Bioenergy crops Miscanthus x giganteus and Panicum virgatum reduce growth and survivorship of Spodoptera frugiperda (Lepidoptera: Noctuidae)

Large-scale cultivation of plants used as biofuels is likely to alter the ecological interactions of current agricultural crops and their insect pests in a myriad of ways. Recent evidence suggests many contemporary maize pests will be able to use potential biofuel crops such as switchgrass, Panicum virgatum L., and miscanthus as hosts. To determine how suitable these biofuels are to the maize, Zea mays L., pest and generalist graminivore, Spodoptera frugiperda (J. E. Smith) (Lepidoptera: Noctuidae), we examined host plant preference and larval performance on foliage grown for commercial biofuel production. Larvae fed leaf tissue from both field- and greenhouse-grown switchgrass and miscanthus were monitored for survival, development, and food use relative to field-grown maize. Survivorship on biofuel crops was high on greenhouse-grown leaf tissue but severely reduced for field-grown switchgrass, and no larvae survived on field-grown miscanthus. Larvae fed field-grown tissue had larger head capsules yet achieved lower pupal weights because the increased toughness of the leaf tissue prevented the assimilation of nitrogen. Given that larvae overwhelmingly preferred maize to other biofuel crop species and that survival and performance were dramatically reduced on biofuel crop species, it is likely that biofuel crops, as grown for field cultivation, will suffer reduced damage from maize pests such as S. frugiperda because of reduced suitability.     

Regulation of clonal growth and dynamics of Panicum virgatum (Poaceae) in tallgrass prairie: effects of neighbor removal and nutrient addition

Clones of the perennial grass Panicum virgatum were studied on the Konza Prairie in northeast Kansas to determine the effects of neighbors, nutrient availability, and physiological integration on ramet population dynamics and clonal growth and architecture. Opposite halves of established clones in the field were subjected differentially to treatments including neighbor removal and nitrogen addition, with intact or severed rhizome connections between halves. Neighborhood competition strongly influenced clone architecture and expansion rates. Removal of neighbors resulted in a >;95% increase in radial clone expansion, intraclonal ramet densities, ramet population growth rates, ramet biomass, and percent of stems flowering, averaged over a 4-year period relative to halves or clones with intact competitors. Plant responses suggest that effects of interclonal neighbors are mediated through alteration of the light environment in the clone canopy and water availability. Addition of nitrogen did not affect lateral spread or clone structure, but resulted in significant increases in ramet size, flowering, and seed production. ANOVA revealed no significant effect of rhizome severing or treatment × severing interactions, suggesting that the size of the integrated physiological unit is much smaller than clone size and/or that physiological integration had no effect on clone responses to environmental heterogeneity at the scale of the diameter of established clones.     

The role of jasmonic acid and ethylene crosstalk in direct defense of Nicotiana attenuata plants against chewing herbivores

We examined performance of herbivores on plants lacking either jasmonate (JA, asLOX3) or ethylene (ET, mETR1) signaling or both (mETR1asLOX3). Plant defenses against Manduca sexta caterpillars were strongly impaired in JA-deficient asLOX3 plants; however, making asLOX3 plants ethylene insensitive did not further increase the performance of the larvae on a mETR1asLOX3 genetic cross. This result demonstrates the dominant role of JA over ET in the regulation of plant defenses against herbivores. However, ET-insensitivity combined with otherwise normal levels of JA in mETR1 plants promoted faster caterpillar growth, which correlated with reduced accumulation of the alkaloidal direct defense nicotine in mETR1 compared to WT plants. Our data points to an important accessory function of ET in the activation of JA-regulated plant defenses against herbivores at the level of alkaloid biosynthesis in the roots and/or accumulation in the leaves.Key words: herbivory, jasmonic acid and ethylene crosstalk, Nicotiana attenuata, nicotine, trypsin proteinase inhibitors (TPIs)     

Seasonal Changes in Shoot and Root Nitrogen Distribution in Switchgrass (Panicum virgatum)

Switchgrass is a promising bioenergy source that is perennial, productive, native to a broad geographic region, and can grow on marginal, nitrogen (N)-poor soils. Understanding N dynamics in switchgrass is critical to predicting productivity, conserving N, and optimizing the timing of harvest. We examined seasonal changes in N distribution in above- and belowground tissues in switchgrass to quantify N retranslocation rates. Above- and belowground biomass from three sites (two in PA and one in NE) were collected and analyzed for biomass growth and N concentrations at 30-day intervals from June through October. Total living plant mass ranged from 10.3?±?2.4 standard error (SE) to 14.9?±?2.5 SE Mg ha^?1. Belowground mass comprised 52–57 % of total mass. Blades had the highest N concentration during summer, ranging from 6 to 22 g kg^?1 N. Aboveground N concentrations decreased from September until autumn senescence, whereas belowground N concentration increased from August until senescence. Across the sites, total N retranslocated from aboveground to belowground components between September and October averaged 16.5?±?7.1 (SE)?kg ha^?1 N representing 26.7 % of the average maximum N content of aboveground biomass. Based on N fertilizer costs, delayed harvest would conserve some N and provide financial savings on fertilizer ($9 ha^?1) if harvest occurs after senescence but before overwinter biomass loss. However, biomass losses of even 10 % will negate potential economic savings accrued from N retention. To maximize environmental and economic savings from N retranslocation and to simultaneously minimize harvest losses, it would be optimal to harvest switchgrass as soon as possible after complete senescence.     

Exsertion,flowering, and shedding in Panicum maxicum (Poaceae)

Within-panicle flowering, exsertion, and seed ripening were investigated in four clones of P. maximum. Flowering, exsertion, and seed ripening were fast processes (<;10 d for each process). Dates and timing were stable within each clone and should allow definition of the harvest date for each genotype and for each harvest method. Clones showed two main flowering patterns differing in the number of flowered spikelets at the date of maximal flowering and in the time of flowering. Seed set was similar to that of allogamous and anemophilous plants. It was nevertheless lower in clones with a high degree of flowering-shedding overlap. In the absence of flowering-shedding overlap, the time for a panicle to flower and to produce seeds was shorter than the time between two emergences of panicles on a tiller. Reproduction timing seemed controlled to minimize competition between panicles on a tiller.     

Silica in grasses as a defence against insect herbivores: contrasting effects on folivores and a phloem feeder

1. Silica, deposited as opaline phytoliths in the leaves of grasses, constitutes 2-5% of dry leaf mass, yet its function remains unclear. It has been proposed that silica may act as an antiherbivore defence by increasing the abrasiveness and reducing the digestibility of grass leaves, although there is little direct experimental evidence to support this. 2. We investigated the effects of manipulated silica levels on the abrasiveness of the leaves of five grass species. We also examined the effects of silica levels on the feeding preferences, growth performance and digestion efficiency of two folivorous insects and one phloem-feeding insect. 3. Silica addition resulted in increases to leaf abrasiveness in four of the five grass species studied. Silica addition also deterred feeding by both folivores and reduced their growth rates and digestion efficiency. 4. These effects resulted in lower pupal mass of the lepidopteron larvae Spodoptera exempta and compensatory feeding by the orthopteran, Schistocerca gregaria. In contrast, silica had no effects on the feeding preference or the population growth of the phloem feeder, Sitobion avenae. 5. Our results demonstrate that silica is an effective defence against folivorous insects, both as a feeding deterrent, possibly mediated by increased abrasiveness, and as a digestibility reducer. The effects of silica on pupal mass and development time may impact on herbivore fitness and exposure to natural enemies. 6. These results are the first demonstration of a direct effect of silica on the abrasiveness of grasses and the adverse impact of silica on herbivore preference and performance.     

Non-protein amino acids in plant defense against insect herbivores: representative cases and opportunities for further functional analysis

Chemical defense against herbivores is of utmost importance for plants. Primary and secondary metabolites, including non-protein amino acids, have been implicated in plant defense against insect pests. High levels of non-protein amino acids have been identified in certain plant families, including legumes and grasses, where they have been associated with resistance to insect herbivory. Non-protein amino acids can have direct toxic effects via several mechanisms, including misincorporation into proteins, obstruction of primary metabolism, and mimicking and interfering with insect neurological processes. Additionally, certain non-protein amino acids allow nitrogen to be stored in a form that is metabolically inaccessible to herbivores and, in some cases, may act as signals for further plant defense responses. Specialized insect herbivores often possess specific mechanisms to avoid or detoxify non-protein amino acids from their host plants. Although hundreds of non-protein amino acids have been found in nature, biosynthetic pathways and defensive functions have been elucidated in only a few cases. Next-generation sequencing technologies and the development of additional plant and insect model species will facilitate further research on the production of non-protein amino acids, a widespread but relatively uninvestigated plant defense mechanism.     

Distribution, morphological description, and molecular characterization of Xiphinema and Longidorous spp. associated with plants (Miscanthus spp. and Panicum virgatum) used for biofuels

Longidorid plant pathogens, specifically Longidorus and Xiphinema , are known to reduce the biomass and seed yields of several monocotyledonous host plants and may serve as vectors for certain plant viruses. The occurrence and densities of species in these genera in the Midwestern United States may be of concern in the production of two biofuel plant species – Miscanthus ( Miscanthus × giganteus ) and Panicum virgatum . Here we present results from a morphological and molecular survey of plant parasitic nematodes in which two species of Xiphinema ( X. americanum and X. rivesi ) and one species of Longidorus ( L. breviannulatus ) were detected. A total of 73 soil samples were collected from 19 sites (nine Miscanthus and 10 P. virgatum ) in three states during 2008. Xiphinema spp. was found in 36% of the Miscanthus plots and in 26% of the P. virgatum plots with a population density range of 22–857/100 cm³ soil. L. breviannulatus was detected in association with Miscanthus plot in Havana, IL, in large numbers (54/100 cm³ soil). The occurrence of this species was associated with severe damage to the fibrous root system, including stunting and necrosis. Short notes, measurements, illustrations and molecular data are given to the species level.     

Thermal requirements for seed germination in Miscanthus compared with Switchgrass (Panicum virgatum), Reed canary grass (Phalaris arundinaceae), Maize (Zea mays) and perennial ryegrass (Lolium perenne)

The high establishment costs of Miscanthus by clonal propagation are a barrier to widespread deployment. Direct sowing is the cheapest method, but limited field trials have given generally poor results. Miscanthus, a perennial grass with C₄ photosynthesis has tropical origins, but is found growing both at high latitudes (>40°) and altitudes (>1000 m) in Asia. In this paper, we investigate if significant variation in the thermal requirements for germination exist in 10 Miscanthus sinensis half‐sib families and compare these with Panicum virgatum (Switchgrass – Trailblazer), Phalaris arundinaceae (Reed canary grass – P10) and Lolium perenne (perennial ryegrass cv AberDart) and maize (Zea mays cv Aviso). The comparisons were made on a thermal gradient bar with a controlled temperature oscillating ± 5 °C on a 12 h cycle and germination was monitored daily for 35 days at mean temperatures ranging from 5.3 to 26.5 °C. Base temperatures were calculated below which germination of at least 50% of viable seeds ceased. Base temperatures were lowest for L. perenne and Zea mays at 3.4 and 4.5 °C respectively; for different Miscanthus half‐sib families base temperatures ranged between 9.7 and 11.6 °C and these were higher than maize and switchgrass which share C₄ photosynthesis with Miscanthus. Parameters derived from germination and temperature were used to predict germination patterns in Europe based on historical climate data. We predict that seed establishment of Miscanthus in spring time is unlikely to be viable in Northern Europe under present climatic conditions without crop management practices aimed at raising soil temperature, and that useful variation in thermal requirement for germination in Miscanthus is available which should facilitate seed germination in other regions.     

Internode structure and cell wall composition in maturing tillers of switchgrass (Panicum virgatum. L)

This work examined cell composition gradients in maturing tillers of switchgrass (Panicum virgatum L.) with the aim of developing baseline information on this important forage and biomass crop. Flowering tillers were collected from plants raised from seeds in a greenhouse and field, harvested at soil level and separated into internodes beginning with the node subtending the peduncle. Internodes were analyzed using microscopy, by fiber digestion, high-performance liquid chromatography and by gas chromatography-mass spectrometry to obtain anatomical and compositional data. Microscopy demonstrated the development and maturation of cortical fibers which eventually became confluent with the fiber sheath surrounding vascular bundles in the lower internodes. Detergent fiber analysis indicated increasing cellulose and lignin contents and decreases in cell solubles and hemicelluloses with increasing distance of the internodes from the top of the plant. Soluble phenolics were greatest in amounts and complexity in top internodes. The lower internodes contained greater levels of wall-bound phenolic acids, principally as 4-coumarate and ferulate.     

Inflorescence, spikelet, and floral development in Panicum maximum and Urochloa plantaginea (Poaceae)

Inflorescence development in Panicum maximum and Urochloa plantaginea was comparatively studied with scanning electron and light microscopy to test the transfer of P. maximum to Urochloa and to look for developmental features applicable to future cladistic studies of the phosphoenol pyruvate carboxykinase (PCK) subtype of C(4) photosynthesis clade (P. maximum and some species of Brachiaria, Chaetium, Eriochloa, Melinis, and Urochloa). Eleven developmental features not discernable in the mature inflorescence were found: direction of branch differentiation; origins of primary branches; apical vs. intercalary development of the main axis; direction of spikelet differentiation; direction of glume, lemma and palea differentiation; position of the lower glume (in some cases); size of the floret meristem; pattern of distal floret development; pattern of gynoecium abortion; differential pollen development between proximal and distal floret; and glume elongation. Inflorescence homologies between P. maximum and U. plantaginea are also clarified. Panicum maximum and U. plantaginea differ not only in their mature inflorescence structure but also in eight fundamental developmental features that exclude P. maximum from Urochloa. The following developmental events are related to sex expression: size of floret meristem, gynoecium abortion, pollen development delay in the proximal floret, glume elongation and basipetal floret maturation at anthesis.     

Relative Performance of Non-Local Cultivars and Local,Wild Populations of Switchgrass (Panicum virgatum) in Competition Experiments

The possibility of increased invasiveness in cultivated varieties of native perennial species is a question of interest in biofuel risk assessment. Competitive success is a key factor in the fitness and invasive potential of perennial plants, and thus the large-scale release of high-yielding biomass cultivars warrants empirical comparisons with local conspecifics in the presence of competitors. We evaluated the performance of non-local cultivars and local wild biotypes of the tallgrass species Panicum virgatum L. (switchgrass) in competition experiments during two growing seasons in Ohio and Iowa. At each location, we measured growth and reproductive traits (plant height, tiller number, flowering time, aboveground biomass, and seed production) of four non-locally sourced cultivars and two locally collected wild biotypes. Plants were grown in common garden experiments under three types of competition, referred to as none, moderate (with Schizachyrium scoparium), and high (with Bromus inermis). In both states, the two “lowland” cultivars grew taller, flowered later, and produced between 2x and 7.5x more biomass and between 3x and 34x more seeds per plant than local wild biotypes, while the other two cultivars were comparable to wild biotypes in these traits. Competition did not affect relative differences among biotypes, with the exception of shoot number, which was more similar among biotypes under high competition. Insights into functional differences between cultivars and wild biotypes are crucial for developing biomass crops while mitigating the potential for invasiveness. Here, two of the four cultivars generally performed better than wild biotypes, indicating that these biotypes may pose more of a risk in terms of their ability to establish vigorous feral populations in new regions outside of their area of origin. Our results support an ongoing assessment of switchgrass cultivars developed for large-scale planting for biofuels.     

The Mycorrhizal Fungus, Sebacina vermifera, Enhances Seed Germination and Biomass Production in Switchgrass (Panicum virgatum L)

Seed dormancy and slow seedling establishment are two major concerns in switchgrass (Panicum virgatum L.) production, often resulting in a poor stand with reduced productivity. Studies were conducted to investigate the stability of artificial associations between switchgrass and the ectomycorrhizal fungus, Sebacina vermifera, and to evaluate the potential benefits of this novel association in seed germination and biomass production. All six strains of S. vermifera tested had a high frequency of colonization on switchgrass roots of a synthetic cultivar NF/GA-993. The positive effects of the associations were reflected in plant height, root length, and biomass production. Inoculated plants produced as much as 75%, 113%, and 18% more shoot biomass than un-inoculated control plants in the first, second, and third harvest, respectively, with no consequent reduction in root biomass. Further, culture filtrates from some strains of S. vermifera increased seed germination in the switchgrass cultivar Kanlow by 52% over the control (p?<?0.05). This study illustrates the great potential of microbial associations to increase biomass production and productivity of switchgrass.     

Characterization and adaptive evolution of {alpha}-tubulin genes in the Miscanthus sinensis complex (Poaceae)

To investigate the organization of and mode of selection in the α-tubulin genes, full-length α-tubulin genes were cloned from four intraspecific taxa of Miscanthus sinensis and its close relatives M. floridulus and M. condensatus using standard polymerase chain reaction (PCR) and rapid amplification of genomic ends (RAGE)-PCR strategies. Genealogical analysis of angiosperms recovered a monophyletic group of Miscanthus α-tubulin genes, which is homologous to the tua5 locus of maize. Two clusters of nearly equal frequency revealed paraphyly within each Miscanthus taxon. Between-cluster recombination was frequent. Additional evidence for co-occurrence of two haplotypes within individuals and a large-scale crossover all suggested a likely allelic relationship between the Miscanthus clusters. Given a long between-species divergence time in Miscanthus, wide occurrence of the trans-species polymorphisms in α-tubulin genes and the approximately equal frequency of each allelic type make it extremely unlikely that α-tubulin diversity has been maintained under neutrality. Balancing selection may have contributed to such an apportioning of genetic variability as well as to high levels of genetic variation in α-tubulin and higher substitution rates at synonymous sites of exons than at intron bases of M. sinensis. In addition, certain effects of demographic oscillation may have distorted the scenario of a functional locus operating under balancing selection.