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DOE OSTI2024

Identification of pleiotropic loci mediating structural and non-structural carbohydrate accumulation within the sorghum bioenergy association panel using high-throughput markers

Molecular characterization of diverse germplasm can contribute to breeding programs by increasing genetic gain for sorghum [ Sorghum bicolor (L.) Moench] improvement. Identifying novel marker-trait associations and candidate genes enriches the existing genomic resources and can improve bioenergy-related traits using genomic-assisted breeding. In the current scenario, identifying the genetic loci underlying biomass and carbon partitioning is vital for ongoing efforts to maximize each carbon sink’s yield for bioenergy production. Here, we have processed a high-density genomic marker (22 466 550) data based on whole-genome sequencing (WGS) using a set of 365 accessions from the bioenergy association panel (BAP), which includes ~19.7 million (19 744 726) single nucleotide polymorphism (SNPs) and 2.7 million (~2 721 824) insertion deletions (indels). A set of high-quality filtered SNP (~5.48 million) derived markers facilitated the assessment of population structure, genetic diversity, and genome-wide association studies (GWAS) for various traits related to biomass and its composition using the BAP. The phenotypic traits for GWAS included seed color (SC), plant height (PH), days to harvest (DTH), fresh weight (FW), dry weight (DW), brix content % (BRX), neutral detergent fiber (NDF), acid detergent fiber (ADF), non-fibrous carbohydrate (NFC), and lignin content. Several novel loci and candidate genes were identified for bioenergy-related traits, and some well-characterized genes for plant height ( Dw1 and Dw2 ) and the YELLOW SEED1 locus ( Y1 ) were validated. We further performed a multi-variate adaptive shrinkage analysis to identify pleiotropic QTL, which resulted in several shared marker-trait associations among bioenergy and compositional traits. Significant marker-trait associations with pleiotropic effects can be used to develop molecular markers for trait improvement using a marker-assisted breeding approach. Significant nucleotide diversity and heterozygosity were observed between photoperiod-sensitive and insensitive individuals of the panel. This diverse bioenergy panel with genomic resources will provide an excellent opportunity for further genetic studies, including selecting parental lines for superior hybrid development to improve biomass-related traits in sorghum.

bioenergy association panel↗

DOE OSTI2022

What is Water's Role in a Carbon Neutral Future? A Summary of Findings from a Webinar Series

There has been ever-growing interest and engagement regarding net-zero and carbon neutrality goals, with many nations committing to steep emissions reductions by mid-century. Although water plays critical roles in various sectors, there has been a distinct gap in discussions to date about the role of water in the transition to a carbon neutral future. To address this need, a webinar was convened in April 2022 to gain insights into how water can support or influence active strategies for addressing emissions activities across energy, industrial, and carbon sectors. The webinar presentations and discussions highlighted various nuances of direct and indirect water use both within and across technology sectors (Figure ES-1). For example, hydrogen and concrete production, water for mining, and inland waterways transportation are all heavily influenced by the energy sources used (fossil fuels vs. renewable sources) as well as local resource availabilities. Algal biomass, on the other hand, can be produced across diverse geographies (terrestrial to sea) in a range of source water qualities, including wastewater and could also support pollution remediation through nutrient and metals recovery. Finally, water also influences carbon dynamics and cycling within natural systems across terrestrial, aquatic, and geologic systems. These dynamics underscore not only the critical role of water within the energy-water nexus, but also the extension into the energy-watercarbon nexus.

54 ENVIRONMENTAL SCIENCES↗

DOE OSTI2022

Value proposition of coatings or new alloys on hammer wear

The goal of this Case Study was to elucidate the value of improving of the life of parts that wear within a system in terms of the additional material cost that it takes to reach the level of improvement. We recognize that the failure limits and system performance are representative of a single system that may or may not exist in the real world, however, our goal of this project was not to provide an answer for a specific system or to provide a full understanding the economics of all potential systems. The analysis was performed using relative changes from the base alloy cost and considered relative improvements in part life in order to generalize the comparison without referencing specific alloys or coatings that might be employed. Ultimately, this work provides a first check of the potential for improving wear characteristics of grinder hammers to provide a meaningful and impactful benefit to biomass preprocessing and conversion systems. Key takeaways include: • Increasing the life of hammers by 3× provides a delivered feedstock cost benefit of approximately $\$2.25$/dry ton assuming that the relative cost of the hammer material of construction is not increased. • The feasible area of relative hammer cost increase ranges from 110% to 122% of the relative life increase, i.e., a 3× life increase can cost 3.66× the cost of the original hammers • Beyond a 3× increase in relative hammer life, little economic benefit is realized • Even when moisture impacts are included, the relationship between relative hammer cost and relative hammer life was reduced only slightly to a range of 108% to 116% • From the simulation we would expect delivered feedstock costs to be within ±$1.10/dry ton, based on the uncertainty analysis

09 BIOMASS FUELS↗

DOE OSTI2024

Fluidized-Bed Gasification of Coal-Biomass-Plastics for Hydrogen Production

Coal is one of the most abundant fossil energy resources in the United States and in the world. The recoverable reserves in the United States are estimated to be about 252 billion tons – more than 350 years of supply at current rates of usage. However, the share of coal in total primary energy consumption in the US has been declining over the years. The decline of coal is mainly attributed to cheap natural gas and precipitous declines in the cost of electricity production from renewable technologies such as wind and solar. Coal can potentially be used if it is coupled with carbon-neutral feedstock such as biomass-agricultural residues, forest biomass, and forest residues. Co-gasification of coal and biomass can become a negative carbon emission technology if the carbon dioxide (CO 2 ) is captured and sequestered. Although the biomass gasification process has a lot of similarities to coal gasification, the large-scale adaptation of power production sourced from biomass has not come to fruition. The main reason is that the power production from biomass is still expensive when compared with natural gas or coal power technologies. To address the feedstock cost, one approach is to use low-cost feedstocks, such as municipal solid wastes (MSW) or plastics, for gasification. Gasification involves the partial oxidation of coal and/or biomass feedstocks to produce a combustible fuel called synthesis gas (syngas) which is composed of carbon monoxide (CO), hydrogen (H 2 ), CO 2 , methane (CH 4 ), nitrogen (N 2 ), water (H 2 O), and other compounds that might be considered as contaminants. Raw syngas from gasification must go through multiple steps to produce high-purity hydrogen. The specific steps depend upon the quality (gas composition, contaminants, and their concentration) and condition (pressure and temperature) of syngas. The long-term goal of the project was to utilize coal and plastics together with biomass to produce energy and fuels using a gasification platform while reducing greenhouse gas emissions. The main objective of this research was to examine gasification performance in a laboratory-scale fluidized-bed gasifier for hydrogen production. The specific objectives of the research were to: (i) study coal-plastic-biomass mixture flowability for consistent feeding in the gasifier; (ii) understand the gasification behavior of the mixture in steam and oxygen environments; (iii) perform thermal property characterization of ash and slag from the mixture feedstock and refractory-ash interface of the mixture under gasification conditions; and (iv) develop process models to determine the technology needed for syngas cleanup and contaminants. The study found that there was no apparent segregation when biomass, coal, and waste plastics were mixed together during feeding. Although there were differences in hydrogen production when individual feedstock were fed, the hydrogen concentration remained almost constant with various blends. Therefore, blending waste plastics with biomass and coal, which are all abundant, is a better approach for energy production. Results of the techno-economic analysis suggested that integration of advanced gasification (GTI’s R-GAS™) and syngas cleanup and conditioning technologies (RTI’s WDP and AFWGS) for clean hydrogen production resulted in substantial benefits, including significant capital cost and operating cost reductions. Advanced technologies resulted in 16% reduction in the hydrogen production cost (COH) from 2.94 $\$$/kg to 2.47 $\$$/g, with further scope for optimization and cost reduction. These advanced technologies also result in lower emissions, and improved energy efficiency.

01 COAL, LIGNITE, AND PEAT↗

NASA NTRS2020

High productivity in hybrid-poplar plantations without isoprene emission to the atmosphere

Hybrid-poplar tree plantations provide a source for biofuel and biomass, but they also increase forest isoprene emissions. The consequences of increased isoprene emissions include higher rates of tropospheric ozone production, increases in the lifetime of methane, and increases in atmospheric aerosol production, all of which affect the global energy budget and/or lead to the degradation of air quality. Using RNA interference (RNAi) to suppress isoprene emission, we show that this trait, which is thought to be required for the tolerance of abiotic stress, is not required for high rates of photosynthesis and woody biomass production in the agroforest plantation environment, even in areas with high levels of climatic stress. Biomass production over 4 y in plantations in Arizona and Oregon was similar among genetic lines that emitted or did not emit significant amounts of isoprene. Lines that had substantially reduced isoprene emission rates also showed decreases in flavonol pigments, which reduce oxidative damage during extremes of abiotic stress, a pattern that would be expected to amplify metabolic dysfunction in the absence of isoprene production in stress-prone climate regimes. However, compensatory increases in the expression of other proteomic components, especially those associated with the production of protective compounds, such as carotenoids and terpenoids, and the fact that most biomass is produced prior to the hottest and driest part of the growing season explain the observed pattern of high biomass production with low isoprene emission. Our results show that it is possible to reduce the deleterious influences of isoprene on the atmosphere, while sustaining woody biomass production in temperate agroforest plantations.

oxidative stress↗

DOE OSTI2024

Microwave-Assisted dry reforming of toluene as a model tar compound using low-cost iron catalyst for syngas clean-up

Gasification of waste feedstock such as biomass, waste plastics suffers from high tar yields during hydrogen-rich syngas production. The presence of tars result in lower quality syngas, lower syngas yields, reactor blockage, reactor down time, and costly maintenance. Therefore, removal of tar from syngas during gasification is essential. Catalytic reforming of tars via in-situ syngas cleanup is an effective way of mitigating tars. This work explores the possibility of microwave-assisted catalytic dry reforming of tars for syngas production. Further, due to its chemical complexity, toluene, which is one of the main tar constituents, could be used as a model tar compound. Toluene dry reforming was studied using the Fe/Al 2 O 3 catalyst under CO 2 under the temperature range of 400–700 °C. The toluene reforming reaction was conducted using microwave and conventional thermal reactors. Under microwave irradiation, CO 2 and toluene conversions are boosted to 80% at 500 °C. Hydrogen and carbon monoxide yields were approximately five times and ten times higher in the microwave reactor at 500 °C, respectively, compared to the productions obtained in the conventional fixed-bed reactor at 700 °C. Filamentous carbon was also produced as a valuable side product to improve the economy of this process and such value-added carbon was only observed in the microwave reactor. Three reaction pathways were observed during microwave reaction: the toluene decomposition produces an initial hydrogen and carbon deposit on the catalyst; the formation of methane and benzene suggests toluene hydrodemethylation as a secondary reaction; and toluene hydrogenolysis forms light alkanes such as methane, and through reforming reaction under CO 2 to syngas.

10 SYNTHETIC FUELS↗

DOE OSTI2023

Pilot-Scale Continuous Plug-Flow Hydrothermal Liquefaction of Food Waste for Biocrude Production

Pilot-scale hydrothermal liquefaction (HTL) of biowaste is a critical step toward commercialization of the HTL technology. Despite many HTL studies conducted with wet biomass, including food waste, few were performed with a pilot-scale continuous plug-flow reactor (PFR), with the biocrude yield and quality analysis based on dewatering (ASTM D2892 Annex X1). This paper describes the development and performance evaluation of a mobile pilot-scale HTL continuous PFR, with a processing capacity of 60 L/h of wet feedstock and 6 L/h of biocrude production. The reactor system was designed for reaction conditions of up to 325 °C and 17.25 MPa. The reactor has a volume of 28.88 L with an additional counterflow heat exchanger volume of 18.07 L. Two types of food wastes, from a food processing plant and grocery store, were processed at 280 °C for 30 min, producing biocrude oil yields of 52.19 and 47.06 wt %, energy recoveries of 68.17 and 70.77%, and carbon recoveries of 66.91 and 64.78%, respectively. Furthermore, due to its high feedstock capacity and reaction volume, large amounts of biocrude oil and post-HTL wastewater (PHW) were obtained from this pilot-scale reactor to allow downstream research on upgrading biocrude oil for transportation fuel as well as PHW treatment and nutrient recovery.

09 BIOMASS FUELS↗

NASA NTRS2020

The Global Ecosystem Dynamics Investigation: High-Resolution Laser Ranging of the Earth’s Forests and Topography

Obtaining accurate and widespread measurements of the vertical structure of the Earth’s forests has been a longsought goal for the ecological community. Such observations are critical for accurately assessing the existing biomass of forests, and how changes in this biomass caused by human activities or variations in climate may impact atmospheric CO2 concentrations. Additionally, the three-dimensional structure of forests is a key component of habitat quality and biodiversity at local to regional scales. The Global Ecosystem Dynamics Investigation (GEDI) was launched to the International Space Station in late 2018 to provide high-quality measurements of forest vertical structure in temperate and tropical forests between 51.6 N & S latitude. The GEDI instrument is a geodetic-class laser altimeter/waveform lidar comprised of 3 lasers that produce 8 transects of structural information. Over its two-year nominal lifetime GEDI is anticipated to provide over 10 billion waveforms at a footprint resolution of 25 m. These data will be used to derive a variety of footprint and gridded products, including canopy height, canopy foliar profiles, Leaf Area Index (LAI), sub-canopy topography and biomass. Additionally, data from GEDI are used to demonstrate the efficacy of its measurements for prognostic ecosystem modeling, habit and biodiversity studies, and for fusion using radar and other remote sensing instruments. GEDI science and technology are unique: no other space-based mission has been created that is specifically optimized for retrieving vegetation vertical structure. As such, GEDI promises to advance our understanding of the importance of canopy vertical variations within an ecological paradigm based on structure, composition and function.

Dubayah, Ralph↗

DOE OSTI2025

Filer City Biomass Carbon Removal and Storage (BiCRS) Net-Negative Study

NorthStar Clean Energy Company (NorthStar) conducted the Filer City Biomass Carbon Removal and Storage (BiCRS) Net-Negative Study to develop a conceptual design and cost estimate for retrofitting the TES Filer City Station with post-combustion carbon capture technology in Mainstee Michigan. The Filer City BiCRS Net-Negative Study allowed NorthStar and team to confirm the commercial feasibility of a post-combustion carbon capture system applied to biomass-fired boilers. At the time of the study, there were no operating facilities in the world with carbon capture applied to flue gas from woody biomass as proposed for Filer City. Due to the solvent-agnostic nature of B&W’s SolveBright™ technology, the team determined that multiple amines, both traditional and proprietary, would be capable of 95% CO 2 capture efficiency with the steam available from Filer City’s existing boilers after modifications to burn 100% biomass. Based on the cost estimates produced, NorthStar can now confirm the Filer City BiCRS Project is economically viable with a combination of tax credits available from the Inflation Reduction Act of 2022 and high-quality Carbon Dioxide Removal Credits sold on the Voluntary Carbon Market. The Filer City BiCRS project is uniquely positioned to become one of the first projects to capture and sequester large volumes of CO 2 from a biogenic source, removing existing CO 2 from the atmosphere. The unlimited version of the Filer City Biomass Carbon Removal and Storage (BiCRS) Net-Negative Study Final Technical Report is attached.

01 COAL, LIGNITE, AND PEAT↗

DOE OSTI2023

Coupled wave-current modeling for hydrodynamic load analysis of macroalgae cultivation farms

Large-scale cultivation of macroalgae is one of the most promising biofuel sources that could reduce our consumption of fossil fuels and would be particularly economically competitive when grown for the co-production of additional goods such as food and textiles. Current production of biofuel from algal biomass is limited by labor costs and lack of data on potential impacts of cultivation and harvesting on marine and coastal environments as well as a comprehensive characterization of the impact of environmental conditions on macroalgal feedstock and their bioproduct and biofuel yield and quality. Awarded by the United States (U.S.) Department of Energy Advanced Research Projects Agency-Energy (ARPA-E) Macroalgae Research Inspiring Novel Energy Resources (MARINER) program, which seeks to enable the U.S. as a global leader in the production of marine biomass, the Marine Biological Laboratory (MBL) has been developing a test system for tropical seaweed cultivation in the Gulf of Mexico and the Caribbean. An integral step in designing macroalgae cultivation farms is to adequately characterize the hydrodynamic climate at potential growth sites. In regions like the Gulf of Mexico and the Caribbean, which are highly exposed to tropical cyclones, it is critical to not only consider the day-to-day hydrodynamic conditions but to also assess the risk of extreme sea states to ensure the survivability of future macroalgae farms. Under these considerations, the Pacific Northwest National Laboratory (PNNL) is providing modeling support for MBL to inform the design and siting of the farm systems that have been proposed for their ARPA-E MARINER project. This report describes the development of a high-resolution coupled storm surge and wave model to simulate the hydrodynamics and wave climate at proposed macroalgae cultivation sites selected by MBL in Florida and Puerto Rico. Model results, including model validation, water level, current distributions, and sea states, are discussed for both selected sites in Florida and Puerto Rico coast. These simulations provide an accurate insight into the hydrodynamic conditions that a macroalgae farm is likely to experience during its operational lifetime, including current information to support fine-scale hydrodynamic load modeling, risk analysis and system design.

09 BIOMASS FUELS↗

DOE OSTI2025

Porous carbon from lignocellulosic biomass with emphasis on corn plant waste residue for energy storage

The rising global demand for sustainable energy storage materials has driven the search for environmentally friendly and cost-effective electrode options. Hydrothermal conversion of lignocellulosic biomass has gained attention due to its low energy requirements and operation at relatively low temperatures, presenting a green alternative to traditional thermochemical methods. The resulting solid product, hydrochar, has been used as an adsorbent and soil amendment; however, chemical/thermal treatment significantly enhances its physical properties. These structural modifications transform hydrochar into an effective porous carbon electrode, offering abundant sites for electrolyte ion transport, critical for high-performance devices like supercapacitors and batteries. This review first discusses various waste biomass and sustainable feedstocks available globally. It compares two primary thermochemical conversion techniques, pyrolysis and hydrothermal carbonization/liquefaction, and examines their respective solid products, biochar and hydrochar, analyzing differences in their physical and chemical characteristics. The focus is placed on hydrochar, summarizing activation methods to produce porous carbon suitable for energy storage applications. Additionally, this review will include a dedicated section on the application of porous carbon derived from corn plant waste residue, considering that corn is one of the most abundant crops grown worldwide, which makes it an important and promising source for sustainable porous carbon production. The role of machine learning models in optimizing hydrothermal processes to produce high-quality hydrochar is also discussed, emphasizing how data-driven approaches can streamline process development. Finally, the review identifies the current challenges and prospects for lignocellulosic biomass-derived porous carbon as a sustainable electrode material in next-generation energy storage technologies.

25 ENERGY STORAGE↗

DOE OSTI2022

Excessive Aurantiochytrium acetophilum docosahexaenoic acid supplementation decreases growth performance and breast muscle mass of broiler chickens

We report Docosahexaenoic acid (DHA) is an n-3 polyunsaturated fatty acid with health-promoting potential. This study was to investigate effects of supplemental DHA from Aurantiochytrium acetophilum on growth performance, health status, meat quality, and protein synthesis signaling of broiler chickens. Day-old male chicks were housed in an environmental control room (6 cages/treatment, 8 chicks/cage), and fed a corn-soybean meal basal diet supplemented with the DHA-rich A. acetophilum biomass (Heliae, Gilbert, AZ) at 0, 1, 2, and 4% (0, 1.7, 3.4 and 6.8 g DHA/kg diet) for 6 weeks. Growth performance was measured weekly. Blood samples were collected at weeks 3 and 6 (2 chicks/cage). Four tissues were sampled (2 chicks/cage) for biochemical and meat quality analyses. Data were analyzed by one-way ANOVA and regression. Compared with the control, the 4% A. acetophilum diet decreased (p < 0.05) body weight gain (19%) and gain to feed ratio (19%) during weeks 4–6. The A. acetophilum supplementation dose-dependently decreased (p < 0.05, R 2 = 0.21–0.54) plasma alanine amino transferase activity and glucose concentrations, but had little effect on plasma activity of alkaline phosphatase or concentrations of inorganic phosphorus and uric acid at weeks 3 and 6. Compared with the control, the 4% A. acetophilum diet decreased (p < 0.05) breast muscle weight by 21%, and down-regulated (p < 0.05) mRNA levels of mammalian target of rapamycin and ribosomal s6 protein (S6), and protein levels of phosphorylated S6 to S6 and phosphorylated S6 kinase beta 1 to S6 kinase beta 1. The A. acetophilum supplementation linearly increased (p < 0.01) lipid peroxidation (R 2 = 0.62–0.90) and hardness and chewiness (R 2 = 0.34–0.44) of breast and thigh muscles. In conclusion, supplemental 4% (6.8 g DHA/kg), but not 1 or 2%, of A. acetophilum impaired growth performance, breast muscle mass accumulation, and(or) protein synthesis signaling of broilers.

59 BASIC BIOLOGICAL SCIENCES↗

DOE OSTI2021

Multi-Omics Driven Metabolic Network Reconstruction and Analysis of Lignocellulosic Carbon Utilization in Rhodosporidium toruloides

An oleaginous yeast Rhodosporidium toruloides is a promising host for converting lignocellulosic biomass to bioproducts and biofuels. In this work, we performed multi-omics analysis of lignocellulosic carbon utilization in R. toruloides and reconstructed the genome-scale metabolic network of R. toruloides . High-quality metabolic network models for model organisms and orthologous protein mapping were used to build a draft metabolic network reconstruction. The reconstruction was manually curated to build a metabolic model using functional annotation and multi-omics data including transcriptomics, proteomics, metabolomics, and RB-TDNA sequencing. The multi-omics data and metabolic model were used to investigate R. toruloides metabolism including lipid accumulation and lignocellulosic carbon utilization. The developed metabolic model was validated against high-throughput growth phenotyping and gene fitness data, and further refined to resolve the inconsistencies between prediction and data. We believe that this is the most complete and accurate metabolic network model available for R. toruloides to date.

09 BIOMASS FUELS↗

DOE OSTI2022

Particle scale impacts on deconstruction energy of pine residues

The goal of this Case Study was to quantify the impacts of variable moisture and ash on hammer mill throughput and energy consumption and on generation of fines that are not able to be fed to conversion, as compared to a status quo Base Case system. Also considered was convertible carbon content (minimum carbon specification) and maximum ash content and the delivered feedstock cost impacts of not being able to feed residue not meeting both specifications to the conversion reactor. Laboratory data on the impacts of input particle size and moisture content on the exit particle size were received from FCIC Subtask 5.2 from their single particle impact population balance modeling study. Additional throughput and energy consumption data were obtained from FCIC Subtask 5.2 for the same grinder with a 6 mm screen in place. These data were utilized to develop the necessary response surface equations to perform throughput analysis using discrete event simulation. Because the ash contents in the separated fines had not been analyzed in the laboratory at the time of the model runs, we chose to assume that the ash distributed proportionally with total mass into the overs and unders in the disk screen following grinding. Key takeaways from this Case Study are that it is significantly more cost effective to hammer mill the residue prior to drying, even though the grinder throughput is lower and energy consumption is higher versus drying first before grinding. An effect of dry grinding versus high moisture grinding is the production of higher amounts of fines during dry grinding, leading to significantly more of the ground feedstock being rejected by conversion for being below a minimum particle size. With wet grinding the system is still able to produce more preprocessed feedstock meeting the minimum particle size specification even though the instantaneous throughput is lower than for the case of grinding dry feedstock. Additionally, even without the higher fines production from dry grinding, the status quo would still be more costly than wet grinding because the material is rejected after the drying energy has already been input for the dry grinding case. Finally, significant reductions in drying energy are obtained by drying after grinding, and those reductions are of far greater magnitude than the grinding energy increase.

09 BIOMASS FUELS↗

NASA NTRS2010

Biomass Burning Emissions from Fire Remote Sensing

Knowledge of the emission source strengths of different (particulate and gaseous) atmospheric constituents is one of the principal ingredients upon which the modeling and forecasting of their distribution and impacts depend. Biomass burning emissions are complex and difficult to quantify. However, satellite remote sensing is providing us tremendous opportunities to measure the fire radiative energy (FRE) release rate or power (FRP), which has a direct relationship with the rates of biomass consumption and emissions of major smoke constituents. In this presentation, we will show how the satellite measurement of FRP is facilitating the quantitative characterization of biomass burning and smoke emission rates, and the implications of this unique capability for improving our understanding of smoke impacts on air quality, weather, and climate. We will also discuss some of the challenges and uncertainties associated with satellite measurement of FRP and how they are being addressed.

Ichoku, Charles↗

DOE OSTI2021

Field evaluation of transgenic hybrid poplars with desirable wood properties and enhanced growth for biofuel production by bicistronic expression of PdGA20ox1 and PtrMYB3 in wood-forming tissue

Background: To create an ideotype woody bioenergy crop with desirable growth and biomass properties, we utilized the viral 2A-meidated bicistronic expression strategy to express both PtrMYB3 ( MYB46 ortholog of Populus trichocarpa , a master regulator of secondary wall biosynthesis) and PdGA20ox1 (a GA20-oxidase from Pinus densiflora that produces gibberellins) in wood-forming tissue (i.e., developing xylem). Results: Transgenic Arabidopsis plants expressing the gene construct DX15::PdGA20ox1-2A-PtrMYB3 showed a significant increase in both stem fresh weight (threefold) and secondary wall thickening (1.27-fold) relative to wild-type (WT) plants. Transgenic poplars harboring the same gene construct grown in a greenhouse for 60 days had a stem fresh weight up to 2.6-fold greater than that of WT plants. In a living modified organism (LMO) field test conducted for 3 months of active growing season, the stem height and diameter growth of the transgenic poplars were 1.7- and 1.6-fold higher than those of WT plants, respectively, with minimal adverse growth defects. Although no significant changes in secondary wall thickening of the stem tissue of the transgenic poplars were observed, cellulose content was increased up to 14.4 wt% compared to WT, resulting in improved saccharification efficiency of the transgenic poplars. Moreover, enhanced woody biomass production by the transgenic poplars was further validated by re-planting in the same LMO field for additional two growing seasons. Conclusions: Taken together, these results show considerably enhanced wood formation of our transgenic poplars, with improved wood quality for biofuel production.

09 BIOMASS FUELS↗

DOE OSTI2021

Developing Cost Effective Biological Removal Technology for Selenium and Nitrate from Flue Gas Desulfurization Wastewater from an Existing Power Facility

Funding from this project supported new research in environmental biotechnology at WVSU Energy and Environmental Science Institute (WVSUEESI) to develop biological treatment techniques for FGD wastewater using photosynthetic organisms algae and plants. We tested the possibility of using algae and plants in treating FGD wastewater. Results suggested that algae and plants could able to perform normal photosynthesis under low concentrations of selenium. However, higher concentrations of selenium are lethal on photosynthesis. The total protein data indicated that FGD wastewater induced the accumulation of complete proteins in the duckweeds compared to control plants. It appears that nitrate, nitrates, and other compounds in FGD wastewater promote total protein accumulation. The FGD wastewater treatment has triggered Triacylglycerols (TAG) and carbohydrate accumulation in duckweed. We studied the differential gene expression using RNA-Sequencing data. Gene expression analysis revealed that glycolysis, fatty acid, triacylglycerol biosynthesis, and starch degradation were up-regulated; on the contrary, gluconeogenesis, photosynthesis, and starch biosynthesis genes were down-regulated by FGD wastewater treatment. We confirmed the expression of some of these genes using q-RT PCR analysis. We demonstrated that when FGD gypsum was blended with the coal mine soil to grow the bioenergy crop, Camelina, it could grow and produce seed. Moreover, Camelina is a non-edible oilseed crop and has heavy metal sequestering properties, making it the right reclamation choice. Besides, this project provided the opportunity for WVSU undergraduates to integrate hands-on research and education. Funding from this grant helped us to enhance student hands-on experience in water quality and environmental science and participation in STEM research and education.

09 BIOMASS FUELS↗

DOE OSTI2022

Techno-economic assessment for the production of algal fuels and value-added products: opportunities for high-protein microalgae conversion

Abstract Background Microalgae possess numerous advantages for use as a feedstock in producing renewable fuels and products, with techno-economic analysis (TEA) frequently used to highlight the economic potential and technical challenges of utilizing this biomass in a biorefinery context. However, many historical TEA studies have focused on the conversion of biomass with elevated levels of carbohydrates and lipids and lower levels of protein, incurring substantial burdens on the ability to achieve high cultivation productivity rates relative to nutrient-replete, high-protein biomass. Given a strong dependence of algal biomass production costs on cultivation productivity, further TEA assessment is needed to understand the economic potential for utilizing potentially lower-cost but lower-quality, high-protein microalgae for biorefinery conversion. Results In this work, we conduct rigorous TEA modeling to assess the economic viability of two conceptual technology pathways for processing proteinaceous algae into a suite of fuels and products. One approach, termed mild oxidative treatment and upgrading (MOTU), makes use of a series of thermo-catalytic operations to upgrade solubilized proteins and carbohydrates to hydrocarbon fuels, while another alternative focuses on the biological conversion of those substrates to oxygenated fuels in the form of mixed alcohols (MA). Both pathways rely on the production of polyurethanes from unsaturated fatty acids and valorization of unconverted solids for use as a material for synthesizing bioplastics. The assessment found similar, albeit slightly higher fuel yields and lower costs for the MA pathway, translating to a residual solids selling price of $899/ton for MA versus $1033/ton for MOTU as would be required to support a $2.50/gallon gasoline equivalent (GGE) fuel selling price. A variation of the MA pathway including subsequent upgrading of the mixed alcohols to hydrocarbon fuels (MAU) reflected a required solids selling price of $975/ton. Conclusion The slight advantages observed for the MA pathway are partially attributed to a boundary that stops at oxygenated fuels versus fungible drop-in hydrocarbon fuels through a more complex MOTU configuration, with more comparable results obtained for the MAU scenario. In either case, it was shown that an integrated algal biorefinery can be economical through optimal strategies to utilize and valorize all fractions of the biomass.

09 BIOMASS FUELS↗