Pavilion Lake Research Articles

The eukaryome of modern microbialites reveals distinct colonization across aquatic ecosystems

Protists are less studied for their role and diversity in ecosystems. Notably, protists have played and still play an important role in microbialites. Microbialites, or lithified microbial mats, represent the oldest evidence of fossil biofilms (~3.5 Gyr). Modern microbialites may offer a unique proxy to study the potential role of protists within a geological context. We examined protist diversity in freshwater (Kelly and Pavilion Lake in British Columbia, Canada) and marine (Highborne Cay, Bahamas) to hypersaline (Shark Bay, Australia) microbialites to decipher their geomicrobiological role. The freshwater microbialite communities were clearly distinct from their marine and hypersaline counterparts. Chlorophytes had higher numerical abundance in freshwater microbialites; whereas pennate diatoms dominated numerically in marine microbialites. Despite the differences, protists across ecosystems may have adopted similar roles and functions. We suggest a consistent biogeochemical role of protists across microbialites globally; but that salinity may shape protist composition and evolution in these ecosystems.

Biosignatures Associated with Freshwater Microbialites.

Freshwater microbialites (i.e., lithifying microbial mats) are quite rare in northern latitudes of the North American continent, with two lakes (Pavilion and Kelly Lakes) of southeastern BC containing a morphological variety of such structures. We investigated Kelly Lake microbialites using carbon isotope systematics, phospholipid fatty acids (PLFAs) and quantitative PCR to obtain biosignatures associated with microbial metabolism. δ13CDIC values (mean δ13CDIC −4.9 ± 1.1‰, n = 8) were not in isotopic equilibrium with the atmosphere; however, they do indicate 13C-depleted inorganic carbon into Kelly Lake. The values of carbonates on microbialite surfaces (δ13C) fell within the range predicted for equilibrium precipitation from ambient lake water δ13CDIC (−2.2 to −5.3‰). Deep microbialites (26 m) had an enriched δ13Ccarb value of −0.3 ± 0.5‰, which is a signature of photoautotrophy. The deeper microbialites (>20 m) had higher biomass estimates (via PLFAs), and a greater relative abundance of cyanobacteria (measured by 16S copies via qPCR). The majority of PLFAs constituted monounsaturated and saturated PLFAs, which is consistent with gram-negative bacteria, including cyanobacteria. The central PLFA δ13C values were highly depleted (−9.3 to −15.7‰) relative to δ13C values of bulk organic matter, suggesting a predominance of photoautotrophy. A heterotrophic signature was also detected via the depleted iso- and anteiso-15:0 lipids (−3.2 to −5.2‰). Based on our carbonate isotopic biosignatures, PLFA, and qPCR measurements, photoautotrophy is enriched in the microbialites of Kelly Lake. This photoautotrophy enrichment is consistent with the microbialites of neighboring Pavilion Lake. This indication of photoautotrophy within Kelly Lake at its deepest depths raises new insights into the limits of measurable carbonate isotopic biosignatures under light and nutrient limitations.

Early-earth nonprotein amino acid metabolites in modern cyanobacterial microbialites

Cyanobacteria are among the earth’s oldest known living groups of organisms and can form layered accretions called microbialites, found in both the fossil record and existing lakes. Studies of cyanobacterial biochemical processes help to understand the evolution of life on earth. The conserved metabolism of cyanobacterial species includes the biosynthesis of unusual nonprotein amino acids such as N-(2-aminoethyl)glycine, hypothesized to have constituted an early form of genetic information in cells. Pavilion Lake in British Columbia, Canada, hosts a population of unique, actively growing microbialites covered in biofilms dominated by cyanobacteria. We hypothesized that the living microbial communities produce dinitrogenous nonprotein amino acids, such as N-(2-aminoethyl)glycine and its structural isomers β-N-methylamino-l-alanine, 2,4-diaminobutyric acid and β-aminomethyl-l-alanine. We analyzed samples in sediment traps collected between 2007 and 2014 in depths ranging from 11 to 46 m. N-(2-aminoethyl)glycine, 2,4-diaminobutyric acid and β-aminomethyl-l-alanine were found in highest concentration in the shallowest microbialite biofilms with a maximum of 22 ng/g, 33 ng/g and 0.4 ng/g, respectively. In contrast, the concentration of β-N-methylamino-l-alanine was highest in collections between 18 and 26 m depths and only β-N-methylamino-l-alanine was found in the deepest water collections. These data provide evidence indicating that the production of these nonprotein amino acids is highly conserved through the evolution of cyanobacteria and suggest that the nitrogen-rich metabolites may have had both an important role in ancient and modern cyanobacterial metabolism. Further research will determine the role of N-(2-aminoethyl)glycine and its isomers in early life metabolism and their current function in photosynthetic cells.

The Complete Genome and Physiological Analysis of the Eurythermal Firmicute Exiguobacterium chiriqhucha Strain RW2 Isolated From a Freshwater Microbialite, Widely Adaptable to Broad Thermal, pH, and Salinity Ranges.

Members of the genus Exiguobacterium are found in diverse environments from marine, freshwaters, permafrost to hot springs. Exiguobacterium can grow in a wide range of temperature, pH, salinity, and heavy-metal concentrations. We characterized Exiguobacterium chiriqhucha strain RW2 isolated from a permanently cold freshwater microbialite in Pavilion Lake, British Columbia using metabolic assays, genomics, comparative genomics, phylogenetics, and fatty acid composition. Strain RW2 has the most extensive growth range for temperature (4–50°C) and pH (5–11) of known Exiguobacterium isolates. Strain RW2 genome predicts pathways for wide differential thermal, cold and osmotic stress using cold and heat shock cascades (e.g., csp and dnaK), choline and betaine uptake/biosynthesis (e.g., opu and proU), antiporters (e.g., arcD and nhaC Na+/K+), membrane fatty acid unsaturation and saturation. Here, we provide the first complete genome from Exiguobacterium chiriqhucha strain RW2, which was isolated from a freshwater microbialite. Its genome consists of a single 3,019,018 bp circular chromosome encoding over 3,000 predicted proteins, with a GC% content of 52.1%, and no plasmids. In addition to growing at a wide range of temperatures and salinities, our findings indicate that RW2 is resistant to sulfisoxazole and has the genomic potential for detoxification of heavy metals (via mercuric reductases, arsenic resistance pumps, chromate transporters, and cadmium-cobalt-zinc resistance genes), which may contribute to the metabolic potential of Pavilion Lake microbialites. Strain RW2 could also contribute to microbialite formation, as it is a robust biofilm former and encodes genes involved in the deamination of amino acids to ammonia (i.e., L-asparaginase/urease), which could potentially boost carbonate precipitation by lowering the local pH and increasing alkalinity. We also used comparative genomic analysis to predict the pathway for orange pigmentation that is conserved across the entire Exiguobacterium genus, specifically, a C30 carotenoid biosynthesis pathway is predicted to yield diaponeurosporene-4-oic acid as its final product. Carotenoids have been found to protect against ultraviolet radiation by quenching reactive oxygen, releasing excessive light energy, radical scavenging, and sunscreening. Together these results provide further insight into the potential of Exiguobacterium to exploit a wide range of environmental conditions, its potential roles in ecosystems (e.g., microbialites/microbial mats), and a blueprint model for diverse metabolic processes.

The Complete Genome and Physiological Analysis of the Microbialite-Dwelling Agrococcus pavilionensis sp. nov; Reveals Genetic Promiscuity and Predicted Adaptations to Environmental Stress.

Members of the bacterial genus Agrococcus are globally distributed and found across environments so highly diverse that they include forests, deserts, and coal mines, as well as in potatoes and cheese. Despite how widely Agrococcus occurs, the extent of its physiology, genomes, and potential roles in the environment are poorly understood. Here we use whole-genome analysis, chemotaxonomic markers, morphology, and 16S rRNA gene phylogeny to describe a new isolate of the genus Agrococcus from freshwater microbialites in Pavilion Lake, British Columbia, Canada. We characterize this isolate as a new species Agrococcus pavilionensis strain RW1 and provide the first complete genome from a member of the genus Agrococcus. The A. pavilionensis genome consists of one chromosome (2,627,177 bp) as well as two plasmids (HC-CG1 1,427 bp, and LC-RRW783 31,795 bp). The genome reveals considerable genetic promiscuity via mobile elements, including a prophage and plasmids involved in integration, transposition, and heavy-metal stress. A. pavilionensis strain RW1 differs from other members of the Agrococcus genus by having a novel phospholipid fatty acid iso-C15:1Δ4, β-galactosidase activity and amygdalin utilization. Carotenoid biosynthesis is predicted by genomic metabolic reconstruction, which explains the characteristic yellow pigmentation of A. pavilionensis. Metabolic reconstructions of strain RW1 genome predicts a pathway for releasing ammonia via ammonification amino acids, which could increase the saturation index leading to carbonate precipitation. Our genomic analyses suggest signatures of environmental adaption to the relatively cold and oligotrophic conditions of Pavilion Lake microbialites. A. pavilionensis strain RW1 in modern microbialites has an ecological significance in Pavilion Lake microbialites, which include potential roles in heavy-metal cycling and carbonate precipitation (e.g., ammonification of amino acids and filamentation which many trap carbonate minerals).

Microbial diversity and biomarker analysis of modern freshwater microbialites from Laguna Bacalar, Mexico.

Laguna Bacalar is a sulfate-rich freshwater lake on the Yucatan Peninsula that hosts large microbialites. High sulfate concentrations distinguish Laguna Bacalar from other freshwater microbialite sites such as Pavilion Lake and Alchichica, Mexico, as well as from other aqueous features on the Yucatan Peninsula. While cyanobacterial populations have been described here previously, this study offers a more complete characterization of the microbial populations and corresponding biogeochemical cycling using a three-pronged geobiological approach of microscopy, high-throughput DNA sequencing, and lipid biomarker analyses. We identify and compare diverse microbial communities of Alphaproteobacteria, Deltaproteobacteria, and Gammaproteobacteria that vary with location along a bank-to-bank transect across the lake, within microbialites, and within a neighboring mangrove root agglomeration. In particular, sulfate-reducing bacteria are extremely common and diverse, constituting 7%-19% of phylogenetic diversity within the microbialites, and are hypothesized to significantly influence carbonate precipitation. In contrast, Cyanobacteria account for less than 1% of phylogenetic diversity. The distribution of lipid biomarkers reflects these changes in microbial ecology, providing meaningful biosignatures for the microbes in this system. Polysaturated short-chain fatty acids characteristic of cyanobacteria account for <3% of total abundance in Laguna Bacalar microbialites. By contrast, even short-chain and monounsaturated short-chain fatty acids attributable to both Cyanobacteria and many other organisms including types of Alphaproteobacteria and Gammaproteobacteria constitute 43%-69% and 17%-25%, respectively, of total abundance in microbialites. While cyanobacteria are the largest and most visible microbes within these microbialites and dominate the mangrove root agglomeration, it is clear that their smaller, metabolically diverse associates are responsible for significant biogeochemical cycling in this microbialite system.

Metagenomic Analysis Suggests Modern Freshwater Microbialites Harbor a Distinct Core Microbial Community.

Modern microbialites are complex microbial communities that interface with abiotic factors to form carbonate-rich organosedimentary structures whose ancestors provide the earliest evidence of life. Past studies primarily on marine microbialites have inventoried diverse taxa and metabolic pathways, but it is unclear which of these are members of the microbialite community and which are introduced from adjacent environments. Here we control for these factors by sampling the surrounding water and nearby sediment, in addition to the microbialites and use a metagenomics approach to interrogate the microbial community. Our findings suggest that the Pavilion Lake microbialite community profile, metabolic potential and pathway distributions are distinct from those in the neighboring sediments and water. Based on RefSeq classification, members of the Proteobacteria (e.g., alpha and delta classes) were the dominant taxa in the microbialites, and possessed novel functional guilds associated with the metabolism of heavy metals, antibiotic resistance, primary alcohol biosynthesis and urea metabolism; the latter may help drive biomineralization. Urea metabolism within Pavilion Lake microbialites is a feature not previously associated in other microbialites. The microbialite communities were also significantly enriched for cyanobacteria and acidobacteria, which likely play an important role in biomineralization. Additional findings suggest that Pavilion Lake microbialites are under viral selection as genes associated with viral infection (e.g CRISPR-Cas, phage shock and phage excision) are abundant within the microbialite metagenomes. The morphology of Pavilion Lake microbialites changes dramatically with depth; yet, metagenomic data did not vary significantly by morphology or depth, indicating that microbialite morphology is altered by other factors, perhaps transcriptional differences or abiotic conditions. This work provides a comprehensive metagenomic perspective of the interactions and differences between microbialites and their surrounding environment, and reveals the distinct nature of these complex communities.

Carbonate fabrics in the modern microbialites of Pavilion Lake: two suites of microfabrics that reflect variation in microbial community morphology, growth habit, and lithification.

Modern microbialites in Pavilion Lake, BC, provide an analog for ancient non-stromatolitic microbialites that formed from insitu mineralization. Because Pavilion microbialites are mineralizing under the influence of microbial communities, they provide insights into how biological processes influence microbialite microfabrics and mesostructures. Hemispherical nodules and micrite-microbial crusts are two mesostructures within Pavilion microbialites that are directly associated with photosynthetic communities. Both filamentous cyanobacteria in hemispherical nodules and branching filamentous green algae in micrite-microbial crusts were associated with calcite precipitation at microbialite surfaces and with characteristic microfabrics in the lithified microbialite. Hemispherical nodules formed at microbialite surfaces when calcite precipitated around filamentous cyanobacteria with a radial growth habit. The radial filament pattern was preserved within the microbialite to varying degrees. Some subsurface nodules contained well-defined filaments, whereas others contained only dispersed organic inclusions. Variation in filament preservation is interpreted to reflect differences in timing and amount of carbonate precipitation relative to heterotrophic decay, with more defined filaments reflecting greater lithification prior to degradation than more diffuse filaments. Micrite-microbial crusts produce the second suite of microfabrics and form in association with filamentous green algae oriented perpendicular to the microbialite surface. Some crusts include calcified filaments, whereas others contained voids that reflect the filamentous community in shape, size, and distribution. Pavilion microbialites demonstrate that microfabric variation can reflect differences in lithification processes and microbial metabolisms as well as microbial community morphology and organization. Even when the morphology of individual filaments or cells is not well preserved, the microbial growth habit can be captured in mesoscale microbialite structures. These results suggest that when petrographic preservation is extremely good, ancient microbialite growth structures and microfabrics can be interpreted in the context of variation in community organization, community composition, and lithification history. Even in the absence of distinct microbial microfabrics, mesostructures can capture microbial community morphology.

Reconstruction of limnology and microbialite formation conditions from carbonate clumped isotope thermometry.

Quantitative tools for deciphering the environment of microbialite formation are relatively limited. For example, the oxygen isotope carbonate-water geothermometer requires assumptions about the isotopic composition of the water of formation. We explored the utility of using 'clumped' isotope thermometry as a tool to study the temperatures of microbialite formation. We studied microbialites recovered from water depths of 10-55 m in Pavilion Lake, and 10-25 m in Kelly Lake, spanning the thermocline in both lakes. We determined the temperature of carbonate growth and the (18)O/(16)O ratio of the waters that microbialites grew in. Results were then compared to current limnological data from the lakes to reconstruct the history of microbialite formation. Modern microbialites collected at shallow depths (11.7 m) in both lakes yield clumped isotope-based temperatures of formation that are within error of summer water temperatures, suggesting that clumped isotope analyses may be used to reconstruct past climates and to probe the environments in which microbialites formed. The deepest microbialites (21.7-55 m) were recovered from below the present-day thermoclines in both lakes and yield radioisotope ages indicating they primarily formed earlier in the Holocene. During this time, pollen data and our reconstructed water (18)O/(16)O ratios indicate a period of aridity, with lower lake levels. At present, there is a close association between both photosynthetic and heterotrophic communities, and carbonate precipitation/microbialite formation, with biosignatures of photosynthetic influences on carbonate detected in microbialites from the photic zone and above the thermocline (i.e., depths of generally <20 m). Given the deeper microbialites are receiving <1% of photosynthetically active radiation (PAR), it is likely these microbialites primarily formed when lower lake levels resulted in microbialites being located higher in the photic zone, in warm surface waters.

Phylogenetic diversity of a microbialite reef in a cold alkaline freshwater lake.

A culture-independent multidomain survey of biodiversity in microbialite structures within the cold alkaline Pavilion Lake (British Columbia, Canada) revealed a largely homogenous community at depths from 10 to 30 m. Real-time quantitative PCR was used to demonstrate that bacteria comprised approximately 80%-95% of recoverable phylotypes. Archaeal phylotypes accounted for <5% of the community in microbialites exposed to the water column, while structures in sediment contact supported 4- to 5-fold higher archaeal abundance. Eukaryal phylotypes were rare and indicated common aquatic diatoms that were concluded not to be part of the microbialite community. Phylogenetic analysis of rRNA genes from clone libraries (N = 491) revealed that alphaproteobacterial phylotypes were most abundant. Cyanobacterial phylotypes were highly diverse but resolved into 4 dominant genera: Acaryochloris, Leptolyngbya, Microcoleus, and Pseudanabaena. Interestingly, microbialite cyanobacteria generally affiliated phylogenetically with aquatic and coral cyanobacterial groups rather than those from stromatolites. Other commonly encountered bacterial phylotypes were from members of the Acidobacteria, with relatively low abundance of the Betaproteobacteria, Chloroflexi, Nitrospirae, and Planctomycetes. Archaeal diversity (N = 53) was largely accounted for by Euryarchaeota, with most phylotypes affiliated with freshwater methanogenic taxa.

Prokaryote populations of extant microbialites along a depth gradient in Pavilion Lake, British Columbia, Canada

Pavilion Lake in British Columbia, Canada, is home to modern-day microbialites that are actively growing at multiple depths within the lake. While microbialite morphology changes with depth and previous isotopic investigations suggested a biological role in the formation of these carbonate structures, little is known about their microbial communities. Microbialite samples acquired through the Pavilion Lake Research Project (PLRP) were first investigated for phototrophic populations using Cyanobacteria-specific primers and 16S rRNA gene cloning. These data were expounded on by high-throughput tagged sequencing analyses of the general bacteria population. These molecular analyses show that the microbial communities of Pavilion Lake microbialites are diverse compared to non-lithifying microbial mats also found in the lake. Phototrophs and heterotrophs were detected, including species from the recently described Chloroacidobacteria genus, a photoheterotroph that has not been previously observed in microbialite systems. Phototrophs were shown as the most influential contributors to community differences above and below 25 meters, and corresponding shifts in heterotrophic populations were observed at this interface as well. The isotopic composition of carbonate also mirrored this shift in community states. Comparisons to previous studies indicated this population shift may be a consequence of changes in lake chemistry at this depth. Microbial community composition did not correlate with changing microbialite morphology with depth, suggesting something other than community changes may be a key to observed variations in microbialite structure.

Autotrophic and heterotrophic associated biosignatures in modern freshwater microbialites over seasonal and spatial gradients

Phospholipid fatty acid (PLFA) profiles and isotopic biosignatures associated with autotrophic and heterotrophic microbial processes in freshwater microbialites exposed to seasonal and spatial gradients in Pavilion Lake, British Columbia were investigated. The PLFA biosignature profiles of the microbialite associated microbial communities were dominated by saturated and monoenoic PLFAs and showed no resolvable response to variation in light or temperature down to a water depth of 33m and across seasons. Microbialite mean δ13Corg values (−26.0±3.8‰) and Δδ13CDIC-org discrimination of ca. 25‰ supported non-CO2 limited photosynthesis. More abundant and 13C-depleted PLFAs (Δδ13C 7–14‰ vs. bulk organic matter) were indicative of autotrophic microbes. Less abundant and 13C-depleted PLFAs (Δδ13C 3–4‰) were indicative of heterotrophic organisms, particularly branched (iso/anteiso15:0 and 10me16:0) PLFAs. Dark coloured microbialites from the bottom of the lake (below 46m water depth) had comparatively low biomass and a higher proportion of branched PLFAs, including biomarkers for sulfate reducing bacteria. Bulk δ13Ccarb values of microbialite carbonate at 6 and 11m water depth were up to ca. 2‰ more 13C enriched than the value predicted for precipitation from ambient dissolved inorganic carbon and had increased biomass in summer, indicating a preserved biosignature of photosynthetic activity. Other δ13Ccarb values were generally within the range predicted for equilibrium precipitation. Estimated precipitation temperature values from δ18Ocarb were consistent with measured late summer water values. While both autotrophic and heterotrophic processes occurred at all depths, preservation of an enriched 13C biosignature was only detected at shallow depths where photosynthetic activity and biomass production were relatively high.

Microstructure variability in freshwater microbialites, Pavilion Lake, Canada

Calcite microbialites in Pavilion Lake, British Columbia, exhibit a diverse range in macro-morphology, biomass abundance, porosity, and mineral content. To evaluate the role of microorganisms in their formation, samples collected from a range of depths were examined by scanning electron microscopy (SEM) and synchrotron radiation-based micro-X-ray fluorescence (μ-XRF) spectroscopy to characterize both their outer surfaces as well as internal structures. Observed trends in both surface colonization as well as microbialite framework with increasing lake depth include decreasing microbial abundance on outer surfaces as well as increasing ratios of carbonate:biomass in the microbialites. Microscopic investigations of the interiors show bacteria and algae entrapped within calcite, with this calcite exhibiting micropores and casts similar in size and shape to microorganisms. Based on these observations, it is hypothesized that microbialite development in Pavilion Lake initiates calcite precipitation in phototrophic microbial mats, i.e., combined phototrophy and heterotrophy, followed by heterotrophic oxidation of organic matter leading to eventual carbonate infilling of the microbial–mineral matrix. In addition, an observed shift from cyanobacteria to algae with increasing lake depth suggests variability in contemporary conditions controlling microbialite growth and diagenesis. High photosynthetic growth rates at shallower depths result in significant porosity and friability due to biomass accumulation outpacing carbonate precipitation. At intermediate depths, lower light levels and slower growth rates of phototrophs lead to a greater proportion of the microbialite matrix being in-filled by carbonate. Carbonates precipitate initially within the bacteria-EPS matrix, with abundant uncalcified algae maintaining microbialite porosity. In the deepest waters, the presence of only sparse algal colonization as well as fine-grained, laminated metal-rich sediments covering microbialites suggests that present-day insolation levels are too low to support the development of photosynthetic microbial mats. As a consequence, heterotrophic carbonate precipitation has progressively in-filled these microbialite interiors to create lithified calcite fabrics that exhibit minimal porosity but preserve the casts of microorganisms as biosignatures. While the origin of microbialites in Pavilion Lake remains unknown, current observations provide valuable information in evaluating how environmental conditions influence microbialite growth in a freshwater, lacustrine environment.

First Draft Genome Sequence from a Member of the Genus Agrococcus , Isolated from Modern Microbialites

We report the first draft genome sequence from a member of the genus Agrococcus, isolated from cold thrombolytic microbialites within Pavilion Lake, British Columbia, Canada. The draft genome assembly for Agrococcus pavilionensis strain RW-1 has a size of 2,878,403 bp with a G+C content of 72.56%.

Magnetotactic bacteria from Pavilion Lake, British Columbia.

Pavilion Lake is a slightly alkaline, freshwater lake located in British Columbia, Canada (50°51'N, 121°44'W). It is known for unusual organosedimentary structures, called microbialites that are found along the lake basin. These deposits are complex associations of fossilized microbial communities and detrital- or chemical-sedimentary rocks. During the summer, a sediment sample was collected from near the lake's shore, approximately 25–50 cm below the water surface. Magnetotactic bacteria (MTB) were isolated from this sample using a simple magnetic enrichment protocol. The MTB isolated from Pavilion Lake belonged to the Alphaproteobacteria class as determined by nucleotide sequences of 16S rRNA genes. Transmission electron microscopy (TEM) revealed that the bacteria were spirillum-shaped and contained a single chain of cuboctahedral-shaped magnetite (Fe3O4) crystals that were approximately 40 nm in diameter. This discovery of MTB in Pavilion Lake offers an opportunity to better understand the diversity of MTB habitats, the geobiological function of MTB in unique freshwater ecosystems, and search for magnetofossils contained within the lake's microbialites.

Pavilion lake microbialites: morphological, molecular and biochemical evidence for a cold-water transition to colonial aggregates.

The presence of microbialite structures in a freshwater, dimictic mid-latitude lake and their establishment after the last ice age about 10,000 years ago is puzzling. Freshwater calcite microbialites at Pavilion Lake, British Columbia, Canada, consist of a complex community of microorganisms that collectively form large, ordered structured aggregates. This distinctive assemblage of freshwater calcite microbialites was studied through standard microbial methods, morphological observations, phospholipid fatty acid (PLFA) analysis, DNA sequencing and the identification of quorum sensing molecules. Our results suggest that the microbialites may represent a transitional form from the exclusively prokaryotic colonial precursors of stromatolites to the multicellular organismic aggregates that give rise to coral reefs.

Reusable science tools for analog exploration missions: xGDS Web Tools, VERVE, and Gigapan Voyage

The Exploration Ground Data Systems (xGDS) project led by the Intelligent Robotics Group (IRG) at NASA Ames Research Center creates software tools to support multiple NASA-led planetary analog field experiments. The two primary tools that fall under the xGDS umbrella are the xGDS Web Tools (xGDS-WT) and Visual Environment for Remote Virtual Exploration (VERVE). IRG has also developed a hardware and software system that is closely integrated with our xGDS tools and is used in multiple field experiments called Gigapan Voyage. xGDS-WT, VERVE, and Gigapan Voyage are examples of IRG projects that improve the ratio of science return versus development effort by creating generic and reusable tools that leverage existing technologies in both hardware and software.xGDS Web Tools provides software for gathering and organizing mission data for science and engineering operations, including tools for planning traverses, monitoring autonomous or piloted vehicles, visualization, documentation, analysis, and search. VERVE provides high performance three dimensional (3D) user interfaces used by scientists, robot operators, and mission planners to visualize robot data in real time. Gigapan Voyage is a gigapixel image capturing and processing tool that improves situational awareness and scientific exploration in human and robotic analog missions. All of these technologies emphasize software reuse and leverage open source and/or commercial-off-the-shelf tools to greatly improve the utility and reduce the development and operational cost of future similar technologies.Over the past several years these technologies have been used in many NASA-led robotic field campaigns including the Desert Research and Technology Studies (DRATS), the Pavilion Lake Research Project (PLRP), the K10 Robotic Follow-Up tests, and most recently we have become involved in the NASA Extreme Environment Mission Operations (NEEMO) field experiments. A major objective of these joint robot and crew experiments is to improve NASAs understanding of how to most effectively execute and increase science return from exploration missions. This paper focuses on an integrated suite of xGDS software and compatible hardware tools: xGDS Web Tools, VERVE, and Gigapan Voyage, how they are used, and the design decisions that were made to allow them to be easily developed, integrated, tested, and reused by multiple NASA field experiments and robotic platforms.

Photosynthetic isotope biosignatures in laminated micro-stromatolitic and non-laminated nodules associated with modern, freshwater microbialites in Pavilion Lake, B.C.

The influence of microbial activity on carbonate precipitation was investigated within micro-stromatolitic nodules associated with modern, freshwater microbialites located in Pavilion Lake, B.C. Observed carbonate δ 13C values enriched by up to + 3.6‰ as compared to predicted abiotic carbonate δ 13C values from measured dissolved inorganic carbon (mean − 1.2‰, n = 13) were consistent with microbial photosynthetic influence on in situ precipitation within the nodule microenvironment. Estimated carbonate precipitation temperatures within the nodules based on δ 18O were consistent with recorded summertime temperatures, indicative of precipitation during the period of highest levels of photosynthetic activity. Low δ 13C values of organic matter within the nodules (− 30.6 to − 21.1‰) and an average inorganic to organic carbon Δ δ 13C value of 26.8‰ reflected the preferential uptake of 12C during non-CO 2 limited photosynthesis, supporting the generation of 13C-enriched DIC. Microelectrode profiles through the nodules showed oxygen supersaturation of up to ∼ 275%, elevated pH compared to ambient water and a lack of any observable dissolved sulphide, Mn or Fe further indicated that photosynthetic activity was the predominant metabolic process within the nodule during light exposure. Microbial phospholipid fatty acid profiles of the nodule communities were indicative of bacteria rather than eukaryotes and PLFA δ 13C values were depleted relative to the bulk cell by 2.6–6.6‰, consistent with a predominance of photosynthetic microbes. Scanning electron microscopy images of the relationship between carbonate minerals and filaments indicated that carbonate precipitation had occurred in situ due to microbial influences on the geochemistry within the nodule microenvironment rather than due to cell surface effects or trapping and binding. The observation of photosynthetically induced 13C-enrichment of in situ precipitated carbonate within the nodule microenvironment is thus a biosignature of the activity of these surface communities and is consistent with the hypothesized role of biology in the formation of microbialites.

Constraining carbon sources and growth rates of freshwater microbialites in Pavilion Lake using 14C analysis

This study determined the natural abundance isotopic compositions ((13)C, (14)C) of the primary carbon pools and microbial communities associated with modern freshwater microbialites located in Pavilion Lake, British Columbia, Canada. The Delta(14)C of dissolved inorganic carbon (DIC) was constant throughout the water column and consistent with a primarily atmospheric source. Observed depletions in DIC (14)C values compared with atmospheric CO(2) indicated effects due either to DIC residence time and/or inputs of (14)C-depleted groundwater. Mass balance comparisons of local and regional groundwater indicate that groundwater DIC could contribute a maximum of 9-13% of the DIC. (14)C analysis of microbial phospholipid fatty acids from microbialite communities had Delta(14)C values comparable with lake water DIC, demonstrating that lake water DIC was their primary carbon source. Microbialite carbonate was also primarily derived from DIC. However, some depletion in microbialite carbonate (14)C relative to lake water DIC occurred, due either to residence time or mixing with a (14)C-depleted carbon source. A detrital branch covered with microbialite growth was used to estimate a microbialite growth rate of 0.05 mm year(-1) for the past 1000 years, faster than previous estimates for this system. These results demonstrate that the microbialites are actively growing and that the primary carbon source for both microbial communities and recent carbonate is DIC originating from the atmosphere. While these data cannot conclusively differentiate between abiotic and biotic formation mechanisms, the evidence for minor inputs of groundwater-derived DIC is consistent with the previously hypothesized biological origin of the Pavilion Lake microbialites.

Limnology of Pavilion Lake, B. C., Canada Characterization of a microbialite forming environment

The objectives of this study are two-fold: (1) to describe and quantify the seasonal physical and chemical limnological properties of Pavilion Lake, a microbialite-rich lake in British Columbia, Canada, and (2) to gain a broader limnological context of Pavilion Lake by examining the limnology and hydrology of the lakes and groundwater systems in the area (∼30 km radius). Pavilion Lake is a dimictic lake with annual ice-cover. It is a hard water (mean CaCO 3 = 181.8 mg L -1), ultra-oligotrophic (mean Total Phosphorus [TP] = 3.3 μg L -1) lake, that is groundwater fed most likely through diffuse, low velocity local and regional sources. Principal Components Analysis (PCA) separated Pavilion Lake from the other groundwater, stream and lake samples along a conductivity and pH gradient on Axis 1 (λ 1 =0.392), and a nutrient (Total Nitrogen [TN], TP) and K +, Mg 2+, Si gradient on the second axis (λ 2 = 0.160). Pavilion Lake has the lowest Ca 2+ and TP concentrations, and the highest Na + concentrations and optical transmissivity amongst all sampled sites. Furthermore, the lake is characterized by low sedimentation rates. These characteristics are potentially important factors in supporting the past and on-going development of microbialites within the lake. Our study provides a limnochemical reference to consider in the conceptualization of ideal environments supporting large-scale microbialites. © E. Schweizerbart'sche Verlagsbuchhandlung 2009.