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All sourcesNASA NTRSDOE OSTI
DOE OSTI2021

Data for Integrating Transcriptomic and Metabolomic Analysis of the Oleaginous Yeast Rhodosporidium toruloides IFO0880 During Growth under Different Carbon Sources

Rhodosporidium toruloides is an oleaginous yeast capable of producing a variety of biofuels and bioproducts from diverse carbon sources. Despite numerous studies showing its promise as a platform microorganism, little is known about its metabolism and physiology. In this work, we investigated the central carbon metabolism in R. toruloides IFO0880 using transcriptomics and metabolomics during growth on glucose, xylose, acetate, or soybean oil. These substrates were chosen because they can be derived from plants. Significant changes in gene expression and metabolite concentrations were observed during growth on these four substrates. We mapped these changes onto the governing metabolic pathways to better understand how R. toruloides reprograms its metabolism to enable growth on these substrates. One notable finding concerns xylose metabolism, where poor expression of xylulokinase induces a bypass leading to arabitol production. Collectively, these results further our understanding of central carbon metabolism in R. toruloides during growth on different substrates. They may also help guide the metabolic engineering and development of better models of metabolism for R. toruloides .

Conversion↗
DOE OSTI2025

Atomic-resolution imaging as a mechanistic tool for studying single-site heterogeneous catalysis

Heterogeneous catalysts dominate the chemical industry but typically feature diverse, incompletely defined active sites. Thus, describing structure-activity relationships, unlike homogeneous catalysts, remains challenging. In contrast, molecularly defined single-site heterogeneous catalysts (SSHCs), using appropriate tools, are poised to address these challenges and provide new avenues for catalysis research and development. The present study explores eco-friendly H 2 production mediated by discrete MoO 2 sites supported on carbon nanohorns (CNHs) and active for alcohol dehydrogenation. While informative, detailed ensemble EXAFS/XANES/XPS, kinetic measurements, and DFT analysis alone cannot provide a full molecular picture of the reaction pathway. Here, using single-molecule atomic-resolution time-resolved electron microscopy (SMART-EM), we propose the identification of four key catalytic intermediates anchored to CNHs and uncover a new reaction pathway involving alkoxide/hemiacetal equilibration and acetal oligomerization. Furthermore, these intermediates are inferred through a combination of theory and SMART-EM, showcasing the potential of SMART-EM as a complementary tool for exploring mechanistic hypotheses in catalysis.

36 MATERIALS SCIENCE↗
DOE OSTI2022

Life cycle analysis of polylactic acids from different wet waste feedstocks

Producing a valuable chemical product through diversion of wet wastes can simultaneously resolve the problems associated with increasing wastes and greenhouse gas emissions from conventional chemical production processes. In this work, we investigated the life-cycle greenhouse gas emissions, water, and fossil-fuel consumption for waste-derived polylactic acids (PLA) from three different waste feedstocks, namely wastewater sludge, food waste, and swine manure, using the Greenhouse Gases, Regulated Emissions, and Energy Use in Technologies (GREET) model. The decarbonization potential of replacing fossil-based resins with the waste-derived polymer was also investigated. The results show that swine manure-to-PLA pathway was the least carbon intensive (—1.4 kgCO 2 e/kg) among the three waste-to-PLA pathways on a cradle-to-grave basis, followed by the food waste case (—1.3 kgCO 2 e/kg) and then by the wastewater sludge case (0.6 kgCO 2 e/kg). In the baseline scenario, all three waste-to-PLA pathways were less carbon intensive than both fossil-based PET and HDPE on a cradle-to-grave basis: 66% (vs. PET) and 56% (vs. HDPE), 171 and 192%, 181 and 205% reduction in GHG emissions for wastewater sludge-, food waste-, and swine manure-to-PLA pathway, respectively. For all sensitivity cases investigated, the food waste- and swine manure-to-PLA pathways were significantly less carbon intensive than their fossil-counterparts. In terms of the annual decarbonization potential of replacing fossil-based PET or HDPE, the wastewater sludge- and food waste-pathway showed higher mitigation potential than the swine manure-pathway: i) 18–28 kilotons CO 2 e-reduction per year for wastewater sludge pathway; ii) 23–26 kTCO 2 e-reduction/yr for food waste pathway; and iii) about 5 kTCO 2 e-reduction/yr for swine manure pathway depending on the type of conventional resin replaced. However, given the abundant availability of the swine manure feedstocks across the United States, the decarbonization potential of swine manure-based pathway can also increase as the plant capacity or the number of plants grow.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗
DOE OSTI2021

Comprehensive analysis of differentially expressed mRNA and circRNA in Ankylosing spondylitis patients’ platelets

Highlights: • 4996 mRNAs and 2942 circRNAs were identified differentially expressed in platelets of AS patients. • Several platelet-derived immune mediators were upregulated to regulate the interaction between platelets and immune cells. • Two downregulated circRNA were identified, and the corresponding circRNA-miRNA-mRNA regulatory network was constructed. • mRNAs and circRNAs found herein could be used as diagnostic biomarkers or new therapeutic targets. Ankylosing spondylitis (AS) is a chronic inflammatory disease significantly decreasing the quality of life. Platelets play an important and active role in the development of AS. Accumulating evidence demonstrated platelets contain diverse RNA repository inherited from megakaryocytes or microvesicles. Platelet RNAs are dynamically affected by pathological conditions and could be used as diagnostic or prognostic biomarkers. However, the role of the platelet RNAs in AS is elusive. In this study, we compared mRNA and circRNA profiles in platelets between AS patients and healthy controls using RNA sequencing and bioinformatic analysis, and found 4996 mRNAs and 2942 circRNAs were differently expressed. The significantly over-expressed mRNAs in AS patients are involved in platelet activity, gap junction, focal adhesion, rap1 and toll and Imd signaling pathway. The previous identified platelet-derived immune mediators such as P2Y1, P2Y12, PF4, GPIbα, CD40L, ICAM2, CCL5 (RANTES), TGF-β (TGF-β1 and TGF-β2) and PDGF (PDGFB and PDGFA) are also included in these over expressed mRNAs, implying these factors may trigger inflammatory cascades and promote the development of AS. Additionally, we found two down-regulated circRNA (circPTPN22 and circFCHSD2) from the intersection analyses of platelets and spinal ligament tissues of AS patients. The circRNA-miRNA-mRNA regulatory network of these two circRNAs was constructed, and the target mRNAs were enriched in Th17 cell differentiation, inflammatory bowel disease, cell adhesion molecules, cytokine-cytokine receptor interaction, Jak-STAT and Wnt signaling pathway, all these pathways participate in the bone remodeling and pro/anti-inflammatory immune regulation in AS. Then, qRT-PCR was performed to validate the expression of selected key mRNAs and circRNAs and the results demonstrated that the expression levels of P2Y12, GPIbα, circPTPN22 and circFCHSD2 were consistent with the sequencing analysis. In addition, the high expression of five predicted miRNAs interacting with circPTPN22 and circFCHSD2 were also detected in AS by qRT-PCR. Taken together, our study presents a comprehensive overview of mRNAs and circRNAs in platelets in AS patients and offers new insight into the mechanisms of platelet involving in the pathogenesis of AS. The mRNAs and circRNAs identified in this study may serve as candidates for diagnosis and targeted treatment of AS.

60 APPLIED LIFE SCIENCES↗
DOE OSTI2025

Design of diverse, functional mitochondrial targeting sequences across eukaryotic organisms using variational autoencoder

Mitochondria play a key role in energy production and metabolism, making them a promising target for metabolic engineering and disease treatment. However, despite the known influence of passenger proteins on localization efficiency, only a few protein-localization tags have been characterized for mitochondrial targeting. To address this limitation, we leverage a Variational Autoencoder to design novel mitochondrial targeting sequences. In silico analysis reveals that a high fraction of the generated peptides (90.14%) are functional and possess features important for mitochondrial targeting. We characterize artificial peptides in four eukaryotic organisms and, as a proof-of-concept, demonstrate their utility in increasing 3-hydroxypropionic acid titers through pathway compartmentalization and improving 5-aminolevulinate synthase delivery by 1.62-fold and 4.76-fold, respectively. Moreover, we employ latent space interpolation to shed light on the evolutionary origins of dual-targeting sequences. Overall, our work demonstrates the potential of generative artificial intelligence for both fundamental research and practical applications in mitochondrial biology.

59 BASIC BIOLOGICAL SCIENCES↗
DOE OSTI2024

Data for "Design of Diverse, Functional Mitochondrial Targeting Sequences Across Eukaryotic Organisms Using Variational Autoencoder"

Mitochondria play a key role in energy production and metabolism, making them a promising target for metabolic engineering and disease treatment. However, despite the known influence of passenger proteins on localization efficiency, only a few protein-localization tags have been characterized for mitochondrial targeting. To address this limitation, we leverage a Variational Autoencoder to design novel mitochondrial targeting sequences. In silico analysis reveals that a high fraction of the generated peptides (90.14%) are functional and possess features important for mitochondrial targeting. We characterize artificial peptides in four eukaryotic organisms and, as a proof-of-concept, demonstrate their utility in increasing 3-hydroxypropionic acid titers through pathway compartmentalization and improving 5-aminolevulinate synthase delivery by 1.62-fold and 4.76-fold, respectively. Moreover, we employ latent space interpolation to shed light on the evolutionary origins of dual-targeting sequences. Overall, our work demonstrates the potential of generative artificial intelligence for both fundamental research and practical applications in mitochondrial biology.

AI/ML↗
DOE OSTI2026

Impacts of Regional Coordination on Transmission Needs for Power System Resource Adequacy [Slides]

Resource adequacy (RA) is an important component of bulk power system reliability that addresses whether there are sufficient resources available to meet electricity demand at all times. RA analysis is used to assess whether the planned power system will provide reliable electricity to consumers while accounting for equipment outages, weather variability, and load uncertainty. Coordination between regions can enable resource sharing to meet RA needs if sufficient transmission capacity exists. Transmission's role in RA coordination can be particularly pronounced for large power systems like the U.S. electricity grid which contains geographically diverse demand and weather-dependent resources. Depending on the level of coordination desired, existing inter-regional transmission capacity may not be sufficient. This study is designed to assess optimal pathways for the development of inter-regional transmission in the U.S. under varying levels of RA coordination. Results can inform long-term grid infrastructure planning and provide insights into potential benefits of greater coordination for generation and transmission planning between regional U.S. power systems.

29 ENERGY PLANNING, POLICY, AND ECONOMY↗
DOE OSTI2021

Diverse ecophysiological adaptations of subsurface Thaumarchaeota in floodplain sediments revealed through genome-resolved metagenomics

Abstract The terrestrial subsurface microbiome contains vastly underexplored phylogenetic diversity and metabolic novelty, with critical implications for global biogeochemical cycling. Among the key microbial inhabitants of subsurface soils and sediments are Thaumarchaeota, an archaeal phylum that encompasses ammonia-oxidizing archaea (AOA) as well as non-ammonia-oxidizing basal lineages. Thaumarchaeal ecology in terrestrial systems has been extensively characterized, particularly in the case of AOA. However, there is little knowledge on the diversity and ecophysiology of Thaumarchaeota in deeper soils, as most lineages, particularly basal groups, remain uncultivated and underexplored. Here we use genome-resolved metagenomics to examine the phylogenetic and metabolic diversity of Thaumarchaeota along a 234 cm depth profile of hydrologically variable riparian floodplain sediments in the Wind River Basin near Riverton, Wyoming. Phylogenomic analysis of the metagenome-assembled genomes (MAGs) indicates a shift in AOA population structure from the dominance of the terrestrial Nitrososphaerales lineage in the well-drained top ~100 cm of the profile to the typically marine Nitrosopumilales in deeper, moister, more energy-limited sediment layers. We also describe two deeply rooting non-AOA MAGs with numerous unexpected metabolic features, including the reductive acetyl-CoA (Wood-Ljungdahl) pathway, tetrathionate respiration, a form III RuBisCO, and the potential for extracellular electron transfer. These MAGs also harbor tungsten-containing aldehyde:ferredoxin oxidoreductase, group 4f [NiFe]-hydrogenases and a canonical heme catalase, typically not found in Thaumarchaeota. Our results suggest that hydrological variables, particularly proximity to the water table, impart a strong control on the ecophysiology of Thaumarchaeota in alluvial sediments.

59 BASIC BIOLOGICAL SCIENCES↗
DOE OSTI2024

Multicriteria-Based Selection of Microalgae Biorefineries: Biomass Composition Defines the Most Suitable Product Portfolios

The supply of microalgae-derived biofuels and bioproducts will be vital in a global-scale bioeconomy. As the diversity of microalgae strains and their compositional plasticity may yield a wide array of products of market interest, the design of effective biorefineries is a central aspect in the path to making microalgae-based products available in the market. In this way, the conversion of microalgae biomass should be planned to employ mature technologies, while maximizing economic and environmental benefits obtained from a diverse product portfolio. This study applied sequential, hybrid Multicriteria Decision Analyses (MCDA) to aid the decision-making process of outlining the most suitable biorefining pathways for specific compositional profiles. For this, the methodology simultaneously considered multiple technical, economic, and environmental criteria, such as market aspects for the main algae-derived products, potential reduction in greenhouse gas (GHG) emissions provided by the biobased alternatives, and technology readiness level of conversion routes, among others. The framework was tested using productivity and compositional data for 13 high-productivity summer strains cultivated under varying nutrient availability. The analysis pointed to compositional profile being a key driver in defining the core biorefining strategy of algae biomass, with lipid-to-carbohydrate ratios higher than roughly 1 warranting the preferential processing of lipids into hydrocarbon fuels with the remainder of the algae biomass compounds being sent to higher-value applications, such as carboxylic acids and renewable thermoplastic substitutes. An in-depth process simulation, techno-economic assessment (TEA), and life-cycle analysis (LCA) effort was carried out as a closing step to corroborate the results stemming from the proposed framework. This study validates the use of MCDAs as a screening method prior to implementing more time-consuming analysis techniques and makes a compelling case for this approach as a product selection tool on a wide range of algae species, thus helping establish species-agnostic (but composition-driven) biorefineries.

biofuels↗
DOE OSTI2025

National User Resource for Biological Accelerator Mass Spectrometry (Final Report)

The National User Resource for Biological Accelerator Mass Spectrometry (User Resource) will provide isotopic analysis (primarily radiocarbon or 14C) by accelerator mass spectrometry (AMS) for NIH- funded researchers across the United States and will be the only User Resource of its type in the United States. The User Resource will provide measurement capability and expertise to a research community that requires highly sensitive, quantitative isotope analyses. Since commissioning a new accelerator mass spectrometer in June 2014, we have measured over 4000 samples a year for collaborators and service users. The User Resource will enable us to continue to meet these research needs, as well as provide for new users whose research programs would benefit from AMS as a measurement tool. The User Resource’s forte will be ultra-high sensitivity quantitation of radiocarbon and selected other radioisotopes for research studies where isotopes are required. Radioisotope labeling studies have been and will continue to be an important tool for addressing many complex biomedical science problems. AMS is a specialized and unique type of mass spectrometry that provides absolute quantitation of radiocarbon and other relevant radioisotopes with extreme sensitivity, having limits of detection in real samples on the order of a few attomol/mg of sample at measurement precisions of ~3%. It is the only instrumental method capable of quantifying radioisotope-labeled agents routinely in real-world samples with such precision and sensitivity. The sensitivity of AMS allows for the quantification of radiolabeled metabolites in extremely complex matrices of cells and organisms at very low concentrations and in small samples. AMS allows studies to be conducted without perturbing metabolism leading to more relevant quantification of metabolic rates and pathways. In addition, it enables quantification of pharmacokinetic and metabolic properties of toxicants at environmentally relevant concentrations in model systems as well as the ability to quantify pharmacokinetics and other molecular endpoints directly in humans. Such quantitative assessments can 1) improve risk assessment for toxicants, 2) address safety and efficacy considerations for therapeutic entities, 3) deepen understanding of xenobiotic and intermediary metabolism, 4) help understand the interactions between critical molecular pathways, and 5) improve efforts to model and predict various metabolic and biological states. These capabilities have been applied in a number of areas including research in carcinogenesis, toxicology, nutrition, pharmacology/drug development and basic biological science. As a NIGMS National Resource the National User Resource for Biological Accelerator Mass Spectrometry will help NIH funded scientists achieve a deeper understanding of the etiology of human health concerns by (1) enabling the quantification of pharmacokinetics and other molecular endpoints directly in humans; (2) offering the ability to conduct quantitative studies using biologics such as proteins or lipids, and thereby reducing the amount of radioisotope usage in biomedical labs; and (3) enabling more relevant studies of metabolic pathways in health and disease through the use of much lower, more biologically-relevant, concentrations of metabolic substrates in cells and intact organisms. Such studies support NIGMS’s basic biomedical research areas that contribute to the understanding of fundamental cellular and physiological principles and enable research supported by the Biophysics, Biomedical Technology, and Computational Biosciences (BBCB); Genetics and Molecular, Cellular, and Developmental Biology (GMCDB); Pharmacology, Physiology, Biological Chemistry (PPBC) and Training, Workforce Development, and Diversity (TWD) Divisions. Over the next five years, our goals are to: 1. Improve the efficiency of operation for AMS measurements through installation of new interfaces to our AMS systems, technical modifications to improve gas accepting ion source efficiency and upgrading our data analysis software for improved ease of use and data reporting. 2. Increase the accessibility and visibility of ultra-sensitive 14C measurements for the biomedical research community by training of new investigators and expanding our national user base. 3. Provide high throughput, ultra-sensitive 14C analysis for the NIGMS and NIH user community.

47 OTHER INSTRUMENTATION↗
DOE OSTI2024

Discovering Innovations in Stress Tolerance through Comparative Gene Regulatory Network Analysis and Cell-Type Specific Expression Maps (Final Technical Report with Cover Page)

Through this grant, we developed a comparative framework to elucidate the mechanisms behind variations in environmental stress responses among a diverse group of species within the Brassicaceae family. Our focus was on the differences in physiological and transcriptomic responses to abscisic acid (ABA), a hormone associated with water stress. We examined the differential growth responses of four Brassicaceae species, finding that most exhibited reduced root growth correlated with smaller meristem size. In contrast, Schrenkiella parvula showed accelerated growth due to increased root cell elongation. We employed RNA sequencing to analyze the transcriptional responses to ABA across these species, and innovative bioinformatics techniques were used to pinpoint biological pathways with significant divergence. Additionally, we utilized DAP-seq to map the gene regulatory networks associated with ABAresponsive transcription factors, revealing that variations in the regulation of growth hormone biosynthesis play a critical role in the distinct ABA effects on root growth among the species. This research sets a new standard for comparative physiology by integrating comparative genomics and transcriptomics to uncover pathway divergences.

59 BASIC BIOLOGICAL SCIENCES↗
DOE OSTI2025

Understanding and Harnessing the Robustness of Undomesticated Yarrowia lipolytica Strains for Biosynthesis of Designer Bioesters (Final Report)

This project seeks to elucidate and harness the exceptional robustness of novel and undomesticated Y. lipolytica isolates, which were identified from a genetic diversity screening for compatibility with bioenergy development. Bioenergy-relevant isolates were further developed as microbial platforms for efficient conversion of undetoxified biomass hydrolysates into designer bio-esters continuously recovered by solvent extraction. The project has three major goals. Goal 1. Elucidate and enhance the endogenous metabolism of Y. lipolytica for superior growth, sugar utilization, and lipid accumulation in undetoxified biomass hydrolysates under hypoxic conditions. Goal 2. Understand and enhance the underlying mechanism of exceptional tolerance of Y. lipolytica to organic solvents. Goal 3. Elucidate and rewire endogenous metabolism of the most robust Y. lipolytica strains for effective conversion of accumulating lipids to designer bio-esters. Significant progress has been made toward completing all research goals. We elucidated and optimized the robustness of Y. lipolytica by utilizing mixed C5 and C6 sugars in switchgrass hydrolysates (SGH) for lipid production (Aim 1). We conducted extensive omics analysis to investigate how genetic diversity among Yarrowia strains, derived from natural isolates or developed through adaptive laboratory evolution, influences lipid production when utilizing SGH. In Aim 2, novel mechanisms and underlying genetics were discovered that enabled Yarrowia strains to thrive in cultures containing high ionic liquid (IL) concentrations. Amongst other novel findings, it was found that sterols strengthened cell membranes to confer IL toxicity resistance, specifically via increased ergosterol content upon exposure to IL. In Aim 3, mechanistic studies elucidated how Y. lipolytica utilized intracellular lipids and alkanes/alkenes, leading to our discovery of novel enzymes and pathways for making short-chain esters. Most notably, thermostable chloramphenicol transferases were repurposed to function as alcohol acetyltransferases in Y. lipolytica, as well as the Gram-negative and Gram-positive bacteria Escherichia coli and thermophile Clostridium thermocellum, respectively.

09 BIOMASS FUELS↗
DOE OSTI2023

Ex vivo to in vivo model of malignant peripheral nerve sheath tumors for precision oncology

Malignant peripheral nerve sheath tumors (MPNST) are aggressive soft tissue sarcomas that often develop in patients with neurofibromatosis type 1 (NF1). To address the critical need for novel therapeutics in MPNST, we aimed to establish an ex vivo 3D platform that accurately captured the genomic diversity of MPNST and could be utilized in a medium-throughput manner for drug screening studies to be validated in vivo using patient-derived xenografts (PDX). Genomic analysis was performed on all PDX-tumor pairs. Selected PDX were harvested for assembly into 3D microtissues. Based on prior work in our labs, we evaluated drugs (trabectedin, olaparib, and mirdametinib) ex vivo and in vivo. For 3D microtissue studies, cell viability was the endpoint as assessed by Zeiss Axio Observer. For PDX drug studies, tumor volume was measured twice weekly. Bulk RNA sequencing was performed to identify pathways enriched in cells. We developed 13 NF1-associated MPNST-PDX and identified mutations or structural abnormalities in NF1 (100%), SUZ12 (85%), EED (15%), TP53 (15%), CDKN2A (85%), and chromosome 8 gain (77%). We successfully assembled PDX into 3D microtissues, categorized as robust (>90% viability at 48 h), good (>50%), or unusable (<50%). We evaluated drug response to “robust” or “good” microtissues, namely MN-2, JH-2-002, JH-2-079-c, and WU-225. Drug response ex vivo predicted drug response in vivo, and enhanced drug effects were observed in select models. These data support the successful establishment of a novel 3D platform for drug discovery and MPNST biology exploration in a system representative of the human condition.

60 APPLIED LIFE SCIENCES↗
DOE OSTI2026

Machine learning-based bias-corrected future projections of ozone concentrations from a chemistry-climate model

Reliable projection of future near-surface ozone is crucial for air quality management and health risk assessment. However, potential biases in spatial distribution, magnitude and trends in ozone concentrations simulated by global chemistry-climate models limit their applicability in regional-scale evaluations. In this study, LightGBM, a machine learning (ML) algorithm is applied to correct biases in CESM2-simulated ozone concentrations over China, the United States and Europe and calibrate future ozone projections under two diverse Shared Socioeconomic Pathways (SSP1-2.6 and SSP5-8.5) scenarios from 2020 to 2060. The ML-based correction significantly improves the spatial distribution and reduces the model bias by 40%–60%. It also reverses the potentially incorrect trend of ozone change under SSP1-2.6 in eastern China. When applying ML-based bias correction to CESM2 future projections, warm season mean ozone concentrations decrease across China, the United States, and Europe by –13.5, –17.9, and –13.7 µg/m³, respectively, between 2020 and 2060 in SSP1-2.6, while they increase by 9.4, 2.0, and 5.2 µg/m³ in SSP5-8.5. Decomposition analysis show that changes in anthropogenic emissions dominate future ozone changes in both scenarios, while strong climate penalty from ozone changes occurs in polluted eastern China and climate benefit is found in western China, the United States and Europe under SSP5-8.5. These findings demonstrate the value of combining ML with chemistry-climate models to produce more accurate air quality projections, thereby informing more effective and region-specific environmental protection strategies.

Chemistry Model↗
DOE OSTI2023

Sequential removal of cation/H + exchangers reveals their additive role in elemental distribution, calcium depletion and anoxia tolerance

Multiple Arabidopsis H + /Cation exchangers (CAXs) participate in high-capacity transport into the vacuole. Previous studies have analysed single and double mutants that marginally reduced transport; however, assessing phenotypes caused by transport loss has proven enigmatic. Here, in this study, we generated quadruple mutants (cax1-4: qKO) that exhibited growth inhibition, an 85% reduction in tonoplast-localised H + /Ca transport, and enhanced tolerance to anoxic conditions compared to CAX1 mutants. Leveraging inductively coupled plasma mass spectrometry (ICP-MS) and synchrotron X-ray fluorescence (SXRF), we demonstrate CAX transporters work together to regulate leaf elemental content: ICP-MS analysis showed that the elemental concentrations in leaves strongly correlated with the number of CAX mutations; SXRF imaging showed changes in element partitioning not present in single CAX mutants and qKO had a 40% reduction in calcium (Ca) abundance. Reduced endogenous Ca may promote anoxia tolerance; wild-type plants grown in Ca-limited conditions were anoxia tolerant. Sequential reduction of CAXs increased mRNA expression and protein abundance changes associated with reactive oxygen species and stress signalling pathways. Multiple CAXs participate in postanoxia recovery as their concerted removal heightened changes in postanoxia Ca signalling. This work showcases the integrated and diverse function of H + /Cation transporters and demonstrates the ability to improve anoxia tolerance through diminishing endogenous Ca levels.

36 MATERIALS SCIENCE↗
DOE OSTI2021

Igiugig Site Visit Report

The National Renewable Energy Laboratory (NREL) team conducted a site visit during January 23-25, 2019 with the Igiugig Village Council (IVC) and other stakeholders to assist the community of Igiugig in refining their long-term energy strategy. The agenda included a tour of the village, data collection, presentations, long-term visioning exercises, identification of energy scenarios, and a school presentation. Participants in the site visit activities spanned a range of local, regional, government, and industry participation. There were 24 attendees at the all-day community workshop on Thursday, January 24th, with participants from the IVC, NREL, Bristol Bay Native Association (BBNA), Bristol Bay Native Corporation (BBNC), Lake and Peninsula Borough, Alaska Energy Authority, Intergrid, ORPC, Deer Stone Consulting, the Southwest Alaska Municipal Conference, and University of Alaska Fairbanks - Alaska Center for Energy and Power. Igiugig is a well-organized rural Alaskan community, with a leadership team that appears to have broad community support. The use of consensus-based decision making is one tangible example of how the leadership team actively invokes and promotes community-focused thinking. The NREL team observed anecdotal evidence of how this approach seems to be strengthening the community: active participation in the visioning exercise, a student hosted fund-raiser dinner at the school, a clean and organized landfill, and well-maintained roads and buildings. Igiugig has wind, solar, and river hydrokinetic resources readily available within the community. Wind has shown to be a promising resource in the region, and has been integrated into microgrids around the state, but the community has had mixed success with wind technologies: some devices failed quickly and others continue to operate. The economics of solar energy are improving in Alaska, and economical solar projects are being installed around the state. Considering that economic activity in Igiugig peaks during the summer sport fishing season, solar could prove to be a valuable supplement to the electrical system. Igiugig has been a test site for two different river hydrokinetic devices, and they have began a third project to operate ORPC's RivGen device, which is delivering valuable device performance data, operations and maintenance experience, and design refinement information. Already, this device has made over 7-million revolutions, and delivered over 8 MWh of power to the community. Igiugig's river resource is fairly unique because it is available year-round and has the potential to provide reliable base-load power for months at a time. A preliminary investigation of this diverse resource mix suggests that Igiugig could achieve very high levels (70% or more) of annual renewable generation contributions. A critical step in pursuing this path is identifying the mix of energy assets (generation and storage) that best meets the community's budget, needs, and goals. The assets that a community installs early in their grid-modernization initiative can constrain the options that are economical at later stages, which may lead to sub-optimal solutions. This is where technical and economic analysis of potential scenarios (i.e., distinct mixes of energy assets) can be useful in identifying the most promising pathways so that a community can make informed and strategic decisions about the assets they install. These analyses are most accurate and informative when they are based on actual technology performance and cost data. As the RivGen project continues to operate and generate this data, we will be better prepared to evaluate the technology's long-term viability and to identify research areas that would improve it. Igiugig's energy projects are at the cutting edge of two intersecting technology areas: 1) deploying an operational river hydrokinetic turbine, and 2) integrating renewable energy sources to achieve very high percent renewables. If these projects are successful, the lessons learned and technologies developed could be valuable for other microgrids around the world. Igiugig's unique resource mix make it an ideal location for this work, and the community's organizational strength make it an ideal partner in this ground-breaking work. Ongoing support for technical assistance to manage and address technical challenges along the way will maximize the probability of project success.

13 HYDRO ENERGY↗
DOE OSTI2021

Revisiting a ‘simple’ fungal metabolic pathway reveals redundancy, complexity and diversity

Next to d-glucose, the pentoses l-arabinose and d-xylose are the main monosaccharide components of plant cell wall polysaccharides and are therefore of major importance in biotechnological applications that use plant biomass as a substrate. Pentose catabolism is one of the best-studied pathways of primary metabolism of Aspergillus niger, and an initial outline of this pathway with individual enzymes covering each step of the pathway has been previously established. However, although growth on l-arabinose and/or d-xylose of most pentose catabolic pathway (PCP) single deletion mutants of A. niger has been shown to be negatively affected, it was not abolished, suggesting the involvement of additional enzymes. Detailed analysis of the single deletion mutants of the known A. niger PCP genes led to the identification of additional genes involved in the pathway. These results reveal a high level of complexity and redundancy in this pathway, emphasizing the need for a comprehensive understanding of metabolic pathways before entering metabolic engineering of such pathways for the generation of more efficient fungal cell factories.

59 BASIC BIOLOGICAL SCIENCES↗
DOE OSTI2025

Supply Chain Energy and Greenhouse Gas Analysis Using the Materials Flows Through Industry (MFI) Tool: Examination of Alternative Technology Scenarios for the U.S. Chemical Sector

Chemical manufacturing is a large and diverse sector of the U.S. economy, with products, fuels, and a wide assortment of materials used daily by both the public and businesses. Currently, several of the largest volume chemicals produced in the United States rely on fossil fuels as a feedstock, energy source, or both. The list of chemicals includes steam cracking products such as ethylene, propylene, benzene, and xylenes as well as products such as ammonia and methanol. The focus for this work is on platform chemicals that are both produced in the largest volume and have a high potential for subsequent processing into more specialized products. In this study, we explore several new pathways that reduce the overall energy consumption and greenhouse gas (GHG) emissions for each product. These pathways include energy efficiency measures applied to existing production methods, the use of bio‐based fuels and/or feedstocks as new production methods, and electrification of high‐energy‐input stages within current production methods. Scenarios for energy demand and GHG reduction were conducted with the National Renewable Energy Laboratory's Materials Flows through Industry tool. Projections of the energy demand and GHG emissions in 2030 and 2050 are included, using grid composition projections from the NREL ReEDS model. The alternative scenarios selected showcase the effect of realistic changes the industry could make, focusing on technologies with a high level of technical readiness.

29 ENERGY PLANNING, POLICY, AND ECONOMY↗