Bio-markers and the search for extinct life on Mars.

Geologic and climatologic studies suggest that conditions on early Mars were similar to early Earth. Because life on Earth is believed to have originated during this early period (3.5 billion years ago), the Martian environment could have also been conducive to the origin of life. To investigate this possibility we must first define the attributes of an early Martian biota. Then, specific geographic locations on Mars must be chosen where life may have occurred (i.e. areas which had long standing water), and within these distinct locations search for key signatures or bio-markers of a possible extinct Martian biota. Some of the key signatures or bio-markers indicative of past biological activity on Earth may be applicable to Mars including: reduced carbon and nitrogen compounds, CO3(2-), SO4(2-), NO3-, NO2- [correction of NO2(2)], Mg, Mn, Fe, and certain other metals, and the isotopic ratios of C, N and S. However, we must also be able to distinguish abiotic from biologic origins for these bio-markers. For example, abiotically fixed N2 would form deposits of NO3- and NO2-, whereas biological processes would have reduced these to ammonium containing compounds, N2O, or N2, which would then be released to the atmosphere. A fully equipped Mars Rover might be able to perform analyses to measure most of these biomarkers while on the Martian surface.     

Early martian environments: the Antarctic and other terrestrial analogs.

The comparability of the early environments of Mars and Earth, and the biological evolution which occurred on early earth, motivates serious consideration of the possibility of an early martian biota. Environments which could have contained this early martian life and which may presently contain evidence of this former life include aquatic, ice, soil, and rock habitats. Several analogs of these potential early martian environments, which can provide useful information in searching for extinct life on Mars, are currently available for study on Earth. These terrestrial analogs include the perennially ice-covered lakes and sandstone rocks in the Polar Deserts of Antarctica, surface of snowfields and glaciers, desert soils, geothermal springs, and deep subsurface environments.     

Exobiology and future Mars missions: The search for Mars'' earliest biosphere

The primordial Mars may have possessed a thick carbon dioxide atmosphere, with liquid water common on the surface, similar in many ways to the primordial Earth. During this epoch, billions of years ago, the surface of Mars could have been conducive to the origin of life. It is possible that life evolved on Mars to be later eliminated as the atmospheric pressure dropped. Analysis of the surface of Mars for the traces of this early martian biota could provide many insights into the phenomenon of life and its coupling to planetary evolution.     

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.     

Life on Mars? II. Physical restrictions.

The primary physical factors important to life's evolution on a planet include its temperature, pressure and radiation regimes. Temperature and pressure regulate the presence and duration of liquid water on the surface of Mars. The prolonged presence of liquid water is essential for the evolution and sustained presence of life on a planet. It has been postulated that Mars has always been a cold dry planet; it has also been postulated that early mars possessed a dense atmosphere of CO2 (> or = 1 bar) and sufficient water to cut large channels across its surface. The degree to which either of these postulates is true correlates with the suitability of Mars for life's evolution. Although radiation can destroy living systems, the high fluxes of UV radiation on the martian surface do not necessarily stop the origin and early evolution of life. The probability for life to have arisen and evolved to a significant degree on Mars, based on the postulated ranges of early martian physical factors, is almost solely related to the probability of liquid water existing on the planet for at least hundreds of millions to billions of years.     

Planetary Protection Policy (U.S.A.).

Through existing treaty obligations of the United States, NASA is committed to exploring space while avoiding biological contamination of the planets, and to the protection of the Earth against harm from materials returned from space. Because of the similarities between Mars and Earth, plans for the exploration of Mars evoke discussions of these Planetary Protection issues. US Planetary Protection Policy will be focused on the preservation of these goals in an arena that will change with the growth of scientific knowledge about the martian environment. Early opportunities to gain the appropriate data will be used to guide later policy implementation. Because human presence on Mars will result in the end of Earth's separation from the martian environment, it is expected that precursor robotic missions will address critical planetary protection concerns before humans arrive.     

Possible complex organic compounds on Mars.

It is suggested that primitive Mars had somehow similar environments as primitive Earth. If life was born on the primitive earth using organic compounds which were produced from the early Earth environment, the same types of organic compounds were also formed on primitive Mars. Such organic compounds might have been preserved on Mars still now. We are studying possible organic formation on primitive and present Mars. A gaseous mixture of CO2, CO, N2 and H2O with various mixing ratios were irradiated with high energy protons (major components of cosmic rays). Hydrogen cyanide and formaldehyde were detected among volatile products, and yellow-brown-colored water-soluble non-volatile substances were produced, which gave amino acids after acid-hydrolysis. Major part of "amino acid precursors" were not simple molecules like aminonitriles, but complex compounds which eluted earlier than free amino acids in cation-exchange HPLC. These organic compounds should be major targets in the future Mars mission. Strategy for the detection of the complex organics on Mars will be discussed.     

Biological nitrogen fixation under primordial Martian partial pressures of dinitrogen.

Early Earth and early Mars were similar enough such that past geochemical and climatic conditions on Mars may have also been favorable for the origin of life. However, one of the most striking differences between the two planets was the low partial pressure of dinitrogen (pN2) on early Mars (18 mb). On Earth, nitrogen is a key biological element and in many ecosystems the low availability of fixed nitrogen compounds is the main factor limiting growth. Biological fixation of dinitrogen on Earth is a crucial source of fixed nitrogen. Could the low availability of dinitrogen in the primordial Martian atmosphere have prevented the existence, or evolution of Martian microbiota? Azotobacter vinelandii and Azomonas agilis were grown in nitrogen free synthetic medium under various partial pressures of dinitrogen ranging from 780-0 mb (total atmosphere=1 bar). Below 400 mb the biomass, cell number, and growth rate decreased with decreasing pN2. Both microorganisms were capable of growth at a pN2 as low as 5 mb, but no growth was observed at a pN2 < or = 1 mb. The data appear to indicate that biological nitrogen fixation could have occurred on primordial Mars (pN2=18 mb) making it possible for a biotic system to have played a role in the Martian nitrogen cycle. It is possible that nitrogen may have played a key role in the early evolution of life on Mars, and that later a lack of available nitrogen on that planet (currently, pN2=0.2 mb) may have been involved in its subsequent extinction.     

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.     

Spectroscopic observation of Mars

Visible and near-infrared reflectance spectroscopy has proven a powerful tool for exploring the geology of Mars. Most of this data has been obtained from Earth, but the technique is ideally suited to orbital application, as proposed for the U.S. Mars Geoscience/Climatology Orbiter mission. Spectral reflectance in the near-UV and visible is highly diagnostic of ferric-iron mineralogy, and has shown that Fe³⁺ in the ubiquitous bright dust and soil is amorphous or poorly-crystalline. Other iron-oxide minerals, indicative of other modes or episodes of crustal alteration, may be found in spatially localized deposits. Clay minerals (hydroxylated silicates) have diagnostic vibrational absorptions throughout the near-infrared. While some form of bound water and/or OH has been known on Mars for many years, a new result presented here is the identification of structural OH in a dilute or poorly crystalline magnesian clay. Salts such as carbonates, sulfates, and nitrates have not yet been detected in martian soils but have diagnostic spectral features in the 3- to 4-μm region, best suited to Mars-orbital observation. Analysis of reflectance spectra of low-albedo regions is a primary source of evidence for a basaltic or ultramafic crust, with identification of abundant clinopyroxene and possible detection of other mafic minerals. The distinctive near-infrared spectral shape of dark regions indicates that the dark materials commonly consist of relatively unaltered rocks or rock fragments very thinly coated by (or mixed with) bright oxidized material similar to the global dust. Visible and near-infrared reflectance spectroscopy is also a sensitive technique for detecting and analyzing water ice, as has been demonstrated on Mars by observations of the north polar cap.     

Life on Mars? I. The chemical environment.

The origin of life at its abiotic evolutionary stage, requires a combination of constituents and environmental conditions that enable the synthesis of complex replicating macromolecules from simpler monomeric molecules. It is very likely that the early stages of this evolutionary process have been spontaneous, rapid and widespread on the surface of the primitive Earth, resulting in the formation of quite sophisticated living organisms within less than a billion years. To what extent did such conditions prevail on Mars? Two companion-papers (Life on Mars? I and II) will review and discuss the available information related to the chemical, physical and environmental conditions on Mars and assess it from the perspective of potential exobiological evolution.     

Selection of sterilization methods for planetary return missions.

Two tasks must be accomplished to provide planetary protection for Mars return missions: (1) sterilization of the scientific module to be landed on Mars and (2) reliable sterilization of all material returned to Earth, while ensuring the scientific integrity of martian samples. This paper examines similarity and differences between these two tasks, and includes a discussion of technological implementation conditions and the nature of terrestrial and hypothesized martian microflora. The feasibility of a number of chemical and physical (ultraviolet and ionizing radiation and heating) methods of sterilization for use on the ground and onboard are discussed and compared. A combination of different methods will probably be selected as the most appropriate for ensuring planetary protection on the return mission.     

Simulation of the environmental climate conditions on martian surface and its effect on Deinococcus radiodurans

The resistance of terrestrial microorganisms under the thermo-physical conditions of Mars (diurnal temperature variations, UV climate, atmospheric pressure and gas composition) at mid-latitudes was studied for the understanding and assessment of potential life processes on Mars. In order to accomplish a targeted search for life on other planets, e.g. Mars, it is necessary to know the limiting physical and chemical parameters of terrestrial life. Therefore the polyextremophile bacterium Deinococcus radiodurans was chosen as test organism for these investigations. For the simulation studies at the Planetary and Space Simulation Facilities (PSI) at DLR, Cologne, Germany, conditions that are present during the southern summer at latitude of 60° on Mars were applied.We could simulate several environmental parameters of Mars in one single experiment: vacuum/low pressure, anoxic atmosphere and diurnal cycles in temperature and relative humidity, energy-rich ultraviolet (UV) radiation as well as shielding by different martian soil analogue materials. These parameters have been applied both single and in different combinations in laboratory experiments. Astonishingly the diurnal Mars-like cycles in temperature and relative humidity affected the viability of D. radiodurans cells quite severely. But the martian UV climate turned out to be the most deleterious factor, though D. radiodurans is red-pigmented due to carotenoids incorporated in its cell wall, which have been assigned not only a possible role as free radical scavenger but also as a UV-protectant. An additional UV-protection was accomplished by mixing the bacteria with nano-sized hematite.     

Science strategy for human exploration of Mars.

The scientific objectives of Mars exploration can be framed within the overarching theme of exploring Mars as another home for life, both for evidence of past or present life on Mars, and as a potential future home for human life. The two major areas of research within this theme are: 1) determining the relationship between planetary evolution, climate change, and life, and 2) determining the habitability of Mars. Within this framework, this paper discusses the exploration objectives for exobiology, climatology and atmospheric science, geology, and martian resource assessment. Human exploration will proceed in four major phases: 1) Precursor missions which will obtain environmental knowledge necessary for human exploration, 2) Emplacement phase which includes the first few human landings where crews will explore the local area of the landing site; 3) Consolidation phase missions where a permanent base will be constructed and crews will be capable of detailed exploration over regional scales; 4) Utilization phase, in which a continuously occupied permanent Mars base exists and humans will be capable of detailed global exploration of the martian surface. The phases of exploration differ primarily in the range and capabilities of human mobility. In the emplacement phase, an unpressurized rover, similar to the Apollo lunar rover, will be used and will have a range of a few tens of kilometers. In the Consolidation phase, mobility will be via a pressurized all-terrain vehicle capable of expeditions from the base site of several weeks duration. In the Utilization phase, humans will be capable of several months long expeditions to any point on the surface of Mars using a suborbital rocket equipped with habitat, lab, and return vehicle. Because of human mobility limitations, it is important to extend the range and duration of exploration in all phases by using teleoperated rover vehicles. Site selection for human missions to Mars must consider the multi-decade time frame of these four phases. We suggest that operations in the first two phases be focused in the regional area containing the Coprates Quadrangle and adjacent areas.     

Did life exist on Mars? Search for organic and inorganic signatures, one of the goals for “SAM” (sample analysis at Mars)

Observation of Mars shows signs of a past Earth-like climate, and, in that case, there is no objection to the possible development of life, in the underground or at the surface, as in the terrestrial primitive biosphere. Sample analysis at Mars (SAM) is an experiment which may be proposed for atmospheric, ground and underground in situ measurements. One of its goals is to bring direct or indirect information on the possibility for life to have developed on Mars, and to detect traces of past or present biological activity. With this aim, it focuses on the detection of organic molecules: volatile organics are extracted from the sample by simple heating, whereas refractory molecules are made analyzable (i.e. volatile), using derivatization technique or fragmentation by pyrolysis. Gaseous mixtures thus obtained are analyzed by gas chromatography associated to mass spectrometry. Beyond organics, carbonates and other salts are associated to the dense and moist atmosphere necessary to the development of life, and might have formed and accumulated in some places on Mars. They represent another target for SAM. Heating of the samples allows the analysis of structural gases of these minerals (CO₂ from carbonates, etc.), enabling to identify them. We also show, in this paper, that it may be possible to discriminate between abiotic minerals, and minerals (shells, etc.) created by living organisms.     

Detection of martian amino acids by chemical derivatization coupled to gas chromatography: in situ and laboratory analysis.

If there is, or ever was, life in our solar system beyond the Earth, Mars is the most likely place to search for. Future space missions will have then to take into account the detection of prebiotic molecules or molecules of biological significance such as amino acids. Techniques of analysis used for returned samples have to be very sensitive and avoid any chemical or biological contamination whereas in situ techniques have to be automated, fast and low energy consuming. Several possible methods could be used for in situ amino acid analyses on Mars, but gas chromatography would likely be the most suitable. Returned samples could be analyzed by any method in routine laboratory use such as gas chromatography, already successfully performed for analyses of organic matter including amino acids from martian meteorites. The derivatization step, which volatilizes amino acids to perform both in situ and laboratory analysis by gas chromatography, is discussed here.     

Atmospheric photochemistry above possible martian hot spots

Considering the possibility of outgassing from some localized sources on Mars, we have developed a one-dimensional photochemical model that includes methane (CH₄), sulfur dioxide (SO₂) and hydrogen sulfide (H₂S). Halogens were considered but were found to have no significant impact on the martian atmospheric chemistry. We find that the introduction of methane into the martian atmosphere results in the formation of mainly formaldehyde (CH₂O), methyl alcohol (CH₃OH) and ethane (C₂H₆), whereas the introduction of the sulfur species produces mainly sulfur monoxide (SO) and sulfuric acid (H₂SO₄). Depending upon the flux of the outgassed molecules from possible hot spots, some of these species and the resulting new molecules may be detectable locally, either by remote sensing (e.g., with the Planetary Fourier Spectrometer on Mars Express) or in situ measurements.     

A review of observed variability in the dayside ionosphere of Mars

Recent measurements by Mars Global Surveyor and Mars Express have greatly increased the number of observations of the martian dayside ionosphere available for study. Together with earlier measurements from the Viking era, these datasets have been used to investigate variations in well-known properties of the martian dayside ionosphere and to discover new ionospheric features. The dayside ionosphere includes the main peak, called the M2 layer, and a lower layer, called the M1 layer. In the topside, above the M2 layer, electron densities exponentially decrease with increasing altitude.     

Planetary protection program for Mars 94/96 mission.

Mars surface in-situ exploration started in 1975 with the American VIKING mission. Two probes landed on the northern hemisphere and provided, for the first time, detailed information on the martian terrain, atmosphere and meteorology. The current goal is to undertake larger surface investigations and many projects are being planned by the major Space Agencies with this objective. Among these projects, the Mars 94/96 mission will make a major contributor toward generating significant information about the martian surface on a large scale. Since the beginning of the Solar System exploration, planets where life could exist have been subject to planetary protection requirements. Those requirements accord with the COSPAR Policy and have two main goals: the protection of the planetary environment from influence or contamination by terrestrial microorganisms, the protection of life science, and particularly of life detection experiments searching extra-terrestrial life, and not life carried by probes and spacecrafts. As the conditions for life and survival for terrestrial microorganisms in the Mars environment became known, COSPAR recommendations were updated. This paper will describe the decontamination requirements which will be applied for the MARS 94/96 mission, the techniques and the procedures which are and will be used to realize and control the decontamination of probes and spacecrafts.     

Phosphorus as a potential guide in the search for extinct life on Mars.

In contrast to the search for extant organisms, the quest for fossil remains of life on Mars need not be guided by the presence of water and organic compounds on the present surface. An appropriate tracer might be the element phosphorus which is a common constituent of living systems. Utilizing terrestrial analogues, it should preferentially exist in the form of sedimentary calcium phosphate (phosphorites), which would have readily resisted changing conditions on Mars. Moreover, higher ratios of P/Th in phosphorites in comparison to calcium phosphates from magmatic rocks give us the possibility to distinguish them from inorganically formed phosphorus deposits at or close to the Martian surface. Identification of anomalous phosphorus enrichments by remote sensing or in situ analysis could be promising approaches for selecting areas preferentially composed of rocks with remains of extinct life.