Christopher S. Romanek

Search for Past Life on Mars: Possible Relic Biogenic Activity in Martian Meteorite ALH84001

Fresh fracture surfaces of the martian meteorite ALH84001 contain abundant polycyclic aromatic hydrocarbons (PAHs). These fresh fracture surfaces also display carbonate globules. Contamination studies suggest that the PAHs are indigenous to the meteorite. High-resolution scanning and transmission electron microscopy study of surface textures and internal structures of selected carbonate globules show that the globules contain fine-grained, secondary phases of single-domain magnetite and iron sulfides. The carbonate globules are similar in texture and size to some terrestrial bacterially induced carbonate precipitates. Although inorganic formation is possible, formation of the globules by biogenic processes could explain many of the observed features, including the PAHs. The PAHs, the carbonate globules, and their associated secondary mineral phases and textures could thus be fossil remains of a past martian biota.

Elongated prismatic magnetite crystals in ALH84001 carbonate globules: Potential Martian magnetofossils

Using transmission electron microscopy (TEM), we have analyzed magnetite (Fe3O4) crystals acid-extracted from carbonate globules in Martian meteorite ALH84001. We studied 594 magnetites from ALH84001 and grouped them into three populations on the basis of morphology: 389 were irregularly shaped, 164 were elongated prisms, and 41 were whisker-like. As a possible terrestrial analog for the ALH84001 elongated prisms, we compared these magnetites with those produced by the terrestrial magnetotactic bacteria strain MV-1. By TEM again, we examined 206 magnetites recovered from strain MV-1 cells. Natural (Darwinian) selection in terrestrial magnetotactic bacteria appears to have resulted in the formation of intracellular magnetite crystals having the physical and chemical properties that optimize their magnetic moment. In this study, we describe six properties of magnetite produced by biologically controlled mechanisms (e.g., magnetotactic bacteria), properties that, collectively, are not observed in any known population of inorganic magnetites. These criteria can be used to distinguish one of the modes of origin for magnetites from samples with complex or unknown histories. Of the ALH84001 magnetites that we have examined, the elongated prismatic magnetite particles (similar to 27% of the total) are indistinguishable from the MV-1 magnetites in five of these six characteristics observed for biogenically controlled mineralization of magnetite crystals.

Global Occurrence of Archaeal amoA Genes in Terrestrial Hot Springs

ABSTRACT Despite the ubiquity of ammonium in geothermal environments and the thermodynamic favorability of aerobic ammonia oxidation, thermophilic ammonia-oxidizing microorganisms belonging to the crenarchaeota kingdom have only recently been described. In this study, we analyzed microbial mats and surface sediments from 21 hot spring samples (pH 3.4 to 9.0; temperature, 41 to 86°C) from the United States, China, and Russia and obtained 846 putative archaeal ammonia monooxygenase large-subunit (amoA) gene and transcript sequences, representing a total of 41 amoA operational taxonomic units (OTUs) at 2% identity. The amoA gene sequences were highly diverse, yet they clustered within two major clades of archaeal amoA sequences known from water columns, sediments, and soils: clusters A and B. Eighty-four percent (711/846) of the sequences belonged to cluster A, which is typically found in water columns and sediments, whereas 16% (135/846) belonged to cluster B, which is typically found in soils and sediments. Although a few amoA OTUs were present in several geothermal regions, most were specific to a single region. In addition, cluster A amoA genes formed geographic groups, while cluster B sequences did not group geographically. With the exception of only one hot spring, principal-component analysis and UPGMA (unweighted-pair group method using average linkages) based on the UniFrac metric derived from cluster A grouped the springs by location, regardless of temperature or bulk water pH, suggesting that geography may play a role in structuring communities of putative ammonia-oxidizing archaea (AOA). The amoA genes were distinct from those of low-temperature environments; in particular, pair-wise comparisons between hot spring amoA genes and those from sympatric soils showed less than 85% sequence identity, underscoring the distinctness of hot spring archaeal communities from those of the surrounding soil system. Reverse transcription-PCR showed that amoA genes were transcribed in situ in one spring and the transcripts were closely related to the amoA genes amplified from the same spring. Our study demonstrates the global occurrence of putative archaeal amoA genes in a wide variety of terrestrial hot springs and suggests that geography may play an important role in selecting different assemblages of AOA.

Nonmarine Crenarchaeol in Nevada Hot Springs

ABSTRACT Glycerol dialkyl glycerol tetraethers (GDGTs) are core membrane lipids of the Crenarchaeota. The structurally unusual GDGT crenarchaeol has been proposed as a taxonomically specific biomarker for the marine planktonic group I archaea. It is found ubiquitously in the marine water column and in sediments. In this work, samples of microbial community biomass were obtained from several alkaline and neutral-pH hot springs in Nevada, United States. Lipid extracts of these samples were analyzed by high-performance liquid chromatography-mass spectrometry and by gas chromatography-mass spectrometry. Each sample contained GDGTs, and among these compounds was crenarchaeol. The distribution of archaeal lipids in Nevada hot springs did not appear to correlate with temperature, as has been observed in the marine environment. Instead, a significant correlation with the concentration of bicarbonate was observed. Archaeal DNA was analyzed by denaturing gradient gel electrophoresis. All samples contained 16S rRNA gene sequences which were more strongly related to thermophilic crenarchaeota than to Cenarchaeum symbiosum, a marine nonthermophilic crenarchaeon. The occurrence of crenarchaeol in environments containing sequences affiliated with thermophilic crenarchaeota suggests a wide phenotypic distribution of this compound. The results also indicate that crenarchaeol can no longer be considered an exclusive biomarker for marine species.

Truncated hexa-octahedral magnetite crystals in ALH84001: Presumptive biosignatures

McKay et al. [(1996) Science 273, 924–930] suggested that carbonate globules in the meteorite ALH84001 contained the fossil remains of Martian microbes. We have characterized a subpopulation of magnetite (Fe3O4) crystals present in abundance within the Fe-rich rims of these carbonate globules. We find these Martian magnetites to be both chemically and physically identical to terrestrial, biogenically precipitated, intracellular magnetites produced by magnetotactic bacteria strain MV-1. Specifically, both magnetite populations are single-domain and chemically pure, and exhibit a unique crystal habit we describe as truncated hexa-octahedral. There are no known reports of inorganic processes to explain the observation of truncated hexa-octahedral magnetites in a terrestrial sample. In bacteria strain MV-1 their presence is therefore likely a product of Natural Selection. Unless there is an unknown and unexplained inorganic process on Mars that is conspicuously absent on the Earth and forms truncated hexa-octahedral magnetites, we suggest that these magnetite crystals in the Martian meteorite ALH84001 were likely produced by a biogenic process. As such, these crystals are interpreted as Martian magnetofossils and constitute evidence of the oldest life yet found.

Chemical, mineralogical and isotope behavior, and phase transformation during the precipitation of calcium carbonate minerals from intermediate ionic solution at 25°C

A set of time-series experiments was performed in which CaCO3 was precipitated (25°C, 1 atm) from aliquots of an NaHCO3 (0.330 M)/CaCl2 (0.0023 M) solution in a closed system to evaluate the effect of phase transformation on calcium carbonate precipitation kinetics and isotope behavior. Monohydrocalcite precipitated initially at a solution saturation state (Ωmhc) slightly greater than one, whereas calcite crystallized later at a significant higher saturation state (Ωcl ≅ 14.4). The precipitation of calcite promoted the dissolution of monohydrocalcite at a rate that exceeded calcite precipitation, producing anomalous behavior in the chemical and isotope composition of the system. The carbon isotope fractionation factors (103 lnα) for monohydrocalcite-HCO3(aq)- and monohydrocalcite-CO2(g) were 0.36 ± 0.01‰ and 8.35 ± 0.01‰, respectively. The oxygen isotope fractionation factor for monohydrocalcite-H2O was 27.8 ± 0.1‰. The carbon isotope fractionation factors for calcite-HCO3 (aq)− and calcite-CO2(g) were 0.94 ± 0.06‰ and 8.93 ± 0.06‰, respectively, whereas the oxygen isotope fractionation factor for calcite-H2O(l) was 28.0 ± 0.2‰. A carbon isotope fractionation factor of 0.58 ± 0.07‰ was determined for the mineral pair calcite-monohydrocalcite, but no fractionation was observed for oxygen isotopes over time steps when both minerals co-precipitated. Fractionation factors for calcite were independent of precipitation rate over the range in rates of 103.96 to 105.63 μmol/m2h. These results extend the upper limit of characterization for the relationship between precipitation rate and isotope partitioning of carbon between calcite, HCO3 (aq)− and CO2(g), and quantitatively document for the first time the independence between precipitation rate and oxygen isotope partitioning in the calcite-H2O(l) system.

Magnetofossils from Ancient Mars: a Robust Biosignature in the Martian Meteorite ALH84001

Evidence of biogenic activity on Mars has profound scientific implications for our understanding of the origin of life on Earth and the presence and diversity of life within the Cosmos. Analysis of the Martian meteorite Allan Hills 84001 (ALH84001) revealed several lines of evidence that has led some investigators to suggest that microbial life existed on Mars approximately 4 billion years ago (45). One of the strongest lines of evidence is the presence of tens-of-nanometer-size magnetite (Fe3O4) crystals found within carbonate globules and their associated rims in the meteorite (57, 58). Approximately one-quarter of these magnetites have remarkable morphological and chemical similarities to magnetite particles produced by magnetotactic bacteria, which occur in aquatic habitats on Earth. Moreover, these types of magnetite particles are not known or expected to be produced by abiotic means either through geological processes or synthetically in the laboratory. We have therefore argued that these Martian magnetite crystals are in fact magnetofossils (57, 58). If this is true, such magnetofossils would constitute evidence of the oldest life forms known. In this respect, we note there is now considerable uncertainty concerning when the earliest terrestrial life forms existed. Until recently, results from the ~3.5-billion-year-old Apex cherts of the Warrawoona group in western Australia held this record (52), although this work is now in question (12).

Factors Controlling the Distribution of Archaeal Tetraethers in Terrestrial Hot Springs

ABSTRACT Glycerol dialkyl glycerol tetraethers (GDGTs) found in hot springs reflect the abundance and community structure of Archaea in these extreme environments. The relationships between GDGTs, archaeal communities, and physical or geochemical variables are underexamined to date and when reported often result in conflicting interpretations. Here, we examined profiles of GDGTs from pure cultures of Crenarchaeota and from terrestrial geothermal springs representing a wide distribution of locations, including Yellowstone National Park (United States), the Great Basin of Nevada and California (United States), Kamchatka (Russia), Tengchong thermal field (China), and Thailand. These samples had temperatures of 36.5 to 87°C and pH values of 3.0 to 9.2. GDGT abundances also were determined for three soil samples adjacent to some of the hot springs. Principal component analysis identified four factors that accounted for most of the variance among nine individual GDGTs, temperature, and pH. Significant correlations were observed between pH and the GDGTs crenarchaeol and GDGT-4 (four cyclopentane rings, m/z 1,294); pH correlated positively with crenarchaeol and inversely with GDGT-4. Weaker correlations were observed between temperature and the four factors. Three of the four GDGTs used in the marine TEX86 paleotemperature index (GDGT-1 to -3, but not crenarchaeol isomer) were associated with a single factor. No correlation was observed for GDGT-0 (acyclic caldarchaeol): it is effectively its own variable. The biosynthetic mechanisms and exact archaeal community structures leading to these relationships remain unknown. However, the data in general show promise for the continued development of GDGT lipid-based physiochemical proxies for archaeal evolution and for paleo-ecology or paleoclimate studies.

Influence of feeding ecology on blood mercury concentrations in four species of turtles

Mercury is a relatively well-studied pollutant because of its global distribution, toxicity, and ability to bioaccumulate and biomagnify in food webs: however, little is known about bioaccumulation and toxicity of Hg in turtles. Total Hg (THg) concentrations in blood were determined for 552 turtles representing four different species (Chelydra serpentina, Sternotherus odoratus, Chrysemys picta, and Pseudemys rubriventris) from a Hg-contaminated site on the South River (VA, USA) and upstream reference sites. Methylmercury and Se concentrations also were determined in a subset of samples. Because the feeding ecology of these species differs drastically, stable isotopes of carbon (delta13C) and nitrogen (delta15N) were employed to infer the relationship between relative trophic position and Hg concentrations. Significant differences were found among sites and species, suggesting that blood can be used as a bioindicator of Hg exposure in turtles. We found differences in THg concentrations in turtles from the contaminated site that were consistent with their known feeding ecology: C. serpentina > or = S. odoratus > C. picta > P. rubriventris. This trend was generally supported by the isotope data, which suggested that individual turtles were feeding at more than one trophic level. Methylmercury followed similar spatial patterns as THg and was the predominant Hg species in blood for all turtles. Blood Se concentrations were low in the system, but a marginally positive relationship was found between THg and Se when species were pooled. The blood THg concentrations for the turtles in the present study are some of the highest reported in reptiles, necessitating further studies to investigate potential adverse effects of these high concentrations.

Stable isotope signature of philopatry and dispersal in a migratory songbird

Stable isotope analysis is widely promoted as a practical method for tracing the geographic origins of migratory birds. However, the extent to which geospatial patterns of isotope ratios in avian tissues are influenced by age-specific, altitudinal, and temporal factors remains largely unexplored. We measured carbon (13C/12C) and nitrogen (15N/14N) isotope ratios in feathers of black-throated blue warblers (Dendroica caerulescens) breeding along a relatively steep altitudinal gradient in the Appalachian Mountains to evaluate the effects of altitude and year on the isotopic signatures of yearling (first breeding season) and older males (>2 years). Breeding males (n = 302) collected during 7 consecutive years exhibited significant age-specific and altitudinal effects in δ13C values and age-specific and temporal effects in δ15N values. The δ13C values of older males increased with altitude at the rate of ≈1.3‰ per 1,000 m, suggesting a high degree of year-to-year philopatry to narrow altitudinal zones, if not to breeding territories. In contrast, absence of altitudinal patterns in yearlings most likely reflects natal dispersal. Carbon isotope variation (δ13C = −26.07 to −20.86‰) observed along a single altitudinal transect (755 m) nearly brackets the range of δ13C values recorded in feathers across the North American breeding range of the warbler from Georgia to New Brunswick (11° of latitude) and from New Brunswick to Michigan (22° of longitude). These data indicate that age-specific and altitudinal effects must be considered when using δ13C values to delineate the geographic origin of avian species with large altitudinal and latitudinal ranges.