The transgenic maize ( Zea mays L.) event MON 88017 produces the Bacillus thuringiensis Berliner (Bt) toxin Cry3Bb1 to provide protection from western corn rootworm ( Diabrotica virgifera virgifera LeConte) larval feeding. In response to reports of reduced performance of Cry3Bb1‐expressing maize at two locations in Illinois, we conducted a two‐year experiment at these sites to characterize suspected resistance, as well as to evaluate root injury and adult emergence. Single‐plant bioassays were performed on larvae from each population that was suspected to be resistant. Results indicate that these populations had reduced mortality on Cry3Bb1‐expressing maize relative to susceptible control populations. No evidence of cross‐resistance between Cry3Bb1 and Cry34/35Ab1 was documented for the Cry3Bb1‐resistant populations. Field studies were conducted that included treatments with commercially available rootworm Bt hybrids and their corresponding non‐Bt near‐isolines. When compared with their near‐isolines, larval root injury and adult emergence were typically reduced for hybrids expressing Cry34/35Ab1 either alone or in a pyramid. In many instances, larval root injury and adult emergence were not significantly different for hybrids expressing mCry3A or Cry3Bb1 alone when compared with their non‐Bt near‐isolines. These findings suggest that Cry34/35Ab1‐expressing Bt maize may represent a valuable option for maize growers where Cry3Bb1 resistance is either confirmed or suspected. Consistent trends in adult size (head capsule width and dry mass) for individuals recovered from emergence cages were not detected during either year of this experiment. Because of the global importance of transgenic crops for managing insect pests, these results suggest that improved decision‐making for insect resistance management is needed to ensure the durability of Bt maize.

The western corn rootworm, Diabrotica virgifera virgifera LeConte, is considered one of the most significant insect pests of maize in North America. Larvae of other secondary subterranean pests such as grape colaspis, Colaspis brunnea (F.), and Japanese beetle, Popillia japonica Newman, can also injure maize seedlings and cause yield loss. In the past decade, maize hybrids containing Bt proteins have been used to manage the western corn rootworm; additionally, seeds are commonly treated with a neonicotinoid and fungicide combination to control secondary pests. Recently, soil‐applied insecticides have been used in conjunction with rootworm Bt hybrids (and seed‐applied insecticides) in areas with perceived risk for increased rootworm larval or secondary pest damage. We conducted a series of trials from 2009 to 2011 that examined multiple rootworm Bt hybrids and their near‐isolines, along with two soil‐applied insecticides, to determine whether the Bt plus insecticide combination resulted in an increased level of efficacy or yield. We also sampled for Japanese beetle and grape colaspis larvae to determine their potential for reducing yield. Densities of secondary pests in our trials were low and likely had no effect on maize yield. The addition of a soil‐applied insecticide to rootworm Bt hybrids improved efficacy only once across 17 location‐years, when overall corn rootworm injury was highest; an improvement in yield was never observed. Our results suggest that the use of a soil‐applied insecticide with a rootworm Bt hybrid should only be considered in scenarios with potentially significant rootworm larval populations. However, potential negative consequences related to trait durability when soil insecticides are used with rootworm Bt maize should be considered.

Maize, Zea mays L., is an economically important crop grown throughout the world. Corn rootworm, Diabrotica spp. (Coleoptera: Chrysomelidae), larvae constitute a significant economic threat to maize production in the United States, where yield losses and management costs associated with corn rootworm species exceed $1 billion annually. Furthermore, the introduction of the western corn rootworm, D. virgifera virgifera LeConte, into maize‐producing regions of Europe has made managing corn rootworm larval injury an international concern. Larvae injure maize plants by feeding on root tissue and are the primary target of management activities. Products commonly used to protect root systems from injury include chemical insecticides (seed or soil applied) and genetically modified maize hybrids expressing toxins derived from Bacillus thuringiensis Berliner (Bt). The confirmation of field‐evolved resistance to various Bt toxins in populations of the western corn rootworm presents a significant management challenge. We performed a meta‐analysis to provide a broad understanding of the relative efficacy of the primary products currently being used to manage corn rootworm larval injury, including insecticidal seed treatments, soil insecticides and Bt hybrids (with and without the addition of soil insecticide). Our analysis is unique in the breadth of locations and years included – we analysed 135 individual trials conducted from 2003 through 2014 at multiple sites in both Illinois and Nebraska. Panel data were produced by pairing the mean node‐injury rating for each treatment of a given trial with the mean node‐injury rating for untreated maize. Linear regression models were developed to estimate the relationship between the potential for corn rootworm larval injury and product performance. For a given level of injury potential, the parameters estimated reveal differences in the degree of root protection offered by the various product categories analysed. Implications for developing long‐term, integrated, and sustainable practices for managing this important pest of maize are discussed.

Corn rootworm, Diabrotica spp., larvae represent a significant and widespread economic threat to corn, Zea mays (L.), production in the United States, where control costs and yield losses associated with these insect pests exceed $1billion annually. Preventing root injury and associated yield loss caused by corn rootworm larvae may be accomplished by the independent use of planting time soil insecticides or transgenic Bt hybrids. However, recent reports of both confirmed and suspected Bt resistance in corn rootworm populations throughout the Corn Belt have led to significant interest in the use of these two management tactics simultaneously. Although this approach has been investigated to some extent previously, information is lacking on how the use of a soil insecticide in tandem with a Bt seed blendBt and refuge (non-Bt) seed mixed into a single productmay affect root protection and yield. We describe an experiment including six trial sites conducted over a three-year period where various seed blends and soil insecticide/seed blend combinations were evaluated. The predominant species contributing to root injury across all sites was the western corn rootworm (Diabrotica virgifera virgifera LeConte). A weighted technique is presented for evaluating root injury for seed blends that offers a reliable estimate of product performance. The addition of a soil insecticide to the seed blend treatments never resulted in significantly improved root protection and failed to provide a consistent yield benefit. Our results suggest that a soil insecticide/seed blend combination approach is not warranted. Additionally, a subanalysis performed on individual refuge and nearby Bt root systems for seed blend treatments provides insight into the spatial characteristics of root injury in seed blend scenarios.

Western corn rootworm is an important corn pest in the U.S. Some farmers noted unexpected corn rootworm injury of transgenic hybrids as early as 2008; however, the full extent of product performance is still not fully understood. We conducted telephone focus groups with farmers in 2013 to gain their perspective of current and future issues for corn rootworm. Respondents were surprised how quickly corn rootworm injury escalated in their fields and were disappointed with incorrect diagnoses from consultants and seed companies. Most participating farmers saw university Extension as an unbiased source of information.

The western corn rootworm, Diabrotica virgifera virgifera LeConte, is one of the most economically important insect pests threatening the production of corn, Zea mays (L.), in the United States. Throughout its history, this insect has displayed considerable adaptability by overcoming a variety of pest management tactics, including the cultural practice of annual crop rotation. Since first reported in Illinois in the late 1980s, populations of the rotation‐resistant western corn rootworm have spread over a wide area of the eastern Corn Belt. Currently, little information is available concerning the interaction of rotation resistance with the use of genetically modified corn expressing insecticidal toxins from Bacillus thuringiensis Berliner (Bt), a popular tactic for preventing larval injury and its associated yield loss. The goal of this greenhouse experiment was to determine whether rotation‐resistant and rotation‐susceptible western corn rootworm larvae differ with respect to survival or development when exposed to single‐ or dual‐toxin (pyramided) Bt corn. Individual corn plants were infested with 225 near‐hatch eggs at the V5 (five leaf collar) growth stage. Larvae developed undisturbed on the root systems for 17 days, after which they were recovered using Berlese–Tullgren funnels. Surviving larvae were counted to estimate mortality, and head capsule widths were measured to assess development. Rotation‐resistant and rotation‐susceptible larvae had statistically similar mean levels of mortality and head capsule widths when exposed to both single‐toxin (Cry3Bb1 or Cry34/35Ab1) and pyramided (Cry3Bb1+ Cry34/35Ab1) Bt corn, suggesting that these two populations do not differ with respect to survival or development when exposed to Bt corn. Additionally, the statistically similar mean levels of mortality for larvae exposed to single‐toxin and pyramided Bt corn suggest that pyramided Bt hybrids containing the Cry3Bb1 and Cry34/35Ab1 toxins do not result in additive mortality for western corn rootworm larvae. Implications for management of this economically important pest are discussed.

The ability of the biomass crop Miscanthus (Miscanthus x giganteus Greef and Deuter ex Hodkinson and Renvoize) to support larval development for both United States and European populations of the western corn root-worm, Diabrotica virgifera virgifera LeConte, suggests an avenue for interactions with corn (Zea mays L.). To provide context to survival of D.v. virgifera on Miscanthus, adult activity and oviposition of Diabrotica spp. were monitored in central Illinois in 2010-2011 in Miscanthus, corn and switchgrass (Panicum virgatum L.). For D. v. virgifera, vial traps within corn plots captured 3-10 times as many adults as in Miscanthus or switchgrass, while soil samples showed females laid approximately 10 times as many eggs in corn as in the perennial grasses. Adult southern corn root-worms, Diabrotica undecimpunctata howardi Barber, were the most abundant species in 2010 and clearly preferred switchgrass as an adult habitat, with vial traps in switchgrass capturing 5-10 times as many D.u. howardi as those in corn or Miscanthus. Based on the small production areas for Miscanthus and switchgrass (and low use of both by D. v. virgifera), it seems likely that there are no current impacts of these perennial grasses on pest status of Diabrotica spp. in corn or other crops. However, adaptations by Diabrotica spp. to pest management practices suggest they could be a source for interactions between biomass and food or feed crops. Early-season soil samples did not recover eggs of D.u. howardi, but their use of switchgrass as an adult habitat suggests additional research in areas where switchgrass may be grown near peanuts, alfalfa or other hosts may be needed. Also, investigation of other candidate bioenergy crops known to support Diabrotica spp. larval development is needed to better understand the possible effects of a changing agricultural landscape on corn rootworms.

A new marafivirus tentatively named Switchgrass mosaic virus (SwMV) was recently detected in Panicum virgatum L. (switchgrass). The virus is related to Maize rayado fino virus (MRFV). As Miscanthus and other grass species are grown alongside switchgrass in our experimental fields, there has been concern that the virus could be transmitted to these plants. To investigate this possibility, leaf samples from Miscanthus spp. and other grasses rotated with switchgrass in a field planting were tested for SwMV by reverse-transcriptase polymerase chain reaction (RT-PCR). Switchgrass mosaic virus was detected in leaf samples of Miscanthusxgiganteus, M. sinensis, M. transmorrisonensis, Saccharum ravennae, Andropogon virginicus and Sorghastrum nutans. The nucleotide sequences of the amplicons from these samples were 99% identical to SwMV in switchgrass. This is the first report of SwMV detection in Miscanthus spp. and the other grasses, apart from switchgrass.

Switchgrass mosaic virus (SwMV) was identified in switchgrass (Panicum virgatum) and was proposed as a new marafivirus based on its genome sequence and comparison with its closest relative, Maize rayado fino virus (MRFV), a type member of the genus Marafivirus. MRFV only infects maize (Zea mays) and its wild relatives, and is naturally transmitted by a corn leafhopper, Dalbulus maidis. Proving that SwMV is transmitted by a different species of leafhopper than D. maidis would provide additional evidence that SwMV is a new species of the genus Marafivirus. In 2010, three leafhopper species were identified in switchgrass fields near Champaign, Illinois - Graminella aureovittata, Graminella mohri and Flexamia atlantica. Switchgrass mosaic virus was detected by two independent RT-PCR assays in 100% of G. mohri and F. atlantica, and 95% of G. aureovittata. Twenty specimens of each species were evaluated. Of the three leafhoppers, only G. aureovittata transmitted the virus to Cave in Rock' switchgrass plants in a growth chamber. Transmission efficiency was 80% and only one of the eight SwMV-infected plants displayed mosaic/yellow streak symptoms. Switchgrass mosaic virus was detected in 78% and 83% of the switchgrass plants in the two fields from which the leafhoppers were collected. The detection of a leafhopper vector of SwMV will facilitate the transmission of the virus and the study of its impact on switchgrass biomass yield.

The corn rootworm complex (Coleoptera: Chrysomelidae) constitutes a significant threat to maize production in the United States, and more recently, in Europe. We conducted an analysis of readily available field trial data to validate an existing damage function for corn rootworm larvae. We used a nested error component model with unbalanced panel data to describe the relationship between yield loss and root injury caused by these insects. These data were collected by personnel with the Insect Management and Insecticide Evaluation Programme (Department of Crop Sciences, University of Illinois) and represent 19 location-years. To our knowledge, this is the largest data set used to estimate a damage function for corn rootworm larvae. Unlike many experiments examining the relationship between root injury and yield loss caused by corn rootworm larvae, the data set used for our analysis includes many Bt maize hybrids. Our model suggests that for each node of roots injured by corn rootworm larvae, a yield loss of approximately 15% can be expected. Statistically significant variance components included an effect of location and experimental error. We speculate that variation in weather across experimental sites was the principal factor contributing to the significant effect of location. The substantial experimental error observed for our model highlights the limitations of utilizing a multi-year, geographically diverse damage function for predicting yield loss because of root injury on a small scale. We discuss major factors contributing to the variance components estimated by our model and suggest techniques for improving future analyses of the damage function for corn rootworm larvae.

Injury from stem-boring caterpillars has been observed on the perennial grass Miscanthus x giganteus Greef and Deuter ex Hodkinson and Renvoize in both its native and introduced ranges. Because some species causing stem injury in the United States have not been identified, potential biomass reductions to M. X giganteus were measured using southwestern corn borer, Diatraea grandiosella Dyar (Crambidae), an insect pest of several related crops within the Andropogoneae. Results indicate D. grandiosella is capable of survival on whorl and stem tissue from hatch to 21 d in the laboratory, and field infestations with third instars support adult development, as exuviae were found during dissection of injured tillers. Relative to uninfested controls, M. X giganteus tillers with stem injury yielded 12-30% less dry mass in four infestations over 2009-2010. As in some D. grandiosella hosts, data indicate decreased susceptibility to stem-boring as tillers increase in size or age. Regressions of residuals (observed - predicted mass) for injured M. X giganteus tillers onto the cumulative length of tunnels per tiller also showed significant negative slopes (i.e., decreasing tiller mass with increasing tunnel length). Although D. grandiosella survival appeared low in both laboratory and field trials, results indicate that M. X giganteus productivity could become limited by other stem-boring caterpillars known to attack Andropogoneae, including the following: Elasmopalpus lignosellus (Zeller) (Pyralidae), Diatraea saccharalis (F.) (Pyralidae), and Eoreuma loftini (Dyar) (Crambidae). For perennial grasses grown exclusively for biomass, certain management strategies for stem borers or other pests may be uneconomical or impractical, suggesting long-term investment in breeding for host plant resistance may be needed.

Soybean aphids, Aphis glycines Matsumura, and soybean cyst nematodes, Heterodera glycines Ichinohe, are economic pests of soybean, Glycine max (L.) Merr., in the north‐central United States. Combined, these pests may account for 20–50% of yield reductions in a soybean crop. Only limited information is available concerning the interaction of these two pests on soybean production. During the summers of 2006 and 2007, we conducted a field‐experiment near Urbana, IL, to evaluate the effect of resistant and susceptible soybean lines on the development and reproduction of both pests in combination with each other. We also examined how each pest, as well as their interaction, affected the yield of susceptible and resistant soybean lines. Soybean plants grown within caged plots were infested with soybean aphids and soybean cyst nematodes; cumulative aphid days and soybean cyst nematode egg densities were determined at the end of each growing season. Soybean aphids were able to survive on all four soybean lines in both years of this study; however, aphid‐resistant lines generally had fewer cumulative aphid days than aphid‐susceptible lines. Likewise, nematode‐resistant lines typically had fewer eggs than nematode‐susceptible lines. During both years, we failed to observe a significant interaction between these two pests on the reproduction of one another. Yield data from 2006 was inconclusive; however, results from 2007 suggest that yield‐loss when soybean aphids and soybean cyst nematodes occur jointly is not significantly greater than when these two pests occur independently. The relationship between these two pests, and our inability to observe an interaction, are discussed.

A 2-year field experiment was conducted in northern Illinois to evaluate the effects of host plant resistance and an insecticidal seed treatment (thiamethoxam) on soybean aphids, Aphis glycines Matsumura and their predators. Densities of soybean aphids varied between the 2 years of the experiment. During both years, resistant plants experienced fewer cumulative aphid days than susceptible plants. Populations of soybean aphids on resistant plants rarely exceeded the economic injury level of 250 soybean aphids per plant. The use of thiamethoxam reduced cumulative aphid days in 2007, but not in 2008. Although soybean aphids reached densities that were sufficient to cause yield-loss for untreated and susceptible plants, no yield-benefit was associated with using the two management tactics in either year. This latter finding suggests that densities of soybean aphids need to be greater and sustained for a longer period of time than what we observed if the two management tactics are expected to provide a yield-benefit. Monitoring natural enemies revealed that densities of key aphidophagous predators were relatively unaffected by host plant resistance or thiamethoxam; the effect of these management tactics on densities of predators, as well as the effectiveness of the method used to sample predators, is discussed.

Miscanthus sinensis Anderss., a perennial grass, is native to eastern Asia. It has been widely grown as an ornamental in temperate regions of the world, including the United States, and recently has become an important component of public and private sector bioenergy feedstock Miscanthus selection programs. In August 2008, stem rot and blight was observed on M. sinensis plants in two irregular patches, ~2 to 2.5 × 1 to 1.5 m each in a trial plot that was preceded by corn, at the University of Illinois Energy Farm near Urbana, IL. At the time of the observation, most plants were dead and the wilted tillers had black, soft rotted basal stems. A few plants were stunted and the crowns of the tillers had black-to-brown soft rot. Some tillers' leaves were dead and others had turned light brown. Sample tissue fragments were surface disinfested in 0.5% NaOCl and plated on 1% water agar (WA). After 3 days of incubation in the dark at 23°C, colonies were transferred to corn meal agar (CMA), potato dextrose agar (PDA), or 10% V8 juice agar and incubated at 23°C under continuous white light for up to 2 weeks. Morphological characteristics of the isolates correspond to those originally described for Pythium sylvaticum W.A. Campb. & J.W. Hendrix (1). The mycelia grew and covered the 10-cm-diameter plates within 5 days. On PDA, the culture was a creamy white mycelial mat of coenocytic hyphae. The isolates produced only globose, terminal or intercalary hyphal swellings ranging from 28 to 48 μm in diameter, but no oogonia were produced on any of the three growth media. No zoospores were produced when agar blocks bearing mycelium were flooded with distilled water or 1% soil water. Sequence analysis was performed with the internal transcribed spacer (ITS) region of the rDNA amplified with primer pair ITS1/ITS4 (3) and the mitochondrially encoded cytochrome c oxydase subunit II (cox II) gene using primers FM58/FM66 (2). The resulting 871-bp ITS nucleotide sequence (Accession No. HM991706) was identical among all three isolates analyzed and 99% identical (100% coverage) to ITS sequences of multiple isolates of P. sylvaticum in GenBank. Likewise, the 544-bp cox II sequence (Accession No. HQ454429) was 99% identical (97% coverage) to cox II sequences of multiple isolates of P. sylvaticum. Six pots of M. sinensis seedlings were inoculated by placing two CMA plugs of a 2-week-old culture of isolate F71 at the crown. The control pots were mock inoculated with sterile CMA plugs. The plants were incubated at ~90% relative humidity (RH) and 25°C day and 22°C night for 3 days, and thereafter left on the greenhouse bench at ~65% RH with alternating 9 h of darkness and 15 h of light. Three weeks after inoculation, two of the inoculated seedlings wilted, others were stunted with leaves wilting from the tip downwards and the stems rotting from the crown upward. A thick mat of mycelia was seen on the rotted basal stems. No symptoms were observed in the control. P. sylvaticum was reisolated from both the rotted basal stems and the wilted foliage. To our knowledge, this is the first report of P. sylvaticum on M. sinensis. Infestation of farm soils with P. sylvaticum could limit M. sinensis biomass production significantly by limiting seedling establishment. References: (1) W. A. Campbell and F. F. Hendrix. Mycologia 59:274, 1967. (2) F. M. Martin. Mycologia 92:711, 2000. (3) T. J. White et al. Page 38 in: PCR Protocols: A Guide to Methods and Applications. M. A. Innis et al., eds. Academic Press, San Diego, 1990.

To help assess the potential for damage by armyworms [Mythimna (Pseudaletia) unipuncta (Haworth) (Lepidoptera: Noctuidae)] to switchgrass (Panicum virgatum L.) and surrounding crops, survival and development were evaluated for larvae reared on leaves of switchgrass, corn (Zea mays L.), and miscanthus (Miscanthus x giganteus Greef and Deuter ex Hodkinson and Renvoize). Additional tests assessed the relationship between leaf position and the concentration of saponins (plant compounds which can provide protection from insect herbivores) and examined the effect of defoliation on switchgrass dry mass. Survival to adulthood was similar when larvae were reared on field-grown leaves of switchgrass and corn. However, lower larval mass (10 d) and delayed development of M. unipuncta (to pupation, adult emergence) suggest switchgrass is an inferior host relative to corn. When fed field-grown miscanthus, no larvae survived 10 d. Few differences were noted between switchgrass and corn grown under controlled (laboratory) conditions, but M. unipuncta survival seemed to decline rapidly when larvae were fed the fourth and fifth leaves of switchgrass. Switchgrass leaf samples collected from different leaf positions and stages of tiller maturity showed up to 10-fold differences in the concentration of the saponin protodioscin, with the greatest concentrations in the fourth and fifth leaves. However, other saponins showed an opposite pattern, indicating the role of protodioscin on insect development should be tested in isolation (e.g., by addition of the purified compound to an artificial diet). Defoliation trials indicated that extremely high M. unipuncta populations may be necessary to cause any significant reduction in switchgrass biomass. Collectively, results suggest M. unipuncta may not present a significant risk to biomass production in switchgrass, but that the spring emergence of switchgrass provides an alternate host for M. unipuncta before colonizing annual food and feed crops.