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Stepping into the Midwest Bioeconomy: Stakeholder Engagement and Geospatial Tools to Assist in Perennial Bioenergy Crop Decision Making and Entrepreneurship
This project, “Ecosystem Services and Farm Entrepreneurship Technical Assistance,” was a three-year project originally planned for FY22–FY24. Due to a late start and a few extensions, it is being completed in early FY25. This project explored opportunities to support the deployment of a bioeconomy with a circular, more sustainable supply chain. Using a tool developed by Argonne to identify agricultural areas suitable for use in the bioeconomy, we sought to create opportunities in the bioeconomy as biomass producers, bioenergy users, and environmental entrepreneurs. We proposed to focus at the beginning on enhancing the tool’s capabilities, while engaging with key stakeholders to improve and expand the tool’s functionality for all potential stakeholders in the bioeconomy. We believe that expanding our tools and technologies, coupled with conversations in agricultural spaces, will be needed as we continue to explore how best to offer farmers whole-of supply-chain opportunities to participate in the bioeconomy. Through this project we have continued to gain a better understanding of the ways in which farmers, landowners, bioenergy users, and environmental entrepreneurs may approach the bioeconomy. In addition, as we improve our analytic toolkit, we can continue to refine our communication and the ways in which we can valuate the bioeconomy. Refining these tools allows us to dive deeper into conversations around plausible policies and drivers for future bioeconomy investment and engagement by stakeholders. By working with farmers and agricultural landowners to enable a sustainable bioeconomy business model, enhance their energy options, and recover resources from their waste streams, this project directly responds to the Bioenergy Technology Office’s (BETO) priorities of building a resilient energy economy. It addresses BETO’s focus on fostering the development and adoption of energy technologies that enable the conversion of waste to energy, efficient land use, and robust job creation. By establishing a technical assistance program that develops capabilities and practices in agricultural areas to implement a bioeconomy future, this program will develop an important linkage between technology being developed at U.S. Department of Energy national laboratories and the agricultural communities of the Midwest. This project focuses on farmers with lower productivity farmland. Because less productive lands create a more difficult revenue stream for conventional crops, these farmers may therefore be more open to alternative agricultural land management regimes. Consequently, the technical assistance program and the methodologies for targeting perennial bioenergy crop application on marginal land provide a distinct opportunity to engage with and invest in the bioeconomy in these economically stressed areas. Stakeholders in this project include farmers and landowners, local conservation organizations (NRCS, SWCS, etc.), universities, non-profit environmental and agricultural entities, farm consultants, environmental regulators, and industry, including the industries working on conversion technologies, anaerobic digestion, pyrolysis, and biochar generation, and the companies interested in trading or purchasing/supporting the valuation of ecosystem services (ES).
Long‐term yields in annual and perennial bioenergy crops in the Midwestern United States
Abstract Many yield predictions in perennial bioenergy species have been made based on data collected during the establishment phase of growth or a limited number of long‐term studies. Few studies compare multiple perennial crops with the dominant agricultural vegetation of the landscape over long time periods. Here, we present the results of 11 years of perennial crop management on fertile agricultural soils in central Illinois, compared with conventional row crop maize/soybean ( Zea mays L., Glycine max L.) production. We examined the long‐term productivity and drought susceptibility of Miscanthus x giganteus Greef et. Deu. ex. Hodkinson et Renvoize (miscanthus), Panicum virgatum L., Cave‐in‐Rock cultivar (switchgrass), and a native prairie mix, in contrast to annual maize/soybean agriculture. Long‐term yields for miscanthus and switchgrass failed to reach initial predictions made during the establishment phase; however, in miscanthus, the 11 th year of production shows little progressive yield loss with age, exceeding the modeled limit for the onset of age‐related decline. Harvest timing and differences in yields from hand and machine harvests in perennial crops likely contribute to overestimates of potential yields. Application of fertilizer to mature miscanthus resulted in significant increases in yield after a severe drought, though modeled effects of management and drought in miscanthus point to a more complex mechanism for yield response.
Frontiers and opportunities in bioenergy crop microbiome research networks
Researchers from across the four U.S. Department of Energy Bioenergy Research Centers engaged in a microbiome workshop that focused on identifying challenges and collaboration opportunities to better understand bioenergy-relevant plant–microbe interactions. The virtual workshop included hands-on educational sessions and a keynote address on current best practices in microbiome science and community microbiome standards, as well as breakout sessions aimed at identifying microbiome-related data and measurements that should be prioritized, opportunities for and barriers to integrating plant metabolites to microbiome research, and strategies for more effectively integrating microbiome data and processes into existing models. Based on participant discussion, key findings of the workshop were the need to prioritize scaling data sharing across BRCs and the broader research community and securing collaborative infrastructure in the areas of microbiome-ecosystem modeling and molecular plant-microbe interactions. This workshop review highlights additional main findings from this event, to encourage cross-site and more holistic meta-analyses while promoting wide scientific community engagement across plant microbiome sciences.
Soil pore characteristics and the fate of new switchgrass-derived carbon in switchgrass and prairie bioenergy cropping systems
Abstract Monoculture switchgrass and restored prairie are promising perennial feedstock sources for bioenergy production on the lands unsuitable for conventional agriculture. Such lands often display contrasting topography that influences soil characteristics and interactions between plant growth and soil C gains. This study aimed at elucidating the influences of topography and plant systems on the fate of C originated from switchgrass plants and on its relationships with soil pore characteristics. For that, switchgrass plants were grown in intact soil cores collected from two contrasting topographies, namely steep slopes and topographical depressions, in the fields in multi-year monoculture switchgrass and restored prairie vegetation. The 13 C pulse labeling allowed tracing the C of switchgrass origin, which X-ray computed micro-tomography enabled in-detail characterization of soil pore structure. In eroded slopes, the differences between the monoculture switchgrass and prairie in terms of total and microbial biomass C were greater than those in topographical depressions. While new switchgrass increased the CO 2 emission in depressions, it did not significantly affect the CO 2 emission in slopes. Pores of 18–90 µm Ø facilitated the accumulation of new C in soil, while > 150 µm Ø pores enhanced the mineralization of the new C. These findings suggest that polyculture prairie located in slopes can be particularly beneficial in facilitating soil C accrual and reduce C losses as CO 2 .
Application of Time-Domain 1 H NMR for Investigating Dynamics of Vegetative Lipids in Bioenergy Crops at Different Developmental Stages
Sweet sorghum is typically cultivated for the food and fodder market. Recently, sweet sorghum varieties are being metabolically transitioned to enhance energy density by accumulating oil droplets in their vegetative tissues for bioenergy applications. Owing to the high biomass yield of sorghum, the transgenic lines can compete with oil-seed crops for biodiesel yield per unit area. In the initial phase of transgenic development, a high-throughput phenotyping method can bridge the gap between the production pipeline and analysis to improve the efficiency of the process. To meet the requirement, the present study extends the application of time-domain 1 H-NMR spectroscopy for rapid quantification and characterization of the total in-situ lipids of sweet sorghum ‘ramada’ to lay the groundwork for analyzing the upcoming large quantity of transgenic samples. NMR technology has been successfully established for analyzing lipid contents of vegetative tissues of non-transgenic variety. The multiexponential analysis of spin-lattice (T 1 ) relaxation spectra obtained from TD-NMR aided the investigation of the dynamics of the free and bound lipid fraction with plant development. The total lipid concentration of bagasse and leaves of non-transgenic sweet sorghum remained unchanged throughout the plant development. Leaves displayed a higher percentage of bound lipids as compared to bagasse. Here, a significant variation in the lipid concentration of juice was observed at the different growth stages with a maximum lipid accumulation of 1.21 ± 0.04% w/w at the boot stage that decreased with further maturity of the plant.
Data for Application of Time-Domain 1H NMR for Investigating Dynamics of Vegetative Lipids in Bioenergy Crops at Different Developmental Stages
Sweet sorghum is typically cultivated for the food and fodder market. Recently, sweet sorghum varieties are being metabolically transitioned to enhance energy density by accumulating oil droplets in their vegetative tissues for bioenergy applications. Owing to the high biomass yield of sorghum, the transgenic lines can compete with oil-seed crops for biodiesel yield per unit area. In the initial phase of transgenic development, a high-throughput phenotyping method can bridge the gap between the production pipeline and analysis to improve the efficiency of the process. To meet the requirement, the present study extends the application of time-domain 1H-NMR spectroscopy for rapid quantification and characterization of the total in-situ lipids of sweet sorghum ‘ramada’ to lay the groundwork for analyzing the upcoming large quantity of transgenic samples. NMR technology has been successfully established for analyzing lipid contents of vegetative tissues of non-transgenic variety. The multiexponential analysis of spin-lattice (T1) relaxation spectra obtained from TD-NMR aided the investigation of the dynamics of the free and bound lipid fraction with plant development. The total lipid concentration of bagasse and leaves of non-transgenic sweet sorghum remained unchanged throughout the plant development. Leaves displayed a higher percentage of bound lipids as compared to bagasse. A significant variation in the lipid concentration of juice was observed at the different growth stages with a maximum lipid accumulation of 1.21 ± 0.04% w/w at the boot stage that decreased with further maturity of the plant.
A 13‐Year Record Indicates Differences in the Duration and Depth of Soil Carbon Accrual Among Potential Bioenergy Crops
Six years after replacing a maize/soybean cropping system, perennial grasses miscanthus (Miscanthus × giganteus) and switchgrass (Panicum virgatum), and a 28-species restored prairie increased particulate organic carbon in surface soils without increasing soil organic carbon (SOC). To resolve potential changes in the quantity and distribution of SOC, soils were resampled after seven to thirteen years to measure bulk density, carbon (C) content, and stable C isotopes to a depth of 1 m. SOC stocks increased between 1.75 and 2.5 Mg ha −1 year −1 in all perennial crops between 2008 and 2016 (nine growing seasons). Despite relatively low litter inputs and belowground biomass, the highest rate of SOC accrual was in restored prairie (2.5 Mg ha −1 year −1 ), followed by miscanthus (2.0 Mg ha −1 year −1 ) and switchgrass (1.75 Mg ha −1 year −1 ). The change in SOC in maize/soybean was not significant. After 2016, total SOC decreased in maize/soybean and miscanthus, resulting in slower overall rates of SOC accumulation over the full sampling period for miscanthus (0.8 Mg ha −1 year −1 ). The rate of SOC accumulation was greatest below 50 cm depth for restored prairie and switchgrass but in the top 10 cm for miscanthus. Stable isotope analysis showed 13 C enrichment in all depths of switchgrass soils, an indication of new organic C accumulation, but mixed results in all other crops. Planting perennial crops on land formerly in an annual maize/soybean cropping system can slow or reverse soil carbon losses, with the greatest increases in SOC from species-rich prairie.
Harnessing plant‐based platform for low‐cost cellulosic sugar recovery from bioenergy crops
The price of pure cellulase enzyme for the recovery of fermentable cellulosic sugars is one of the major challenges that limit the commercialization of second-generation biofuels and bio-based products. This work shows a means to greatly reduce the cost of cellulases. The abundant capacity of plants to synthesize and hyperaccumulate transgenic proteins in their plastids has been demonstrated for two gene constructs using either NPT II or Tet C to regulate the expression of bacterial cellulase Cel6A. Previously, the expression of Cel6A to 20–35% of total soluble leaf protein in tobacco was shown in two consecutive field trials. Moving forward, the key challenge was the extraction of active enzymes from the transgenic tobacco leaves hyperaccumulating bacterial cellulases in a cost-effective manner. The study showed that unpurified crude extracts of NPT II and Tet C transgenic tobacco leaves recovered ~23% and ~29% w/w cellulosic sugars from energycane bagasse, respectively. A supplementation of as low as 25% of purified commercial cellulase improved the glucose recovery by 2.9 times (~85% w/w) compared to 100% crude extract, which is comparable to the glucose recovery obtained by commercial cellulases, thereby suggesting a reduced requirement of commercial cellulases leading to a cost reduction of 75% for biorefineries. Assuming a stable 40% cellulase yield in total soluble protein under field conditions and multiple harvests (one to three) a year, the study estimates that the potential cost for saccharification of 1 t of lignocellulosic biomass can be reduced to 67–200 USD by using crude leaf extracts of transgenic tobacco.
Exploring the role of plant lysin motif receptor-like kinases in regulating plant-microbe interactions in the bioenergy crop Populus
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Multifunctional landscapes for dedicated bioenergy crops lead to low-carbon market-competitive biofuels
Switchgrass is a promising feedstock for cellulosic biorefineries, due to its ability to maintain comparatively high biomass yields across a wide range of soil and climatic conditions. However, there is an incomplete understanding of the economic and environmental tradeoffs associated with cultivating switchgrass on low-productivity land for conversion to biofuels. This study surveys prior literature and demonstrates a new integrated assessment framework, including agroecosystem, ecosystem services valuation, technoeconomic, and life-cycle assessment models, to quantify and contextualize the economic and environmental impacts of switchgrass cultivation on marginal land with downstream conversion to biofuels. Monetizing and incorporating the value of ecosystem services, such as improved water quality benefits from nitrate and sediment reductions, climate change mitigation benefits from CO 2 emission reduction, and recreational and pollination benefits from increased biodiversity, the modeled multifunctional landscape reduces the ethanol production cost by 33.3–58.9 cents/$\scriptsize{L}$-gasoline-equivalent ($\$$1.3–2.2/gge). Planting switchgrass in low productivity land improves soil health, resulting in the carbon footprint reduction credit of 12.8–20.2 gCO 2e /MJ. For an improved switchgrass-to-ethanol conversion pathway with the maximum benefits from ecosystem services, the minimum ethanol selling price and carbon footprint of ethanol, respectively, could reach to 31 cents/$\scriptsize{L}$-gasoline-equivalent (47% reduction relative to average gasoline price) and 3 gCO 2e /MJ (97% reduction relative to gasoline). In conclusion, this low carbon renewable ethanol leads to substantial State and/or Federal policy incentives (~$\$$1/$\scriptsize{L}$-gasoline-equivalent) providing a large benefit to biorefinery operators, farmers, and the public as a whole.
Effects of nitrogen fertilization and bioenergy crop type on spatial distributions of extracellular hydrolases associated with nitrogen and phosphorus acquisition
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Anti-fungal bioactive terpenoids in the bioenergy crop switchgrass (Panicum virgatum) may contribute to ecotype-specific microbiome composition
Abstract Plant derived bioactive small molecules have attracted attention of scientists across fundamental and applied scientific disciplines. We seek to understand the influence of these phytochemicals on rhizosphere and root-associated fungi. We hypothesize that – consistent with accumulating evidence that switchgrass genotype impacts microbiome assembly – differential terpenoid accumulation contributes to switchgrass ecotype-specific microbiome composition. An initial in vitro Petri plate-based disc diffusion screen of 18 switchgrass root derived fungal isolates revealed differential responses to upland- and lowland-isolated metabolites. To identify specific fungal growth-modulating metabolites, we tested fractions from root extracts on three ecologically important fungal isolates – Linnemania elongata, Trichoderma sp. and Fusarium sp. Saponins and diterpenoids were identified as the most prominent antifungal metabolites. Finally, analysis of liquid chromatography-purified terpenoids revealed fungal inhibition structure – activity relationships (SAR). Saponin antifungal activity was primarily determined by the number of sugar moieties – saponins glycosylated at a single core position were inhibitory whereas saponins glycosylated at two core positions were inactive. Saponin core hydroxylation and acetylation were also associated with reduced activity. Diterpenoid activity required the presence of an intact furan ring for strong fungal growth inhibition. These results inform future breeding and biotechnology strategies for crop protection with reduced pesticide application.
Selective designer hydrolysates from Miscanthus bioenergy crop for sustainable utilization
Designer enzymatic hydrolysates deliver targeted, optimized fermentable sugars while reducing the formation of inhibitory by-products. Herein, five distinct enzymatic hydrolysates (EH) were generated from Miscanthus biomass using alkaline-ethanol fractionation and hydrothermal-mechanical refining (HMR) pretreatment approaches. Noticeably, alkali-ethanol fractionated hemicellulose A (HMA) and hemicellulose B (HMB) hydrolysis resulted in xylose-rich enzymatic hydrolysates with 104.8 ± 5.6 g L −1 and 173.4 ± 4.6 g L −1 xylose, respectively. All five hydrolysates encompassing varied carbon to nitrogen ratios were evaluated for lipid production using an engineered oleaginous yeast, R. toruloides 880-ADS. Microbial fermentation of xylose-rich hydrolysates, viz. HMR-EH2, HMA-EH, and HMB-EH, resulted in lipid yields ranging from 0.05 to 0.12 g g −1 sugar. Compositional and 2D NMR analyses revealed that HMA-EH residues are highly lignin-enriched while preserving the non-condensed structure conducive to valorization. Overall, the findings highlighted a novel, selective trade-off approach for maximizing lipid titers from designer hydrolysates, corroborating complete resource recovery for establishing integrated biorefineries.
Stochastic and deterministic processes shape bioenergy crop microbiomes along a vertical soil niche
Abstract Sustainable biofuel cropping systems aim to address climate change while meeting energy needs. Understanding how soil and plant‐associated microbes respond to these different cropping systems is key to promoting agriculture sustainability and evaluating changes in ecosystem functions. Here, we leverage a long‐term biofuel cropping system field experiment to dissect soil and root microbiome changes across a soil‐depth gradient in poplar, restored prairie and switchgrass to understand their effects on the microbial communities. High throughput amplicon sequencing of the fungal internal transcribed spacer (ITS) and prokaryotic 16S DNA regions showed a common trend of root and soil microbial community richness decreasing and evenness increasing with depth. Ecological niche (root vs. soil) had the strongest effect on community structure, followed by depth, then crop. Stochastic processes dominated the structuring of fungal communities in deeper soil layers while operational taxonomic units (OTUs) in surface soil layers were more likely to co‐occur and to be enriched by plant hosts. Prokaryotic communities were dispersal limited at deeper depths. Microbial networks showed a higher density, connectedness, average degree and module size in deeper soils. We observed a decrease in fungal‐fungal links and an increase of bacteria–bacteria links with increasing depth in all crops, particularly in the root microbiome.
DayCent MUVP model calibrated for the simulation of bioenergy crops grown on set-aside land in the US
DayCent MUVP version (Methanogenesis, UV litter degradation and Photosynthesis). DAYCENT is the daily time-step version of the CENTURY biogeochemical model (Parton et al., 1994). DAYCENT simulates fluxes of C and N among the atmosphere, vegetation, and soil (Del Grosso et al., 2001a; Parton et al., 1998). Key submodels include soil water content and temperature by layer, plant production and allocation of net primary production (NPP), decomposition of litter and soil organic matter, mineralization of nutrients, N gas emissions from nitrification and denitrification, and CH4 oxidation in non-saturated soils.