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At least 181 records · Page 10
All sourcesNASA NTRSDOE OSTI
DOE OSTI2023

Improved Biofuel Production through Discovery and Engineering of Terpene Metabolism in Switchgrass

Project Objectives - Of the myriad specialized metabolites that plants deploy to adapt to environmental challenges, terpenes form the largest group. In many major crops, unique terpene blends serve as key stress defenses that directly impact plant fitness and yield. In addition, terpenes, such as bisabolene and pinene, are used for producing renewable biofuels. Essential to advancing a broader use of terpenes for biofuel feedstock engineering is a system-wide knowledge of the diverse biosynthetic machinery and defensive potential of often species-specific terpene blends. The proposed project would merge genome-wide enzyme discovery with comparative –omics, protein structural and plant microbiome studies to define the biosynthesis and stress-defensive functions of the switchgrass (Panicum virgatum) terpene network. These insights would be combined with developing and applying non-transgenic genome editing tools to design plants with desirable terpene blends for higher productivity and biofuel production on marginal lands. As a dedicated lignocellulosic feedstock for U.S. biofuel production with high net energy yield, stress tolerance, and available genome resources, switchgrass is well-suited for devising new avenues for biofuel production. Project Description – The diversity of plant terpene defenses is governed by species-specific families of terpene synthase (TPS) and cytochrome P450 monooxygenase (P450) enzymes. Mining of the switchgrass genome (genotype Alamo) identified ~100 TPS and P450 candidate genes, and combinatorial biochemical analysis of synthesized TPSs and P450s revealed more than a dozen enzymes with common and novel activities. In addition, several identified terpene metabolites and the corresponding transcripts were up-regulated in response to abiotic stressors. These findings demonstrate a unique switchgrass terpene network with probable importance to abiotic stress tolerance, thus providing a large chemical portfolio for optimizing crop resistance, yield, and biofuel composition. Leveraging these preliminary data, we propose to generate a genome-wide map of the switchgrass terpene metabolic network through multi-gene co-expression analyses that allow the efficient cross-validation of TPS and P450 functions. Key enzymes would further be applied to structure-function studies via X-ray protein crystallography, homology modeling and site-directed mutagenesis to gain mechanistic insight into the catalytic specificity of switchgrass terpene metabolism and provide gene and amino acid targets for genome editing. In tandem with terpene pathway discovery, system-wide metabolomics, transcriptomics and proteomics studies in switchgrass accessions of contrasting drought tolerance would define the role of switchgrass terpene metabolism in conferring abiotic stress resilience. Metabolic changes would be assessed in a combined approach of targeted (terpenes) and untargeted metabolite profiling using a high-resolution LC-MS/MS approach, differential gene expression analyses through multiplexed Illumina RNA sequencing, and quantitative analysis of high-priority pathway enzymes using multiple reaction monitoring (MRM). Drawing on these insights, knock-down/out mutants of stress-associated pathway nodes would be generated by optimizing transient virus-induced gene silencing (VIGS) and CRISPR/Cas9 systems under control of the Tobacco Rattle Virus (TRV). The resulting mutant lines would then be analyzed for stress susceptibility and the impact on the root microbiome to define gene functions in planta. Knowledge of terpene pathways, enzyme mechanisms and bioactivities would be applied to enhance switchgrass stress resilience and to tailor-make terpene blends for biofuel production. Here, TRV-enabled CRISPR/Cas9 genome editing, including allele-specific knock-out of redundant genes, engineering of enzyme specificity via structure-guided point mutations, and overexpression of terpene genes relevant to stress-protection or biofuel production, would be used to increase metabolic flux toward desired pathways. Broader Impacts - Integrating the system-wide discovery, mechanistic analysis and non-transgenic genome engineering of the switchgrass terpene network aligns the required steps to unlock the chemical potential of this important metabolite class to generate crops that are more resistant to stress and provide advanced biofuel production in light of rising climate pressures as foreseeable challenges for bioenergy crop cultivation. The proposed project would further offer interdisciplinary student training through active involvement in the project and integration of research concepts and outcomes into newly-developed graduate and undergraduate courses on Plant Biotechnology.

09 BIOMASS FUELS↗
DOE OSTI2025

Co-Firing Switchgrass and Waste Coal in A Power Plant: A Techno-Economic and Life Cycle Evaluation for The Ohio River Valley (SWITCH) (Final Technical Report for Ohio State/FE0032204)

Abandoned coal mine lands (AMLs) represent one of the most persistent environmental challenges in the United States. Prior to the enactment of the Surface Mining Control and Reclamation Act (SMCRA) in 1977, coal mining operations were not legally required to reclaim disturbed lands, leaving behind approximately 500,000 AML sites nationwide. These sites pose severe environmental and health risks, including acid mine drainage, soil and water contamination, and spontaneous combustion of waste coal piles. Millions of Americans live within one mile of these AMLs, underscoring the urgency of remediation. Traditional reclamation practices, such as planting cool-season grasses, often fail to fully restore ecological function or leverage the economic potential of these lands. This project addressed these challenges by developing integrated strategies for resource recovery, land reclamation, and sustainable energy production. This project evaluated an integrated strategy to convert this liability into an opportunity by recovering waste coal and co-firing it with switchgrass (Panicum virgatum L.) cultivated on reclaimed or marginal AML areas in existing coal-fired power plants. Switchgrass not only provides a renewable feedstock but also aids in land reclamation and carbon sequestration. 1) Remote Sensing and Machine Learning for Waste Coal Identification Using Sentinel-2 satellite imagery and supervised classification, we applied four machine learning models to detect historical waste coal piles. Random Forest achieved the highest accuracy (precision: 86%, recall: 77%). Time-series analysis revealed gradual vegetation recovery since 1986, indicating natural reclamation processes in historical sites, while active mining areas showed ongoing disturbance. This workflow enables scalable monitoring and prioritization of reclamation efforts. 2) UAS-Based Stockpile Volume Estimation To quantify recoverable waste coal, we evaluated Unmanned Aerial Systems (UAS) equipped with Light Detection and Ranging (LiDAR) and multispectral sensors. Structure-from-Motion (SfM) photogrammetry combined with interpolated Digital Terrain Models (DTMs) achieved strong agreement with LiDAR reference volumes (Root Mean Square Error (RMSE) ≈147 m 3 , Mean Absolute Percentage Error (MAPE) ≈2%). Sensitivity analysis confirmed that spatial resolution significantly influences accuracy, emphasizing the need for high-resolution data for precise volume estimation. This approach offers a scalable, cost-effective, and accurate alternative to conventional ground-based surveys. 3) Switchgrass Cultivation for Bioenergy and Water Quality Improvement We assessed the hydrological and water quality impacts of converting AMLs to switchgrass production areas using the Soil and Water Assessment Tool (SWAT). Results showed that converting 10% of the watershed area into the switchgrass production zone reduced streamflow by 3.1%, total suspended solids by 18.1%, total nitrogen by 7.6%, and total phosphorus by 6.2%, while achieving biomass yields of 8.6–9.2 metric tons per hectare. These findings highlight switchgrass as a dual-benefit strategy for land reclamation and bioenergy feedstock production. 4) Integrated Co-Firing and CCS for Carbon-Negative Power Generation We modeled co-firing scenarios using the Power Plant Flexible Model (PPFM) to evaluate plant efficiency, greenhouse gas (GHG) emissions, and levelized cost of electricity (LCOE). Without carbon capture and storage (CCS), increasing switchgrass co-firing ratios reduced LCOE from $\$$150/MWh at 0% biomass to $\$$110/MWh at full substitution. Under CCS, costs remained higher (~$\$$250/MWh at 0% biomass) but decreased to $\$$200/MWh at 100% biomass, while enabling net-zero or carbon-negative electricity due to switchgrass sequestration benefits. Although CCS introduced efficiency penalties, pairing it with biomass co-firing offset these impacts and maximized climate benefits. Overall, optimizing co-firing ratios between 60-100%, supported by reliable logistics and storage strategies, emerged as a practical pathway to balance affordability, sustainability, and net-zero or negative GHG emissions while promoting productive reuse of AMLs.

01 COAL, LIGNITE, AND PEAT↗
DOE OSTI2024

RWRtoolkit Narrative

This narrative presents a demonstration of RWRtools methods to the important bioenergy feedstock Panicum virgatum (Switchgrass) by leveraging networks from the model plant Arabidopsis thaliana. We used a genome-wide association study (GWAS) to identify single nucleotide polymorphisms (SNPs) associated with Switchgrass shoot biomass. We then assigned these SNPs to Switchgrass genes, and using Phytozome (Goodstein et al., 2012), assigned these 38 Switchgrass GWAS genes to 32 Arabidopsis orthologs. These 32 genes are the starting set of genes (i.e. the seed gene set) for this demo narrative.

59 BASIC BIOLOGICAL SCIENCES↗
DOE OSTI2024

Heteronuclear single quantum coherence (HSQC) NMR spectra of lignin isolated from switchgrass residues after fermentation with milling

Here we present a curated dataset of two-dimensional heteronuclear single quantum coherence (HSQC) nuclear magnetic resonance (NMR) spectra of lignin isolated from a herbaceous energy crop (Panicum virgatum L.). The lowland variant “Timber” switchgrass from Ernst seeds was used. The switchgrass was knife milled and passed through a 2 mm sieve prior to consolidated bioprocessing (CBP) process. Switchgrass was suspended in Milli-Q water and autoclaved for 90 min on liquid cycles. The residues after autoclaving were then subjected to CBP using coculture of Clostridium thermocellum (C. thermocellum DSM 1313 (LL1004)) and Thermoanaerobacterium thermosaccarolyticum ( T. thermosaccharolyticum HG-8 ATCC 31960 (LL1244)). Once-fermented and twice-fermented (FF) switchgrass were subjected to ball and disc milling in bioreactors at 55 °C and 60, 48 grams/L solids loadings for primary and secondary fermentation respectively. When fermentations were completed, the residual solids were rinsed with milli-Q water. Lignin was isolated from the pretreated residues after ball-milling in a porcelain jar with ceramic balls via Retsch PM 200 at 600 rpm for 2 h followed by enzymatic hydrolysis in acetate buffer (pH 4.8, 50 °C) for 48 h. The dry lignin samples were dissolved in deuterated dimethyl sulfoxide (d6) and characterized using 13C–1H HSQC in a Bruker Avance III HD 500-MHz NMR spectrometer. A standard Bruker pulse sequence (hsqcetgpsisp.2) was used on a Prodigy platform cryoprobe. The spectra were acquired with the following acquisition conditions: 230 ppm spectral width in F1 (13C) dimension with 256 data points and 12 ppm spectral width in F2 (1H) dimension with 2048 data points, a 90° pulse, with a C–H coupling constant of 145 Hz, a 1.0 s pulse delay, and 64 scans. Spectra were processed using the Bruker TopSpin 3.6 software.

Lignin, HSQC, Switchgrass, CBP , Ball mill, Disc m↗
DOE OSTI2026

Root of the matter-Impacts of harvest frequency on soil C and nutrients in switchgrass

We evaluated how harvest frequency affected plant C allocation and soil-water nutrient concentrations in a switchgrass (Panicum virgatumL.) system on the Eastern Shore of Maryland, U.S.A. Switchgrass established in 2023 was harvested once (1Cut), twice (2Cut), or three times (3Cut) during the 2024 growing season to represent potential feedstock-management regimes for anaerobic digestion. Aboveground biomass was measured at each harvest, and root biomass distribution, root C stocks, and soil properties were assessed to 60 cm after the growing season. During 2025, rainfall-synchronized soil water collected with tension lysimeters was analyzed for dissolved inorganic nitrogen (DIN) and inorganic phosphate (Pi) concentrations.

09 BIOMASS FUELS↗
DOE OSTI2024

Perennializing marginal croplands: going back to the future to mitigate climate change with resilient biobased feedstocks

Managing annual row crops on marginally productive croplands can be environmentally unsustainable and result in variable economic returns. Incorporating perennial bioenergy feedstocks into marginally productive cropland can engender ecosystem services and enhance climate resiliency while also diversifying farm incomes. We use one of the oldest bioenergy-specific field experiments in North America to evaluate economically and environmentally sustainable management practices for growing perennial grasses on marginal cropland. This long-term field trial called 9804 was established in 1998 in eastern Nebraska and compared the productivity and sustainability of corn (Zea mays L.)—both corn grain and corn stover—and switchgrass (Panicum virgatum L.) bioenergy systems under different harvest strategies and nitrogen (N) fertilizer rates. This experiment demonstrated that switchgrass, compared to corn, is a reliable and sustainable bioenergy feedstock. This experiment has been a catalyst for other bioenergy projects which have also expanded our understanding of growing and managing bioenergy feedstocks on marginal cropland. We (1) synthesize research from this long-term experiment and (2) provide perspective concerning both the knowledge gained from this experiment and knowledge gaps and how to fill them as well as the role switchgrass will play in the future of bioenergy.

09 BIOMASS FUELS↗
DOE OSTI2025

Drought adaptation index (DAI) based on BLUP as a selection approach for drought-resilient switchgrass germplasm

This study introduces a Drought Adaptation Index (DAI), derived from Best Linear Unbiased Prediction (BLUP), as a method to assess drought resilience in switchgrass (Panicum virgatum L.). A panel of 404 genotypes was evaluated under drought-stressed (CV) and well-watered (UC) conditions over four consecutive years (2019–2022). BLUP-estimated biomass yields were used to calculate the DAI, which enabled classification of genotypes into four adaptation groups: very well-adapted, well-adapted, adapted, and unadapted. The DAI was compared with conventional drought tolerance indices, including the Stress Susceptibility Index (SSI), Stress Tolerance Index (STI), Geometric Mean Productivity (GMP), and Yield Stability Index (YSI). Correlation analyses demonstrated strong agreement between DAI and these indices, supporting its validity and consistency. Biplot analyses using the Genotype plus Genotype-by-Environment Interaction (GGE) and Additive Main Effects and Multiplicative Interaction (AMMI) models revealed significant genotype-by-environment interactions (GEI) and identified J222.A, J463.A, and J295.A. A as high-performing genotypes, with J222.A exhibiting greater yield stability across treatments and years. Additionally, DAI isoline curves provided a graphical representation of differential genotype performance under drought and control conditions. These visualizations aided in distinguishing genotypes with stable and superior biomass yield across contrasting environments. Overall, the BLUP-based DAI is a robust and practical selection tool that improves the accuracy of identifying drought-resilient, high-yielding switchgrass genotypes. Its integration into breeding programs offers a comprehensive framework for improving biomass productivity and stress adaptation under variable climatic conditions. The application of DAI supports the development of climate-resilient cultivars and contributes to sustainable bioenergy and forage production systems.

BLUP↗
DOE OSTI2021

Plant Growth Promotion Diversity in Switchgrass-Colonizing, Diazotrophic Endophytes

Endophytic nitrogen-fixing (diazotrophic) bacteria are essential members of the microbiome of switchgrass ( Panicum virgatum ), considered to be an important commodity crop in bioenergy production. While endophytic diazotrophs are known to provide fixed atmospheric nitrogen to their host plant, there are many other plant growth-promoting (PGP) capabilities of these organisms to be demonstrated. The diversity of PGP traits across different taxa of switchgrass-colonizing endophytes is understudied, yet critical for understanding endophytic function and improving cultivation methods of important commodity crops. Here, we present the isolation and characterization of three diazotrophic endophytes: Azospirillum agricola R1C, Klebsiella variicola F10Cl, and Raoultella terrigena R1Gly. Strains R1C and F10Cl were isolated from switchgrass and strain R1Gly, while isolated from tobacco, is demonstrated herein to colonize switchgrass. Each strain exhibited highly diverse genomic and phenotypic PGP capabilities. Strain F10Cl and R1Gly demonstrated the highest functional similarity, suggesting that, while endophyte community structure may vary widely based on host species, differences in functional diversity are not a clearly delineated. The results of this study advance our understanding of diazotrophic endophyte diversity, which will allow us to design robust strategies to improve cultivation methods of many economically important commodity crops.

59 BASIC BIOLOGICAL SCIENCES↗
DOE OSTI2023

Comparative feedstock analysis in Setaria viridis L. as a model for C 4 bioenergy grasses and panicoid crop species

Second generation feedstocks for bioethanol will likely include a sizable proportion of perennial C4 grasses, principally in the Panicoideae clade. The Panicoideae contain agronomically important annual grasses including Zea mays L. (maize), Sorghum bicolor (L.) Moench (sorghum), and Saccharum officinarum L. (sugar cane) as well as promising second generation perennial feedstocks including Miscanthus × giganteus and Panicum virgatum L. (switchgrass). The underlying complexity of these polyploid grass genomes is a major limitation for their direct manipulation and thus driving a need for rapidly cycling comparative model. Setaria viridis (green millet) is a rapid cycling C4 panicoid grass with a relatively small and sequenced diploid genome and abundant seed production. Stable, transient, and protoplast transformation technologies have also been developed for Setaria viridis making it a potentially excellent model for other C4 bioenergy grasses. Here, the lignocellulosic feedstock composition, cellulose biosynthesis inhibitor response and saccharification dynamics of Setaria viridis are compared with the annual sorghum and maize and the perennial switchgrass bioenergy crops as a baseline study into the applicability for translational research. A genome-wide systematic investigation of the cellulose synthase-A genes was performed identifying eight candidate sequences. Two developmental stages; (a) metabolically active young tissue and (b) metabolically plateaued (mature) material are examined to compare biomass performance metrics.

09 BIOMASS FUELS↗
DOE OSTI2023

Seasonal decline in leaf photosynthesis in perennial switchgrass explained by sink limitations and water deficit

Leaf photosynthesis of perennial grasses usually decreases markedly from early to late summer, even when the canopy remains green and environmental conditions are favorable for photosynthesis. Understanding the physiological basis of this photosynthetic decline reveals the potential for yield improvement. We tested the association of seasonal photosynthetic decline in switchgrass (Panicum virgatum L.) with water availability by comparing plants experiencing ambient rainfall with plants in a rainfall exclusion experiment in Michigan, USA. For switchgrass exposed to ambient rainfall, daily net CO 2 assimilation ( A' net ) declined from 0.9 mol CO 2 m -2 day -1 in early summer to 0.43 mol CO 2 m -2 day -1 in late summer (53% reduction; P<0.0001). Under rainfall exclusion shelters, soil water content was 73% lower and A' net was 12% and 26% lower in July and September, respectively, compared to those of the rainfed plants. Despite these differences, the seasonal photosynthetic decline was similar in the season-long rainfall exclusion compared to the rainfed plants; A' net in switchgrass under the shelters declined from 0.85 mol CO 2 m -2 day -1 in early summer to 0.39 mol CO 2 m -2 day -1 (54% reduction; P<0.0001) in late summer. These results suggest that while water deficit limited A' net late in the season, abundant late-season rainfalls were not enough to restore A' net in the rainfed plants to early-summer values suggesting water deficit was not the sole driver of the decline. Alongside change in photosynthesis, starch in the rhizomes increased 4-fold (P<0.0001) and stabilized when leaf photosynthesis reached constant low values. Additionally, water limitation under shelters had no negative effects on the timing of rhizome starch accumulation, and rhizome starch content increased ~6-fold. These results showed that rhizomes also affect leaf photosynthesis during the growing season. Towards the end of the growing season, when vegetative growth is completed and rhizome reserves are filled, diminishing rhizome sink activity likely explained the observed photosynthetic declines in plants under both ambient and reduced water availability.

59 BASIC BIOLOGICAL SCIENCES↗
DOE OSTI2023

Perennial grass root system specializes for multiple resource acquisitions with differential elongation and branching patterns

Roots optimize the acquisition of limited soil resources, but relationships between root forms and functions have often been assumed rather than demonstrated. Furthermore, how root systems co-specialize for multiple resource acquisitions is unclear. Theory suggests that trade-offs exist for the acquisition of different resource types, such as water and certain nutrients. Measurements used to describe the acquisition of different resources should then account for differential root responses within a single system. To demonstrate this, we grew Panicum virgatum in split-root systems that vertically partitioned high water availability from nutrient availability so that root systems must absorb the resources separately to fully meet plant demands. We evaluated root elongation, surface area, and branching, and we characterized traits using an order-based classification scheme. Plants allocated approximately 3/4th of primary root length towards water acquisition, whereas lateral branches were progressively allocated towards nutrients. However, root elongation rates, specific root length, and mass fraction were similar. Our results support the existence of differential root functioning within perennial grasses. Similar responses have been recorded in many plant functional types suggesting a fundamental relationship. Root responses to resource availability can be incorporated into root growth models via maximum root length and branching interval parameters.

59 BASIC BIOLOGICAL SCIENCES↗
DOE OSTI2023

Field performance of switchgrass plants engineered for reduced recalcitrance

Switchgrass ( Panicum virgatum L.) is a promising perennial bioenergy crop that achieves high yields with relatively low nutrient and energy inputs. Modification of cell wall composition for reduced recalcitrance can lower the costs of deconstructing biomass to fermentable sugars and other intermediates. We have engineered overexpression of OsAT10 , encoding a rice BAHD acyltransferase and QsuB , encoding dehydroshikimate dehydratase from Corynebacterium glutamicum , to enhance saccharification efficiency in switchgrass. These engineering strategies demonstrated low lignin content, low ferulic acid esters, and increased saccharification yield during greenhouse studies in switchgrass and other plant species. In this work, transgenic switchgrass plants overexpressing either OsAT10 or QsuB were tested in the field in Davis, California, USA for three growing seasons. No significant differences in the content of lignin and cell wall-bound p -coumaric acid or ferulic acid were detected in transgenic OsAT10 lines compared with the untransformed Alamo control variety. However, the transgenic overexpressing QsuB lines had increased biomass yield and slightly increased biomass saccharification properties compared to the control plants. This work demonstrates good performance of engineered plants in the field, and also shows that the cell wall changes in the greenhouse were not replicated in the field, emphasizing the need to validate engineered plants under relevant field conditions.

59 BASIC BIOLOGICAL SCIENCES↗
DOE OSTI2022

Sensitive Measures of Soil Health Reveal Carbon Stability Across a Management Intensity and Plant Biodiversity Gradient

Soil carbon (C) is a major driver of soil health, yet little is known regarding how sensitive measures of soil C shift temporally within a single growing season in response to short-term weather perturbations. Our study aimed to i) Examine how long-term management impacts soil C cycling and stability across a management intensity and plant biodiversity gradient and ii) Assess how sensitive soil health indicators change temporally over the course of a single growing season in response to recent weather patterns. Here we quantify a variety of sensitive soil C measures at four time points across the 2021 growing season at the W.K. Kellogg Biological Station’s Long Term Ecological Research Trial (LTER) located in southwest Michigan, USA. The eight systems sampled included four annual soybean ( Glycine max ) systems that ranged in management intensity (conventional, no-till, reduced input, and biologically-based), two perennial biofuel cropping systems (switchgrass ( Panicum virgatum) and hybrid poplars ( Populus nigra x P.maximowiczii )), and two unmanaged systems (early successional system and a mown but never tilled grassland). We found that unmanaged systems with increased perenniality enhanced mineralizable C (Min C) and permanganate oxidizable C (POXC) values. Additionally, all soil health indicators were found to be sensitive to changes in short-term weather perturbations over the course of the growing season. The implications of this study are threefold. First, this study assess indicators of labile and stable C pools over the course of the growing season and reflects the stability of soil C in different systems. Second, POXC, Min C, and ß-glucosidase (GLU) activity are sensitive soil health indicators that fluctuate temporally, which means that these soil health indicators could help elucidate the impact that weather patterns have on soil C dynamics. Lastly, for effective monitoring of soil C, sampling time and frequency should be considered for a comprehensive understanding of soil C cycling within a system.

Martin, Tvisha↗
DOE OSTI2023

Genome-Resolved Metagenomics of Nitrogen Transformations in the Switchgrass Rhizosphere Microbiome on Marginal Lands

Switchgrass (Panicum virgatum L.) remains the preeminent American perennial (C4) bioenergy crop for cellulosic ethanol, that could help displace over a quarter of the US current petroleum consumption. Intriguingly, there is often little response to nitrogen fertilizer once stands are established. The rhizosphere microbiome plays a critical role in nitrogen cycling and overall plant nutrient uptake. We used high-throughput metagenomic sequencing to characterize the switchgrass rhizosphere microbial community before and after a nitrogen fertilization event for established stands on marginal land. We examined community structure and bulk metabolic potential, and resolved 29 individual bacteria genomes via metagenomic de novo assembly. Community structure and diversity were not significantly different before and after fertilization; however, the bulk metabolic potential of carbohydrate-active enzymes was depleted after fertilization. We resolved 29 metagenomic assembled genomes, including some from the ‘most wanted’ soil taxa such as Verrucomicrobia, Candidate phyla UBA10199, Acidobacteria (rare subgroup 23), Dormibacterota, and the very rare Candidatus Eisenbacteria. The Dormibacterota (formally candidate division AD3) we identified have the potential for autotrophic CO utilization, which may impact carbon partitioning and storage. Our study also suggests that the rhizosphere microbiome may be involved in providing associative nitrogen fixation (ANF) via the novel diazotroph Janthinobacterium to switchgrass.

60 APPLIED LIFE SCIENCES↗
DOE OSTI2021

Optimal N Application Rates on Switchgrass for Producers and a Biorefinery

This study analyzes the effects of N fertilizer application rates on profitability of growing switchgrass and using the feedstock in a pyrolysis biorefinery facility to create a source of sustainable aviation fuel (SAF) supply in Tennessee. Switchgrass (Panicum virgatum L.) is a perennial bunchgrass native to North America with traits suitable for biofuel and co-product production. Previous chemical analysis has shown that ash content in switchgrass is related to the amount of nitrogen applied to the field, while at the biorefinery level, the percentage ash content reduces the biorefinery fuel output. To obtain optimal nitrogen (N) application rates for the switchgrass producers and the biorefinery, a two-part analysis is employed. First, a partial budgeting profitability analysis is conducted for this cropping enterprise at the farm-gate level without considering downstream implications of biomass quality, i.e., ash content. Second, the effects of higher ash content as a percentage of the feedstock on biorefinery output are analyzed. Results show farm-gate profit is maximized when N fertilizer is applied at 111 kg/ha, while as a result of increased production levels and decreased percentage ash content, biorefinery profit is maximized when N is applied at 157 kg/ha. Lower ash could lead to premium prices paid to switchgrass producers if higher quality feedstock were to be demanded as part of an integrated biofuel industry.

Robertson, Keven Alan↗
DOE OSTI2021

Impact of Harvest on Switchgrass Leaf Microbial Communities

Switchgrass is a promising feedstock for biofuel production, with potential for leveraging its native microbial community to increase productivity and resilience to environmental stress. Here, we characterized the bacterial, archaeal and fungal diversity of the leaf microbial community associated with four switchgrass (Panicum virgatum) genotypes, subjected to two harvest treatments (annual harvest and unharvested control), and two fertilization levels (fertilized and unfertilized control), based on 16S rRNA gene and internal transcribed spacer (ITS) region amplicon sequencing. Leaf surface and leaf endosphere bacterial communities were significantly different with Alphaproteobacteria enriched in the leaf surface and Gammaproteobacteria and Bacilli enriched in the leaf endosphere. Harvest treatment significantly shifted presence/absence and abundances of bacterial and fungal leaf surface community members: Gammaproteobacteria were significantly enriched in harvested and Alphaproteobacteria were significantly enriched in unharvested leaf surface communities. These shifts were most prominent in the upland genotype DAC where the leaf surface showed the highest enrichment of Gammaproteobacteria, including taxa with 100% identity to those previously shown to have phytopathogenic function. Fertilization did not have any significant impact on bacterial or fungal communities. We also identified bacterial and fungal taxa present in both the leaf surface and leaf endosphere across all genotypes and treatments. These core taxa were dominated by Methylobacterium, Enterobacteriaceae, and Curtobacterium, in addition to Aureobasidium, Cladosporium, Alternaria and Dothideales. Local core leaf bacterial and fungal taxa represent promising targets for plant microbe engineering and manipulation across various genotypes and harvest treatments. Our study showcases, for the first time, the significant impact that harvest treatment can have on bacterial and fungal taxa inhabiting switchgrass leaves and the need to include this factor in future plant microbial community studies.

59 BASIC BIOLOGICAL SCIENCES↗
DOE OSTI2023

Designing Iowa Agricultural Landscapes to Improve Environmental Co-Benefits of Bioenergy Production

Cellulosic bioenergy feedstocks are needed to improve carbon (C) management while provisioning biomass for bioproducts and biofuel. The transition to increased cellulosic biomass production can be guided by land management plans designed to improve economic, environmental, and ecological performance. We constructed a sustainability model to compare landscape designs for biofuel production from corn (Zea mays L.) stover and switchgrass (Panicum virgatum L.) in central Iowa, USA. We used the model to compare environmental and socioeconomic outcomes associated with four landscape management strategies, with and without cellulosic biomass markets. We evaluated (1) a fuelshed area containing over 1.2 million ha (3 million acres) of corn and soybean (Glycine max (L.) Merr.) within 80 km (50 miles) of a commercial-scale cellulosic biorefinery in Nevada, Iowa, and (2) the South Fork watershed containing over 72,000 ha (178,000 acres) of these row crops within eight north central Iowa HUC-12 (hydrologic unit code) watersheds. At both landscape scales, we found that it is possible to achieve multiple environmental and socioeconomic benefits concomitantly with cellulosic biomass production by strategically collecting corn stover and converting the 10% of the lowest-profitability row crop land to perennial switchgrass. Potential benefits from landscape design include increased biodiversity, soil and water quality improvements, increased soil carbon sequestration for climate change mitigation, and reduced fertilizer use and cost. Our model results showed that increasing benefits can accrue when complementary conservation practices (e.g., reduced tillage, use of a rye cover crop) are combined and integrated throughout a fuelshed or watershed area. We conclude that ecologically based landscape designs offer valuable insights about costs and benefits of land management alternatives, with relevance for achieving stakeholder goals.

54 ENVIRONMENTAL SCIENCES↗
DOE OSTI2023

Photosynthesis and rhizome carbohydrate concentrations of switchgrass grown from reserve-depleted rhizomes

A long-standing question in perennial grass breeding and physiology is whether yield improvement strategies could compromise winter survival. Perennial grasses rely on the pool of carbohydrates accumulated in storage organs from the previous growing season for winter maintenance and spring regrowth. Yield improvement strategies could reduce winter survival if they increase biomass and grain yields at the expense of carbon allocation to storage. Therefore, it is crucial to better understand the dependence of regrowth on storage reserves. We experimentally depleted switchgrass ( Panicum virgatum L.) rhizome reserves by storing rhizomes for two weeks at 5 °C (control treatment) and 25 °C (reserve-depleted treatment).

bioenergy↗