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Plants release a wealth of metabolites into the rhizosphere that can shape the composition and activity of microbial communities in response to environmental stress. The connection between rhizodeposition and rhizosphere microbiome succession has been suggested, particularly under environmental stress conditions, yet definitive evidence is scarce. In this study, we investigated the relationship between rhizosphere chemistry, microbiome dynamics, and abiotic stress in the bioenergy crop switchgrass grown in a marginal soil under nutrient-limited, moisture-limited, and nitrogen (N)-replete, phosphorus (P)-replete, and NP-replete conditions. We combined 16S rRNA amplicon sequencing and LC-MS/MS-based metabolomics to link rhizosphere microbial communities and metabolites. We identified significant changes in rhizosphere metabolite profiles in response to abiotic stress and linked them to changes in microbial communities using network analysis. N-limitation amplified the abundance of aromatic acids, pentoses, and their derivatives in the rhizosphere, and their enhanced availability was linked to the abundance of bacterial lineages from Acidobacteria, Verrucomicrobia, Planctomycetes, and Alphaproteobacteria. Conversely, N-amended conditions increased the availability of N-rich rhizosphere compounds, which coincided with proliferation of Actinobacteria. Treatments with contrasting N availability differed greatly in the abundance of potential keystone metabolites; serotonin and ectoine were particularly abundant in N-replete soils, while chlorogenic, cinnamic, and glucuronic acids were enriched in N-limited soils. Serotonin, the keystone metabolite we identified with the largest number of links to microbial taxa, significantly affected root architecture and growth of rhizosphere microorganisms, highlighting its potential to shape microbial community and mediate rhizosphere plant-microbe interactions.

Summary: Dinitrotoluene sulfonates (DNTSes) are highly toxic hazards regulated by the Resource Conservation and Recovery Act (RCRA) in the United States. The trinitrotoluene (TNT) red water formed during the TNT purification process consists mainly of DNTSes. Certain plants, including switchgrass, reed and alfalfa, can detoxify low concentrations of DNTS in TNT red water‐contaminated soils. However, the precise mechanism by which these plants detoxify DNTS remains unknown. In order to aid in the development of phytoremediation resources with high DNTS removal rates, we identified and characterized 1‐hydroxymethyl‐2,4‐dinitrobenzene sulfonic acid (HMDNBS) and its glycosylated product HMDNBS O‐glucoside as the degradation products of 2,4‐DNT‐3‐SO3Na, the major isoform of DNTS in TNT red water‐contaminated soils, in switchgrass via LC–MS/MS‐ and NMR‐based metabolite analyses. Transcriptomic analysis revealed that 15 UDP‐glycosyltransferase genes were dramatically upregulated in switchgrass plants following 2,4‐DNT‐3‐SO3Na treatment. We expressed, purified and assayed the activity of recombinant UGT proteins in vitro and identified PvUGT96C10 as the enzyme responsible for the glycosylation of HMDNBS in switchgrass. Overexpression of PvUGT96C10 in switchgrass significantly alleviated 2,4‐DNT‐3‐SO3Na‐induced plant growth inhibition. Notably, PvUGT96C10‐overexpressing transgenic switchgrass plants removed 83.1% of 2,4‐DNT‐3‐SO3Na in liquid medium after 28 days, representing a 3.2‐fold higher removal rate than that of control plants. This work clarifies the DNTS detoxification mechanism in plants for the first time, suggesting that PvUGT96C10 is crucial for DNTS degradation. Our results indicate that PvUGT96C10‐overexpressing plants may hold great potential for the phytoremediation of TNT red water‐contaminated soils. [ABSTRACT FROM AUTHOR]

Technologies for producing renewable fuels and chemicals rely on the production of stable intermediates. For thermochemical technologies, pyrolysis of biomass produces oils that must compromise between carbon yield and oil quality. Bio-oil extraction has largely focused on regular bio-oils (~ 33 wt% O) and partially deoxygenated oils (< 12 wt% O). Furthermore, it is desired to extract phenolics without direct distillation of bio-oils, which would enable extraction from the heaviest portion of bio-oil. Mid-level oxygen (MLO) bio-oils (16–25 wt% O) produced from switchgrass were characterized for their ability to separate into phenolic-rich fractions. Toluene-soluble portions of the oils underwent NaOH extraction to extract one-ring phenolics, while toluene-insoluble portions were fractionated with iso-propyl alcohol (IPA). While phenolic extraction proceeded without distillation (having been a prerequisite for partially deoxygenated bio-oils), the efficiency of extraction was less than optimal, owing to the presence of other oxygenated compounds in the hydrocarbon-rich fraction. Both IPA-insoluble and IPA-soluble fractions underwent solvent liquefaction reactions with base additives. While using water as a reaction medium produced greater concentrations of phenols than when using methanol, addition of sodium carbonate produced narrower product distributions of phenols and inhibited formation of benzenediols. [ABSTRACT FROM AUTHOR]

The diverse class of plant diterpenoid metabolites serves important functions in mediating growth, chemical defence, and ecological adaptation. In major monocot crops, such as maize (Zea mays), rice (Oryza sativa), and barley (Hordeum vulgare), diterpenoids function as core components of biotic and abiotic stress resilience. Switchgrass (Panicum virgatum) is a perennial grass valued as a stress‐resilient biofuel model crop. Previously we identified an unusually large diterpene synthase family that produces both common and species‐specific diterpenoids, several of which accumulate in response to abiotic stress. Here, we report discovery and functional characterization of a previously unrecognized monofunctional class I diterpene synthase (PvKSL1) via in vivo co‐expression assays with different copalyl pyrophosphate (CPP) isomers, structural and mutagenesis studies, as well as genomic and transcriptomic analyses. In particular, PvKSL1 converts ent‐CPP into ent‐abietadiene, ent‐palustradiene, ent‐levopimaradiene, and ent‐neoabietadiene via a 13‐hydroxy‐8(14)‐ent‐abietene intermediate. Notably, although featuring a distinct ent‐stereochemistry, this product profile is near‐identical to bifunctional (+)‐levopimaradiene/abietadiene synthases occurring in conifer trees. PvKSL1 has three of four active site residues previously shown to control (+)‐levopimaradiene/abietadiene synthase catalytic specificity. However, mutagenesis studies suggest a distinct catalytic mechanism in PvKSL1. Genome localization of PvKSL1 distant from other diterpene synthases, and its phylogenetic distinctiveness from known abietane‐forming diterpene synthases, support an independent evolution of PvKSL1 activity. Albeit at low levels, PvKSL1 gene expression predominantly in roots suggests a role of diterpenoid formation in belowground tissue. Together, these findings expand the known chemical and functional space of diterpenoid metabolism in monocot crops. [ABSTRACT FROM AUTHOR]

Nitrogen (N) availability is a well‐known driver of ecosystem structure and function, but as air quality regulations continue to reduce atmospheric N deposition, there is a need to understand how managed and unmanaged ecosystems respond to widespread decreases in terrestrial N availability. Historical N eutrophication, from pollution or fertilisation, may continue to constrain contemporary responses to decreases in available N because of altered plant and microbial feedbacks. Thus, while certain management practices like prescribed fire remove N from grassland ecosystems, the role of fire supporting ecosystems recovering from chronic N input is unknown. To address this knowledge gap, we ceased a 30‐year N‐fertilisation treatment at a field experiment in a tallgrass prairie ecosystem crossed with burned and fire‐suppressed (unburned) treatments. We established subplots within each previously fertilised, recovering plot, fertilised at the same historical rate (10 g N m−2 year−1 as NH4NO3), to compare plant and soil properties in recovering plots with control (never‐fertilised) and still‐fertilised treatments within different fire regimes. We document different N‐fertilisation legacies among ecosystem properties in burned and unburned prairies recovering from N‐fertilisation. Soil N availability, nitrification and denitrification potentials in recovering plots remained higher than controls for 3–5 years—indicative of positive legacies—in both burned and unburned prairies, but burning did not reduce this legacy. In burned prairies, however, a positive legacy in above‐ground plant production persisted because a more productive grass species (switchgrass) replaced the previously dominant species (big bluestem) even though root C:N, but not soil C:N, increased to return back to control levels. Consequently, the main N loss pathways in burned and unburned prairies (pyrovolatilisation and microbially mediated processes, respectively) led to similar losses of soil total N (20–28 g N m−2) over 5 years. Synthesis: Our results indicate that N eutrophication induces positive legacies of ecosystem functions that can persist for at least half a decade. N‐induced legacies arise because of shifts in soil microbial N‐cycling and plant functional traits. As a result, different management practices may elicit similar trajectories of ecosystem recovery in terms of total and available soil N because of different plant and microbial feedbacks. [ABSTRACT FROM AUTHOR]

This study investigates uncertainties in greenhouse gas (GHG) emission factors related to switchgrass‐based biofuel production in Michigan. Using three life cycle assessment (LCA) databases—US lifecycle inventory (USLCI) database, GREET, and Ecoinvent—each with multiple versions, we recalculated the global warming intensity (GWI) and GHG mitigation potential in a static calculation. Employing Monte Carlo simulations along with local and global sensitivity analyses, we assess uncertainties and pinpoint key parameters influencing GWI. The convergence of results across our previous study, static calculations, and Monte Carlo simulations enhances the credibility of estimated GWI values. Static calculations, validated by Monte Carlo simulations, offer reasonable central tendencies, providing a robust foundation for policy considerations. However, the wider range observed in Monte Carlo simulations underscores the importance of potential variations and uncertainties in real‐world applications. Sensitivity analyses identify biofuel yield, GHG emissions of electricity, and soil organic carbon (SOC) change as pivotal parameters influencing GWI. Decreasing uncertainties in GWI may be achieved by making greater efforts to acquire more precise data on these parameters. Our study emphasizes the significance of considering diverse GHG factors and databases in GWI assessments and stresses the need for accurate electricity fuel mixes, crucial information for refining GWI assessments and informing strategies for sustainable biofuel production. [ABSTRACT FROM AUTHOR]

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 CO2 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 CO2 concentration and light, we measured leaf net photosynthesis of switchgrass under different CO2 concentrations and light levels in 2020 and 2021 for each of the five precipitation treatments. We first evaluated four light and CO2 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 CO2 response curves. Precipitation change did not influence the maximum net photosynthetic rate (Pmax) 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 Pmax of five precipitation treatments was 17.6 μmol CO2 m−2 s−1, and the ambient treatment tended to have a higher Pmax. 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 CO2 response. The ambient treatment had the highest Pmax, largest α, and lowest θ, Rd, and CO2 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. [ABSTRACT FROM AUTHOR]

Perennial ornamental grasses are often recommended for rain gardens, but few data support their use. We conducted two experiments to evaluate the ability of ornamental grass cultivars to grow while subjected to cyclical flooding, submergence, and drought typical of rain gardens. Our objectives were to determine the effects of cyclical flood and drought (Expt. 1) and submergence depth and duration (Expt. 2) on grass growth and survival. Seven cultivars were evaluated during greenhouse trials, including Pixie Fountain tufted hairgrass [Deschampsia cespitosa (L.) P. Beauv.], Northwind switchgrass (Panicum virgatum L.), Red October big bluestem (Andropogon gerardii Vitman), Purpurascens Chinese silvergrass (Miscanthus sinensis Andersson), Blue HeavenVR little bluestem [Schizachyrium scoparium (Michx.) Nash], Blonde Ambition blue grama grass [Bouteloua gracilis (Kunth) Lag. ex Griffiths], and Karl Foerster feather reed grass [Calamagrostis ×acutiflora (Schrad.) DC]. During Expt. 1, grasses underwent four cycles of flooding duration (2 days or 7 days) followed by drought (drying to volumetric soil water contents of 0.14 or 0.07 cm³·cm-3). During Expt. 2, grasses were cyclically submerged at 15 or 30 cm above the soil surface for 2, 4, or 7 days, followed by floodwater removal and drainage for 2 days before being resubmerged. Cyclical submergence continued until the 7-day submergence treatments completed four cycles. Both experiments were replicated in a full factorial randomized complete block design. Controls were included in both experiments. Plants were measured to determine plant height, shoot count, visual damage rating, shoot dry weight, and root dry weight. Floodwater chemistry and soil reducing conditions were measured during Expt. 2. Chinese silvergrass and switchgrass survived cyclical soil flooding/drought and submergence for 7 days at a depth of 30 cm while maintaining acceptable foliar damage. All grasses survived cyclical flood and drought when the soil volumetric water content was maintained at 14%, suggesting they can withstand periodic soil flooding as long as the water is not too deep. As water depth and duration increased from 4 days to 7 days, little bluestem, blue grama grass, and feather reed grass experienced significant foliar damage. Tufted hair grass and big bluestem experienced significant foliar damage when submerged for 2 days. Our results showed that perennial ornamental grasses can tolerate cyclical flood and drought and periodic submergence, but that plant conditions and survival vary, which can inform strategic plant placement within rain gardens, bioretention basins, and other stormwater management systems. [ABSTRACT FROM AUTHOR]

Mitigating the effects of climate stress on crops is important for global food security. The microbiome associated with plant roots, the rhizobiome, can harbor beneficial microbes that alleviate stress, but the factors influencing their recruitment are unclear. We conducted a greenhouse experiment using field soil with a legacy of growing switchgrass and common bean to investigate the impact of short-term drought severity on the recruitment of active bacterial rhizobiome members. We applied 16S rRNA and 16S rRNA gene sequencing for both crops and metabolite profiling for switchgrass. We included planted and unplanted conditions to distinguish environment- versus plant-mediated rhizobiome drivers. Differences in community structure were observed between crops and between drought and watered and planted and unplanted treatments within crops. Despite crop-specific communities, drought rhizobiome dynamics were similar across the two crops. The presence of a plant more strongly explained the rhizobiome variation in bean (17%) than in switchgrass (3%), with a small effect of plant mediation during drought observed only for the bean rhizobiome. The switchgrass rhizobiome was stable despite changes in rhizosphere metabolite profiles between planted and unplanted treatments. We conclude that rhizobiome responses to short-term drought are crop-specific, with possible decoupling of plant exudation from rhizobiome responses. In a study of short-term drought, the authors show an effect of plant mediation on the active rhizosphere microbiome of common bean, but not switchgrass. They conclude that microbiome responses are crop-specific and have possible decoupling of plant exudation and microbiome response. [ABSTRACT FROM AUTHOR]

Background and aims: Root exudation is a key process for plant nutrient acquisition, but the controls on root exudation and its relationship to soil C and N processes in agroecosystems are unclear. We hypothesized that root exudation rates would be related to root morphological traits, N fertilization, and soil moisture. We also anticipated that root exudation would be correlated with bulk soil enzyme activity. Methods: Root exudation, root traits, and bulk soil extracellular enzyme activity were assessed in maize (Zea mays L.), soybean (Glycine max (L.) Merr.), biomass sorghum (Sorghum bicolor (L.) Moench), giant miscanthus (Miscanthus × giganteus), and switchgrass (Panicum virgatum L.). Measurements were taken in situ during two growing seasons with contrasting precipitation regimes, and N fertilization rate was varied in sorghum during one year. Results: Specific root exudation (per unit root surface area) was negatively related to root diameter and was generally higher in annuals than perennials. Sorghum N fertilization did not affect root exudation rates, and soil moisture regime had no effect on annual root exudation rates within maize, sorghum, and miscanthus. Specific root exudation was negatively related to bulk soil C- and N-degrading soil enzyme activities. Conclusion: Intrinsic plant characteristics appeared more important than environmental variables in controlling in situ root exudation rates. The relationships between root diameter, root exudation, and soil C and N processes link root morphological traits to soil functions and demonstrate the potential tradeoffs among plant nutrient acquisition strategies in agroecosystems. [ABSTRACT FROM AUTHOR]

The area of land dedicated to growing maize for bioenergy in the United Kingdom is rapidly expanding. To understand how maize production influences soil carbon (C) dynamics, and whether this is influenced by soil type, we measured net ecosystem exchange (NEE) using the eddy covariance technique over the 2021 growing season. We combined the NEE data with C imports and exports to calculate the net ecosystem productivity (NEP) of two maize crops grown for bioenergy in the United Kingdom, one site on mineral soil and the other on lowland agricultural peat. Maize was similarly productive at both sites—gross primary productivity was 1107 g C m−2 at the site with mineral soil and 1407 g C m−2 at the peat site. However, total ecosystem respiration was considerably higher from the peat site (1198 g C m−2) compared with the mineral soil site (678 g C m−2). After accounting for the removal of C in harvested biomass, both sites were net C sources, but C losses were over two times greater from the peat site (NEP = 290 g C m−2) than the mineral site (NEP = 136 g C m−2). While annual crops may be needed to produce bioenergy in the short term, growing maize for bioenergy in the United Kingdom does not appear to be a viable option for C sequestration over the long term, as it leads to high carbon losses from agroecosystems, especially those on organic soils. Instead, growing perennial bioenergy crops on mineral soils with a low organic C content is a more appropriate option. [ABSTRACT FROM AUTHOR]

Due to the environmental consequences of annual‐dominated cropping systems, there is an increasing need to identify agronomic strategies that incorporate perennial crops. One strategy for increasing perennial cover is through the targeted use of annually harvested perennial food and bioproduct crops in buffer strips, which has the potential to create new revenue streams for farmers and substantially mitigate agricultural nutrient pollution from conventional cropping systems. As buffers are typically installed on marginal land, it is critical to understand how landscape position influences the success of perennial crops. The objectives of this study were to determine the relatively early influence of landscape position on the productivity of a variety of perennial crops and their subsequent soil nutrients and soil water storing capabilities. In this experiment, nine perennial (alfalfa, alsike clover, indiangrass, switchgrass, big bluestem, prairie cordgrass, intermediate wheatgrass, high‐diversity polyculture, low‐diversity polyculture) and two annual (corn, soybean) crops were planted across two landscape positions (hillslope and deposition). Plant biomass, plant tissue nitrogen, soil moisture, and soil NO3‐N and NH4‐N were measured and compared at two different locations in Minnesota. Overall, the polyculture mixes, and to some extent intermediate wheatgrass, performed the best with respect to biomass production while also providing ecosystem services across most soil by landscape position combinations tested in this study. However, there were some important findings specific to each soil and landscape position combination, mainly oriented toward biomass production. We also observed temporal patterns in soil moisture and depth‐related patterns in soil N reductions. This study presents an opportunity to optimize the use of perennial crops on marginal agricultural lands for improved environmental and economic benefit. [ABSTRACT FROM AUTHOR]

Introduction: Melatonin (N-acetyl-5-methoxytryptamine) is a molecule implicated in multiple biological functions, but exerts contrasting effects on plants owing to concentration differences. Hydroxyindole O-methyltransferase (HIOMT), which catalyzes the last step of melatonin synthesis, plays a crucial role in this context. Methods: Transgenic switchgrass overexpressing oHIOMT with different melatonin levels displayed distinct morphological changes in a concentrationdependent manner. In this study, we divided the transgenic switchgrass into two groups: melatonin-moderate transgenic (MMT) plants and melatonin-rich transgenic (MRT) plants. To determine the concentration-dependent effect of melatonin on switchgrass growth and stress resistance, we conducted comparative morphological, physiological, omics and molecular analyses between MMT, MRT and wild-type (WT) plants. Results: We found that oHIOMT overexpression, with moderate melatonin levels, was crucial in regulating switchgrass growth through changes in cell size rather than cell number. Moderate levels of melatonin were vital in regulating carbon fixation, stomatal development and chlorophyll metabolism. Regarding salt tolerance, melatonin with moderate levels activated numerous defense (e.g. morphological characteristics, anatomical structure, antioxidant enzymatic properties, non-enzymatic capacity and Na+/K+ homeostasis). Additionally, moderate levels of oHIOMT overexpression were sufficient to increase lignin content and alter monolignol compositions with an increase in the S/G lignin ratio. Discussion: Taken together, oHIOMT overexpression in switchgrass with different melatonin levels resulted in morphological, anatomical, physiological and molecular changes in a concentration-dependent manner, which characterized by stimulation at low doses and inhibition at high doses. Our study presents new ideas and clues for further research on the mechanisms of the concentration-dependent effect of melatonin. [ABSTRACT FROM AUTHOR]

Abstract Background MicroRNA396 (miR396) plays an important role in the regulation of plant growth and development by repressing the expression level of its target growth-regulating factor (GRF) family genes. In our previous study, we found that overexpression of miR396 negatively regulated both tillering and biomass yield in switchgrass (Panicum virgatum L.). We, therefore, speculated that blocking the expression of miR396 could enhance switchgrass tillering and biomass yield. Here, we produced transgenic switchgrass plants overexpressing a target mimicry form of miR396 (MIM396) in wild type (WT) and Os-MIR319b overexpressing switchgrass plant (with higher enzymatic hydrolysis efficiency, but reduced tillering), in which the expression of miR396 was blocked. The phenotype and biological yields of these plants were analyzed. Results Blocking miR396 to improve its target PvGRFs expression in switchgrass improved the tiller number and dry weight of transgenic plants. Further morphological analysis revealed that MIM396 plants increased the number of aerial branches and basal tillers compared to those of wild-type plants. The enzymatic efficiency of MIM396 plants was reduced; however, the total sugar production per plant was still significantly higher than that of wild-type plants due to the increase in biomass. In addition, blocking miR396 in a transgenic switchgrass plant overexpressing Os-MIR319b (TG21-Ms) significantly increased the PvGRF1/3/5 expression level and tiller number and biomass yield. The miR156-target gene PvSPL4, playing a negative role in aerial and basal buds outgrowth, showed significant downregulated in MIM396 and TG21-Ms. Those results indicate that miR396-PvGRFs, through disrupting the PvSPL4 expression, are involved in miR319-PvPCFs in regulating tiller number, at least partly. Conclusions MIM396 could be used as a molecular tool to improving tiller number and biomass yield in switchgrass wild type and miR319b transgenic plants. This finding may be applied to other graminaceous plants to regulate plant biological yield.

Abstract Monoculture switchgrass and restored prairie are promising perennial feedstock sources for bioenergy production on the lands unsuitable for conventional agriculture. Such lands often display contrasting topography that influences soil characteristics and interactions between plant growth and soil C gains. This study aimed at elucidating the influences of topography and plant systems on the fate of C originated from switchgrass plants and on its relationships with soil pore characteristics. For that, switchgrass plants were grown in intact soil cores collected from two contrasting topographies, namely steep slopes and topographical depressions, in the fields in multi-year monoculture switchgrass and restored prairie vegetation. The 13C pulse labeling allowed tracing the C of switchgrass origin, which X-ray computed micro-tomography enabled in-detail characterization of soil pore structure. In eroded slopes, the differences between the monoculture switchgrass and prairie in terms of total and microbial biomass C were greater than those in topographical depressions. While new switchgrass increased the CO2 emission in depressions, it did not significantly affect the CO2 emission in slopes. Pores of 18–90 µm Ø facilitated the accumulation of new C in soil, while > 150 µm Ø pores enhanced the mineralization of the new C. These findings suggest that polyculture prairie located in slopes can be particularly beneficial in facilitating soil C accrual and reduce C losses as CO2.

SUMMARY: Culm development in grasses can be controlled by both miR156 and cytokinin. However, the crosstalk between the miR156‐SPL module and the cytokinin metabolic pathway remains largely unknown. Here, we found CYTOKININ OXIDASE/DEHYDROGENASE4 (PvCKX4) plays a negative regulatory role in culm development of the bioenergy grass Panicum virgatum (switchgrass). Overexpression of PvCKX4 in switchgrass reduced the internode diameter and length without affecting tiller number. Interestingly, we also found that PvCKX4 was always upregulated in miR156 overexpressing (miR156OE) transgenic switchgrass lines. Additionally, upregulation of either miR156 or PvCKX4 in switchgrass reduced the content of isopentenyl adenine (iP) without affecting trans‐zeatin (tZ) accumulation. It is consistent with the evidence that the recombinant PvCKX4 protein exhibited much higher catalytic activity against iP than tZ in vitro. Furthermore, our results showed that miR156‐targeted SPL2 bound directly to the promoter of PvCKX4 to repress its expression. Thus, alleviating the SPL2‐mediated transcriptional repression of PvCKX4 through miR156 overexpression resulted in a significant increase in cytokinin degradation and impaired culm development in switchgrass. On the contrary, suppressing PvCKX4 in miR156OE transgenic plants restored iP content, internode diameter, and length to wild‐type levels. Most strikingly, the double transgenic lines retained the same increased tiller numbers as the miR156OE transgenic line, which yielded more biomass than the wild type. These findings indicate that the miR156‐SPL module can control culm development through transcriptional repression of PvCKX4 in switchgrass, which provides a promising target for precise design of shoot architecture to yield more biomass from grasses. [ABSTRACT FROM AUTHOR]

Flowering is the change from the vegetative state to the reproductive state. Flowering initiation is regulated by endogenous genetic components and by numerous environmental factors. These stimuli determine the different molecular mechanisms that are required to give way to flowering and vary according to the dose of the stimulus and the crop used. Jatropha curcas is a perennial shrub that produces oil from its seeds of adequate quality to be transformed into biofuels. However, this plant still presents limitations for its establishment as a commercial crop since it produces low seeds, which can be attributed to its low production of female flowers. This plant also presents multiple and asynchronous flowering periods as a consequence of the accession and of the climatic conditions, which implies that the fruits do not ripen synchronously, and therefore the harvests are constant, manual, and expensive. Knowing the mechanisms that regulate the flowering of J. curcas is important to improve its seed production. Therefore, current revision covers the analysis of genes involved in flowering to gain information on the pathways involved in floral transition and in the differentiation of the sexual organs of J. curcas. [ABSTRACT FROM AUTHOR]

In order to obtain high yields of chemicals and fuels from biomass, feedstocks need to be selected that contain high amounts of glucose, xylose, or lignin, depending on the end product. The first objective of this study was to evaluate the biomass composition of 20 samples from 11 broadleaf and grass crop species grown at two distinct sites in Montana. Another objective was to investigate the influence of growing location and seed type to determine which cultivars may be best for renewable chemical production in the northern Great Plains. There was a significant effect due to the growing location as the biomass samples from the site that had higher precipitation exhibited a significantly higher acid-insoluble lignin (23.8%) content compared to the biomass samples from the lower precipitation site (20.0%). Additionally, the 357 napus had a significantly higher amount of glucose (1.8% more) and acid-soluble lignin (0.2% more) than compared to the 940 napus both grown at the Froid site that may be attributed to genetic modifications. In terms of sugar content for potential upgrading, the grasses had significantly more glucose (36.1% total), xylose (24.3%), and arabinose (2.3%) than the Brassicaceae biomasses, but the lignin content was not significantly different. Switchgrass, in particular, had the highest total sugar content of close to 70 wt%. Understanding the impact of growing location on biomass composition is an important consideration for optimizing biomass utilization strategies and developing sustainable bioenergy production systems. [ABSTRACT FROM AUTHOR]

Perennial tall grasses show promise as bioenergy crops due to high productivity and efficient nutrient use. Ongoing research on bioenergy grasses seeks to reduce their reliance on synthetic nitrogen (N) fertilizer, the manufacture of which relies on fossil fuel combustion. Excessive use of fertilizers also causes adverse environmental consequences and leads to the evolutionary loss of plant traits beneficial to sustainable N cycle. Notably, perennial tall grasses have exhibited the potential to maintain high biomass yield without the need for N fertilizer or causing soil N depletion. Perennial grasses can be adept at interacting with their microbial partners to facilitate N acquisition and retention via mechanisms such as biological N fixation and nitrification inhibition. These inherent N management traits should be preserved and optimized at the this early stage of bioenergy grass breeding programs. This review examines the impact of external N on bioenergy grass production and explores the potential of leveraging advantageous N‐cycling attributes of perennial tall grasses, laying groundwork for future management and research efforts. With minimized dependency on external N input, the cultivation of perennial energy grasses will pave the way toward more resilient agricultural systems and play a significant role in addressing key global energy and environmental challenges. [ABSTRACT FROM AUTHOR]

Incorporating native forbs within native warm‐season grass (NWSG) pastures has the potential to benefit cattle, pollinators, and wildlife beyond that of NWSG monocultures. However, when grazing NWSG pastures, rotational stocking is recommended as opposed to continuous stocking. Therefore, to evaluate whether within‐season rest is needed for native pasture sustainability, two NWSG grazing experiments were conducted near Greeneville, TN, 2017–2020, to assess the persistence of native forbs when an 11‐species native forb blend was interseeded into established switchgrass (SG; Panicum virgatum L.) and big bluestem/indiangrass (BBIG; Andropogon gerardii Vitman and Sorghastrum nutans (L.) Nash) pastures. Each experiment was a completely randomized design with four replicates of each within‐season rest grazing treatment (no rest, early rest, middle rest, late rest, and no graze). Within‐season rest was not influential for NWSG tiller density or total forb plant density thus indicating persistence of forbs may not require rotational stocking. Purple prairie clover (Dalea purpurea Vent.) did not establish while Illinois bundleflower (Desmanthus illinoensis (Michx.) MacMill. ex B.L. Rob. & Fernald) was only observed flowering once despite having the greatest seeding rate among forbs. Based on establishment and flowering of the 11 species in the current mixture, interseeding a 6‐species polyculture of black‐eyed Susan (Rudbeckia hirta L.), dixie ticktrefoil (Desmodium tortuosum (Sw.) DC.), eastern purple coneflower (Echinacea purpurea (L.) Moench), lanceleaf coreopsis (Coreopsis lanceolata L.), Maximilian sunflower (Helianthus maximiliani Schrad.), and oxeye sunflower (Heliopsis helianthoides (L.) Sweet) could allow for plant biodiversity while offering floral resources for pollinators during the NWSG grazing season. [ABSTRACT FROM AUTHOR]

Chalcone synthase (CHS) and chalcone isomerase (CHI) catalyze the first two committed steps of the flavonoid pathway that plays a pivotal role in the growth and reproduction of land plants, including UV protection, pigmentation, symbiotic nitrogen fixation, and pathogen resistance. Based on the obtained X-ray crystal structures of CHS, CHI, and chalcone isomerase-like protein (CHIL) from the same monocotyledon, Panicum virgatum, along with the results of the steady-state kinetics, spectroscopic/thermodynamic analyses, intermolecular interactions, and their effect on each catalytic step are proposed. In addition, PvCHI's unique activity for both naringenin chalcone and isoliquiritigenin was analyzed, and the observed hierarchical activity for those type-I and -II substrates was explained with the intrinsic characteristics of the enzyme and two substrates. The structure of PvCHS complexed with naringenin supports uncompetitive inhibition. PvCHS displays intrinsic catalytic promiscuity, evident from the formation of p-coumaroyltriacetic acid lactone (CTAL) in addition to naringenin chalcone. In the presence of PvCHIL, conversion of p-coumaroyl-CoA to naringenin through PvCHS and PvCHI displayed ~400-fold increased Vmax with reduced formation of CTAL by 70%. Supporting this model, molecular docking, ITC (Isothermal Titration Calorimetry), and FRET (Fluorescence Resonance Energy Transfer) indicated that both PvCHI and PvCHIL interact with PvCHS in a non-competitive manner, indicating the plausible allosteric effect of naringenin on CHS. Significantly, the presence of naringenin increased the affinity between PvCHS and PvCHIL, whereas naringenin chalcone decreased the affinity, indicating a plausible feedback mechanism to minimize spontaneous incorrect stereoisomers. These are the first findings from a three-body system from the same species, indicating the importance of the macromolecular assembly of CHS-CHI-CHIL in determining the amount and type of flavonoids produced in plant cells. [ABSTRACT FROM AUTHOR]

Background: MicroRNA396 (miR396) plays an important role in the regulation of plant growth and development by repressing the expression level of its target growth-regulating factor (GRF) family genes. In our previous study, we found that overexpression of miR396 negatively regulated both tillering and biomass yield in switchgrass (Panicum virgatum L.). We, therefore, speculated that blocking the expression of miR396 could enhance switchgrass tillering and biomass yield. Here, we produced transgenic switchgrass plants overexpressing a target mimicry form of miR396 (MIM396) in wild type (WT) and Os-MIR319b overexpressing switchgrass plant (with higher enzymatic hydrolysis efficiency, but reduced tillering), in which the expression of miR396 was blocked. The phenotype and biological yields of these plants were analyzed. Results: Blocking miR396 to improve its target PvGRFs expression in switchgrass improved the tiller number and dry weight of transgenic plants. Further morphological analysis revealed that MIM396 plants increased the number of aerial branches and basal tillers compared to those of wild-type plants. The enzymatic efficiency of MIM396 plants was reduced; however, the total sugar production per plant was still significantly higher than that of wild-type plants due to the increase in biomass. In addition, blocking miR396 in a transgenic switchgrass plant overexpressing Os-MIR319b (TG21-Ms) significantly increased the PvGRF1/3/5 expression level and tiller number and biomass yield. The miR156-target gene PvSPL4, playing a negative role in aerial and basal buds outgrowth, showed significant downregulated in MIM396 and TG21-Ms. Those results indicate that miR396-PvGRFs, through disrupting the PvSPL4 expression, are involved in miR319-PvPCFs in regulating tiller number, at least partly. Conclusions: MIM396 could be used as a molecular tool to improving tiller number and biomass yield in switchgrass wild type and miR319b transgenic plants. This finding may be applied to other graminaceous plants to regulate plant biological yield. [ABSTRACT FROM AUTHOR]

Polyploid plants often exhibit enhanced stress tolerance. Switchgrass is a perennial rhizomatous bunchgrass that is considered ideal for cultivation in marginal lands, including sites with saline soil. In this study, we investigated the physiological responses and transcriptome changes in the octoploid and tetraploid of switchgrass (Panicum virgatum L. 'Alamo') under salt stress. We found that autoploid 8× switchgrass had enhanced salt tolerance compared with the amphidiploid 4× precursor, as indicated by physiological and phenotypic traits. Octoploids had increased salt tolerance by significant changes to the osmoregulatory and antioxidant systems. The salt-treated 8× Alamo plants showed greater potassium (K+) accumulation and an increase in the K+/Na+ ratio. Root transcriptome analysis for octoploid and tetraploid plants with or without salt stress revealed that 302 upregulated and 546 downregulated differentially expressed genes were enriched in genes involved in plant hormone signal transduction pathways and were specifically associated with the auxin, cytokinin, abscisic acid, and ethylene pathways. Weighted gene co-expression network analysis (WGCNA) detected four significant salt stress-related modules. This study explored the changes in the osmoregulatory system, inorganic ions, antioxidant enzyme system, and the root transcriptome in response to salt stress in 8× and 4× Alamo switchgrass. The results enhance knowledge of the salt tolerance of artificially induced homologous polyploid plants and provide experimental and sequencing data to aid research on the short-term adaptability and breeding of salt-tolerant biofuel plants. [ABSTRACT FROM AUTHOR]

Biochar has been considered one of the viable solutions for atmospheric carbon sequestration because of its resistance to mineralization in soils. However, our knowledge of the mineralization rates of biochar in soils is still inconclusive due to the drawbacks of either the study methods or insufficient characterization of the studied materials or both. The objectives of this study were (1) to characterize switchgrass (SG) biochar by multi-element scanning thermal analysis (MESTA), and (2) to determine the mineralization rates of the SG biochar carbon (BC) in soil by the 13C tracer method. The results indicated that the 400 °C and the 600 °C SG biochars were not significantly different in chemical composition or thermal stability. A MESTA thermogram of the SG feedstock can predict the yield and thermal stability of the resulting biochar. The 13C tracer determined the mineralization rates and deduced the corresponding mean residence times (MRTs) of the BC, SG-C, and native SOC to be 347 years, 4 years, and 65 years, respectively. The MRT of BC should be a minimal estimate because of the limited incubation period. Even so, the MRT was already two orders of magnitude greater than that of the SG feedstock indicating the carbon sequestration potential of the biochar in soil. [ABSTRACT FROM AUTHOR]

Inoculation of arbuscular mycorrhizal fungi (AMF) or biochar (BC) application can improve photosynthesis and promote plant growth under saline‐alkali stress. However, little is known about the effects of the two combined on growth and physiological characteristics of switchgrass under saline‐alkali stress. This study examined the effects of four treatments: (1) no AMF inoculation and no biochar addition (control), (2) biochar (BC) alone, (3) AMF (Rhizophagus irregularis, Ri) alone, and (4) the combination of both (BC+Ri) on the plant biomass, antioxidant enzymes, chlorophyll, and photosynthetic parameters of switchgrass under saline‐alkali stress. The results showed that the above‐ground, belowground and total biomass of switchgrass in the BC+Ri treatment group was significantly higher (+136.7%, 120.2% and 132.4%, respectively) than in other treatments compared with Control. BC+Ri treatment significantly increased plant leaves' relative chlorophyll content, antioxidant enzyme activity, and photosynthesis parameters. It is worth noting that the transpiration rate, stomatal conductance, net photosynthetic rate, PSII efficiency and other photosynthetic‐related indexes of the BC+Ri treatment group were the highest (38% to 54% higher than other treatments). The fitting results of light response and CO2 response curves showed that the light saturation point, light compensation point, maximum carboxylation rate and maximum electron transfer rate of switchgrass in the Ri+BC treatment group were the highest. In conclusion, biochar combined with Ri has potential beneficial effects on promoting switchgrass growth under saline‐alkali stress and improving the activity of antioxidant enzymes and photosynthetic characteristics of plants. [ABSTRACT FROM AUTHOR]

As a potential crop in saline-alkali land, the growth of switchgrass could also be threatened by salt stress. Promoting the growth of switchgrass under salt stress by humic acid has great significance in the utilization of saline-alkali land. In this study, a pot experiment was arranged to investigate the responses of photosynthetic and physicochemical characteristics of switchgrass to HA under salt stress. Results showed that humic acid increased the photosynthetic function of switchgrass and enhanced plant height by 41.1% and dry weight by 26.9% under salt stress. Correlation analysis showed that the membrane aquaporin gene PvPIP1, malondialdehyde, ascorbate peroxidase, abscisic acid, polyamine, and jasmonic acid were important factors affecting the photosynthetic function of switchgrass in this study. Meanwhile, HA reduced the content of malondialdehyde, indicating the alleviation of the membrane damage caused by salt stress. On the other hand, HA upregulated the relative expression of the PvPIP1 gene and activated ascorbate peroxidase, abscisic acid, polyamine, and jasmonic acid in switchgrass to resist salt stress. These improved the membrane stability and promoted the photosynthetic activity of switchgrass to enhance the plant's tolerance against salt stress and growth. Results from this study are helpful to the efficient growing of switchgrass and the sustainable development of saline-alkali land. [ABSTRACT FROM AUTHOR]

The production of bioproduct feedstocks such as switchgrass (Panicum virgatum L.) and willow (Salix spp.) on degraded lands provides an opportunity to grow dedicated bioenergy crops with the potential to capture and store carbon in the soil while reducing competition with land for food production. However, how the production of these crops alters plant–soil–microbe interactions that govern soil C accumulation in highly degraded soil is underexplored. The objectives of this study were to examine select biological and chemical properties related to stable soil organic matter (SOM) production from the growth of switchgrass and willow on marginal soil over two growing seasons and whether biochar amendment can positively affect these parameters. To address our objectives, paired former surface mined lands and non‐mine impacted marginal agriculture sites were selected across West Virginia, USA, and biochar and unamended control treatments were imposed. Through the first two growing seasons, microbial activity and demand for carbon (C) increased and was accompanied by a shift in extracellular enzyme investment for decomposition‐associated enzymes. Mineral‐associated organic matter C increased over the two growing seasons, and this increase was greater in the mine sites compared to the agriculture sites. Compared to each site's previous land use, C losses were observed under bioproduct systems in the agriculture, but not the mine sites. Biochar amendments did not impact microbial activity but did increase the C:N of SOM. Overall, our results suggest that the early growth of switchgrass and willow can result in C accumulation in marginal and highly degraded lands. Core Ideas: Switchgrass (Panicum virgatum L.) and willow (Salix spp.) can be produced for bioproducts on highly degraded soil.Microbial demand for carbon (C) increased, while C inputs from the crops were variable over the first two growing seasons.Short‐term C losses were observed as compared to the previous land usein the agriculture, but not the reclaimed mine sites.There is potential for carbon accumulation in Appalachian reclaimed mine and marginal agriculture soil. [ABSTRACT FROM AUTHOR]

Biomass densification can reduce space needs during storage and transportation. It is important to quantify the amount of energy needed to densify biomass. Small bales (0.36 x 0.46 x 0.86 m) of herbaceous biomass crops, corn stover, indiangrass, switchgrass, and wheat straw, were recompressed in a compression chamber to determine their compression behavior in baled form. This chamber had the same cross-sectional area as a small baler. On average, the volume of all bales that were recompressed was reduced by 67.4%. Mean bale densities (d.b.) for corn stover, indiangrass, switchgrass, and wheat straw were increased from 94 to 264 kg m-3, 117 to 352 kg m-3, 139 to 384 kg m-3, and 98 to 277 kg m-3, respectively. The baled crops follow a nonlinear density vs. pressure relationship that can be accurately predicted using an exponential equation. When increasing loading or compression speed, the energy, power, and specific energy required to recompress the baled crops also increased. However, since each crop behaved differently during recompression, the effect of speed on compression pressure and density was dependent on the type of crop. At 60% reduction in bale volume and at the lowest compression speed (2.54 mm s-1), switchgrass required 15.6%, 61.7%, and 55.4% more energy than indiangrass, corn stover, and wheat straw, respectively. There were no significant differences in power required to compress different crops at the slowest speed (α = 0.05), but wheat straw required a lower specific energy of 0.725 kJ kg-1(d.b.) compared to between 0.734 and 0.814 kJ kg-1(d.b.) for the other crops. At the fastest compression speed (106.7 mm s-1), switchgrass required 32.4%, 59.4%, and 73.0% more energy, 12.9%, 20.5%, and 27.0% more power, and 15.7%, 5.4%, and 20.7% more specific energy than indiangrass, corn stover, and wheat straw, respectively. The purpose of this study was to determine the power requirement and energy consumption required to densify small bales of biomass crops using recompression to high enough densities to allow tractor trailers to be fully loaded to legal weight capacities within legal dimensions for highway transportation. Results from this study will be helpful when calculating energy balances related to biomass bale recompression for use as a feedstock in biofuel production and also for structural design and component selection for bale recompression machinery. [ABSTRACT FROM AUTHOR]

Premise: Leaf tensile resistance, a leaf's ability to withstand pulling forces, is an important determinant of plant ecological strategies. One potential driver of leaf tensile resistance is growing season length. When growing seasons are long, strong leaves, which often require more time and resources to construct than weak leaves, may be more advantageous than when growing seasons are short. Growing season length and other ecological conditions may also impact the morphological traits that underlie leaf tensile resistance. Methods: To understand variation in leaf tensile resistance, we measured size‐dependent leaf strength and size‐independent leaf toughness in diverse genotypes of the widespread perennial grass Panicum virgatum (switchgrass) in a common garden. We then used quantitative genetic approaches to estimate the heritability of leaf tensile resistance and whether there were genetic correlations between leaf tensile resistance and other morphological traits. Results: Leaf tensile resistance was positively associated with aboveground biomass (a proxy for fitness). Moreover, both measures of leaf tensile resistance exhibited high heritability and were positively genetically correlated with leaf lamina thickness and leaf mass per area (LMA). Leaf tensile resistance also increased with the growing season length in the habitat of origin, and this effect was mediated by both LMA and leaf thickness. Conclusions: Differences in growing season length may promote selection for different leaf lifespans and may explain existing variation in leaf tensile resistance in P. virgatum. In addition, the high heritability of leaf tensile resistance suggests that P. virgatum will be able to respond to climate change as growing seasons lengthen. [ABSTRACT FROM AUTHOR]

Under changing climatic scenarios, grassland conservation and development have become imperative to impart functional sustainability to their ecosystem services. These goals could be effectively and efficiently achieved with targeted genetic improvement of native grass species. To the best of our literature search, very scant research findings are available pertaining to gene editing of noncultivated grass species (switch grass, wild sugarcane, Prairie cordgrass, Bermuda grass, Chinese silver grass, etc.) prevalent in natural and semi-natural grasslands. Thus, to explore this novel research aspect, this study purposes that gene editing techniques employed for improvement of cultivated grasses especially sugarcane might be used for non-cultivated grasses as well. Our hypothesis behind suggesting sugarcane as a model crop for genetic improvement of noncultivated grasses is the intricacy of gene editing owing to polyploidy and aneuploidy compared to other cultivated grasses (rice, wheat, barley, maize, etc.). Another reason is that genome editing protocols in sugarcane (x = 10-13) have been developed and optimized, taking into consideration the high level of genetic redundancy. Thus, as per our knowledge, this review is the first study that objectively evaluates the concept and functioning of the CRISPR (clustered regularly interspaced short palindromic repeats)/Cas9 technique in sugarcane regarding high versatility, target specificity, efficiency, design simplicity, and multiplexing capacity in order to explore novel research perspectives for gene editing of non-cultivated grasses against biotic and abiotic stresses. Additionally, pronounced challenges confronting sugarcane gene editing have resulted in the development of different variants (Cas9, Cas12a, Cas12b, and SpRY) of the CRISPR tool, whose technicalities have also been critically assessed. Moreover, different limitations of this technique that could emerge during gene editing of non-cultivated grass species have also been highlighted. [ABSTRACT FROM AUTHOR]

Introduction: Enhancing soil cation exchange capacity (CEC) is of paramount importance for sustainable agriculture and ecosystem health. This study investigated the pivotal role of biochar in altering soil CEC and challenges conventional assumptions about universal effects of biochar application. Methods: Contrasting biochar types, one with a low ash content of 4.4% (switchgrass-derived biochar, SGB) and the other with a high ash content of 45.9% (poultry litter-derived biochar, PLB) were used. Two experiments treated with increasing biochar application rates were conducted: one without plants and the other grown with ryegrass. Effective CEC (summation method) was determined by two extraction methods: Mehlich-3 (M3) and ammonium acetate (AA). Results and discussion: The SGB decreased CEC by 27% on average (from both experiments) from the lowest to the highest rate of biochar application, while the PLB significantly increased CEC by 91%. This highlights the critical role of biochar properties in influencing CEC dynamics. In the second experiment, ryegrass cultivation revealed the greater importance of the calcium and magnesium/potassium ratio ([Ca+Mg]/K) in the soil CEC than CEC only for plant growth in biochar-amended soils. The ratios for optimum ryegrass production ranged from 82-86 (M3) and 69-74 (AA), which was translated to 88:11:1 Ca:Mg:K percent base saturation ratios. Moreover, predictive models for estimation of soil CEC after biochar application were successfully developed based on initial soil and biochar CEC. However, M3 was more reliable than AA for such predictions mainly because it was more successful in predicting nutrient availability from biochar. These models offer a promising tool for informed soil management decisions. Conclusion: This research emphasizes the importance of biochar feedstock, elucidates nutrient ratio effects on plant growth, and provides a practical means to anticipate soil CEC changes post-biochar application. [ABSTRACT FROM AUTHOR]

Monoculture switchgrass and restored prairie are promising perennial feedstock sources for bioenergy production on the lands unsuitable for conventional agriculture. Such lands often display contrasting topography that influences soil characteristics and interactions between plant growth and soil C gains. This study aimed at elucidating the influences of topography and plant systems on the fate of C originated from switchgrass plants and on its relationships with soil pore characteristics. For that, switchgrass plants were grown in intact soil cores collected from two contrasting topographies, namely steep slopes and topographical depressions, in the fields in multi-year monoculture switchgrass and restored prairie vegetation. The 13C pulse labeling allowed tracing the C of switchgrass origin, which X-ray computed micro-tomography enabled in-detail characterization of soil pore structure. In eroded slopes, the differences between the monoculture switchgrass and prairie in terms of total and microbial biomass C were greater than those in topographical depressions. While new switchgrass increased the CO2 emission in depressions, it did not significantly affect the CO2 emission in slopes. Pores of 18–90 µm Ø facilitated the accumulation of new C in soil, while > 150 µm Ø pores enhanced the mineralization of the new C. These findings suggest that polyculture prairie located in slopes can be particularly beneficial in facilitating soil C accrual and reduce C losses as CO2. [ABSTRACT FROM AUTHOR]

Improving the precision of aboveground biomass (AGB) estimation in lowland tropical forests is crucial to enhancing our understanding of carbon dynamics and formulating climate change mitigation strategies. This study proposes an AGB estimation method for lowland tropical forests in Xishuangbanna, which include various vegetation types, such as Pinus kesiya var. langbianensis, oak, Hevea brasiliensis, and other broadleaf trees. In this study, 2016 forest management inventory data are integrated with remote sensing variables from Landsat 8 OLI (L8) and Sentinel 2A (S2) imagery to estimate forest AGB. The forest age and aspect were utilized as stratified variables to construct the random forest (RF) models, which may improve the AGB estimation accuracy. The key findings are as follows: (1) through variable screening, elevation was identified as the main factor correlated with the AGB, with texture measures derived from a pixel window size of 7 × 7 perform best for AGB sensitivity, followed by 5 × 5, with 3 × 3 being the least effective. (2) A comparative analysis of imagery groups for the AGB estimation revealed that combining L8 and S2 imagery achieved superior performance over S2 imagery alone, which, in turn, surpassed the accuracy of L8 imagery. (3) Stratified models, which integrated aspect and age variables, consistently outperformed the unstratified models, offering a more refined fit for lowland tropical forest AGB estimation. (4) Among the analyzed forest types, the AGB of P. kesiya var. langbianensis forests was estimated with the highest accuracy, followed by H. brasiliensis, oak, and other broadleaf forests within the RF models. These findings highlight the importance of selecting appropriate variables and sensor combinations in addition to the potential of stratified modeling approaches to improve the precision of forest biomass estimation. Overall, incorporating stratification theory and multi-source data can enhance the AGB estimation accuracy in lowland tropical forests, thus offering crucial insights for refining forest management strategies. [ABSTRACT FROM AUTHOR]

The increasing cost of fossil-based energy sources has driven research in bio-based alternatives, such as perennial grasses for feedstock. The mid-Atlantic receives appreciable summer rainfall that may support a two-cut-per-year−1 harvest. At Virginia State University, a study on annual forage sorghum and two one-year stand perennials, miscanthus, and selected switchgrass ecotypes was carried out. The experimental design was a split-plot with harvest systems and feedstock grass species randomly assigned to the main and sub-plots, respectively. Only perennial grasses were assigned to the two-cut-per-year−1 system. The first cut occurred in early summer, and the second and single cut occurred after the frost-kill. Under the two-cut system, in 2022, the first-cut dry matter (DM) yield ranged from 8.9 Mg ha−1 in Blackwell to 14.7 Mg ha−1 in BoMaster. Additionally, except for BoMaster, the regrowth DM yields were within 10% of the first-cut DM yield. Under the one-cut system, the yield ranged from 10.8 Mg ha−1 in Blackwell to 23.2 Mg ha−1 in sorghum. Under the two-cut system, in 2023, miscanthus produced the greatest first-cut DM yield of 18.4 Mg ha−1, while other perennials averaged 10.1 Mg ha−1. Compared to the first cut, the hot and dry summer significantly reduced regrowth for all feedstock species, with the miscanthus DM yield dropping by 64%. While forage attributes differ among feedstock species, in general, both the first cut and regrowth showed greater crude protein and mineral elements, as well as lower ADF contents compared to a single cut following a killing freeze. Sorghum had better forage quality for the one-cut-per-year−1 feedstock material, and, along with the first cut and regrowth, it may have the potential for use as forage for maintenance energy in animal systems. For perennials, the two-cut-per-year−1 system removed the greatest quantities of nutrients during both years, with the first-cut harvest contributing about 65% of all removed N and K. Sorghum removed the greatest quantities of nutrients compared to the perennial under a one-cut-per-year−1 system. Therefore, while a two-cut-per-year−1 system can result in the greatest DM yields for dual-purpose use, its adoption calls for a critical analysis of economic benefits that considers feedstock bioenergy processing approaches, stand persistence, and fertilizer management strategies to address potential soil fertility depletion due to mineral element mining. [ABSTRACT FROM AUTHOR]

Litter decomposition facilitates the recycling of often limiting resources, which may promote plant productivity responses to diversity, that is, overyielding. However, the direct relationship between decomposition, k, and overyielding remains underexplored in grassland diversity manipulations.We test whether local adaptation of microbes, that is, home‐field advantage (HFA), N‐priming from plant inputs or precipitation drive decomposition and whether decomposition generates overyielding. Within a grassland diversity‐manipulation, altering plant richness (1, 2, 3 and 6 species), composition (communities composed of plants from a single‐family or multiple‐families) and precipitation (50% and 150% ambient growing season precipitation), we conducted a litter decomposition experiment. In spring 2020, we deployed four replicate switchgrass, Panicum virgatum, litter bags (1.59 mm mesh opening), collecting them over 7 months to estimate litter k.Precipitation was a strong, independent driver of decomposition. Switchgrass decomposition accelerated with grass richness and decelerated as phylogenetic dissimilarity from switchgrass increased, suggesting decomposition is fastest at 'home'. However, decomposition slowed with switchgrass density. In plots that contained switchgrass, we observed no relationship between decomposition and fungal saprotroph dissimilarity from switchgrass. However, in plots without switchgrass, decomposition slowed with increasing saprotroph dissimilarity from switchgrass. Combined these findings suggest that HFA is strongest when closely related neighbours, that is, heterospecific neighbours, are present in the community, rather than other individuals of the same species, that is, conspecifics. Legumes accelerated decomposition with more litter N remaining in those plots, suggesting that N‐inputs from planted legumes are priming decomposition of litter C. However, decomposition and overyielding were unrelated in legume communities. While in grass communities, overyielding and decomposition were positively related and the relationship was strongest in plots with low densities of switchgrass, that is, with heterospecific neighbours.Combined these findings suggest that plant species richness and community composition stimulate litter decomposition through multiple mechanisms, including N‐priming, but only HFA from local adaptation of microbes on closely related species correlates with overyielding, likely through resource recycling. Our results link diversity with ecosystem processes facilitating above‐ground productivity. Whether diversity loss will affect litter decomposition, productivity or both is contingent on resident plant traits and whether a locally adapted soil microbiome is maintained. Read the free Plain Language Summary for this article on the Journal blog. [ABSTRACT FROM AUTHOR]

Brazil is among the world's largest producers of green coconut, which contributes to inappropriate disposal and socioenvironmental impacts. Concomitantly, some of its coastal cities produce a great diversity of fish and large amounts of solid waste. This paper reports on the use of samples of fish scales (100FS) and green coconut shells (100GCS) and their mixtures in 75%FS:25%GCS (B25), 50%FS:50%GCS (B50), and 25%FS:75%GCS (B75) proportions and quantification of their Higher Heating Values (HHV) and Lower Heating Values (LHV), and Ultimate (UA) and Proximate Analyses (PA). Their thermal behavior was investigated by thermogravimetry (TG/DTG) and differential scanning calorimetry (DSC), whereas scanning electron microscopy (SEM), energy dispersive spectroscopy (EDS), X-ray diffraction (XRD), and Fourier transformed infrared (FTIR) were employed for analyses of their physicochemical and morphostructural properties. When compared to in natura samples, SEM images of the blends detected a structural disorder and a highly fibrous structure with an elongated chain and surface roughness. HHV were superior in samples with 100GCS (16.64 MJ kg−1), B75 (15.80 MJ kg−1), and B50 (14.98 MJ kg−1), and lower in B25 (14.16 MJ kg−1) and 100FS (13.03 MJ kg−1), with acceptable values for different biomasses. TG/DTG and DSC curves showed similarities among the samples, with the detection of their main thermoconversion stages. According to the data, the samples can be applied as renewable energy sources to mitigate socioecological illnesses and social vulnerabilities resulting from the archaic and inadequate management of solid waste. [ABSTRACT FROM AUTHOR]

Abstract 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 CO2 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 CO2 concentration and light, we measured leaf net photosynthesis of switchgrass under different CO2 concentrations and light levels in 2020 and 2021 for each of the five precipitation treatments. We first evaluated four light and CO2 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 CO2 response curves. Precipitation change did not influence the maximum net photosynthetic rate (Pmax) 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 Pmax of five precipitation treatments was 17.6 μmol CO2 m−2 s−1, and the ambient treatment tended to have a higher Pmax. 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 CO2 response. The ambient treatment had the highest Pmax, largest α, and lowest θ, Rd, and CO2 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.

Abstract This study investigates uncertainties in greenhouse gas (GHG) emission factors related to switchgrass‐based biofuel production in Michigan. Using three life cycle assessment (LCA) databases—US lifecycle inventory (USLCI) database, GREET, and Ecoinvent—each with multiple versions, we recalculated the global warming intensity (GWI) and GHG mitigation potential in a static calculation. Employing Monte Carlo simulations along with local and global sensitivity analyses, we assess uncertainties and pinpoint key parameters influencing GWI. The convergence of results across our previous study, static calculations, and Monte Carlo simulations enhances the credibility of estimated GWI values. Static calculations, validated by Monte Carlo simulations, offer reasonable central tendencies, providing a robust foundation for policy considerations. However, the wider range observed in Monte Carlo simulations underscores the importance of potential variations and uncertainties in real‐world applications. Sensitivity analyses identify biofuel yield, GHG emissions of electricity, and soil organic carbon (SOC) change as pivotal parameters influencing GWI. Decreasing uncertainties in GWI may be achieved by making greater efforts to acquire more precise data on these parameters. Our study emphasizes the significance of considering diverse GHG factors and databases in GWI assessments and stresses the need for accurate electricity fuel mixes, crucial information for refining GWI assessments and informing strategies for sustainable biofuel production.

Arbuscular mycorrhizal fungi (AMF) and biochar application individually can enhance plant tolerance to saline-alkali stress and promote plant growth efficiency. However, little is known about the potential synergistic effects of their combination on improving plant growth and soil quality under saline-alkali stress. This experiment adopted the potted method to explore the effects of four treatments on switchgrass growth and soil quality: biochar (BC), Rhizophagus irregularis (Ri), biochar + Ri (BR) and a control without biochar or Ri (CK). Compared to the CK treatment, the switchgrass biomass increased by 92.4 %, 148.6 %, and 177.3 % in the BC, Ri, and BR treatment groups, respectively. Similarly, the rhizosphere soil quality index increased by 29.33 %, 22.7 %, and 49.1 % in the respective treatment groups. The BR treatment significantly altered the rhizosphere soil microbial composition and diversity. Notably, compared to the other treatments, the archaeal α-diversity in the BR group showed a significant decrease. BR treatment significantly increased the relative abundance of bacteria, fungi and archaea at the genus level (e.g., Bacillus, Trichome and candidatus_methanopenens). Network analysis showed that the complexity and closeness of interactions between different microbial taxa were stronger in the BC, Ri and BR treatments than in the CK treatment, with BR being the more prominent. In summary, biochar combined with Ri has a better effect on promoting the growth of switchgrass under saline-alkali stress, improving the quality of saline-alkali soil, and increasing soil microbial diversity. This study provides a new approach for the efficient development and utilization of saline-alkali land.

In the North-Central United States, lowland ecotype switchgrass can increase yield by up to 50% compared with locally adapted but early flowering cultivars. However, lowland ecotypes are not winter tolerant. The mechanism for winter damage is unknown but previously has been associated with late flowering time. This study investigated heading date (measured for two years) and winter survivorship (measured for three years) in a multi-generation population generated from two winter-hardy lowland individuals and diverse southern lowland populations. Sequencing data (311,776 markers) from 1,306 individuals were used to evaluate genome-wide trait prediction through cross-validation and progeny prediction (n = 52). Genetic variance for heading date and winter survivorship was additive with high narrow-sense heritability (0.64 and 0.71, respectively) and reliability (0.68 and 0.76, respectively). The initial negative correlation between winter survivorship and heading date degraded across generations (F1r = -0.43, pseudo-F2r = -0.28, pseudo-F2 progeny r = -0.15). Within-family predictive ability was moderately high for heading date and winter survivorship (0.53 and 0.52, respectively). A multi-trait model did not improve predictive ability for either trait. Progeny predictive ability was 0.71 for winter survivorship and 0.53 for heading date. These results suggest that lowland ecotype populations can obtain sufficient survival rates in the northern United States with two or three cycles of effective selection. Despite accurate genomic prediction, naturally occurring winter mortality successfully isolated winter tolerant genotypes and appears to be an efficient method to develop high-yielding, cold-tolerant switchgrass cultivars.

Abstract Background Switchgrass (Panicum virgatum L.) is a warm-season perennial (C4) grass identified as an important biofuel crop in the United States. It is well adapted to the marginal environment where heat and moisture stresses predominantly affect crop growth. However, the underlying molecular mechanisms associated with heat and drought stress tolerance still need to be fully understood in switchgrass. The methylation of H3K4 is often associated with transcriptional activation of genes, including stress-responsive. Therefore, this study aimed to analyze genome-wide histone H3K4-tri-methylation in switchgrass under heat, drought, and combined stress. Results In total, ~ 1.3 million H3K4me3 peaks were identified in this study using SICER. Among them, 7,342; 6,510; and 8,536 peaks responded under drought (DT), drought and heat (DTHT), and heat (HT) stresses, respectively. Most DT and DTHT peaks spanned 0 to + 2000 bases from the transcription start site [TSS]. By comparing differentially marked peaks with RNA-Seq data, we identified peaks associated with genes: 155 DT-responsive peaks with 118 DT-responsive genes, 121 DTHT-responsive peaks with 110 DTHT-responsive genes, and 175 HT-responsive peaks with 136 HT-responsive genes. We have identified various transcription factors involved in DT, DTHT, and HT stresses. Gene Ontology analysis using the AgriGO revealed that most genes belonged to biological processes. Most annotated peaks belonged to metabolite interconversion, RNA metabolism, transporter, protein modifying, defense/immunity, membrane traffic protein, transmembrane signal receptor, and transcriptional regulator protein families. Further, we identified significant peaks associated with TFs, hormones, signaling, fatty acid and carbohydrate metabolism, and secondary metabolites. qRT-PCR analysis revealed the relative expressions of six abiotic stress-responsive genes (transketolase, chromatin remodeling factor-CDH3, fatty-acid desaturase A, transmembrane protein 14C, beta-amylase 1, and integrase-type DNA binding protein genes) that were significantly (P

Abstract Background The small YABBY plant-specific transcription factor has a prominent role in regulating plant growth progress and responding to abiotic stress. Results Here, a total of 16 PvYABBYs from switchgrass (Panicum virgatum L.) were identified and classified into four distinct subgroups. Proteins within the same subgroup exhibited similar conserved motifs and gene structures. Synteny analyses indicated that segmental duplication contributed to the expansion of the YABBY gene family in switchgrass and that complex duplication events occurred in rice, maize, soybean, and sorghum. Promoter regions of PvYABBY genes contained numerous cis-elements related to stress responsiveness and plant hormones. Expression profile analysis indicated higher expression levels of many PvYABBY genes during inflorescence development and seed maturation, with lower expression levels during root growth. Real-time quantitative PCR analysis demonstrated the sensitivity of multiple YABBY genes to PEG, NaCl, ABA, and GA treatments. The overexpression of PvYABBY14 in Arabidopsis resulted in increased root length after treatment with GA and ABA compared to wild-type plants. Conclusions Taken together, our study provides the first genome-wide overview of the YABBY transcription factor family, laying the groundwork for understanding the molecular basis and regulatory mechanisms of PvYABBY14 in response to ABA and GA responses in switchgrass.

BackgroundAlthough the genome for the allotetraploid bioenergy crop switchgrass (Panicum virgatum) has been established, limitations in mutant resources have hampered in planta gene function studies toward crop optimization. Virus-induced gene silencing (VIGS) is a versatile technique for transient genetic studies. Here we report the implementation of foxtail mosaic virus (FoMV)-mediated gene silencing in switchgrass in above- and below-ground tissues and at different developmental stages.ResultsThe study demonstrated that leaf rub-inoculation is a suitable method for systemic gene silencing in switchgrass. For all three visual marker genes, Magnesium chelatase subunit D (ChlD) and I (ChlI) as well as phytoene desaturase (PDS), phenotypic changes were observed in leaves, albeit at different intensities. Gene silencing efficiency was verified by RT-PCR for all tested genes. Notably, systemic gene silencing was also observed in roots, although silencing efficiency was stronger in leaves (~ 63-94%) as compared to roots (~ 48-78%). Plants at a later developmental stage were moderately less amenable to VIGS than younger plants, but also less perturbed by the viral infection.ConclusionsUsing FoMV-mediated VIGS could be achieved in switchgrass leaves and roots, providing an alternative approach for studying gene functions and physiological traits in this important bioenergy crop.

Biomass-derived solvents have been proposed as a novel pathway in biorefining for the realization of biofuels and bioproducts derived from lignocellulosic biomass. Cyrene derived from cellulose has recently been shown to have a high potential as a green organic solvent for pretreating poplar biomass. However, due to its high dynamic viscosity nature, high Cyrene concentration could cause negative effects on the sugar release of the pretreated biomass as well as driving up the operational cost of the lignin recovery. In this study, we combine experimental and computational approaches to examine the impact of Cyrene pretreatment with reduced Cyrene concentration under mild conditions on switchgrass lignin. Our experimental studies indicated correlation between pretreatment condition and recovery and structure modification of lignin. Switchgrass lignin extracted by Cyrene pretreatment possessed high preservation of β-O-4 ether inter-unit linkage, which could provide versatility in the integration of downstream lignin valorization into the modern biorefinery industries. Molecular modeling examining the solvation of switchgrass lignin polymer and the disaggregation of low-molecular weight lignin aggregates under pretreatment conditions indicated that a preferential interaction exists between Cyrene and lignin, which likely drives lignin release, and that the disruption of inter-lignin contacts can be modulated as a non-monotonic function of Cyrene : water ratio. Further, while Cyrene–lignin interactions permit the solubilization of lignin, simulations with proxy reactive-species reveal that changes to the diffusion of these reactive proxies and their localization near linkage sites under Cyrene conditions may inhibit chemical processes. The results indicated that loss of pretreatment efficacy caused by low Cyrene concentration could be compensated by prolonged pretreatment time and high catalyst dosage. [ABSTRACT FROM AUTHOR]

Eastern gamagrass (Tripsacum dactyloides) is highly palatable, ideal for grazing and hay production in the United States. It is deep rooted and resilient, tolerant of flooding and drought. Objectives of this study were to develop plant parameters for different ecotypes of this grass. Data collected in field plots of diverse ecotypes included biomass, leaf area index (LAI), light extinction coefficient (k), and radiation use efficiency (RUE). Average LAI was 1.06 and average k was −1.05. The power response of k to LAI offers a new approach to simulating light interception at very low LAI values and throughout the range of LAI values of these ecotypes and similar grass species. The RUE values, which ranged from 1.16 to 4.31 g/MJ, highlight the immense diversity of eastern gamagrass. The high RUE values for the most productive ecotypes emphasizes the importance of this grass species for hay and grazing. While not as large as switchgrass (Panicum virgatum) values, it is still a prominent forage species that even is comparable to maize (Zea mays L.) in productivity when expressed as radiation use efficiency. These results are an important step in developing relationships and parameters to simulate the different ecotypes of this grass with process-based models. [ABSTRACT FROM AUTHOR]

Summary: MiRNAs have been reported to be the key regulators involving a wide range of biological processes in diverse plant species, but their functions in switchgrass, an important biofuel and forage crop, are largely unknown. Here, we reported the novel function of miR528, which has expanded to four copies in switchgrass, in controlling biomass trait of tillering number and regrowth rate after mowing. Blocking miR528 activity by expressing short tandem target mimic (STTM) increased tiller number and regrowth rate after mowing. The quadruple pvmir528 mutant lines derived from genome editing also showed such improved traits. Degradome and RNA‐seq analysis, combined with in situ hybridization assay revealed that up‐regulation of two miR528 targets coding for Cu/Zn‐SOD enzymes, might be responsible for the improved traits of tillering and regrowth in pvmir528 mutant. Additionally, natural variations in the miR528‐SOD interaction exist in C3 and C4 monocot species, implying the distinct regulatory strength of the miR528‐SOD module during monocot evolution. Overall, our data illuminated a novel role of miR528 in controlling biomass traits and provided a new target for genetic manipulation‐mediated crop improvement. [ABSTRACT FROM AUTHOR]

Perennial biomass cropping systems are generally considered to be more environmentally friendly and sustainable compared to annual cropping systems, largely due to reduced input requirements. Nitrogen fertilizer is, by far, the most expensive input into perennial grass-based bioenergy systems, demanding research efforts focused on increasing nitrogen-use efficiency of biomass crops. The purpose of this study was to identify genetic variability for nitrogen-use efficiency and to determine if it is possible to identify genotypes that can produce high biomass yields without nitrogen fertilizer applications. A total of 83 half-sib families were grown in replicated paired plots on two different marginal soils. Paired plots consisted of 0 or 100 kg N ha−1, allowing for a highly precise computation of the direct impact of nitrogen fertilizer. There were strong genotype x location interactions, as well as strong genotype x nitrogen rate interactions at one of the two locations. For both locations, the direct response to nitrogen fertilization on biomass yield was strongly and negatively related to biomass yield without nitrogen fertilizer. Many families had high yield without fertilizer application, but also had non-significant responses to the fertilizer application. However, the ranking of these families was highly inconsistent between the two locations and soil types. These results suggested that certain families were likely responding to favorable associations with nitrogen-fixing microbes and that those associations were inconsistent between the two soil types. Development of new switchgrass cultivars that can form associations with nitrogen-fixing microbes should be conducted under conditions that favor such associations, strictly avoiding application of nitrogen fertilizers. [ABSTRACT FROM AUTHOR]