Infestation Rates and Tiller Morphology Effects by the Switchgrass Moth on Six Cultivars of Switchgrass

Switchgrass (Panicum virgatum L.) is a potential biomass crop for native species-based biofuel systems in North America. A recently identified pest of switchgrass, the switchgrass moth, Blastobasis repartella (Dietz) (Lepidoptera: Coleophoridae), feeds in the basal above-ground internodes and below-ground in the proaxis and rhizomes, causing premature tiller and rhizome loss. Our goal was to determine genetic and temporal variation among six upland cultivars for frequency of tiller infestation by larvae of the switchgrass moth in mature stands in the northern Great Plains and if variation in biomass production was associated with variation in frequency of infestation. Data were collected in 2011 and 2012 for tiller density, biomass, frequency of infestation, number of leaves per healthy and infested tiller, and weights of healthy and infested tillers. Differences were found among cultivars for tiller density, biomass yield, and numbers of leaves per healthy and infested tillers. ‘Summer’, ‘Sunburst’, ‘Pathfinder’, and ‘Cave-In-Rock’ were the highest yielding cultivars. Mean frequency of infestation was different between 2011 (6.7 %) and 2012 (9.6 %). Infested tillers had one less collared leaf than healthy tillers. The weights of healthy tillers were ca. 3× those of infested tillers in both years, suggesting an impact on biomass accumulation and economic value. Levels of infestation were similar for all six cultivars, indicating no feeding preference by the switchgrass moth larva among genetically diverse cultivars of switchgrass. Regression of biomass yield on frequency of infestation showed negative linear relationships for ‘Carthage’ and ‘Kentucky 1625’.     

Nitrogen-Use Dynamics in Switchgrass Grown for Biomass

This work examines N use by switchgrass (Panicum virgatum L.). A study was conducted on two well-established ‘Cave-in-Rock’ switchgrass stands in Blacksburg (37° 11′ N, 80° 25′ W) and Orange (38° 13′ N, 78° 07′ W) Virginia, USA. Plots were fertilized in 2001 (year 1) with 0, 90, 180, or 270 kg N per hectare. No additional N was applied in 2002 (year 2) and 2003 (year 3), and biomass was harvested in July and November for years 2 and 3 (but only in November of year 1). Root and soil samples were collected in May, July, September, and November each year and analyzed for N. Nitrogen fertilization did not increase yields in 2001 (year 1), but it did provide residual benefits in 2002 (year 2) and 2003 (year 3). Root-N levels at 15 cm depth increased with fertilization, fluctuated seasonally between roots and shoots, and root-N was reduced over the course of the study. With two harvests per year, about 100 kg N hectare per year were removed in biomass, even in plots with no N added—suggesting N already present in the soils (at 15 cm depth) contributed to yields; but the soil mineral-N pools were reduced by the end of year 3. Nitrogen-use efficiency, apparent N recovery, and partial factor productivity were reduced with higher N applications. The data support the notion that biomass production can be achieved with minimal N inputs, but stands must be managed to maintain that N reserve over the long term. There is also a need to quantify the N pool to depths greater than 15 cm in other agro-ecoregions.     

Efficient Methods of Estimating Switchgrass Biomass Supplies

Switchgrass (Panicum virgatum L.) is being developed as a biofuel feedstock for the United States. Efficient and accurate methods to estimate switchgrass biomass feedstock supply within a production area will be required by biorefineries. Our main objective was to determine the effectiveness of indirect methods for estimating biomass yields and composition of switchgrass fields. Indirect measurements were conducted in eastern Nebraska from 2003 to 2007 in which switchgrass biomass yields were manipulated using three nitrogen rates (0 kg N ha^-1, 60 kg N ha^-1, and 120 kg N ha^-1) and two harvest periods (August and post-killing frost). A modified Robel pole was used to determine visual obstruction, elongated leaf height, and canopy height measurements. Prediction models from the study showed that elongated leaf height, visual obstruction, and canopy height measurements accounted for >?91%, >?90%, and >?82% of the variation in switchgrass biomass, respectively. Regression slopes were similar by cultivar (“Cave-in-Rock” and “Trailblazer”), harvest period, and across years indicating that a single model is applicable for determining biomass feedstock supply within a region, assuming similar harvesting methods. Sample numbers required to receive the same level of precision were as follows: elongated leaf height<canopy height<visual obstruction. Twenty to 30 elongated leaf height measurements in a field could predict switchgrass biomass yield within 10% of the mean with 95% confidence. Visual obstruction is recommended on switchgrass fields with low to variable stand densities while elongated leaf height measurements would be recommended on switchgrass fields with high, uniform stand densities. Incorporating an ocular device with a Robel pole provided reasonable frequency estimates of switchgrass, broadleaf weeds, and grassy weeds at the field scale.     

The Effect of Drying Rate on the Survival of Three Desiccation-tolerant Angiosperm Species

The effect of drying rate on the survival of three angiospermresurrection plants, Craterostigma wilmsii (homoiochlorophyllous),Xerophyta humilis (poikilochlorophyllous) and Myrothamnus flabellifolius(homoiochlorophyllous) was examined. All species survived slowdrying, but only C. wilmsii was able to survive rapid drying.C. wilmsii was rapidly able to induce protection mechanismssuch as folding of cell walls to prevent mechanical stress andcurling of leaves to minimize light stress, and thus survivedfast drying. Rapid drying of X. humilis andM. flabellifoliusappeared to allow insufficient time for complete induction ofprotection mechanisms. In X. humilis, there was incomplete replacementof water in vacuoles, the photosynthetic apparatus was not dismantled,plasma membrane disruption occurred and quantum efficiency ofphotosystem II (F_V/F_M) did not recover on rehydration. Rapidlydried leaves of M. flabellifolius did not fold tightly againstthe stem and F_V/F_Mdid not recover. Ultrastructural studies showedthat subcellular damage incurred during drying was exacerbatedon rehydration. The three species co-occur in environments inwhich they experience high desiccation pressures. C. wilmsiihas few features to retard water loss and thus the ability forrapid induction of subcellular protection is vital to survival.X. humilis and M. flabellifolius are able to retard water lossand protection is acquired relatively slowly. Copyright 1999Annals of Botany Company Chlorophyll fluorescence, Craterostigma wilmsii, drying rate, Myrothamnus flabellifolius, resurrection plant, ultrastructure, Xerophyta humilis.     

Switchgrass Harvest Progression in the North-Central USA

In the North-Central USA, switchgrass to be used as a biomass feedstock typically will be harvested in the autumn. The accumulated area harvested over the harvest season (defined here as the harvest progression) will influence the size of the machinery fleet and seasonal labor required to complete the majority of the harvest before the first lasting snow. A harvest progression model was developed that uses drying rate, mower and baler productivity, and weather conditions as major inputs. Ten years of weather data (2005–2014) from Wisconsin, Iowa, and Nebraska (WI, IA, NE) were used. Harvest progression was modeled for four harvest systems involving conventional and intensive conditioning both swathed and tedded (CC, IC, CCT, and ICT, respectively) and two dates at which harvest began (1 September and after a killing frost). To reduce risk of exposing crop to prolonged periods of inclement weather, mowers were idled when more than 80 ha were cut but not yet baled. For all sites, the harvest start date and the mower idled constraint had greater impact on harvest progression than the type of harvest system. Harvest progression was greatest when mowing started on 1 September and continued whenever weather permitted (i.e., no mower idled constraint). Compared to the harvest system used today (CC), using the IC system resulted in more area harvested with less crop exposed to rain after cutting and considerably less area left to be baled in the spring. Starting harvest on 1 September, using intensive conditioning, and not idling the mowers might be considered the system that best balances the desire for rapid harvest progression, small equipment fleet size, low-capital expenditures, and maximum labor utilization.     

Topsoil Thickness Influences Nitrogen Management of Switchgrass

Switchgrass (Panicum virgatum L.) is an attractive bioenergy crop option for eroded portions of claypan landscapes where grain crop production is marginally profitable. Topsoil thickness above the claypan, or depth to claypan (DTC), can vary widely within fields, and little information exists on its impacts on N management of switchgrass. Therefore, a study was conducted at the University of Missouri South Farm near Columbia, Missouri, to determine whether topsoil thickness influenced fertilizer N requirements of switchgrass. Switchgrass was planted in 2009 on main plots with a range of DTC classified as exposed (<8 cm), shallow (8–15 cm), moderate (16–30 cm), and deep (>30 cm) and was harvested annually at postdormancy during 2011 to 2015. Three split-plot treatments were 0, 67, or 101 kg N ha^?1 applied annually in May, and a fourth was three intercropped native legumes as the N source. Across years, the legume treatment apparently supplied no N because it produced the same or less switchgrass yield than the nonfertilized treatment. Topsoil proved valuable as switchgrass yield, nutrient removal, and profit usually increased as DTC increased. Fertilization with 101 kg N ha^?1 on exposed, shallow, or moderate DTC and 67 kg N ha^?1 on deep DTC was required to obtain the highest biomass yield, but it also increased nutrient removal. Strikingly, profit across years was negative for the legume treatment and highest with no fertilizer on all DTC classes. Therefore, improvements are needed before intercropped legumes are profitable, and N fertilization may be needed only periodically to maximize switchgrass profit on claypan soils.     

Farm-Scale Production Cost of Switchgrass for Biomass

The economic potential of cellulosic biomass from switchgrass has heretofore been evaluated using estimates of farm costs based on extrapolation from experimental data and budget estimates. The objective of the project reported here was to estimate the cost of production that would be experienced by farmers on commercial production situations. Switchgrass was produced as a biomass crop on commercial-scale fields by ten contracting farmers located from northern North Dakota to southern Nebraska. Results showed a wide range of yields and costs across the five production years and ten sites, with an overall average cost of $65.86 Mg^?1 of biomass dry matter, and annualized yield of 5.0 Mg ha^?1. The low-cost half of the producers were able to produce at an average cost of $51.95 Mg^?1over the 5-year period. When projected to a full 10-year rotation, their cost fell further to $46.26 Mg^?1. We conclude that substantial quantities of biomass feedstock could have been produced in this region at a cost of about $50 Mg^?1 at the farm gate, which translates to about $0.13/l of ethanol. These results provide a more reliable benchmark for current commercial production costs as compared to other estimates, which range from $25 to $100 Mg^?1.     

Transcriptional Analysis of Flowering Time in Switchgrass

Over the past two decades, switchgrass (Panicum virgatum) has emerged as a priority biofuel feedstock. The bulk of switchgrass biomass is in the vegetative portion of the plant; therefore, increasing the length of vegetative growth will lead to an increase in overall biomass yield. The goal of this study was to gain insight into the control of flowering time in switchgrass that would assist in development of cultivars with longer vegetative phases through delayed flowering. RNA sequencing was used to assess genome-wide expression profiles across a developmental series between switchgrass genotypes belonging to the two main ecotypes: upland, typically early flowering, and lowland, typically late flowering. Leaf blades and tissues enriched for the shoot apical meristem (SAM) were collected in a developmental series from emergence through anthesis for RNA extraction. RNA from samples that flanked the SAM transition stage was sequenced for expression analyses. The analyses revealed differential expression patterns between early- and late-flowering genotypes for known flowering time orthologs. Namely, genes shown to play roles in photoperiod response and the circadian clock in other species were identified as potential candidates for regulating flowering time in the switchgrass genotypes analyzed. Based on their expression patterns, many of the differentially expressed genes could also be classified as putative promoters or repressors of flowering. The candidate genes presented here may be used to guide switchgrass improvement through marker-assisted breeding and/or transgenic or gene editing approaches.     

The Biology and Agronomy of Switchgrass for Biofuels

Switchgrass (Panicum virgatum L.)—a perennial, warm-season (C₄) species—evolved across North America into multiple, divergent populations. The resulting natural variation within the species presents considerable morphological diversity and a wide range of adaptation. The species was adopted as a crop—initially as a forage—only in the last 50 yr. Its potential uses have recently been expanded to include biofuels. Management of switchgrass for biofuels is informed by an understanding of the plant's biology. Successful establishment requires attention to seed dormancy and weed control as well as proper depth and date of planting. The plant's growth rate is closely tied to temperature, but timing of reproductive development is linked to photoperiod. Accordingly, the period of vegetative growth can be extended by planting lower-latitude cultivars at higher latitudes. This strategy may provide a yield advantage, but cold tolerance can become limiting. Switchgrass is thrifty in its use of applied N; it appears able to obtain N from sources that other crops cannot tap. The N removed in harvested biomass is often greater than the amount of N applied. In areas with sufficient rainfall, sustainable yields of ～15 Mg ha^?1 yr^?1 may be achievable by applying ～50 kg N ha^?1 yr^?1. Harvesting biomass once per season—after plants have senesced and translocated N into perennial tissues—appears to allow plants to maintain an internal N reserve. Two harvests yr^?1 may increase yields in some cultivars, but a single annual harvest maximizes yields in many cases. If two harvests are taken, more N must be applied to compensate for the N removed in the midseason harvest. Taking more than two harvests yr^?1 often adversely affects long-term productivity and persistence. Switchgrass has potential as a renewable fuel source, but such use will likely require large infrastructural changes; and, even at maximum output, such systems could not provide the energy currently being derived from fossil fuels.     

Grass Invasion into Switchgrass Managed for Biomass Energy

Switchgrass (Panicum virgatum) is a C₄ perennial grass and is the model herbaceous perennial bioenergy feedstock. Although it is indigenous to North American grasslands east of the Rocky Mountains and has been planted for forage and conservation purposes for more than 75 years, there is concern that switchgrass grown as a biofuel crop could become invasive. Our objective is to report on the invasion of C₄ and C₃ grasses into the stands of two switchgrass cultivars following 10 years of management for biomass energy under different N and harvest management regimes in eastern Nebraska. Switchgrass stands were invaded by big bluestem (Andropogon gerardii), smooth bromegrass (Bromus inermis), and other grasses during the 10 years. The greatest invasion by grasses occurred in plots to which 0 N had been applied and with harvests at anthesis. In general, less grass encroachment occurred in plots receiving at least 60 kg of N ha^?1 or in plots harvested after frost. There were differences among cultivars with Cave-in-Rock being more resistant to invasion than Trailblazer. There was no observable evidence of switchgrass from this study invading into border areas or adjacent fields after 10 years of management for biomass energy. Results indicate that switchgrass is more likely to be invaded by other grasses than to encroach into native prairies or perennial grasslands seeded on marginally productive cropland in the western Corn Belt of the USA.     

Effect of Lime Pretreatment on Granulation of Switchgrass

Densification of switchgrass into consistent and high-density solid feedstock will reduce the cost of transport, handling, and storage to produce fuels and chemicals. Development a novel, low-cost densification technology is critical for reducing the delivered cost of feedstock while improving the bulk flow properties of densified products. In this paper, a novel wet granulation technology was proposed to investigate the effect of lime pretreatment on the production of switchgrass granules. Granulation is a process of agglomerating fine powders by wetting powder surfaces with liquid binders and mild application of shear/vibrating forces. Switchgrass was size reduced into fine powders using a knife mill and pretreated with three lime loading rates (0.05, 0.1, 0.2 g/g of biomass) at 121 °C for 30 min and at room temperature (25 °C) for 72 h. The structural modification of pretreated samples was analyzed by scanning electron microscopy and autofluorescence microscopy. Pretreated samples were granulated using a pan granulator with pre-formulated starch binder. Granules made from 20 % (0.2 g/g of biomass) lime loading rate had significantly higher single granule density and angle of repose with lower binder requirement than that of untreated granules. Lime treatment did not significantly increase the bulk density and hardness of granules. Lime-treated granules had significantly higher ash content and lower gross calorific value than that of untreated granules. In overall, lime treatment was not attractive to produce granules for thermochemical conversion platform, but lime-treated granules could be used to produce liquid biofuels and platform chemicals in biochemical conversion platform.     

Managing Spatial and Temporal Switchgrass Biomass Yield Variability

Biorefineries that plan to use switchgrass exclusively will encounter year-to-year variability in feedstock production. The economic success of the biorefinery will depend in part on the ability of the management team to strategically identify land for conversion from current use to the production of switchgrass enabling a flow of feedstock for the life of the biorefinery. The objective of this research is to determine the optimal quality, quantity, and location of land to lease while considering the spatial and temporal variability of switchgrass biomass yield. A calibrated biophysical simulation model was used to simulate switchgrass biomass yields for 50 years based on historical weather data from 1962 to 2011, for three land capability classes for each of 30 counties. Mathematical programming models were constructed and solved to determine the optimal leasing scheme for each of three strategies for a biorefinery that requires 2,000 Mg/day. As expected, a model based on the assumption that the average yield would be obtained in each year finds that production from land identified for leasing would be insufficient to fulfill the biorefinery’s needs in half of the years. In the absence of other sources of biomass, the feedstock shortage would require forced idling of the biorefinery for an average of 29.5 days during these years. Results of a strategy of leasing sufficient land to cover feedstock needs in the worst year from among 50 years for which data are available are compared to that of a strategy enabling year-to-year storage.     

Soil Carbon Storage by Switchgrass Grown for Bioenergy

Life-cycle assessments (LCAs) of switchgrass (Panicum virgatum L.) grown for bioenergy production require data on soil organic carbon (SOC) change and harvested C yields to accurately estimate net greenhouse gas (GHG) emissions. To date, nearly all information on SOC change under switchgrass has been based on modeled assumptions or small plot research, both of which do not take into account spatial variability within or across sites for an agro-ecoregion. To address this need, we measured change in SOC and harvested C yield for switchgrass fields on ten farms in the central and northern Great Plains, USA (930 km latitudinal range). Change in SOC was determined by collecting multiple soil samples in transects across the fields prior to planting switchgrass and again 5 years later after switchgrass had been grown and managed as a bioenergy crop. Harvested aboveground C averaged 2.5?±?0.7 Mg C ha^?1 over the 5 year study. Across sites, SOC increased significantly at 0–30 cm (P?=?0.03) and 0–120 cm (P?=?0.07), with accrual rates of 1.1 and 2.9 Mg C ha^?1 year^?1 (4.0 and 10.6 Mg CO₂ ha^?1 year^?1), respectively. Change in SOC across sites varied considerably, however, ranging from ?0.6 to 4.3 Mg C ha^?1 year^?1 for the 0–30 cm depth. Such variation in SOC change must be taken into consideration in LCAs. Net GHG emissions from bioenergy crops vary in space and time. Such variation, coupled with an increased reliance on agriculture for energy production, underscores the need for long-term environmental monitoring sites in major agro-ecoregions.     

Dihaploid Stocks of Switchgrass Isolated by a Screening Approach

Manipulation of ploidy in switchgrass has potential to accelerate inbred production and to provide insight about genome structure through either sequencing or cytogenetic approaches. We have identified two dihaploid individuals isolated from among the progeny of a controlled cross between two individuals of the cultivars Alamo and Kanlow. The dihaploid lines were initially distinguished from the parental lines by their reduced heterozygosity and were subsequently confirmed through estimation of C values by flow cytometry and chromosome counts of metaphase root tip squash preparations. These plants are functionally sterile, with floral bracts that remain closed and inviable pollen. They can be easily distinguished from tetraploid individuals by their reduced stature, smaller epidermal cell size, and lower number of chloroplasts per guard cell. Aberrant meiosis in these individuals is evidenced by a lack of regular pairing at diakinesis and metaphase I and suggests that the non-homologous genomes are distinct from one another. The reduced genome size of these dihaploids will facilitate basic genome studies and genetic analyses that are impossible or problematic in polyploid accessions.     

A Genomics Approach to Deciphering Lignin Biosynthesis in Switchgrass

It is necessary to overcome recalcitrance of the biomass to saccharification (sugar release) to make switchgrass (Panicum virgatum) economically viable as a feedstock for liquid biofuels. Lignin content correlates negatively with sugar release efficiency in switchgrass, but selecting the right gene candidates for engineering lignin biosynthesis in this tetraploid outcrossing species is not straightforward. To assist this endeavor, we have used an inducible switchgrass cell suspension system for studying lignin biosynthesis in response to exogenous brassinolide. By applying a combination of protein sequence phylogeny with whole-genome microarray analyses of induced cell cultures and developing stem internode sections, we have generated a list of candidate monolignol biosynthetic genes for switchgrass. Several genes that were strongly supported through our bioinformatics analysis as involved in lignin biosynthesis were confirmed by gene silencing studies, in which lignin levels were reduced as a result of targeting a single gene. However, candidate genes encoding enzymes involved in the early steps of the currently accepted monolignol biosynthesis pathway in dicots may have functionally redundant paralogues in switchgrass and therefore require further evaluation. This work provides a blueprint and resources for the systematic genome-wide study of the monolignol pathway in switchgrass, as well as other C4 monocot species.     

Agrobacterium-Mediated Transformation of Switchgrass and Inheritance of the Transgenes

Switchgrass (Panicum virgatum L.) has been developed into an important biofuel crop. Embryogenic calli induced from caryopses or inflorescences of the lowland switchgrass cultivar Alamo were used for Agrobacterium-mediated transformation. A chimeric hygromycin phosphotransferase gene (hph) was used as the selectable marker and hygromycin as the selection agent. Embryogenic calli were infected with Agrobacterium tumefaciens strain EHA105. Calli resistant to hygromycin were obtained after 5 to 8 weeks of selection. Soil-grown transgenic switchgrass plants were obtained 4 to 5 months after Agrobacterium infection. The transgenic nature of the regenerated plants was demonstrated by PCR, Southern blot hybridization analysis, and GUS staining. T1 progeny were obtained after reciprocal crosses between transgenic and untransformed control plants. Molecular analyses of the T1 progeny revealed various patterns of segregation. Transgene silencing was observed in the progeny with multiple inserts. Interestingly, reversal of the expression of the silenced transgene was found in segregating progeny with a single insert.     

Determining Switchgrass Breakeven Prices in a Landscape Design System

Precision agriculture technologies allow producers to identify areas of fields that are underperforming and unprofitable. If these less productive parts of the field could be converted to a bioenergy crop through subfield management strategies (landscape design), there may be potential gains to farmer revenue, biomass availability, and reduced adverse environmental impacts. Switchgrass is considered as a potential energy crop due its ability to thrive in marginal conditions. Previous studies have examined switchgrass production and breakeven costs, but have not looked at how production costs may change when produced in a landscape design situation. Adapting costs to the partial field situation, this paper determines the switchgrass breakeven prices ($ ton^?1) which equate producers’ net revenues in a base case (all corn) and landscape design case. That breakeven price is the price at which the farmer would be indifferent between the base and landscape design cases. We examine the case of a general, 100-acre field in Iowa, with 15 acres converted to switchgrass production, as well as 11 actual fields in Central Iowa where unprofitable subfields are assumed to be converted to switchgrass production, and the remaining portion of the field remains in corn. We find an average switchgrass breakeven price of $173 ton^?1 when land costs are included, and an average of $114 ton^?1 when no land costs are considered. A stochastic analysis to obtain a distribution of switchgrass breakeven prices under uncertainty is performed, producing distributions of switchgrass breakeven prices of $65–$266 ton^?1 and $108– $432 ton^?1 with and without land costs, respectively.     

Linkage Maps of Lowland and Upland Tetraploid Switchgrass Ecotypes

Switchgrass (Panicum virgatum L.) is a native perennial warm season (C₄) grass that has been identified as a promising species for bioenergy research and production. Consequently, biomass yield and feedstock quality improvements are high priorities for switchgrass research. The objective of this study was to develop a switchgrass genetic linkage map using a full-sib pseudo-testcross mapping population derived from a cross between two heterozygous genotypes selected from the lowland cultivar ‘Alamo’ (AP13) and the upland cultivar ‘Summer’ (VS16). The female parent (AP13) map consists of 515 loci in 18 linkage groups (LGs) and spans 1,733 cM. The male parent (VS16) map arranges 363 loci in 17 LGs and spans 1,508 cM. No obvious cause for the lack of one LG in VS16 could be identified. Comparative analyses between the AP13 and VS16 maps showed that the two major ecotypic classes of switchgrass have highly colinear maps with similar recombination rates, suggesting that chromosomal exchange between the two ecotypes should be able to occur freely. The AP13 and VS16 maps are also highly similar with respect to marker orders and recombination levels to previously published switchgrass maps. The genetic maps will be used to identify quantitative trait loci associated with biomass and quality traits. The AP13 genotype was used for the whole genome-sequencing project and the map will thus also provide a tool for the anchoring of the switchgrass genome assembly.