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DOE OSTI2016

Antimicrobial and anti-inflammatory activity of switchgrass-derived extractives

Switchgrass is an increasingly important biofuel crop, but knowledge of switchgrass fungal pathogens is not extensive. The purpose of this research was to identify the fungal pathogens that decrease crop yield of switchgrass grown in Tennessee and to investigate a potential sustainable disease management strategy from a value-added by-product of the switchgrass biofuel conversion process. The specific objectives were 1) to identify and characterize prevalent fungal pathogens of switchgrass in Tennessee, 2) assess switchgrass seed produced in the United States for seedborne fungal pathogens, and 3) evaluate switchgrass extractives for antimicrobial activity against plant pathogens.

09 BIOMASS FUELS↗

DOE OSTI2015

Reduction of ethanol yield from switchgrass infected with rust caused by Puccinia emaculata

Switchgrass ( Panicum virgatum ) is an important biofuel crop candidate thought to have low disease susceptibility. As switchgrass production becomes more prevalent, monoculture and production fields in close proximity to one another may increase the spread and severity of diseases such as switchgrass rust caused by the pathogen Puccinia emaculata . The objective of this research was to examine the impact of rust on ethanol yield in switchgrass. In 2010 and 2012, naturally infected leaves from field-grown Alamo and Kanlow in Knoxville, TN (2010, 2012) and Crossville, TN (2012) were visually categorized as exhibiting low, medium, or high disease based on the degree of chlorosis and sporulation. P. emaculata was isolated from each disease range to confirm infection. Samples from 2010 were acid/heat pretreated and subjected to two runs of simultaneous saccharification and fermentation (SSF) with Saccharomyces cerevisiae D 5 A to measure ethanol yield. Near-infrared spectroscopy (NIRS) was used to estimate ethanol yield for 2012 samples. SSF and NIRS data were analyzed separately using ANOVA. Disease level effects were significant within both models (P < 0.05) and both models explained a large amount of variation in ETOH (SSF: R 2 = 0.99, NIRS: R 2 = 0.99). In the SSF dataset, ethanol was reduced by 35 % in samples exhibiting medium disease symptoms and by 55 % in samples exhibiting high disease symptoms. In the NIRS dataset, estimated ethanol was reduced by 10 % in samples exhibiting medium disease symptoms and by 21 % in samples exhibiting high disease symptoms. Lastly, results indicate that switchgrass rust will likely have a negative impact on ethanol yield in switchgrass grown as a biofuel crop.

09 BIOMASS FUELS↗

DOE OSTI2017

Light and temperature effects on miR156 transgenic switchgrass flowering: A simulated latitudinal study

The control of flowering in perennial grasses is an important trait, especially among biofuel feedstocks. Lignocellulosic biomass may be increased commensurate with decreased or delayed flowering as the plant allocates energy for stems and leaves harvested for bioenergy at the end of the growing season. For transgenic feedstocks, such as switchgrass ( Panicum virgatum L. ) grown in its geographic center of distribution, it is foreseeable that regulators may require greatly decreased gene flow frequencies to enable commercialization. Transgenic switchgrass with various overexpression levels of a rice microRNA gene, miR156, when grown in field conditions, holds promise for decreased flowering, yielding high biomass, and altered cell wall traits, which renders it as a potential crossing partner for further breeding with switchgrass lines for decreased recalcitrance. In the current research, we simulated a latitudinal cline in controlled growth chamber experiments for various individual sites from the tropics to cool-temperate conditions which included weekly average high and low temperatures and day lengths over the switchgrass growing season for each simulated site: Guayaquil, Ecuador; Laredo, Texas, USA; and Brattleboro, Vermont, USA. Flowering and reproduction among transgenic lines with low (T-14 and T-35)--tomoderate (T-27 and T-37) overexpression of miR156 were assessed. Lower simulated latitudes (higher temperatures with low-variant day length) and long growing seasons promoted flowering of the miR156 transgenic switchgrass lines. Tropical conditions rescued the flowering phenotype in all transgenic lines except T-27. Higher numbers of plants in lines T-35 and T-37 and the controls produced panicles, which also occurred earlier in the study as temperatures increased and day length decreased. Line T-14 was the exception as more clonal replicates flowered in the cool-temperate (Vermont) conditions. Increased biomass was found in transgenic lines T-35 and T-37 in tropical conditions. No difference in biomass was found in subtropical (Texas) chambers, and two lines (T-14 and T-35) produced less biomass than the control in cool-emperate conditions. Our findings suggest that switchgrass plants engineered to overexpress miR156 for delayed flowering to promote bioconfinement and biomass production may be used for plant breeding at tropical sites.

59 BASIC BIOLOGICAL SCIENCES↗

DOE OSTI2020

Discrete element modeling of switchgrass particles under compression and rotational shear

Switchgrass is a perennial herbaceous plant regarded as a biomass energy crop in the United States for its highadaptability and yield potential. Processing and handling of switchgrass particles are challenging due to the erratic mechanical and flow behavior originating from their intrinsic particulate properties. Here, we present a bonded-sphere discrete element model designed specifically for switchgrass particles. The model simultaneously captures three key particulate features, i.e., fibrous particle shapes, a wide range of particle sizes, and particle deformability. Realistic yet computationally efficient particle shape templates are created based on the image analysis data of switchgrass specimens. A fitting procedure is proposed to ensure both the particle width and length distributions are captured, a unique requirement for fibrous particles. Two full-scale numerical models, i.e., a uniaxial compression model and a Schulze ring shear model, are developed using information fromphysical experiments. The model is calibrated using experimental data of chopped-small switchgrass specimens, and then, is validated using data of chopped-large specimens in both compression and ring-shear tests. Numerical results show that the numerical models capture bulk densities accurately (with an error of 3%) while slightly underestimate the bulk friction angle. Furthermore, an extensive sensitivity analysis reveals that (1) switchgrass particles with rougher edges (due to different processing techniques) exhibit a higher shear strength and a lower flowability; (2) stiffer particles yield a lower bulk density (up to 21% lower) compared to more deformable particles, indicating particle deformability should be incorporated when modeling biomass flow in a preprocessing system.

09 BIOMASS FUELS↗

DOE OSTI2016

Understanding Multiscale Structural Changes During Dilute Acid Pretreatment of Switchgrass and Poplar

Biofuels produced from lignocellulosic biomass hold great promise as a renewable alternative energy and fuel source. To realize a cost and energy efficient approach, a fundamental understanding of the deconstruction process is critically necessary to reduce biomass recalcitrance. Here, the structural and morphological changes over multiple scales (5–6000 Å) in herbaceous (switchgrass) and woody (hybrid poplar) biomass during dilute sulfuric acid pretreatment were explored using neutron scattering and X-ray diffraction. Switchgrass undergoes a larger increase (20–84 Å) in the average diameter of the crystalline core of the elementary cellulose fibril than hybrid poplar (19–50 Å). Switchgrass initially forms lignin aggregates with an average size of 90 Å that coalesce to 200 Å, which is double that observed for hybrid poplar, 55–130 Å. Switchgrass shows a smooth-to-rough transition in the cell wall surface morphology unlike the diffuse-to-smooth transition of hybrid poplar. Yet, switchgrass and hybrid poplar pretreated under the same experimental conditions result in pretreated switchgrass producing higher glucose yields (~76 wt %) than pretreated hybrid poplar (~60 wt %). This observation shows that other aspects like cellulose allomorph transitions, cellulose accessibility, cellular biopolymer spatial distribution, and enzyme–substrate interactions may be more critical in governing the enzymatic hydrolysis efficiency.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗

DOE OSTI2022

Responsiveness of miscanthus and switchgrass yields to stand age and nitrogen fertilization: A meta‐regression analysis

Abstract Optimal management of the perennial bioenergy crops, miscanthus and switchgrass, requires an understanding of their responsiveness to nitrogen (N) fertilizer at different maturity stages across locations and growing conditions. Earlier studies that have examined the yield response of these crops to N and stand age using field experiments or meta‐analysis techniques provide mixed evidence. We extend earlier studies by applying a multi‐level mixed‐effects (MLME) meta‐regression model to conduct a more extensive multivariate regression of yield response of these crops to N and stand age, while controlling for climate and location conditions and unobserved factors related to study design. Our findings are based on 1403 and 2811 yield observations for miscanthus and switchgrass, respectively, from experiments conducted between 2002 and 2019 across the rainfed region in the United States. We find statistically significant evidence that an additional year of maturity increases miscanthus and switchgrass yields but at a decreasing rate; yields peak at the 7th and 6th year respectively, for the observed range of applied N rates and stands. We also find that an increase in N application increases yield by a statistically significant level, but at a declining rate; the magnitude of the yield response to N is, however, small and varies with the age of the crop. The impact of N is larger on older compared to younger and middle‐aged stands of miscanthus. In contrast, the impact of N on switchgrass is larger on middle‐aged compared to younger and older stands of switchgrass. We do not find a statistically significant effect of soil productivity on yield for either crop. This analysis provides a basis for developing N application recommendations and optimal rotation age for miscanthus and switchgrass and shows that these energy crops can grow just as productively on low productivity land as on high productivity land.

59 BASIC BIOLOGICAL SCIENCES↗

DOE OSTI2018

Gene regulatory networks for lignin biosynthesis in switchgrass (Panicum virgatum )

Summary Cell wall recalcitrance is the major challenge to improving saccharification efficiency in converting lignocellulose into biofuels. However, information regarding the transcriptional regulation of secondary cell wall biogenesis remains poor in switchgrass ( Panicum virgatum ), which has been selected as a biofuel crop in the United States. In this study, we present a combination of computational and experimental approaches to develop gene regulatory networks for lignin formation in switchgrass. To screen transcription factors ( TF s) involved in lignin biosynthesis, we developed a modified method to perform co‐expression network analysis using 14 lignin biosynthesis genes as bait (target) genes. The switchgrass lignin co‐expression network was further extended by adding 14 TF s identified in this study, and seven TF s identified in previous studies, as bait genes. Six TF s (Pv MYB 58/63, Pv MYB 42/85, Pv MYB 4, Pv WRKY 12, Pv SND 2 and Pv SWN 2) were targeted to generate overexpressing and/or down‐regulated transgenic switchgrass lines. The alteration of lignin content, cell wall composition and/or plant growth in the transgenic plants supported the role of the TF s in controlling secondary wall formation. RNA ‐seq analysis of four of the transgenic switchgrass lines revealed downstream target genes of the secondary wall‐related TF s and crosstalk with other biological pathways. In vitro transactivation assays further confirmed the regulation of specific lignin pathway genes by four of the TF s. Our meta‐analysis provides a hierarchical network of TF s and their potential target genes for future manipulation of secondary cell wall formation for lignin modification in switchgrass.

transcription factors↗

DOE OSTI2022

Identification of microRNAs responsive to arbuscular mycorrhizal fungi in Panicum virgatum (switchgrass)

Background: MicroRNAs (miRNAs) are important post-transcriptional regulators involved in the control of a range of processes, including symbiotic interactions in plants. MiRNA involvement in arbuscular mycorrhizae (AM) symbiosis has been mainly studied in model species, and our study is the first to analyze global miRNA expression in the roots of AM colonized switchgrass ( Panicum virgatum ), an emerging biofuel feedstock. AM symbiosis helps plants gain mineral nutrition from the soil and may enhance switchgrass biomass production on marginal lands. Our goals were to identify miRNAs and their corresponding target genes that are controlling AM symbiosis in switchgrass. Results: Through genome-wide analysis of next-generation miRNA sequencing reads generated from switchgrass roots, we identified 122 mature miRNAs, including 28 novel miRNAs. By comparing miRNA expression profiles of AM-inoculated and control switchgrass roots, we identified 15 AM-responsive miRNAs across lowland accession “Alamo”, upland accession “Dacotah”, and two upland/lowland F 1 hybrids. We used degradome sequencing to identify target genes of the AM-responsive miRNAs revealing targets of miRNAs residing on both K and N subgenomes. Notably, genes involved in copper ion binding were targeted by downregulated miRNAs, while upregulated miRNAs mainly targeted GRAS family transcription factors. Conclusion: Through miRNA analysis and degradome sequencing, we revealed that both upland and lowland switchgrass genotypes as well as upland-lowland hybrids respond to AM by altering miRNA expression. We demonstrated complex GRAS transcription factor regulation by the miR171 family, with some miR171 family members being AM responsive while others remained static. Copper miRNA downregulation was common amongst the genotypes tested and we identified superoxide dismutases and laccases as targets, suggesting that these Cu-miRNAs are likely involved in ROS detoxification and lignin deposition, respectively. Other prominent targets of the Cu miRNAs were blue copper proteins. Overall, the potential effect of AM colonization on lignin deposition pathways in this biofuel crop highlights the importance of considering AM and miRNA in future biofuel crop development strategies.

59 BASIC BIOLOGICAL SCIENCES↗

DOE OSTI2021

Expression of a bacterial 3-dehydroshikimate dehydratase (QsuB) reduces lignin and improves biomass saccharification efficiency in switchgrass (Panicum virgatum L.)

Abstract Background Lignin deposited in plant cell walls negatively affects biomass conversion into advanced bioproducts. There is therefore a strong interest in developing bioenergy crops with reduced lignin content or altered lignin structures. Another desired trait for bioenergy crops is the ability to accumulate novel bioproducts, which would enhance the development of economically sustainable biorefineries. As previously demonstrated in the model plant Arabidopsis, expression of a 3-dehydroshikimate dehydratase in plants offers the potential for decreasing lignin content and overproducing a value-added metabolic coproduct (i.e., protocatechuate) suitable for biological upgrading. Results The 3-dehydroshikimate dehydratase QsuB from Corynebacterium glutamicum was expressed in the bioenergy crop switchgrass ( Panicum virgatum L.) using the stem-specific promoter of an O-methyltransferase gene ( pShOMT ) from sugarcane. The activity of pShOMT was validated in switchgrass after observation in-situ of beta-glucuronidase (GUS) activity in stem nodes of plants carrying a pShOMT::GUS fusion construct. Under controlled growth conditions, engineered switchgrass lines containing a pShOMT::QsuB construct showed reductions of lignin content, improvements of biomass saccharification efficiency, and accumulated higher amount of protocatechuate compared to control plants. Attempts to generate transgenic switchgrass lines carrying the QsuB gene under the control of the constitutive promoter pZmUbi-1 were unsuccessful, suggesting possible toxicity issues associated with ectopic QsuB expression during the plant regeneration process. Conclusion This study validates the transfer of the QsuB engineering approach from a model plant to switchgrass. We have demonstrated altered expression of two important traits: lignin content and accumulation of a co-product. We found that the choice of promoter to drive QsuB expression should be carefully considered when deploying this strategy to other bioenergy crops. Field-testing of engineered QsuB switchgrass are in progress to assess the performance of the introduced traits and agronomic performances of the transgenic plants.

59 BASIC BIOLOGICAL SCIENCES↗

DOE OSTI2017

Pollen-mediated gene flow from transgenic to non-transgenic switchgrass (Panicum virgatum L.) in the field

Background: Switchgrass is C4 perennial grass species that is being developed as a cellulosic bioenergy feedstock. It is wind-pollinated and considered to be an obligate outcrosser. Genetic engineering has been used to alter cell walls for more facile bioprocessing and biofuel yield. Gene flow from transgenic cultivars would likely be of regulatory concern. In this study we investigated pollen-mediated gene flow from transgenic to nontransgenic switchgrass in a 3-year field experiment performed in Oliver Springs, Tennessee, U.S.A. using a modified Nelder wheel design. The planted area (0.6 ha) contained sexually compatible pollen source and pollen receptor switchgrass plants. One hundred clonal switchgrass ‘Alamo’ plants transgenic for an orange-fluorescent protein (OFP) and hygromycin resistance were used as the pollen source; whole plants, including pollen, were orange-fluorescent. To assess pollen movement, pollen traps were placed at 10 m intervals from the pollen-source plot in the four cardinal directions extending to 20 m, 30 m, 30 m, and 100 m to the north, south, west, and east, respectively. To assess pollination rates, nontransgenic ‘Alamo 2’ switchgrass clones were planted in pairs adjacent to pollen traps. Results: In the eastward direction there was a 98% decrease in OFP pollen grains from 10 to 100 m from the pollen-source plot (Poisson regression, F1,8 = 288.38, P < 0.0001). At the end of the second and third year, 1,820 F1 seeds were collected from pollen recipient-plots of which 962 (52.9%) germinated and analyzed for their transgenic status. Transgenic progeny production detected in each pollen-recipient plot decreased with increased distance from the edge of the transgenic plot (Poisson regression, F1,15 = 12.98, P < 0.003). The frequency of transgenic progeny detected in the eastward plots (the direction of the prevailing wind) ranged from 79.2% at 10 m to 9.3% at 100 m. Conclusions: In these experiments we found transgenic pollen movement and hybridization rates to be inversely associated with distance. However, these data suggest pollen-mediated gene flow is likely to occur up to, at least, 100 m. This study gives baseline data useful to determine isolation distances and other management practices should transgenic switchgrass be grown commercially in relevant environments.

09 BIOMASS FUELS↗

DOE OSTI2014

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↗

DOE OSTI2019

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↗

DOE OSTI2019

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↗

DOE OSTI2021

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↗

DOE OSTI2018

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↗

DOE OSTI2022

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↗

DOE OSTI2024

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)↗

DOE OSTI2023

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↗