovered in the Halley dust particles on 1 mu scales are indicative of preserved circumstellar grains which survived processing in the interstellar clouds, and may be related to the presolar silicon carbide, diamond and graphite grains recently discovered in carbonaceous chondrites. Less than 0.1% of the bulk mass in the primitive meteorites studied consists of these cosmically important grains. A larger mass fraction (approximately 5%) of chemically heterogeneous organic grains is found in the nucleus of comet Halley. The isotopic anomalies discovered in the PUMA 1 Giotto data in comet Halley are probably also attributable to preserved circumstellar grains. Thus the extent of grain processing in the interstellar environment is much less than predicted by interstellar grain models, and a significant fraction of comet nuclei (approximately 5%) may be in the form of preserved circumstellar matter. Comet nuclei probably formed in much more benign environments than primitive meteorites.     

Chemical composition measurements of the atmosphere of Jupiter with the Galileo Probe mass spectrometer.

The Galileo Probe entered the atmosphere of Jupiter on December 7, 1995. Measurements of the chemical and isotopic composition of the Jovian atmosphere were obtained by the mass spectrometer during the descent over the 0.5 to 21 bar pressure region over a time period of approximately 1 hour. The sampling was either of atmospheric gases directly introduced into the ion source of the mass spectrometer through capillary leaks or of gas, which had been chemically processed to enhance the sensitivity of the measurement to trace species or noble gases. The analysis of this data set continues to be refined based on supporting laboratory studies on an engineering unit. The mixing ratios of the major constituents of the atmosphere hydrogen and helium have been determined as well as mixing ratios or upper limits for several less abundant species including: methane, water, ammonia, ethane, ethylene, propane, hydrogen sulfide, neon, argon, krypton, and xenon. Analysis also suggests the presence of trace levels of other 3 and 4 carbon hydrocarbons, or carbon and nitrogen containing species, phosphine, hydrogen chloride, and of benzene. The data set also allows upper limits to be set for many species of interest which were not detected. Isotope ratios were measured for 3He/4He, D/H, 13C/12C, 20Ne/22Ne, 38Ar/36Ar and for isotopes of both Kr and Xe.