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At least 73 records · Page 4
All sourcesNASA NTRSDOE OSTI
DOE OSTI2021

Elioraea tepida , sp. nov., a Moderately Thermophilic Aerobic Anoxygenic Phototrophic Bacterium Isolated from the Mat Community of an Alkaline Siliceous Hot Spring in Yellowstone National Park, WY, USA

Strain MS-P2 T was isolated from microbial mats associated with Mushroom Spring, an alkaline siliceous hot spring in Yellowstone National Park, WY, USA. The isolate grows chemoheterotrophically by oxygen-dependent respiration, and light stimulates photoheterotrophic growth under strictly oxic conditions. Strain MS-P2 T synthesizes bacteriochlorophyll a and the carotenoid spirilloxanthin. However, photoautotrophic growth did not occur under oxic or anoxic conditions, suggesting that this strain should be classified as an aerobic anoxygenic phototrophic bacterium. Strain MS-P2 T cells are motile, curved rods about 0.5 to 1.0 μm wide and 1.0 to 1.5 μm long. The optimum growth temperature is 45–50 °C, and the optimum pH for growth is circum-neutral (pH 7.0–7.5). Sequence analysis of the 16S rRNA gene revealed that strain MS-P2 T is closely related to Elioraea species, members of the class Alphaproteobacteria, with a sequence identity of 96.58 to 98%. The genome of strain MS-P2 T is a single circular DNA molecule of 3,367,643 bp with a mol% guanine-plus-cytosine content of 70.6%. Based on phylogenetic, physiological, biochemical, and genomic characteristics, we propose this bacteriochlorophyll a-containing isolate is a new species belonging to the genus Elioraea, with the suggested name Elioraeatepida. The type-strain is strain MS-P2 T (= JCM33060 T = ATCC TSD-174 T ).

59 BASIC BIOLOGICAL SCIENCES↗
DOE OSTI2023

Hydrogen Energy Storage System at Borrego Springs Towards an H2 Enabled 100 Renewable Microgrid

San Diego Gas & Electric's Borrego Springs Microgrid is one of the largest microgrids in the USA, serving 2500 residential customers, 300 commercial and industrial customers, and has a peak demand of approximately 14 MW. This microgrid is located at the end of a long transmission line, and is subjected to extreme weather events like storms, wildfires, and flooding, which frequently cause grid outages. The microgrid currently relies on 3.65 MW of diesel-powered generators to provide grid-forming services during these outages, which results in greenhouse gas and criteria pollutant emissions. However, the microgrid has access to approximately 37 MW of installed solar generation and struggles with overgeneration and curtailment. 1.5 MW/4.5 MWh of grid-scale batteries have been installed to capture some the overgeneration and provide resiliency, but a longer-duration low-greenhouse gas emission energy storage solution is needed. In our project, the team will evaluate in the lab and demonstrate in the field a grid-forming fuel cell inverter that can provide grid-forming services while utilizing hydrogen's energy storage scalability. We will first utilize NREL's Renewable Energy Integration and Optimization (REopt) platform to perform analyses on future microgrid scenarios that use hydrogen assets to reduce or eliminate the need for diesel backup generators. We will then evaluate the grid-forming inverter through power hardware-in-the-loop and controller hardware-in-the-loop experiments at ARIES, and de-risk the field deployment and operation of hydrogen assets in the microgrid setting. Finally, this project will demonstrate the operation of the fuel cell inverter and updated microgrid controller by operating in grid-forming mode in the Borrego Springs microgrid.

Borrego Springs↗
DOE OSTI2024

Hydrogen Energy Storage System at Borrego Springs Towards an H2 Enabled 100% Renewable Microgrid

San Diego Gas & Electric's Borrego Springs Microgrid is one of the largest microgrids in the USA, serving 2500 residential customers, 300 commercial and industrial customers, and has a peak demand of approximately 14 MW. This microgrid is located at the end of a long transmission line, and is subjected to extreme weather events like storms, wildfires, and flooding, which frequently cause grid outages. The microgrid currently relies on 3.65 MW of diesel-powered generators to provide grid-forming services during these outages, which results in greenhouse gas and criteria pollutant emissions. However, the microgrid has access to approximately 37 MW of installed solar generation and struggles with overgeneration and curtailment. 1.5 MW/4.5 MWh of grid-scale batteries have been installed to capture some the overgeneration and provide resiliency, but a longer-duration low-greenhouse gas emission energy storage solution is needed. In our project, the team will evaluate in the lab and demonstrate in the field a grid-forming fuel cell inverter that can provide grid-forming services while utilizing hydrogen's energy storage scalability. We will first utilize NREL's Renewable Energy Integration and Optimization (REopt) platform to perform analyses on future microgrid scenarios that use hydrogen assets to reduce or eliminate the need for diesel backup generators. We will then evaluate the grid-forming inverter through power hardware-in-the-loop and controller hardware-in-the-loop experiments at ARIES, and de-risk the field deployment and operation of hydrogen assets in the microgrid setting. Finally, this project will demonstrate the operation of the fuel cell inverter and updated microgrid controller by operating in grid-forming mode in the Borrego Springs microgrid.

blackstart with fuel cells↗
DOE OSTI2022

Improving Abundance Estimates of Spring–Summer Snake River Chinook Salmon for Fisheries Management

Abstract The Columbia River basin is home to a run of spring–summer Chinook Salmon Oncorhynchus tshawytscha that returns to the Snake River drainage of Idaho, Oregon, and Washington in the Pacific Northwest. Historically, the run was one of the more productive throughout the Columbia River basin. However, Snake River spring–summer Chinook Salmon have experienced declines in abundance due to overfishing, habitat degradation, and dams. Several stocks are listed as threatened under the U.S. Endangered Species Act and are supported by mitigation hatcheries funded by Idaho Power Company, the Lower Snake River Compensation Plan, and the Bonneville Power Administration. To maximize tribal and state harvest of returning hatchery adults, minimize impacts on wild fish, and ensure that enough hatchery fish return to meet broodstock needs, careful fisheries management is required. Since 2008, managers have used hatchery adults, PIT-tagged as juveniles and detected at Lower Granite Dam, to generate adult abundance estimates. In season, these estimates inform state and tribal harvest shares and ensure that broodstock needs are met. Postseason, they provide smolt-to-adult survival and return rates. Since 2012, parentage-based tagging (PBT) has provided an alternative method to estimate stock- and age-specific returns at Lower Granite Dam, since returning hatchery adults sampled at Lower Granite Dam can be assigned to their parents. We compared stock-specific abundance estimates between PIT- and PBT-derived methodologies for return years 2016–2019. Across all years, PIT tag estimates accounted for 65% of the PBT-based estimates at Lower Granite Dam across all age-groups and release sites combined. This underrepresentation across all groups equated to 49,833 fish that were not accounted for in PIT tag abundance estimates. It is clear that PBT-based estimates should aide in-season harvest management and postseason run reconstruction to avoid the known bias of estimates from PIT tags, especially during years of low returns when increased accuracy is critical.

Coykendall, D. Katharine (ORCID:0000000211482397)↗
DOE OSTI2023

Remote effects of Tibetan Plateau spring land temperature on global subseasonal to seasonal precipitation prediction and comparison with effects of sea surface temperature: the GEWEX/LS4P Phase I experiment

The prediction skill for precipitation anomalies in late spring and summer months—a significant component of extreme climate events—has remained stubbornly low for years. This paper presents a new idea that utilizes information on boreal spring land surface temperature/subsurface temperature (LST/SUBT) anomalies over the Tibetan Plateau (TP) to improve prediction of subsequent summer droughts/floods over several regions over the world, East Asia and North America in particular. The work was performed in the framework of the GEWEX/LS4P Phase I (LS4P-I) experiment, which focused on whether the TP LST/SUBT provides an additional source for subseasonal-to-seasonal (S2S) predictability. The summer 2003, when there were severe drought/flood over the southern/northern part of the Yangtze River basin, respectively, has been selected as the focus case. With the newly developed LST/SUBT initialization method, the observed surface temperature anomaly over the TP has been partially produced by the LS4P-I model ensemble mean, and 8 hotspot regions in the world were identified where June precipitation is significantly associated with anomalies of May TP land temperature. Consideration of the TP LST/SUBT effect has produced about 25–50% of observed precipitation anomalies in most hotspot regions. The multiple models have shown more consistency in the hotspot regions along the Tibetan Plateau-Rocky Mountain Circumglobal (TRC) wave train. The mechanisms for the LST/SUBT effect on the 2003 drought over the southern part of the Yangtze River Basin are discussed. For comparison, the global SST effect has also been tested and 6 regions with significant SST effects were identified in the 2003 case, explaining about 25–50% of precipitation anomalies over most of these regions. This study suggests that the TP LST/SUBT effect is a first-order source of S2S precipitation predictability, and hence it is comparable to that of the SST effect. With the completion of the LS4P-I, the LS4P-II has been launched and the LS4P-II protocol is briefly presented.

54 ENVIRONMENTAL SCIENCES↗
DOE OSTI2023

Dual carbonate clumped isotopes (Δ 47 -Δ 48 ) constrains kinetic effects and timescales in peridotite-associated springs at the Cedars, Northern California

The Cedars is an area in Northern California with a chain of highly alkaline springs resulting from CO 2 -charged meteorological water interacting with a peridotite body. Serpentinization resulting from this interaction at depth leads to the sequestration of various carbonate minerals into veins accompanied by a release of Ca 2+ and OH – enriched water to the surface, creating an environment which promotes rapid precipitation of CaCO 3 at surface springs. This environment enables us to apply the recently developed Δ 47 -Δ 48 dual clumped isotope analysis to probe kinetic isotope effects (KIEs) and timescales of CO 2 transformation in a region with the potential for geological CO 2 sequestration. Here, we analyzed CaCO 3 recovered from various localities and identified significant kinetic fractionations associated with CO 2 absorption in a majority of samples, characterized by enrichment in Δ 47 values and depletion in Δ 48 values relative to equilibrium. Surface floes exhibited the largest KIEs (ΔΔ 47 : 0.163‰, ΔΔ 48 : -0.761‰). Surface floe samples begin to precipitate out of solution within the first hour of CO 2 absorption, and the dissolved inorganic carbon (DIC) pool requires a residence time of >100 h to achieve isotopic equilibria. The Δ 48 /Δ 47 slope of samples from the Cedars (-3.223 ± 0.519) is within the range of published theoretical values designed to constrain CO 2 hydrolysis-related kinetic fractionation (-1.724 to -8.330). The Δ 47 /δ 18 O slope (-0.009 ± 0.001) and Δ 47 /δ 13 C slope (-0.009 ± 0.001) are roughly consistent with literature values reported from a peridotite in Oman of -0.006 ± 0.002 and -0.005 ± 0.002, respectively. The consistency of slopes in the multi-isotope space suggests the Δ 47 -Δ 48 dual carbonate clumped isotope framework can be applied to study CO 2 -absorption processes in applied systems, including sites of interest for geological sequestration.

58 GEOSCIENCES↗
DOE OSTI2023

Achieving superior mechanical properties: Tailoring multicomponent microstructure in AISI 9254 spring steel through a two-stage Q&P process and nanoscale carbide integration

In the pursuit of lightweight, durable steel, we have successfully developed a multicomponent structure in AISI 9254 spring steel using a two-stage quenching and partitioning (Q&P) process. The primary objective of this process was to engineer an optimized microstructure consisting of nanobainite, martensite, and nano-carbides. Utilizing the insights gained from the results of advanced techniques such as X-ray diffraction (XRD), scanning electron microscopy (SEM), transmission electron microscopy (TEM), and atom probe tomography (APT) performed on the as-received AISI 9254 spring steel, we refined the quenching and partitioning (Q&P) path, leading to the successful establishment of a bainitic transformation for superior mechanical properties. Our tensile tests revealed a high yield strength (≈ 1600 ± 25 MPa) and ultimate tensile strength (≈ 1850 ± 50 MPa), along with considerable elongation (≈ 11.15 ± 0.25%). We also identified that pre-formed martensite lath defects and high silicon content play crucial roles during the Q&P process, preventing carbide coalescence and increasing strain-hardening capacity. Finally, this study demonstrates the potential of a Q&P process to generate high-strength, ductile steel for automotive and aerospace applications.

36 MATERIALS SCIENCE↗
DOE OSTI2026

Tree-ring evidence marks year 2022 as the driest spring season in nearly four centuries in the Western Himalayas

The Hindukush-Karakoram-Himalayan region is a crucial freshwater source for billions across South Asia, yet its climate remains poorly understood due to limited long-term records. Winter and spring precipitation govern snow accumulation and downstream water availability in dry months particularly across the Western Himalayas (WH), but recent decades show intensifying droughts with unclear long-term context. Here, we have reconstructed a nearly four-century-long spring i.e. February to May (FMAM) precipitation for the Lahaul region of the (WH), an area dominated by the Western Disturbances. This record was developed using moisture-sensitive Cedrus deodara (Deodar) tree-rings from three high-elevation sites. A regional composite tree-ring-width chronology, developed through a Nested Principal Component Analysis and modeled with a nonlinear Generalized Additive Model (GAM) that explains 71 % of the variance during the calibration period. We identified the last two decades as the most precipitation deficit phase and the year 2022 showing the driest FMAM on record. The observed rise in the FMAM dry episodes post 1999 CE in our reconstruction, corresponds to the meteorological records. This recent drying is linked to a northward shift of the subtropical westerly jet and reduced moisture transport, both associated with unusual sea surface temperature patterns in the tropical Indian Ocean and the Western Pacific Ocean. Our results provide compelling evidence of long-term hydroclimatic instability in the WH and emphasize the value of tree-ring records in extending precipitation histories beyond the instrumental observations. Such reconstructions can be benchmarks to validate high-resolution climate models and formulate adaptation policies to mitigate future risks.

Cedrus deodara↗
DOE OSTI2025

Sources and Composition of Organic Aerosols in the Central Arctic during Spring and Summer

Organic aerosols (OA) are a major component of Arctic aerosol mass and influence the region’s radiation budget, yet their sources and physicochemical properties remain largely unknown. We investigate OA sources and climate-relevant characteristics over the central Arctic Ocean in spring and summer, by applying positive matrix factorization to aerosol mass spectrometry data from two ship-based expeditions (2018 and 2020), complemented by total and interstitial aerosol measurements during fog periods. Six distinct OA factors were identified: haze-related OA, Arctic oxygenated OA, two mixed-OA types resembling biomass burning and primary marine OA linked to warm-air intrusions, marine OA, and hydrocarbon-related OA. Seasonal transitions strongly shaped OA composition. Following polar sunrise, highly oxygenated OA, likely formed secondarily from photo-oxidized volatile organic compounds, became dominant. After the polar vortex collapsed in May, episodic spikes in marine OA from the marginal ice zone appeared, alongside a reduced influence from Eurasian anthropogenic sources. These transitions influenced OA oxidation state and related properties including volatility, acidity, and hygroscopicity, highlighting the role of the Arctic spring atmosphere as an active photochemical reactor. Overall, OA was highly oxidized, with particles activated in fog during summer showing even greater oxidation, suggesting that central Arctic OA can be highly cloud active.

Arctic haze↗
DOE OSTI2023

Spring Regional Sea Surface Temperatures as a Precursor of European Summer Heatwaves

Different spring and early summer North Atlantic sea surface temperature anomalies (SSTAs) have been shown to precede recent European summer heatwaves (EuSHWs). So far, the limited number of observed events associated with several physical mechanisms has prevented a robust identification of SSTAs as precursors. Here, we extend beyond previous studies by combining 100 historical simulations (1850–2005) of the MPI Grand-Ensemble with an explainable neural-network method. We find that the spring tripolar North Atlantic pattern with positive SSTAs in the Subtropical Gyre is a precursor of EuSHWs. In addition, positive SSTAs west of the Iberian Peninsula, and in the North Sea, the Baltic Sea and the Mediterranean Sea relate to distinct early summer soil moisture anomaly patterns and are precursors of western and southeastern EuSHWs, respectively. While the phase of the tripolar North Atlantic pattern indicates whether a EuSHW might emerge, regional SSTAs indicate the spatial characteristics of EuSHWs.

54 ENVIRONMENTAL SCIENCES↗
DOE OSTI2026

Spring Dust in Colorado Plateau: Transport Pathways and Interannual Variability Derived From Two Decades of MERRA‐2 Reanalysis

Spring dust storms in the U.S. Southwest significantly impact environmental and human systems, yet their climatological patterns and driving mechanisms remain poorly understood. Using two decades of MERRA-2 reanalysis data and self-organizing map clustering, we identified four distinct dust transport pathways from surrounding and remote deserts for the top 10% of spring dust events impacting the Colorado Plateau: (a) intense southwesterly near-surface transport where eastward upward transport is blocked by topography, (b) substantial remote transport plus weaker surface emissions, (c) moderate southwesterly transport with stronger upper-level transport, and (d) rare northwesterly pattern linked to the Great Basin sources, which accounts for 38%, 14%, 37%, and 11% of total events, respectively. The frequency and intensity of these events strongly correlate with large-scale climate variability, with increased dust events during La Niña. Our results provide a robust framework for improving seasonal dust forecasts and understanding future dust dynamics under climate change.

aerosol transport↗
DOE OSTI2020

Polysaccharide niche partitioning of distinct Polaribacter clades during North Sea spring algal blooms

Massive releases of organic substrates during marine algal blooms trigger growth of many clades of heterotrophic bacteria. Algal polysaccharides represent the most diverse and structurally complex class of these substrates, yet their role in shaping the microbial community composition is poorly understood. We investigated, whether polysaccharide utilization capabilities contribute to niche differentiation of Polaribacter spp. (class Flavobacteriia; known to include relevant polysaccharidedegraders) that were abundant during 2009–2012 spring algal blooms in the southern North Sea. We identified six distinct Polaribacter clades using phylogenetic and phylogenomic analyses, quantified their abundances via fluorescence in situ hybridization, compared metagenome-assembled genomes, and assessed in situ gene expression using metaproteomics. Four clades with distinct polysaccharide niches were dominating. Polaribacter 2-a comprised typical first responders featuring small genomes with limited polysaccharide utilization capacities. Polaribacter 3-a were abundant only in 2010 and possessed a distinct sulfated α-glucoronomannan degradation potential. Polaribacter 3-b responded late in blooms and had the capacity to utilize sulfated xylan. Polaribacter 1-a featured high numbers of glycan degradation genes and were particularly abundant following Chattonella algae blooms. These results support the hypothesis that sympatric Polaribacter clades occupy distinct glycan niches during North Sea spring algal blooms.

59 BASIC BIOLOGICAL SCIENCES↗
DOE OSTI2025

Microbiome Comparison and Pathogen Identification for Three Migrating Passerines Captured During Spring Season in Jordan Using 16S rRNA Sequencing

Jordan is located on an important spot along the Mediterranean and Black Sea Flyway. Hundreds of migratory bird species have been identified stopping over in Jordan during spring and autumn migratory seasons. Compared to mammals and economically important birds, the microbiomes of wild bird species are severely understudied. Gut microbial composition is a valuable source of information that reflects food preferences, foraging behavior, and the risk of pathogen transmission to humans and other animals. In this study, we assessed the microbiome composition of three species of migrating passerines (willow warblers, lesser whitethroats, and common reed warblers) captured during the spring migration stopover in Jordan in 2023. A total of 59 fecal samples were selected evenly from the three species and subjected to 16S sequencing and microbiome analysis. Our objectives were to determine the diversity of bacteria in these three species, assess the amount of intra- and inter-specific variation, and detect pathogenic genera and species that could pose health risks to humans, domestic animals, and wildlife. Bacteria mainly belonged to the phyla Proteobacteria (62%), Actinobacteriota (18%), Firmicutes (13%), Cyanobacteria (5%), and Bacteroidota (1%). The results reveal that lesser whitethroats had the greatest variation in bacterial genus richness, Shannon diversity, and microbial composition compared to willow warblers and common reed warblers. The three bird species harbored several pathogenic genera and species, including Campylobacter, Enterococcus, Escherichia-Shigella, Mycoplasma, Rickettsia, Clostridium perfringens, and Vibrio cholerae. We suggest further investigation to understand the relationship between migratory behavior and their gut microbiome. We advocate for the use of advanced molecular techniques to characterize the pathogens found in migratory birds that might have public and environmental health impacts in addition to economic loss.

59 BASIC BIOLOGICAL SCIENCES↗
DOE OSTI2023

Relationships between fluid mixing, biodiversity, and chemosynthetic primary productivity in Yellowstone hot springs

The factors that influence biodiversity and productivity of hydrothermal ecosystems are not well understood. Here we investigate the relationship between fluid mixing, biodiversity, and chemosynthetic primary productivity in three co-localized hot springs (RSW, RSN, and RSE) in Yellowstone National Park that have different geochemistry. All three springs are sourced by reduced hydrothermal fluid, but RSE and RSN receive input of vapour phase gas and oxidized groundwaters, with input of both being substantially higher in RSN. Metagenomic sequencing revealed that communities in RSN were more biodiverse than those of RSE and RSW in all dimensions evaluated. Microcosm activity assays indicate that rates of dissolved inorganic carbon (DIC) uptake were also higher in RSN than in RSE and RSW. Together, these results suggest that increased mixing of reduced volcanic fluid with oxidized fluids generates additional niche space capable of supporting increasingly biodiverse communities that are more productive. These results provide insight into the factors that generate and maintain chemosynthetic biodiversity in hydrothermal systems and that influence the distribution, abundance, and diversity of microbial life in communities supported by chemosynthesis. These factors may also extend to other ecosystems not supported by photosynthesis, including the vast subterranean biosphere and biospheres beneath ice sheets and glaciers.

59 BASIC BIOLOGICAL SCIENCES↗
DOE OSTI2023

Distribution and abundance of tetraether lipid cyclization genes in terrestrial hot springs reflect pH

Many Archaea produce membrane-spanning lipids that enable life in extreme environments. These isoprenoid glycerol dibiphytanyl glycerol tetraethers (GDGTs) may contain up to eight cyclopentyl and one cyclohexyl ring, where higher degrees of cyclization are associated with more acidic, hotter or energy-limited conditions. Recently, the genes encoding GDGT ring synthases, grsAB, were identified in two Sulfolobaceae; however, the distribution and abundance of grs homologs across environments inhabited by these and related organisms remain a mystery. To address this, we examined the distribution of grs homologs in relation to environmental temperature and pH, from thermal springs across Earth, where sequences derive from metagenomes, metatranscriptomes, single-cell and cultivar genomes. The abundance of grs homologs shows a strong negative correlation to pH, but a weak positive correlation to temperature. Archaeal genomes and metagenome-assembled genomes (MAGs) that carry two or more grs copies are more abundant in low pH springs. We also find grs in 12 archaeal classes, with the most representatives in Thermoproteia, followed by MAGs of the uncultured Korarchaeia, Bathyarchaeia and Hadarchaeia, while several Nitrososphaeria encodes >3 copies. Furthermore, our findings highlight the key role of grs-catalysed lipid cyclization in archaeal diversification across hot and acidic environments.

59 BASIC BIOLOGICAL SCIENCES↗
DOE OSTI2024

Visual and genetic stock identification of a test fishery to forecast Columbia River spring Chinook salmon stocks 2 weeks into the future

Abstract Modern fisheries management strives to balance opposing goals of protection for weak stocks and opportunity for harvesting healthy stocks. Test fisheries can aid management of anadromous fishes if they can forecast the strength and timing of an annual run with adequate time to allow fisheries planning. Integration of genetic stock identification (GSI) can further maximize utility of test fisheries by resolving run forecasts into weak‐ and healthy‐stock subcomponents. Using 5 years (2017–2022) of test fishery data, our study evaluated accuracy, resolution, and lead time of predictions for stock‐specific run timing and abundance of Columbia River spring Chinook salmon ( Oncorhynchus tshawytscha ). We determined if this test fishery (1) could use visual stock identification (VSI) to forecast at the coarse stock resolution (i.e., classification of “lower” vs. “upriver” stocks) upon which current management is based and (2) could be enhanced with GSI to forecast at higher stock resolution. VSI accurately identified coarse stocks (83.3% GSI concordance), and estimated a proxy for abundance (catch per unit effort, CPUE) of the upriver stock in the test fishery that was correlated ( R 2 = 0.90) with spring Chinook salmon abundance at Bonneville dam (Rkm 235). Salmon travel rates (~8.6 Rkm/day) provided predictions with 2‐week lead time prior to dam passage. Importantly, GSI resolved this predictive ability as finely as the hatchery broodstock level. Lower river stock CPUE in the test fishery was correlated with abundance at Willamette Falls (Rkm 196, R 2 = 0.62), but could not be as finely resolved as achieved for upriver stocks. We described steps to combine VSI and GSI to provide timely in‐season information and with prediction accuracy of ~12.4 mean absolute percentage error and high stock resolution to help plan Columbia River mainstem fisheries.

Hess, Jon E.↗
DOE OSTI2020

Short-Term Stable Isotope Probing of Proteins Reveals Taxa Incorporating Inorganic Carbon in a Hot Spring Microbial Mat

The upper green layer of the chlorophototrophic microbial mats associated with the alkaline siliceous hot springs of Yellowstone National Park consists of oxygenic cyanobacteria ( Synechococcus spp.), anoxygenic Roseiflexus spp., and several other anoxygenic chlorophototrophs. Synechococcus spp. are believed to be the main fixers of inorganic carbon (C i ), but some evidence indicates that Roseiflexus spp. also contribute to inorganic carbon fixation during low-light, anoxic morning periods. Contributions of other phototrophic taxa have not been investigated. In order to follow the pathway of C i incorporation into different taxa, mat samples were incubated with [ 13 C]bicarbonate for 3 h during the early-morning, low-light anoxic period. Extracted proteins were treated with trypsin and analyzed by mass spectrometry, leading to peptide identifications and peptide isotopic profile signatures containing evidence of 13 C label incorporation. A total of 25,483 peptides, corresponding to 7,221 proteins, were identified from spectral features and associated with mat taxa by comparison to metagenomic assembly sequences. A total of 1,417 peptides, derived from 720 proteins, were detectably labeled with 13 C. Most 13 C-labeled peptides were derived from proteins of Synechococcus spp. and Roseiflexus spp. Chaperones and proteins of carbohydrate metabolism were most abundantly labeled. Proteins associated with photosynthesis, C i fixation, and N 2 fixation were also labeled in Synechococcus spp. Importantly, most proteins of the 3-hydroxypropionate bi-cycle for C i fixation in Roseiflexus spp. were labeled, establishing that members of this taxocene contribute to C i fixation. Other taxa showed much lower [ 13 C]bicarbonate incorporation. IMPORTANCE Yellowstone hot spring mats have been studied as natural models for understanding microbial community ecology and as modern analogs of stromatolites, the earliest community fossils on Earth. Stable-isotope probing of proteins (Pro-SIP) permitted short-term interrogation of the taxa that are involved in the important process of light-driven C i fixation in this highly active community and will be useful in linking other metabolic processes to mat taxa. Here, evidence is presented that Roseiflexus spp., which use the 3-hydroxypropionate bi-cycle, are active in C i fixation. Because this pathway imparts a lower degree of selection of isotopically heavy C i than does the Calvin-Benson-Bassham cycle, the results suggest a mechanism to explain why the natural abundance of 13 C in mat biomass is greater than expected if only the latter pathway were involved. Understanding how mat community members influence the 13 C/ 12 C ratios of mat biomass will help geochemists interpret the 13 C/ 12 C ratios of organic carbon in the fossil record.

59 BASIC BIOLOGICAL SCIENCES↗