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DOE OSTI2024

2023 Billion-Ton Report: An Assessment of U.S. Renewable Carbon Resources - Microalgae

Microalgae is a unique biomass resource that does not need to compete for land and water with other biomass feedstocks because it can be cultivated on low-quality unencumbered land using noncompetitive water types including saline and wastewater. It is included as a complementary resource alongside other biomass feedstocks reported in this study, albeit at higher biomass production costs reflective of more capital-intensive farming operations than typical for terrestrial biomass. Higher biomass costs can be offset by the potential to produce value-added coproducts unique to compositional constituents of microalgae. Relative to the 2016 Billion-Ton Report, this chapter reflects the latest analysis from the 2022 Algae Harmonization Update, which uses the latest parameterized and high-performing saline algal strain, second-generation carbon capture of point-source waste CO 2 , and high-pressure pipeline transport resolved to specific point-source types, saline water sourcing up to 40,000 mg/L total dissolved solids for source and makeup water salinity, blowdown water treatment and recycle, and brine disposal handling. National-scale algal biomass availability potential was calculated at 152 million tons/yr ash-free dry weight (AFDW) (191 million tons/yr dry weight) at an average biomass

09 BIOMASS FUELS↗

DOE OSTI2025

Soil fertility management for sustainable Miscanthus × giganteus production: Increased tiller weight from nitrogen management explains yield gains in aged miscanthus

Aging-related yield decline in Miscanthus × giganteus (miscanthus) remains a major constraint to sustainable biomass production. This study evaluated how nitrogen (N) management and soil fertility influence yield-component traits and productivity in aging miscanthus. Trials were conducted at two sites established in 2008 at the University of Illinois Energy Farm, Urbana, IL. (i) The Sun Grant trial received 0, 60, and 120 kg N ha −1 annually until 2015. Starting 2021, half of each plot received 60 or 120 kg N ha −1 , resulting in six legacy-contemporary treatments: 0N–0N, 0N–120N, 60N–0N, 60N–60N, 120N–0N, 120N–120N. (ii) The Energy Farm trial remained unfertilized until 2014, when one half of each plot received 56 kg N ha −1 , forming two treatments: 0N–0N, 0N–56N. Sun Grant trial results showed N fertilization increased tiller density (tillers m −2 ) and tiller weight (g tiller −1 ) in juvenile to early-mature miscanthus (2011–2015). After N withdrawal, both traits declined (20 % and 40 %), though legacy effects persisted in tiller weight in the aging stands (2020–2023). Contemporary N had little effect on tiller density but increased tiller weight by 34 %–77 %, resulting in 23 %–106 % higher machine-harvested biomass yield in 0–120N, 60-60N, and 120-120N plots. At the Energy Farm trial, 0N–56N plots yielded 59 %–108 % more biomass than 0N–0N. Soil total N increased (Sun Grant: 47 % by 2020; Energy Farm: 58 % by 2023), while Mehlich-3 P (42 %–44 %) and K (21 %–46 %) declined. These findings identify tiller weight as a key determinant of biomass yield in aging miscanthus and highlight the need for P and K management for long-term productivity.

09 BIOMASS FUELS↗

DOE OSTI2026

Biomolecular Films for Direct Air Capture of CO 2

Efficient utilization of CO 2 is amongst the most critical cost drivers in algal biomass production in open pond systems. CO 2 delivery costs represent approximately 20% of the final biomass selling price in algal mass cultivation systems. Technologies that enable direct air capture (DAC) of atmospheric CO 2 to decouple algae cultivation from CO 2 point sources thus present an opportunity to improve the economics and resource potential of algal biomass. Current DAC technologies typically employ amine- or caustic-based absorption, demanding significant water and/or energy inputs and incurring substantial capital expenditures. Conversely, bio-based approaches to DAC offer a means to bypass conventional technoeconomic and sustainability hurdles. We integrate recent advances in computational metabolic modeling, algal genetic engineering, algal cultivation, and algal biomass upgrading to enable directed localization and self-assembly of carbonic anhydrase molecular films to gas-liquid interfaces for enhanced CO 2 capture and conversion.

09 BIOMASS FUELS↗

DOE OSTI2025

Data for Soil Fertility Management for Sustainable Miscanthus × giganteus Production: Increased Tiller Weight from Nitrogen Management Explains Yield Gains in Aged Miscanthus

Aging-related yield decline in Miscanthus × giganteus (miscanthus) remains a major constraint to sustainable biomass production. This study evaluated how nitrogen (N) management and soil fertility influence yield-component traits and productivity in aging miscanthus. Trials were conducted at two sites established in 2008 at the University of Illinois Energy Farm, Urbana, IL. (i) The Sun Grant trial received 0, 60, and 120 kg N ha−1 annually until 2015. Starting 2021, half of each plot received 60 or 120 kg N ha−1, resulting in six legacy-contemporary treatments: 0N–0N, 0N–120N, 60N–0N, 60N–60N, 120N–0N, 120N–120N. (ii) The Energy Farm trial remained unfertilized until 2014, when one half of each plot received 56 kg N ha−1, forming two treatments: 0N–0N, 0N–56N. Sun Grant trial results showed N fertilization increased tiller density (tillers m−2) and tiller weight (g tiller−1) in juvenile to early-mature miscanthus (2011–2015). After N withdrawal, both traits declined (20 % and 40 %), though legacy effects persisted in tiller weight in the aging stands (2020–2023). Contemporary N had little effect on tiller density but increased tiller weight by 34 %–77 %, resulting in 23 %–106 % higher machine-harvested biomass yield in 0–120N, 60-60N, and 120-120N plots. At the Energy Farm trial, 0N–56N plots yielded 59 %–108 % more biomass than 0N–0N. Soil total N increased (Sun Grant: 47 % by 2020; Energy Farm: 58 % by 2023), while Mehlich-3 P (42 %–44 %) and K (21 %–46 %) declined. These findings identify tiller weight as a key determinant of biomass yield in aging miscanthus and highlight the need for P and K management for long-term productivity.

miscanthus↗

DOE OSTI2026

Microalgae Commodities Production with a Direct Air Capture Process (Final Technical Report)

This report documents the work performed under Award Number DE-EE0009276, "Microalgae Commodities Production with a Direct Air Capture Process," to advance the technology for large-scale, low-cost microalgae biomass production in the United States. The project's overarching goal was to decouple microalgae production from enriched CO 2 point sources, such power plant flue gases, which otherwise limit the national resource potential for algal biomass.

09 BIOMASS FUELS↗

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 OSTI2020

The Economic Accessibility of CO2 Sequestration through Bioenergy with Carbon Capture and Storage (BECCS) in the US

Bioenergy with carbon capture and storage (BECCS) is one strategy to remove CO2 from the atmosphere. To assess the potential scale and cost of CO2 sequestration from BECCS in the US, this analysis models carbon sequestration net of supply chain emissions and costs of biomass production, delivery, power generation, and CO2 capture and sequestration in saline formations. The analysis includes two biomass supply scenarios (near-term and long-term), two biomass logistics scenarios (conventional and pelletized), and two generation technologies (pulverized combustion and integrated gasification combined cycle). Results show marginal cost per tonne CO2 (accounting for costs of electricity and CO2 emissions of reference power generation scenarios) as a function of CO2 sequestered (simulating capture of up to 90% of total CO2 sequestration potential) and associated spatial distribution of resources and generation locations for the array of scenario options. Under a near-term scenario using up to 206 million tonnes per year of biomass, up to 181 million tonnes CO2 can be sequestered annually at scenario-average costs ranging from $62 to $137 per tonne CO2; under a long-term scenario using up to 740 million tonnes per year of biomass, up to 737 million tonnes CO2 can be sequestered annually at scenario-average costs ranging from $42 to $92 per tonne CO2. These estimates of CO2 sequestration potential may be reduced if future competing demand reduces resource availability or may be increased if displaced emissions from conventional power sources are included. Results suggest there are large-scale opportunities to implement BECCS at moderate cost in the US, particularly in the Midwest, Plains States, and Texas.

09 BIOMASS FUELS↗

NASA NTRS1990

Use of inedible wheat residues from the KSC-CELSS breadboard facility for production of fungal cellulase

Cellulose and xylan (a hemicellulose) comprise 50 percent of inedible wheat residue (which is 60 percent of total wheat biomass) produced in the Kennedy Space Center Closed Ecological Life Support System (CELSS) Breadboard Biomass Production Chamber (BPC). These polysaccharides can be converted by enzymatic hydrolysis into useful monosaccharides, thus maximizing the use of BPC volume and energy, and minimizing waste material to be treated. The evaluation of CELSS-derived wheat residues for production for cellulase enzyme complex by Trichoderma reesei and supplemental beta-glucosidase by Aspergillus phoenicis is in progress. Results to date are given.

Strayer, R. F.↗

NASA NTRS2016

Combining Livestock Production Information in a Process-Based Vegetation Model to Reconstruct the History of Grassland Management

Grassland management type (grazed or mown) and intensity (intensive or extensive) play a crucial role in the greenhouse gas balance and surface energy budget of this biome, both at field scale and at large spatial scale. However, global gridded historical information on grassland management intensity is not available. Combining modelled grass-biomass productivity with statistics of the grass-biomass demand by livestock, we reconstruct gridded maps of grassland management intensity from 1901 to 2012. These maps include the minimum area of managed vs. maximum area of unmanaged grasslands and the fraction of mown vs. grazed area at a resolution of 0.5deg by 0.5deg. The grass-biomass demand is derived from a livestock dataset for 2000, extended to cover the period 19012012. The grass-biomass supply (i.e. forage grass from mown grassland and biomass grazed) is simulated by the process-based model ORCHIDEE-GM driven by historical climate change, risingCO2 concentration, and changes in nitrogen fertilization. The global area of managed grassland obtained in this study increases from 6.1 x 10(exp 6) km(exp 2) in 1901 to 12.3 x 10(exp 6) kmI(exp 2) in 2000, although the expansion pathway varies between different regions. ORCHIDEE-GM also simulated augmentation in global mean productivity and herbage-use efficiency over managed grassland during the 20th century, indicating a general intensification of grassland management at global scale but with regional differences. The gridded grassland management intensity maps are model dependent because they depend on modelled productivity. Thus specific attention was given to the evaluation of modelled productivity against a series of observations from site-level net primary productivity (NPP) measurements to two global satellite products of gross primary productivity (GPP) (MODIS-GPP and SIF data). Generally, ORCHIDEE-GM captures the spatial pattern, seasonal cycle, and inter-annual variability of grassland productivity at global scale well and thus is appropriate for global applications presented here.

Livestock dataset↗

DOE OSTI2021

Understanding the dynamics of vacuole trafficking and secretion for enhanced plant storage reserves

Vacuoles are essential in plants and serve roles in both protein and organelle degradation for reutilization and storage of high energy molecules such as storage proteins which are critical for seed germination under normal and stress conditions and impact biomass production and nutrition. In addition, plants secrete many different molecules such as cell wall components, lipids, waxes and others that are important for plant protection, structure and biomass. Vacuole accumulation and secretion are closely linked and highly regulated pathways within the endomembrane trafficking network. We have discovered many drug-like synthetic small molecules that can suppress vacuole trafficking and enhance secretion; other chemicals can enhance vacuole trafficking and suppress secretion. The ability of our compounds to modulate these pathways indicates that these pathways are tightly coupled and highly regulated. Thus, to achieve the goals of enhanced energy reserves or enhanced secretion it is essential to understand how this dynamic is controlled. Bioactive chemicals have provided us unique insight into natural mechanisms controlling the dynamics of vacuole trafficking and secretion that was not possible previously. One such compound, ES2, negatively modulates secretion while enhancing vacuole trafficking. This is the first compound discovered that targets an essential secretory complex known as the exocyst and will reveal important regulatory domains controlling secretion and vacuole trafficking. During the current funding period, we determined the structure of the EXO70A1 in plants and identifying its binding site. To understand the dynamics of vacuole trafficking it is key to have compounds that inhibit vacuole trafficking as well as enhancing it (ES2). We have used sophisticated image analysis to identify another novel compound ES17 that enhances secretion while negatively modulating vacuole trafficking (opposite of ES2). We have already made rapid progress. Our preliminary data indicates that ES17 very specifically targets a late stage of vesicle fusion with the tonoplast which is highly regulated. Thus, ES17 is an ideal compound to dissect the regulation of vacuole targeting dynamics. In addition, we have now synthesized a biotinylated version of ES17 that is fully functional, permitting the use of chemical proteomics to identify the cognate target with a high probability of success. We propose to investigate the ES17 target which can negatively modulate vacuole trafficking and combine this with knowledge of the ES2 target which can promote vacuole trafficking. Dissecting the mechanisms controlling the dynamics of secretion vs vacuole accumulation and the use of synthetic modulators will permit us to understand and regulate trafficking to enhance secretion to the plasma membrane and cell wall or the accumulation of biological energy reserves for increased biomass, nutritional value, stress resistance, and the secretion of specialized plant products.

59 BASIC BIOLOGICAL SCIENCES↗

DOE OSTI2024

Nitrogen fertilization effects on aged Miscanthus × giganteus stands: Exploring biomass yield, yield components, and biomass prediction using in–season morphological traits

For sustainable biomass production of Miscanthus × giganteus (hereafter miscanthus), understanding the impact of stand age and nitrogen (N) fertilization on biomass yield is crucial. This study investigated the effects of varying N fertilization rates (0, 56, 112, and 168 kg N ha –1 ) on yield components (tiller height, density, and weight) and their correlations with end-of-season biomass yield in miscanthus. We also explored end-of-season biomass yield prediction using in-season traits (canopy height, leaf area index, and leaf chlorophyll content [LCC]). The study was conducted at two sites in Illinois: a previously unfertilized 10-year-old miscanthus research stand at Urbana and a 16-year-old commercial stand at Pesotum with a history of annual 56N application. Results from 2018 to 2021 in Urbana and 2020 to 2021 in Pesotum showed increased biomass yields with N fertilization, varying by rate, year, and location. Biomass yield in Pesotum peaked at 56N, while in Urbana, it increased significantly at 112 kg N ha –1 . Biomass yield was strongly correlated with tiller height and weight measured at Urbana across N rates. Morphological traits measured every 2–3 weeks during the 2020 and 2021 growing seasons showed that canopy height was the strongest single predictor of miscanthus biomass yield, followed by LCC. Mid-August to September measurements of these traits were the best predictors of biomass yield. Multiple regressions involving the canopy height and LCC further improved yield predictions. We conclude that while N enhances biomass yields of aging miscanthus, the optimum rate depends on the site, environmental conditions, and management history.

59 BASIC BIOLOGICAL SCIENCES↗

DOE OSTI2024

Data from: Biomass yield, yield components and growing season phenotypic measurements of Miscanthus

For sustainable biomass production of Miscanthus × giganteus (hereafter miscanthus), understanding the impact of stand age and nitrogen (N) fertilization on biomass yield is crucial. This study investigated the effects of varying N fertilization rates (0, 56, 112, and 168 kg N ha-1) on yield components (tiller height, density, and weight) and their correlations with end-of-season biomass yield in miscanthus. We also explored end-of-season biomass yield prediction using in-season traits (canopy height, leaf area index (LAI), and leaf chlorophyll content (LCC)). The study was conducted at two sites in Illinois: a previously unfertilized 10-year-old miscanthus research stand at Urbana and a 16-year-old commercial stand at Pesotum with a history of annual 56N application. Results from 2018-2021 in Urbana and 2020-2021 in Pesotum showed increased biomass yields with N fertilization, varying by rate, year, and location. Biomass yield in Pesotum peaked at 56N, while in Urbana, it increased significantly at 112 kg N ha-1. Biomass yield was strongly correlated with tiller height and weight measured at Urbana across N rates. Morphological traits measured every 2-3 weeks during the 2020 and 2021 growing seasons showed that canopy height was the strongest single predictor of miscanthus biomass yield, followed by LCC. Mid-August to September measurements of these traits were the best predictors of biomass yield. Multiple regressions involving the canopy height and LCC further improved yield predictions. We conclude that while N enhances biomass yields of aging miscanthus, the optimum rate depends on the site, environmental conditions, and management.

Aging↗

DOE OSTI2025

Enhanced Production of Algae Lipids and Carbohydrates for Fuel and Polyurethane Precursors

This Final Technical Report summarizes the goals, approach, and outcomes of the project “Enhanced Production of Algae Lipids and Carbohydrates for Fuel and Polyurethane Precursors”. The project addressed the challenge of improving microalgae biomass productivity and simultaneously producing valuable polyurethane precursors (PUPs), from both lipids and carbohydrates, that can be converted into renewable biofuels and bio-based polyurethane (PU) products. By integrating advanced genetic engineering, traditional breeding, high-throughput screening, pilot cultivation, and chemical conversion technologies, the team achieved significant advancements in algae biotechnology.

09 BIOMASS FUELS↗

DOE OSTI2020

Biochemical upgrading of high-protein biomass and grain products

The present invention relates to methods of upgrading biomass to provide useful chemical intermediates, fuels, amino acids, nutrients, etc. In particular examples, the biomass is a by-product of ethanol production and is mainly used as high-protein feed. Described herein are methods for upgrading such biomass, such as by implementing pre-treatment conditions and by employing fermentation conditions including modified organisms.

09 BIOMASS FUELS↗

DOE OSTI2022

Biochemical upgrading of high-protein biomass and grain products

Davis, Ryan Wesley↗

DOE OSTI2026

Linking Plant and Microbial Traits to Soil Carbon for Reliable and Resilient Bioenergy Systems

Bioenergy systems in the United States offer a dual opportunity to supply renewable feedstocks while enhancing ecosystem services such as hydrologic regulation, erosion control, and soil carbon (C) storage. National assessments highlight the potential to grow perennial energy crops to improve soil function and ecosystem resilience. Realizing this potential requires understanding the ecological mechanisms that govern how C is added, transformed, and stabilized in soils. Plant traits determine the quantity, depth, and chemistry of organic inputs, while microbial processes—including carbon use efficiency, necromass formation, and trophic interactions—mediate their transformation and partitioning among soil carbon pools. These biological pathways are shaped by soil physical and chemical properties, including aggregation, texture, and mineralogy, and by environmental drivers such as temperature, moisture, and disturbance, leading to context-dependent outcomes across landscapes. Management practices that diversify feedstocks, minimize disturbance, and maintain soil cover can promote both biomass production and C retention, while microbial amendments and rhizosphere engineering offer emerging, but often context-dependent, tools to optimize plant–microbe interactions. Trade-offs between biomass yield and soil carbon storage may arise when systems favor rapid aboveground productivity at the expense of belowground inputs and microbial processing, underscoring the importance of trait combinations that support both functions. Advances in monitoring, reporting, and verification—spanning precision agriculture, remote sensing, and biosensing—are improving predictive capacity through microbial-explicit process models and model–experiment (ModEx) frameworks. By connecting soil, plant, and microbial processes with advances in modeling and biosensing, this review outlines research priorities focused on trait-based parameterization and ModEx integration. These priorities will support the design of bioenergy systems that are both reliable and resilient, enhancing renewable energy production and ecosystem sustainability.

bioenergy systems↗

NASA NTRS1987

Algal culture studies for CELSS

Microalgae are well-suited as a component of a Closed Environmental Life Support System (CELSS), since they can couple the closely related functions of food production and atmospheric regeneration. The objective was to provide a basis for predicting the response of CELSS algal cultures, and thus the food supply and air regeneration system, to changes in the culture parameters. Scenedesmus growth was measured as a function of light intensity, and the spectral dependence of light absorption by the algae as well as algal respiration in the light were determined as a function of cell concentration. These results were used to test and confirm a mathematical model that describes the productivity of an algal culture in terms of the competing processes of photosynthesis and respiration. The relationship of algal productivity to cell concentration was determined at different carbon dioxide concentrations, temperatures, and light intensities. The maximum productivity achieved by an air-grown culture was found to be within 10% of the computed maximum productivity, indicating that CO2 was very efficiently removed from the gas stream by the algal culture. Measurements of biomass productivity as a function of cell concentration at different light intensities indicated that both the productivity and efficiency of light utilization were greater at higher light intensities.

Radmer, R.↗

DOE OSTI2022

BETO 2021 Peer Review - 1.3.2.001 - Algae Biomass Composition

Addressing critical improvements in biomass productivity and associated biochemical composition is a priority for the economic and sustainable commercial development of biofuels and bioproducts from algae. Capitalizing on pathways that integrate engineering approaches with fundamental biochemistry of photosynthetic organisms will lead to a better understanding of the complex nexus of algae growth rates, productivity and composition. This project focuses on identifying the critical factors for economic development of fuel and bioproduct technologies. Algal compositional characteristics form the foundation of robust economic and business models. This project supports that foundation by developing and validating accurate compositional methods and disseminating them to the greater community. Simultaneously, we build a deep understanding of the dynamic biochemical composition and carbon allocation for biomass value and conversion yields. A co-product portfolio developed under this project demonstrated a 30% increase in intrinsic value. Additionally, an integrated pipeline of molecular diversity mapping for product discovery with quantitative demonstration across species was deployed over the BETO algae program. The advances made here are highly relevant to BETO's multi-year program targets of reducing costs and integrating dynamic biomass composition with conversion processes to provide options for bioproducts, all leveraging the molecular diversity of algae.

algal compositional characteristics↗