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Project Reports

• Biosignatures in Ancient Rocks

  The Biosignatures in Ancient Rocks group investigates the co-evolution of life and environment on early Earth using a combination of geological field work, geochemical analysis, genomics, and numerical simulation.

  ROADMAP OBJECTIVES: 1.1 3.2 4.1 4.2 4.3 5.1 5.2 5.3 6.1 6.2 7.1 7.2

• Habitability of Icy Worlds

  Habitability of Icy Worlds investigates the habitability of liquid water environments in icy worlds, with a focus on what processes may give rise to life, what processes may sustain life, and what processes may deliver that life to the surface. Habitability of Icy Worlds investigation has three major objectives. Objective 1, Seafloor Processes, explores conditions that might be conducive to originating and supporting life in icy world interiors. Objective 2, Ocean Processes, investigates the formation of prebiotic cell membranes under simulated deep-ocean conditions, and Objective 3, Ice Shell Processes, investigates astrobiological aspects of ice shell evolution.

  ROADMAP OBJECTIVES: 1.1 2.1 2.2 3.1 3.2 3.3 3.4 4.1 5.1 6.1 6.2 7.1 7.2

• Survivability of Icy Worlds

  Survivability of Icy worlds (Investigation 2) focuses on survivability. As part of our Survivability investigation, we examine the similarities and differences between the abiotic chemistry of planetary ices irradiated with ultraviolet photons (UV), electrons, and ions, and the chemistry of biomolecules exposed to similar conditions. Can the chemical products resulting from these two scenarios be distinguished? Can viable microbes persist after exposure to such conditions? These are motivating questions for our investigation.

  ROADMAP OBJECTIVES: 2.2 3.2 5.1 5.3 7.1 7.2

• Project 1B: Extracellular Polymeric Substances (EPS) and Bacterial Toxicity of Oxides

  Our interdisciplinary project examined the hypotheses that (1) bacterial cell membranes are ruptured in contact with specific mineral surfaces, (2) biofilm-forming extra-cellular polymeric substances (EPS) may have evolved to shield against membrane rupture (cell lysis), (3) differences in cell-wall structure of Gram-negative and Gram-positive bacteria may influence the susceptibility of cells to toxic minerals and (4) mineral toxicity depends on its surface chemistry and nanoparticle size.

  We have confirmed that the viability of wild-type bacterial cells which make EPS and biofilm is higher than that of mutant-type bacteria when exposed to oxide minerals. The effect is seen for both Gram negative and Gram positive bacteria, where the latter are less susceptible to the toxicity of minerals (Xu et al., 2012, Astrobiology; Zhu et al., in prep.). Thus, the thicker peptidoglycan outermost layer of the Gram positive cell surface provides an additional layer of protection compared to the less rigid, phospholipid outer membrane of the Gram negative bacteria (Zhu et al., in prep.). Most interestingly, EPS production could be induced by exposure to toxic minerals. The toxicity of the minerals depends on their surface chemistry (surface charge, ability to generate photocatalytic reactive oxygen species (ROS)) and size (Xu et al., in review).

  By understanding the mechanisms for membranolysis, especially under the extreme conditions of high radiation and heavy impacts during early planetary history, the project addresses the NASA Astrobiology Institute’s (NAI) Roadmap goals of understanding the origins of cellularity, the evolution of mechanisms for survival at environmental limits, and preservation of biosignatures, and NASA’s Strategic Goal of advancing scientific knowledge of the origin and evolution of the Earth’s biosphere and the potential for life elsewhere.

  ROADMAP OBJECTIVES: 3.4 5.1

• Ecology of Extreme Environments: Characterization of Energy Flow, Bioenergetics, and Biodiversity in Early Earth Analog Ecosystems

  The distribution of organisms and their metabolic functions on Earth is rooted, at least in part, to the numerous adaptive radiations that have resulted in the ability to occupy new ecological niches through evolutionary time. Such responses are recorded in extant organismal geographic distribution patterns (e.g., habitat range), as well as in the genetic record of organisms. The extreme variation in the geochemical composition of present day hydrothermal environments is likely to encompass many of those that were present on early Earth, when key metabolic processes are thought to have evolved. Environments such Yellowstone National Park (YNP), Wyoming harbor >12,000 geothermal features that vary widely in temperature and geochemical composition. Such environments provide a field laboratory for examining the tendency for guilds of organisms to inhabit particular ecological niches and to define the range of geochemical conditions tolerated by that functional guild (i.e., habitat range or zone of habitability). In this aim, we are examining the distribution and diversity of genes that encode for target metalloproteins in YNP environments that harbor geochemical properties that are thought to be similar to those that characterize early Earth. Using a number of newly developed computational approaches, we have been able to deduce the primary environmental parameters that constrain the distribution of a number of functional processes and which underpin their diversity. Such information is central to constraining the parameter space of environment types that are likely to have facilitated the emergence of these metal-based biocatalysts.

  ROADMAP OBJECTIVES: 3.2 3.3 3.4 4.1 4.2 5.1 5.2 5.3

• Bacterial Steroids and Triterpenoids

  Methylococcus capsulatus is one of a handful of bacteria that are capable of producing both sterols and the sterol-like hopanoid lipids. In this project, we are studying the biosynthesis and function of both sterols and hopanoids of M. capsulatus in order to gain insight into the evolutionary and functional significance of these molecules in the bacterial domain.

  ROADMAP OBJECTIVES: 5.1 5.3

• Molecular Evolution: A Top Down Approach to Examine the Origin of Key Biochemical Processes

  The emergence of metalloenzymes capable of activating substrates such as CO, N2, and H2, were significant advancements in biochemical reactivity and in the evolution of complex life. Examples of such enzymes include [FeFe]- and [NiFe]-hydrogenase that function in H2 metabolism, Mo-, V-, and Fe-nitrogenases that function in N2 reduction, and CO dehydrogenases that function in the oxidation of CO. Many of these metalloenzymes have closely related paralogs that catalyze distinctly different chemistries, an example being nitrogenase and its closely related paralog protochlorophyllide reductase that functions in the biosynthesis of bacteriochlorophyll (photosynthesis). By specifically focusing on the origin and subsequent evolution of these metallocluster biosynthesis proteins in relation to paralogous proteins that have left clear evidence in the geological record (photosynthesis and the rise of O2), we have been able to obtain significant insight into the origin and evolution of these functional processes, and to place these events in evolutionary time.

  The genomes of extant organisms provide detailed histories of key events in the evolution of complex biological processes such as CO, N2, and H2 metabolism. Advances in sequencing technology continue to increase the pace by which unique (meta)genomic data is being generated. This now makes it possible to seamlessly integrate genomic information into an evolutionary context and evaluate key events in the evolution of biological processes (e.g., gene duplications, fusions, and recruitments) within an Earth history framework. Here we describe progress in using such approaches in examining the evolution of CO, N2, and H2 metabolism.

  ROADMAP OBJECTIVES: 3.2 4.1 5.1

• Biosignatures in Relevant Microbial Ecosystems

  PSARC is investigating microbial life in some of Earth’s most mission-relevant modern ecosystems. These environments include the extremely salty Dead Sea, the impact-fractured crust of the Chesapeake Bay impact structure, methane seeps on the ocean floor, deep ice in the Greenland ice sheet, and oxygen-free waters including deep subsurface groundwater. We target environments that, when studied, provide fundamental information that can serve as the basis for future solar system exploration. Combining our expertise in molecular biology, geochemistry, microbiology, and metagenomics, and in collaboration with some of the planet’s most extreme explorers, we are deciphering the microbiology, fossilization processes, and recoverable biosignatures from these mission-relevant environments.

  ROADMAP OBJECTIVES: 4.1 4.3 5.1 5.2 5.3 6.1 7.1 7.2

• Project 1D: Establishing Biogenicity and Environmental Setting of Precambrian Kerogen and Microfossils

  This study demonstrates new abilities to use in situ measurements of carbon isotope ratios in microfossil kerogen as a biosignature and to establish taxonomic and micro-structural correlations.

  ROADMAP OBJECTIVES: 2.1 4.1 5.1 5.2 6.1 7.1

• Developing New Biosignatures

  The Developing New Biosignatures project is aimed at creating innovative approaches for the analyses of cells and other organic material, finding ways in which metal abundances and isotope systems reflect life, and developing creative approaches for using environmental DNA to study present and past life.

  ROADMAP OBJECTIVES: 4.1 5.1 7.1

• Project 5: Geological-Biological Interactions

  This project seeks to better understand the interplay between microbes and extreme environments. Towards this end our NAI supported scientists study hot spring environments, both continental and sub marine, environments of active serpentinization where pH may exceed 11, and in the high Arctic. We use molecular, isotopic, and molecular biological approaches to get at the core of the relationship between the microbial world and the natural energy provided by geological processes.

  ROADMAP OBJECTIVES: 4.1 5.1 6.1 6.2 7.1

• Geochemical Signals for Low Oxygen Worlds

  We are studying the physiology of sulfate reducing bacteria, organisms that perform a key microbial metabolism in anoxic worlds. By calibrating microbial sulfur isotope effects, we can infer the redox level of paleoenvironments in the geologic past by studying sedimentary records. The sulfur cycle is intimately linked to the redox budget of the Earth’s surface, such that this study will help inform us about the evolution of aerobic environments, a key process that set the stage for animal evolution. Similarly, we also are studying the role of oxygen in controlling the budget and transformations of nitrogen in the ocean. Nitrogen is a critical nutrient limiting marine production, and the balance of its redox cycling controls how much nitrogen is added or removed from the ocean by redox-sensitive processes.

  ROADMAP OBJECTIVES: 1.1 4.1 4.2 5.1 7.1

• Postdoctoral Fellow Report: Steven Mielke

  S. P. Mielke completed an NAI NASA Postdoctoral Program (NPP) fellowship during September 1, 2011 to February 29, 2012. His postdoctoral research has provided the basis for the project: “The Long-Wavelength Limit for Oxygenic Photosynthesis.” He continues this research as a Research Associate at Rockefeller University.

  ROADMAP OBJECTIVES: 3.2 4.2 5.1 5.3 6.2 7.2

• Geochemical Signatures of Multicellular Life

  Sterols are essential membrane components of eukaryotes but their structural diversity varies across different eukaryotic lineages. Our research aimed to determine if are any systematic variations in sterol structures between Metazoa and their immediate unicellular relatives. We found that there is a stepwise reduction in production of 24-alkyl sterol on the path toward eumetazoa suggesting an evolutionary preference for C27 sterols among animals and their kin. A major exception to this finding are the Demosponges which produce a structurally diverse array of sterols.
  We also completed a study of sediments and oils in the South Oman Salt Basin that reported an array of unusual hydrocarbon patterns that are likely to be biosignatures for early metazoa.

  ROADMAP OBJECTIVES: 4.2 5.1

• The Subglacial Biosphere – Insights Into Life-Sustaining Strategies in an Extraterrestrial Analog Environment

  Sub-ice environments are prevalent on Earth today and are likely to have been more prevalent the Earth’s past during episodes of significant glacial advances (e.g., snow-ball Earth). Numerous metabolic strategies have been hypothesized to sustain life in sub-ice environments. Common among these hypotheses is that they are all independent of photosynthesis, and instead rely on chemical energy. Recently, we demonstrated the presence of an active assemblage of methanogens in the subglacial environment of an Alpine glacier (Boyd et al., 2010). The distribution of methanogens is narrowly constrained, due in part to the energetics of the reactions which support this functional class of organism (namely carbon dioxide reduction with hydrogen and acetate fermentation). Methanogens utilize a number of metalloenzymes that have active site clusters comprised of a unique array of metals. During the course of this study, we identified other features that were suggestive of other active and potentially relevant metabolic strategies in the subglacial environment, such as nitrogen cycling. The goals of this project are 1) identifying a suite of biomarkers indicative of biological CH4 production 2). quantifying the flux of CH4 from sub-ice systems and 3). developing an understanding how life thrives at the thermodynamic limits of life. This project represents a unique extension of the ABRC and bridges the research goals of several nodes, namely the JPL-Icy Worlds team and the ASU-Follow the Elements team.

  ROADMAP OBJECTIVES: 2.1 2.2 5.1 5.2 5.3 6.1 6.2 7.1 7.2

• The Long Wavelength Limit for Oxygenic Photosynthesis

  Photosynthesis produces signs of life (biosignatures) on a planetary scale: atmospheric oxygen and the reflectance signature of photosynthetic pigments. Oxygenic photosynthesis is therefore a primary target in NASA’s search for life on habitable planets in other solar systems. An unanswered question is what the upper limit is to the photon wavelength at which oxygenic photosynthesis can remain viable. On other planets that have a parent star very different spectrally from our Sun, can we expect oxygen from plants of different colors from those on Earth?

  The cyanobacterium, Acaryochloris marina serves as a model organism for oxygenic photosynthesis adapted to low light and red-shifted light environments similar to what may be found on habitable planets orbiting M stars. Until A. marina was discovered in 1996, all known oxygenic photosynthesis relied on the pigment chlorophyll a (Chl a). A. marina instead uses chlorophyll d, which can absorb the far-red and near-infrared light in A. marina’s habitat. We use photoacoustics in the lab to measure the energy storage efficiency of A. marina with lasers, and molecular electrostatics modeling to surmise how replacement of Chl a by Chl d in A. marina affects arrangements within the photosystem molecules. We are finding that A. marina can perform oxygenic photosynthesis quite efficiently in its unique light niche.

  ROADMAP OBJECTIVES: 3.2 4.2 5.1 5.3 6.2 7.2

• Viral Ecology and Evolution

  This project is aimed at probing the occurrence and evolution of archaeal viruses in the extreme environments in the thermal areas in Yellowstone National Park. Viruses are the most abundant life-like entities on the planet and are likely a major reservoir of genetic diversity for all life on the planet and these studies are aimed at providing insights into the role of viruses in the evolution of early life on Earth.

  ROADMAP OBJECTIVES: 5.1 5.2 5.3 6.1 6.2

• Project 2F: Potential for Lithotrophic Microbial Oxidation of Fe(II) in Basalt Glass

  Ferrous iron (Fe(II)) can serve as an energy source for a wide variety of chemolithotrophic microorganisms (organisms that gain energy from metabolism of inorganic compounds). Fe(II) oxidation may have played a role in past (and possibly, present) life on Mars, whose crust is rich in primary Fe(II)-bearing silicate minerals, as well as Fe-bearing clay minerals formed during weathering of primary silicates. This project examined the potential for microbial oxidation of Fe(II) in basaltic glass. Recent research suggests that near‐surface hydrothermal venting may have occurred during past periods of active volcanic/tectonic activity on Mars. Such activities could have produced basalt glass phases that might have served as energy sources for chemolithotrophic microbial activity. Previous and ongoing NAI‐supported studies have shown that an established chemolithoautotrophic Fe(II)‐oxidizing, nitrate‐reducing culture can grow by oxidation of Fe(II) insoluble Fe(II)‐bearing phyllosilicate phases such as biotite and smectite. The initial goal of this project was to determine whether or not this culture is capable of oxidizing Fe(II) in basalt glass. In addition we tested basaltic glass oxidation by a culture of Desulfitobacterium frappieri, as previous studies demonstrated that D. frappieri is capable of nitrate-dependent oxidation of structural Fe(II) in smectite. Finally, in situ and enrichment culturing experiments were conducted to determine whether indigenous Fe(II)-oxidizing organisms in a groundwater iron seep were capable of colonization and oxidation of basaltic glass. The results of these experiments showed that while the various cultures were readily capable of smectite oxidation with nitrate, none were able to carry-out significant oxidation of Fe(II) in basalt glass. We speculate that Fe(II) atoms in the amorphous glass are somehow occluded and therefore not accessible to outer membrane cytochrome systems thought to be involved in extracellular Fe(II) oxidation.

  ROADMAP OBJECTIVES: 2.1 5.1 5.3

• Interdisciplinary Studies of Earth’s Seafloor Biosphere

  The remote deep sediment-buried ocean basaltic crust is Earth’s largest aquifer and host to the least known and potentially one of the most significant biospheres on Earth. CORK observatories have provided unparalleled access to this remote environment. They are enabling groundbreaking research in crustal fluid flow, (bio)geochemical fluid/crustal alteration, and the emerging field of deep crustal biosphere

  ROADMAP OBJECTIVES: 4.1 4.2 5.1 5.2 5.3 6.1 6.2 7.1 7.2

• Understanding Past Earth Environments

  For much of the history Earth, life on the planet existed in an environment very different than that of modern-day Earth. Thus, the ancient Earth represents a planet with a biosphere that is both dramatically different than the one in which we live, but that is also accessible to detailed study. As such, it serves as a model for what types of biospheres we may find on other planets. A particular focus of our work was on the “Early Earth” (formation through to about 500 million years ago), a timeframe poorly represented in the geological and fossil records but comprises the majority of Earth’s history. We have studied the composition and pressure of the ancient atmosphere; modeled the effects of clouds on such a planet; studied the sulfur, oxygen and nitrogen cycles; and explored atmospheric formation of molecules that were likely important to the origins of life on Earth.

  ROADMAP OBJECTIVES: 1.1 1.2 4.1 4.2 5.1 5.2 6.1

• Measuring Interdisciplinarity Within Astrobiology Research

  To integrate the work of the diverse scientists working on astrobiology, we have harvested and analyzed thousands of astrobiology documents to reveal areas of potential connection. This framework allows us to identify crossover documents that guide scientists quickly across vast interdisciplinary libraries, suggest productive interdisciplinary collaborations, and provide a metric of interdisciplinary science.

  ROADMAP OBJECTIVES: 1.1 1.2 2.1 2.2 3.1 3.2 3.3 3.4 4.1 4.2 4.3 5.1 5.2 5.3 6.1 6.2 7.1 7.2

• Stoichiometry of Life, Task 2a: Field Studies – Yellowstone National Park

  Our stoichiometry studies are determining the relationships between the elemental compositions of organisms and the elemental compositions of their environments. We experimentally determine how changes in element availability (N, P, Fe) affect the community structure in hot spring ecosystems. We also use stable isotopes (¹⁵N and ¹³C) to trace which metabolisms actively utilize N and C and where in cells these elements are used. Recently, our team has shown for the first time that nitrogen (N₂) fixation can occur at temperatures >85oC (Loiacono et al. 2012). We are also developing robust environmental sensors for hot springs that reveal chemical and thermal gradients at scales similar to the observed spatial distributions in hot spring microbial communities.

  ROADMAP OBJECTIVES: 5.1 5.2 5.3 6.1 6.2 7.2

• Stoichiometry of Life, Task 2b: Field Studies – Cuatro Cienegas

  Cuatro Cienegas is a unique biological preserve in México (state of Coahuila) in which there is striking microbial diversity, potentially related to extreme scarcity of phosphorus. We aim to understand this relationship via field sampling of biological and chemical characteristics and a series of enclosure and whole-pond fertilization experiments. These studies help in identifying the element signatures that microbes develop when key nutrient elements are scarce. Furthermore, the chemical and physical environments of the desert aquatic habitats at Cuatro Cienegas are analogous to those that may have existed on Mars during times in its past when it was losing its own surface water. Thus, these data may help in interpreting information about element signatures obtained from the Curiosity rover as it explores Gale Crater.

  ROADMAP OBJECTIVES: 5.1 5.2 5.3 6.1 6.2

• Project 4A: Field Analog Geology and Astrobiology in Support of Mars Science Laboratory and Future Mars Surface Missions

  In 2011 we have characterized the mineralogy, organic compounds and microbiology of selected sample sites from desert areas of Utah in the vicinity of MDRS in Hanksville (Foing et al. 2011). The samples were partly analyzed in situ and later distributed to the various laboratories for post-analysis. Among the important findings of this field research campaign in the Utah desert are the diversity in the mineralogical composition of soil samples even when collected in close proximity, the low abundances of detectable polycyclic aromatic hydrocarbons (PAHs) and amino acids, and the presence of biota of all three domains of life with significant heterogeneity (Ehrenfreund et al., 2011). As a follow up study EuroMoonMars campaigns in February-March 2012 collected new samples from the area around the Mars Desert Research Station (MDRS) in Utah, (Canyonlands area), a region known for its geomorphological and geochemical similarity to Mars.

  ROADMAP OBJECTIVES: 2.1 5.1

• Stoichiometry of Life, Task 3b: Ancient Records – Genomic

  The goal of Task 3b is to bring the enormous and ever-increasing repository of genomic data, both from single organisms and natural environments, to bear on understanding the history of life on Earth. Team members bring together innovative, integrative methods for understanding the interaction and feedback between life and environment, in particular how nutrient and energy limitations shape evolution. These efforts are focused not only on ancient records, but also are playing an important role in understanding how life and environment co-evolve on the modern Earth.

  ROADMAP OBJECTIVES: 5.1 5.2 5.3

• Unicellular Protists of the Neoproterozoic

  We investigated 1) how microbial processes shape some sedimentary rocks, 2) how microbial processes influence the isotopic composition of sulfur-rich minerals that are used to understand the evolution of oxygen and the cycling of carbon in the past, 3) searched for fossils of organisms that lived between 716 and 635 million years ago, surviving times when ice covered entire oceans, even at the equator and 4) used these fossils, recovered from limestone rocks, to understand the cycling of carbon during this unusual time.

  ROADMAP OBJECTIVES: 4.1 4.2 5.1 6.1 7.1

• Project 4E: Preliminary Studies of Fe Isotope Biogeochemistry in Fe-Rich Yellowstone National Park Hot Springs

  This preliminary project provided background information for future studies of the structure, function, and signatures (living and non-living) of Fe redox-based microbial life in the volcanic terrain of Yellowstone National Park (YNP). The focus on Fe redox-based systems stems from our expanding knowledge of the wide range of microbial energy metabolisms that are known to be associated with Fe redox transformations on Earth and potentially on other planets. Moreover, Fe redox transformations provide the potential for generation of mineralogical, isotopic, and organic biosignatures of past and present microbial life, which represent premier targets for testing the hypothesis that life currently exists or existed in the past on Mars. Preliminary data on Fe geochemistry and isotopic composition, and microbial community composition, was obtained for two contrasting Fe-rich springs in YNP: Chocolate Pots (CP), a warm, circumneutral environment that has formed on top of the Pleistocene-age Lava Creek Tuff, where a mixture of Fe-rich acid-sulfate geothermal fluids and neutral-pH groundwater from the Gibbon River catchment emerge to the surface; and The Gap site, a hot, acid-sulfate spring in the Norris Basin which supports active chemolithotrophic Fe(II) oxidation, analogous to other hot spring environments in YNP. The geochemical data demonstrated significant changes in aqueous Fe abundance and/or speciation along the flow paths at both sites, leading to accumulation of abundant Fe(III) oxides as well as aqueous Fe(III) at the acidic Gap site. A distinct separation in Fe isotope composition between aqueous Fe and deposited Fe(III) oxides (mainly amorphous Fe-Si coprecipitates) was also detected, with the oxide enriched in ⁵⁶Fe relative to ⁵⁴Fe as expected for redox-driven Fe isotope fractionation. However, the degree of fractionation was less the value of ca. 3 ‰ expected in closed system at isotope equilibrium. We suggest that internal regeneration of Fe(II) via dissimilatory Fe(III) reduction could enrich the aqueous Fe(II) pool in the heavy isotope, leading a much lower degree of Fe isotope fractionation – and hence a fundamentally different pattern of Fe isotope fractionation – than would occur in a strictly Fe(II) oxidation-driven reaction system. In support of this argument, an initial set of culturing experiments designed to recovery thermophilic Fe(III)-reducing organisms from CP and Gap materials resulted in the recovery of active Fe(III) reducers from both sites. In addition, preliminary pyrosequencing of ¹⁶S rRNA genes recovered from Gap solids provide evidence for Fe(III) reduction potential by the resident microflora. Particularly in the case of the Gap, sequences related to known Archaeal fermenters and elemental S/Fe(III) oxide reducers were abundant.

  ROADMAP OBJECTIVES: 2.1 5.1 5.3