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The Hunt for Green Every April: Factors Affecting Fitness in Switchgrass

This grant funded work was undertaken to develop fundamental biological knowledge of the factors affecting the complex plant trait “fitness” in switchgrass (Panicum virgatum L.), a plant being developed as a biomass crop. Using a diverse range of latitudinally-adapted switchgrass plants, genomic, molecular and physiological studies were performed to track a number of different aspects of plant genetics and physiology over the course of the growing season. Work was performed on both genetically unrelated and genetically related plants. Plants were established in the field from seedlings raised in a greenhouse, or from clones present in other field nurseries. Field grown plants were used as the source of all tissues. The three objectives of this proposal were:(1) Transcript Profiling, Metabolomics, and C and N Partitioning and Recycling in Crowns and Rhizomes of Switchgrass over two growing seasons; (2) Gene Profiling During Regreening and Dormancy of Bulked Segregants; (3) Extent of Linkage Disequilibrium in Populations for Adaptation and Fitness Traits Being Developed for Central and Northern USA, that Show Significant Heterosis. Objective 1 results: Plants were labeled using 13 CO2 (a stable isotope) using an acrylic chamber constructed specifically for this purpose. Plants became labeled with 13 C and label decayed in aerial tissues over the course of the growing season. Varying amounts of 13 C were recovered in the rhizomes. These data are being analyzed. Plants were also labeled with 15 N-urea. Plants absorbed significant amounts of label that was remobilized to the growing shoots. N-dynamics would suggest that a portion of the 15 N absorbed into the crowns and rhizomes is sequestered below ground. Variable amounts of 15 N were translocated from the shoots to the roots over the course of the growing season. Polar metabolites extracted from a diverse array of rhizomes were analyzed using GCMS. Data indicated that there was a significant shift in metabolite pools over the course of the growing season, and differences in the levels of specific metabolites could be linked to the progression of dormancy. Several metabolites that accumulate in dormant rhizomes were identified. Some of these metabolites could be potentially linked to winter-survival of switchgrass. Extensive high-throughput sequencing was conducted on crown and rhizome samples collected from field grown plants. Initial work was performed on a Roche 454 system. All later work was performed on an Illumina sequencing-by-synthesis system. Some of these datasets have been published as peer-reviewed papers, other data are currently being analyzed and being readied for publication. Objective 2 results: Genetically related but phenotypically divergent plants from an octaploid switchgrass population were grown in a replicated field nursery. Rhizomes were harvested at four different times over the course of the growing season from plants with high winter survival and those with lower winter survival. RNA-Seq was performed on harvested materials. Initial analysis suggests that plants with lowered winter survival experience a greater level of cellular stress in dormant tissues. This aspect of plant function is being probed in greater depth. Objective 3 results: A total of 592 individual clones with three clonal replications in a randomized complete block design from each of five populations used in Objective 1 studies were rated for heading date in 2012 and 2014, green-up day of year in 2013, anthesis date in 2012, and yield in 2012, they were also subjected to NIR spectroscopy to derive cell wall composition estimates based on prior NIR calibrations. Plants were genotyped via a genotyping by sequencing (GBS) approach from reduced representation libraries constructed with adaptors that identified each individual. Libraries generated with the restriction enzyme PstI and called SNPs using Samtools after alignment to version 1.1 of the switchgrass genome sequence. A total of approximately 40,000 SNPs were found. These were then further filtered to eliminate markers with a minor allele frequency of < 0.05. The results of population analysis using STRUCTURE with expected population sizes or cluster numbers (K), clearly shows the hybrid composition of the KxS population and discriminated easily between upland (Summer) and lowland (Kanlow) populations. Under an assumption of 5 distinct populations there were detectable differences in allele frequencies between subpopulations within the three Kanlow populations particularly with respect to Kanlow EM and Kanlow base. We detected 110 SNPs with an allele frequency difference of ≥ 0.2 between Kanlow EM and Kanlow base populations, while 120 SNPs showed an allele frequency difference of ≥ 0.15 between Kanlow N1 and Kanlow base populations. These data are being readied for publication.

59 BASIC BIOLOGICAL SCIENCES↗

Soil microclimates influence annual carbon loss via heterotrophic soil respiration in maize and switchgrass bioenergy cropping systems

Heterotrophic soil respiration (R H ) is the main pathway of carbon (C) loss from litter and soil organic matter, and thus R H partially determines ecosystem C storage. Because R H is sensitive to soil temperature and moisture, aboveground factors that influence soil microclimate, such as plant structure and residue management, may in turn affect belowground C loss via R H , but this relationship has not been quantified. Here, e examined multiyear soil microclimate differences to 1-m depth, measured seasonal trends of R H , and parameterized crop-specific microclimate-RH models to quantify the effect of soil microclimate differences on annual R H in temperate no-till maize and switchgrass bioenergy cropping systems. Summertime soil temperatures were typically warmer in maize compared to switchgrass, likely resulting from lower leaf area index (LAI) in maize. In contrast, winter soil temperatures were usually warmer in switchgrass than maize, due in part to more consistent snow retention within the switchgrass litter stubble. Daily soil temperature ranges were less extreme in the perennial switchgrass system compared to the annual no-till maize system. Soil moisture near the soil surface was usually lower in maize than switchgrass, but the opposite was true below about 50 cm. R H showed strong seasonal trends, with warmer and drier soil conditions generally leading to higher R H in both crops. Modeled scenarios indicated that the differences in crop-specific soil microclimates accounted for 4 to 17% of the annual R H flux, with the dominant soil microclimate effects on R H occurring during the summer. So, the soil microclimate serves as a strong coupling between aboveground properties and belowground C loss via R H in temperate agroecosystems. Agricultural management practices such as planting date, plant density, and residue management could be targeted to promote soil microclimates that reduce R H , thereby reducing gaseous belowground C losses and potentially enhancing ecosystem C storage.

59 BASIC BIOLOGICAL SCIENCES↗

Influences of nitrogen fertilization and climate regime on the above-ground biomass yields of miscanthus and switchgrass: A meta-analysis

Perennial grasses are touted as sustainable feedstocks for energy production. Such benefits, however, may be offset if excessive nitrogen (N) fertilization leads to economic and environmental issues. Moreover, as yields respond to changes in climate, nutrient requirements will change, and thus guidance on minimal N inputs is necessary to ensure sustainable bioenergy production. Here, a pairwise meta-analysis was conducted to investigate the effects of N fertilization (amount and duration) and climate on the above-ground biomass yields of miscanthus ( Miscanthus x giganteus ) and switchgrass ( Panicum virgatum L. ). Both regression models and meta-analyses showed that switchgrass was more responsive to N than miscanthus, although both showed significant and positive N effects. Meta-analysis further showed that the positive growth response of miscanthus to N application increased with N addition rates of 60–300 kg N ha -1 year -1 , but the magnitude of the response decreased with the number of years of fertilization (duration). N effects on switchgrass biomass increased and peaked at rates of 120–160 kg N ha -1 year -1 and 5–6 years of N inputs, but diminished for rates >300 kg N ha -1 year -1 and >7 years. Meta-analysis further revealed that the influences of N on switchgrass increased with both mean annual temperature and precipitation. Miscanthus yields were less responsive to climate than switchgrass yields. This meta-analysis helps fill a gap in estimation of biofeedstock yields based on N fertilization and could help better estimate minimum N requirements and soil management strategies for miscanthus and switchgrass cultivation across climatic conditions, thereby improving the efficiency and sustainability of bioenergy cropping systems.

09 BIOMASS FUELS↗

Pore architecture and particulate organic matter in soils under monoculture switchgrass and restored prairie in contrasting topography

Bioenergy cropping systems can substantially contribute to climate change mitigation. However, limited information is available on how they affect soil characteristics, including pores and particulate organic matter (POM), both essential components of the soil C cycle. The objective of this study was to determine effects of bioenergy systems and field topography on soil pore characteristics, POM, and POM decomposition under new plant growth. We collected intact soil cores from two systems: monoculture switchgrass (Panicum virgatum L.) and native prairie, at two contrasting topographical positions (depressions and slopes), planting half of the cores with switchgrass. Pore and POM characteristics were obtained using X-ray computed micro-tomography (μCT) (18.2 µm resolution) before and after new switchgrass growth. Diverse prairie vegetation led to higher soil C than switchgrass, with concomitantly higher volumes of 30–90 μm radius pores and greater solid-pore interface. Yet, that effect was present only in the coarse-textured soils on slopes and coincided with higher root biomass of prairie vegetation. Surprisingly, new switchgrass growth did not intensify decomposition of POM, but even somewhat decreased it in monoculture switchgrass as compared to non-planted controls. Our results suggest that topography can play a substantial role in regulating factors driving C sequestration in bioenergy systems.

59 BASIC BIOLOGICAL SCIENCES↗

Microtopography‐induced transient waterlogging affects switchgrass (Alamo) growth in the lower coastal plain of North Carolina, USA

Abstract Very limited information is currently available on growth responses of switchgrass (lowland cultivars) to transient waterlogging in lowland or poorly drained areas. This study investigated impacts of microtopography‐induced transient waterlogging on switchgrass (Alamo cultivar) growth, represented by leaf‐level gas exchange and biomass yield, in an established experimental field located in the Atlantic coastal plain of North Carolina, USA . Intensive leaf‐level gas exchange measurements were conducted on switchgrass at paired spots with distinct elevations in three sub‐blocks. Aboveground biomass was randomly collected across the study field to explore the potential impacts of the transient waterlogging on biomass yield. The sum of excess water ( SEW ) was calculated based on measured instantaneous water table depth to generalize the relationship between biomass yield and intensity of transient waterlogging. Results showed significant ( P ≤ 0.0001) treatment effects on leaf‐level gas exchange, characterized by evident reduction in both CO 2 assimilation rate and stomatal conductance when water table was at or near the soil surface at low positions. Negative impacts of transient waterlogging on leaf‐level gas exchange became more evident with the increasing of elevation differences between paired subplots. Stomatal closure was found to be the main mechanism responsible for the decline of net assimilation under transient waterlogging. Aboveground biomass yields of switchgrass showed relatively high spatial variability and were positively and linearly correlated with microtopography (represented by elevation in the analysis) ( P < 0.03, R 2 > 0.77). Further analysis showed that biomass yields were negatively correlated with SEW ( P < 0.001, R 2 > 0.6) with an exponential relationship. Results of this study strongly demonstrated transient waterlogging could negatively affect switchgrass growth by suppressing leaf‐level gas exchange rates and ultimately reducing biomass yield. Findings from this study have critical implications for evaluating the economic viability of growing switchgrass on marginal lands that are subject to transient waterlogging stresses.

Tian, Shiying↗

Determining spatially varying profit‐maximizing management practices for miscanthus and switchgrass production in the rainfed United States

Abstract Determining optimal management practices for the profitable production of perennial energy crops is critical for scaling up production beyond experimental levels. Although many experimental field studies have examined the effects of management practices on the performance of miscanthus and switchgrass, there are no recommendations for economically optimal nitrogen (N) application rates and how they should vary spatially and with the age of the energy crop as well as on optimal rotation age of the energy crop to maximize profits. We develop a modeling framework to determine economically optimal crop management decisions and simulate the variability under various scenarios for miscanthus and switchgrass production across 2287 counties in the rainfed United States. We find that profit‐maximizing N recommendations for these crops vary across maturity stages and regions and can increase the landowner's profits compared with a uniform N rate across ages and regions. We also find that the optimal rotation for these crops is shorter than the productive physical lifespan (15–20 and 10 years for miscanthus and switchgrass, respectively). Specifically, the N rate that maximizes the economic returns is negligible for miscanthus and 111 kg ha −1 for switchgrass production at age 2. The mean profit‐maximizing N rate increases with age for miscanthus, peaking at 151 kg ha −1 at age 11 before declining to 114 kg ha −1 at the optimal rotation age of 13 years while that for switchgrass is 150 kg ha −1 for middle‐aged stands and declines to 114 kg ha −1 at the optimal rotation of 8–9 years. We find that miscanthus is the most profitable energy crop in the northern region of the rainfed United States while switchgrass is most profitable in the south of the rainfed United States. Our findings are useful for improving assessments of the profitability of energy crops and guiding future management decisions by landowners.

54 ENVIRONMENTAL SCIENCES↗

Rhizosphere microbial community structure in high-producing, low-input switchgrass families

Switchgrass ( Panicum virgatum L.) is a native, low-input North American perennial crop primarily grown for bioenergy, livestock forage, and industrial fiber. To achieve no-input switchgrass production that meets biomass needs, several switchgrass genotypes have been identified that have a low or negative response to nitrogen fertilizer, i . e ., the biomass accumulation with added nitrogen is less than or equal to that when grown without nitrogen. In order to improve the viability of low-input switchgrass production, a more detailed understanding of the biogeochemical mechanisms active in these select genotypes is needed. 16S and ITS amplicon sequencing and qPCR of key functional genes were applied to switchgrass rhizospheres to elucidate microbial community structure in high-producing, no-input switchgrass families. Rhizosphere microbial community structure differed strongly between sites, and nitrogen responsiveness.

Stonoha-Arther, Christina (ORCID:0000000263964507)↗

AmeriFlux FLUXNET-1F US-AR1 ARM USDA UNL OSU Woodward Switchgrass 1

This is the AmeriFlux Management Project (AMP) created FLUXNET-1F version of the carbon flux data for the site US-AR1 ARM USDA UNL OSU Woodward Switchgrass 1. This is the FLUXNET version of the carbon flux data for the site US-AR1 ARM USDA UNL OSU Woodward Switchgrass 1 produced by applying the standard ONEFlux (1F) software. Site Description - The ARM USDA UNL OSU Woodward Switchgrass 1 tower is located on public land owned by the USDA-ARS Southern Plains Range Research Station in Woodward, Oklahoma. The site is on a former native prairie that is in the process of changing to switchgrass. A second companion site (ARM USDA UNL OSU Woodward Switchgrass 2) is on a former wheat field. In Spring 2009, the former native prairie site was burned, cattle were put on the pasture to graze down emergent grass, and broadleaf herbicide was sprayed. In Summer 2009, the cattle were removed from the pasture, and the site was sprayed with herbicide to kill all grass. In Spring 2010, prior to the planting of switchgrass, final herbicide was sprayed to kill cheat grass and to control broadleaf plants.

Billesbach, Dave↗

AmeriFlux FLUXNET-1F US-AR2 ARM USDA UNL OSU Woodward Switchgrass 2

This is the AmeriFlux Management Project (AMP) created FLUXNET-1F version of the carbon flux data for the site US-AR2 ARM USDA UNL OSU Woodward Switchgrass 2. This is the FLUXNET version of the carbon flux data for the site US-AR2 ARM USDA UNL OSU Woodward Switchgrass 2 produced by applying the standard ONEFlux (1F) software. Site Description - The ARM USDA UNL OSU Woodward Switchgrass 2 tower is located on public land owned by the USDA-ARS Southern Plains Range Research Station in Woodward, Oklahoma. The site is on a former wheat field that is in the process of changing to switchgrass. A companion site (ARM USDA UNL OSU Woodward Switchgrass 1) is on a former native prairie. Previous wheat was planted in Fall 2008. In Spring 2009, herbicide was applied to kill the wheat prior to switchgrass planting. Later in the year, the site was sprayed with post-emergence herbicide. In 2010, fertilization occurred before herbicide was sprayed for broadleaf control.

Billesbach, Dave↗

Identification of Adaptive Fungal Pathogen Resistance Loci in Switchgrass (Final Report)

Switchgrass (Panicum virgatum) has great potential as a sustainable bioenergy and bioproducts feedstock. However, there are a number of challenges that must be overcome to achieve this goal. One of the most significant challenges to switchgrass productivity are fungal pathogens, especially rusts and smuts. The primary goals of our project were to characterize the distribution of fungal pathogens on switchgrass across central North America and to identify genetic loci that control resistance to those pathogens. The research was primarily conducted with large common garden plantings of a switchgrass, distributed along a latitudinal gradient from South Texas to South Dakota. By conducting this research simultaneously at 10 field locations, we were able establish the relative contributions of genes and the environment to pathogen resistance. Through this research, we identified multiple genomic regions (loci) that contribute to rust resistance and found that the effects of these loci on resistance were highly influenced by field site location. Our research revealed that most of the rust damage to switchgrass from Texas to the northern United States is caused by a single fungal species, Puccinia novopanici. Therefore, genetic variation within this species and/or local environmental conditions are responsible for the inconsistent efficacy of pathogen resistance loci. We are currently investigating these two non-mutually exclusive hypotheses. In addition to our studies of rust pathogens, we were able to identify resistance loci for the head smut fungal pathogen Tilletia maclaganii. The funding from this grant facilitated a collaboration with the DOE Joint Genome Institute to develop a reference genome for T. maclaganii. A reference genome will make population genetic studies of this smut fungus possible, which in turn will facilitate the development of strategies to mitigate its impact. Overall, the funded research has greatly advanced our understanding of the genetic basis of fungal pathogen resistance switchgrass. This knowledge will help facilitate the breeding of more resistant cultivars and thus, increase the productivity of swichgrass generally.

09 BIOMASS FUELS↗

Data from: Temporal dynamics of free‐living nitrogen fixation in the switchgrass rhizosphere

Here we present data associated with the manuscript, Temporal dyanmics of free-living nitrogen fixation in the switchgrass rhizosphere. Free-living nitrogen fixation (FLNF) represents an important terrestrial N source and is gaining interest for its potential to contribute plant available N to bioenergy cropping systems. Switchgrass, a cellulosic bioenergy crop, may be reliant on FLNF when particularly when grown on low N marginal lands. These potential contributions of FLNF to switchgrass and the controls on this process are not well understood. We evaluated drivers of FLNF rates and N-fixing microbial community composition in field-grown switchgrass systems over two years with high temporal sampling. We found climate variables to be strong drivers of FLNF in switchgrass systems, compared to other environmental and biological factors including soil nutrients and N-fixing microbial community composition. Increased soil moisture availability tended to promote FLNF, but extreme rainfall events appeared to be detrimental. These climate-related responses suggest FLNF-derived N contributions may be reduced under projected climate shifts. We also found a significant, but weak correlation between N-fixing microbial community composition and FLNF rates including an observed shift in community composition between 2017 and 2018 and a similarly significant difference in FLNF rates between years. Lastly, we found that seasonal FLNF N contributions, based on measurement with high temporal resolution, may meet up to 80% of switchgrass N demands.

09 BIOMASS FUELS↗

Gasification performance of switchgrass pretreated with torrefaction and densification

The purpose of this study was to investigate gasification performance of four switchgrass pretreatments (torrefaction at 230 and 270 °C, densification, and combined torrefaction and densification) and three gasification temperatures (700, 800 and 900 °C). Gasification was performed in a fixed-bed externally heated reactor with air as an oxidizing agent. Switchgrass pretreatment and gasification temperature had significant effects on gasification performance such as gas yields, syngas lower heating value (LHV), and carbon conversion and cold gas efficiencies. With an increase in the gasification temperature, yields of H2 and CO, syngas LHV, and gasifier efficiencies increased whereas CH4, CO2 and N2 yields decreased. Among all switchgrass pretreatments, gasification performance of switchgrass with combined torrefaction and densification was the best followed by that of densified, raw and torrefied switchgrass. Gasification of combined torrefied and densified switchgrass resulted in the highest yields of H2 (0.03 kg/kg biomass) and CO (0.72 kg/kg biomass), highest syngas LHV (5.08 MJ m-3), CCE (92.53%), and CGE (68.40%) at the gasification temperature of 900 °C.

09 BIOMASS FUELS↗

The productivity of perennial crops miscanthus and switchgrass is more resistant to vapor pressure deficit stress than maize

Rising atmospheric vapor pressure deficit (D) with warming is an increasingly important driver of crop productivity loss, yet the relative sensitivity of conventional seed crops and alternative perennial crops remains poorly resolved. In this study, we combined a 9-year eddy covariance carbon flux record (2008 – 2016) with 600+ midday leaf water potential (ψ L ) measurements (2024 – 2025) from adjacent maize (Zea mays), miscanthus (Miscanthus x giganteus), and switchgrass (Panicum virgatum) plots to: (1) quantify the limitations of elevated D on crop-specific productivity and (2) evaluate how these responses vary as a function of soil moisture (Θ) status. We found that maize strictly regulated ψ L and its gross primary productivity (GPP) was strongly reduced by increasing D. In contrast, miscanthus and switchgrass allowed larger ψ L declines and sustained higher GPP under similar moisture constraints. D exerted a larger limitation on maize GPP than Θ, but D-driven declines were less severe when accompanied by high Θ. GPP sensitivity to D was also influenced by Θ for miscanthus and switchgrass, but the buffering effect was stronger compared to maize. Specifically, high D paired with high Θ were the most productive conditions for the perennials, reflecting temperature-driven gains in photosynthetic efficiency when evaporative stress was mitigated by sufficient soil water supply. Overall, miscanthus and switchgrass displayed greater resistance to atmospheric drought and more stable GPP across hydroclimate variability than maize. These findings highlight that maize is more vulnerable to future projections of rising D than miscanthus or switchgrass.

drought↗

Hemicellulose characterization of deuterated switchgrass

This paper describes the structural characterization of hemicellulose isolated from hydroponically grown switchgrass in H 2 O medium (protiated) or 50% D 2 O medium (deuterated) through compositional analysis, GPC, FTIR, 13 C and 1 H/ 13 C HSQC NMR. 4-O-methyl glucuronoarabinoxylan (GAX), the major hemicellulose in switchgrass isolated from deuterated switchgrass, had structural properties similar to hemicellulose isolated from protiated switchgrass. Both had comparable arabinose to xylose ratio (0.25) and molecular weight (47–50 kDa). Finally, structural similarities show that deuterated switchgrass hemicellulose can be used as a model carbohydrate polymer in neutron scattering, or pharmaceutical studies due to their immunomodulatory activity and gastroprotective effects.

59 BASIC BIOLOGICAL SCIENCES↗

Natural variability and antioxidant properties of commercially cultivated switchgrass extractives

Optimum utilization of lignocellulosic bioenergy feedstock, such as switchgrass, via recovery of value-added phytochemicals and reduction of waste streams will increase the economic feasibility as well as the sustainability of integrated biorefineries. Switchgrass contains significant amount of overlooked non-structural components, i.e. extractives, which encompasses phenolic derivatives with potential antimicrobial and antioxidant properties. As a perennial crop, it is essential to assess the natural variability of switchgrass extractives content and composition in order to determine the feasibility of its extraction and utilization. In this study, three switchgrass cultivars were harvested from four commercial farms in their second and third growing seasons. Their respective extractive fractions were characterized for the presence of free sugars, total phenolics (TPC), hydroxycinnamic acid (HCAs) derivatives, total flavonols, and other important phytochemicals. Biomass harvested in the third growing season displayed greater amounts of TPC, HCAs, and total flavonols. Environmental conditions such as total precipitation, and cultural practices including harvest time, played important roles in the accumulation of TPC. As a result, the switchgrass extractives also exhibited DPPH (2,2-diphenyl-1-picrylhydrazyl) radical scavenging activity with a positive correlation between inhibiting concentration (IC 50 ) value and total flavonols content.

59 BASIC BIOLOGICAL SCIENCES↗

Nitrogen rate and landscape impacts on life cycle energy use and emissions from switchgrass-derived ethanol

Switchgrass-derived ethanol has been proposed as an alternative to fossil fuels to improve sustainability of the US energy sector. In this study, life cycle analysis (LCA) was used to estimate the environmental benefits of this fuel. To better define the LCA environmental impacts associated with fertilization rates and farm-landscape topography, results from a controlled experiment were analyzed. Data from switchgrass plots planted in 2008, consistently managed with three nitrogen rates (0, 56, and 112 kg N ha –1 ), two landscape positions (shoulder and footslope), and harvested annually (starting in 2009, the year after planting) through 2014 were used as input into the Greenhouse gases, Regulated Emissions and Energy use in transportation (GREET) model. Simulations determined nitrogen (N) rate and landscape impacts on the life cycle energy and emissions from switchgrass ethanol used in a passenger car as ethanol–gasoline blends (10% ethanol:E10, 85% ethanol:E85s). Results indicated that E85s may lead to lower fossil fuels use (58 to 77%), greenhouse gas (GHG) emissions (33 to 82%), and particulate matter (PM2.5) emissions (15 to 54%) in comparison with gasoline. However, volatile organic compounds (VOCs) and other criteria pollutants such as nitrogen oxides (NOx), particulate matter (PM10), and sulfur dioxides (SO x ) were higher for E85s than those from gasoline. Nitrogen rate above 56 kg N ha –1 yielded no increased biomass production benefits; but did increase (up to twofold) GHG, VOCs, and criteria pollutants. Lower blend (E10) results were closely similar to those from gasoline. The landscape topography also influenced life cycle impacts. Biomass grown at the footslope of fertilized plots led to higher switchgrass biomass yield, lower GHG, VOCs, and criteria pollutants in comparison with those at the shoulder position. Lastly, results also showed that replacing switchgrass before maximum stand life (10–20 years.) can further reduce the energy and emissions reduction benefits.

09 BIOMASS FUELS↗

Temporal dynamics of free‐living nitrogen fixation in the switchgrass rhizosphere

Abstract Free‐living nitrogen fixation (FLNF) represents an important terrestrial N source and is gaining interest for its potential to contribute plant available N to bioenergy cropping systems. Switchgrass, a cellulosic bioenergy crop, may be particularly reliant on FLNF when grown on low N systems, like marginal lands. However, the potential contributions of FLNF to switchgrass as well as the controls on this process are not well understood. In this study, we evaluated drivers of FLNF rates and N‐fixing microbial (diazotrophic) community composition in field‐grown switchgrass systems over two growing seasons with high temporal sampling. We found that climate variables are strong drivers of FLNF rates in switchgrass systems, compared to other environmental and biological factors including soil nutrients and diazotrophic community composition. Increased soil moisture availability generally promoted FLNF rates, but extreme rainfall events were detrimental. These climate‐related responses suggest a potential for loss of FLNF‐derived N contributions under projected climate shifts. We found a significant, but weak correlation between diazotrophic community composition and FLNF rates. We also observed a significant shift in the diazotrophic community composition between 2017 and 2018 and similarly measured a significant difference in FLNF rates between growing seasons. Lastly, we found that seasonal FLNF N contributions, based on measurement with high temporal resolution, has the potential to meet up to 80% of switchgrass N demands suggesting that FLNF measurements extrapolated from fewer time points or locations may underestimate these potential N contributions.

59 BASIC BIOLOGICAL SCIENCES↗

Photosynthetic responses of switchgrass to light and CO 2 under different precipitation treatments

Switchgrass ( Panicum virgatum L .) is a prominent bioenergy crop with robust resilience to environmental stresses. However, our knowledge regarding how precipitation changes affect switchgrass photosynthesis and its responses to light and CO 2 remains limited. To address this knowledge gap, we conducted a field precipitation experiment with five different treatments, including −50%, −33%, 0%, +33%, and +50% of ambient precipitation. To determine the responses of leaf photosynthesis to CO 2 concentration and light, we measured leaf net photosynthesis of switchgrass under different CO 2 concentrations and light levels in 2020 and 2021 for each of the five precipitation treatments. We first evaluated four light and CO 2 response models (i.e., rectangular hyperbola model, nonrectangular hyperbola model, exponential model, and the modified rectangular hyperbola model) using the measurements in the ambient precipitation treatment. Based on the fitting criteria, we selected the nonrectangular hyperbola model as the optimal model and applied it to all precipitation treatments, and estimated model parameters. Overall, the model fit field measurements well for the light and CO 2 response curves. Precipitation change did not influence the maximum net photosynthetic rate ( P max ) but influenced other model parameters including quantum yield ( α ), convexity ( θ ), dark respiration ( Rd ), light compensation point ( LCP ), and saturated light point ( LSP ). Specifically, the mean P max of five precipitation treatments was 17.6 μmol CO 2 m −2 s −1 , and the ambient treatment tended to have a higher P max . The +33% treatment had the highest α , and the ambient treatment had lower θ and LCP , higher Rd , and relatively lower LSP . Furthermore, precipitation significantly influenced all model parameters of CO 2 response. The ambient treatment had the highest P max , largest α , and lowest θ , R d , and CO 2 compensation point LCP . Overall, this study improved our understanding of how switchgrass leaf photosynthesis responds to diverse environmental factors, providing valuable insights for accurately modeling switchgrass ecophysiology and productivity.

09 BIOMASS FUELS↗