Cross-pollination of nontransgenic corn ears with transgenic Bt corn: efficacy against lepidopteran pests and implications for resistance management

The efficacy of nontransgenic sweet corn, Zea mays L., hybrids cross-pollinated by Bacillus thuringiensis (Bt) sweet corn hybrids expressing Cry1Ab toxin was evaluated in both field and laboratory studies in Minnesota in 2000. Non-Bt and Bt hybrids (maternal plants) were cross-pollinated with pollen from both non-Bt and Bt hybrids (paternal plants) to create four crosses. Subsequent crosses were evaluated for efficacy in the field against European corn borer, Ostrinia nubilalis (Hübner), and corn earworm, Helicoverpa zea (Boddie), and in laboratory bioassays against O. nubilalis. Field studies indicated that crosses with maternal Bt plants led to low levels of survival for both O. nubilalis and H. zea compared with the non-Bt x non-Bt cross. However, the cross between non-Bt ears and Bt pollen led to survival rates of 43 and 63% for O. nubilalis and H. zea larvae, respectively. This intermediate level of survival also was reflected in the number of kernels damaged. Laboratory bioassays for O. nubilalis, further confirmed field results with larval survival on kernels from the cross between non-Bt ears and Bt pollen reaching 60% compared with non-Bt crossed with non-Bt. These results suggest that non-Bt refuge plants, when planted in proximity to Bt plants, and cross-pollinated, can result in sublethal exposure of O. nubilalis and H. zea larvae to Bt and may undermine the high-dose/refuge resistance management strategy for corn hybrids expressing Cry1Ab.     

Modeling the impact of cross-pollination and low toxin expression in corn kernels on adaptation of European corn borer (Lepidoptera: Crambidae) to transgenic insecticidal corn

We used a mathematical model with processes reflecting larval mortality resulting from feeding on cross-pollinated ears or Bt ears of corn to analyze the risk of evolution of Cry-toxin resistance in Ostrinia nubilalis (Hübner). In the simulations, evolution of resistance was delayed equally well by both seed mixtures and blocks with the same proportion of refuge. Our results showed that Bt-pollen drift has little impact on the evolution of Bt resistance in O. nubilalis. However, low-toxin expression in ears of transgenic corn can reduce the durability of transgenic corn expressing single toxin, whereas durability of pyramided corn hybrids is not significantly reduced. The toxin-survival rate of heterozygous larvae in Bt-corn ears expressing one or two proteins has more impact on evolution of Bt resistance in O. nubilalis than the parameters related to larval movement to Bt ears or the toxin-survival rate of the homozygous susceptible larvae in Bt ears. Bt resistance evolves slower when toxin mortality is distributed across the first two larval stadia than when only the first instars are susceptible to Bt toxins. We suggest that stakeholders examine toxin-survival rates for insect pests and take into account that instars may feed on different parts of Bt corn.     

Quantitative assessment of biotic mortality factors of the European corn borer (Lepidoptera: Crambidae) in field corn.

Five treatments were used to exclude naturally occurring predators and parasitoids, based on body size and flight ability, to assess their effect on Ostrinia nubilalis (Hübner) populations on corn plaits. Two initial O. nubilalis egg densities (one egg mass and three egg masses per plant) were assigned to each treatment. Egg predation was higher in uncaged treatments than in caged treatments. Flying insect predators, primarily Coleomegilla maculata DeGeer (Coleoptera: Coccinellidae), reduced egg densities by 50%. Thirty-five to 84% of O. nubilalis larvae were infected with Nosema pyrausta (Paillot) (Microspora: Nosematidae). The incidence of Beauveria bassiana (Balsamo) Vuillemin (Deuteromycotina: Hyphomycetes), ranged from 0 to 21%, whereas larval parasitism, mainly by Macrocentrus cingulum Reinhard (Hymenoptera: Braconidae) ranged from 0 to 31%. In contrast to previous studies, this 3-yr field study documents that egg predation and larval infections of O. nubilalis were significant and consistent biotic mortality factors.     

Influence of plant severing on movement of Ostrinia nubilalis larvae in Zea mays hybrid seed production fields

Genetically engineered corn hybrids that contain a cry gene from the bacterium Bacillus thuringiensis Berliner (Bt) are gaining popularity for controlling the corn pest Ostrinia nubilalis (Hübner). Continuous use of Bt corn, however, could select for O. nubilalis that are resistant to this corn. Monitoring for insect resistance is important, because it could help maintain the Bt technology. A possible monitoring method is to collect larval insects in commercial drying bins after harvest from Bt seed production fields. A drawback to this method is that these collections may be contaminated by insects that moved as later instars from severed non-Bt male rows into the adjacent Bt female rows. These larvae have little to no exposure to Bt toxin, resulting in possible "false positives." The objectives of this study were to first find which combination of planting and severing dates produces the least number of larvae that move from non-Bt male plants to Bt female plants and to assess O. nubilalis larval movement from severed non-Bt male rows to Bt female rows. Field studies in 2002 and 2003 were designed to simulate a hybrid seed production field. Results suggest that movement of O. nubilalis larvae from male corn is minimized when corn is planted early and male plants are severed by 2 wk post-anthesis. This reduces the likelihood of false positives by reducing the number of susceptible larvae moving between Bt and non-Bt plants. Also, larvae moved to all four female rows that were adjacent to the severed rows, but there were significantly more larvae found in the closest row compared with the other three. These results could be used to develop a monitoring program to find O. nubilalis larvae with resistance to Bt corn in field populations of O. nubilalis.     

Growth, development, and survival of Nosema pyrausta-infected European corn borers (Lepidoptera: Crambidae) reared on meridic diet and Cry1Ab

Transgenic corn, Zea mays L., hybrids expressing crystal protein endotoxin genes from Bacillus thuringiensis Berliner are an increasingly popular tactic for managing the European corn borer, Ostrinia nubilalis (Hübner), in North America. O. nubilalis populations also are often vulnerable to the ubiquitous entomopathogenic microsporidium Nosema pyrausta (Paillot). We examined the effect of feeding meridic diet incorporated with purified Cry1Ab on growth, development, and survival of Nosema-infected and uninfected neonate O. nubilalis. Infected larvae developed more slowly than uninfected larvae. Increasing the concentration of Cry1Ab in diet reduced larval development, and this effect was amplified by microsporidiosis. Infected larvae weighed significantly less than uninfected larvae. The relationship among Nosema infection, Cry1Ab concentration, and larval weight was fitted to an exponential function. The LC50 of infected larvae was one-third that of uninfected larvae, indicating that infected larvae are more vulnerable to toxin. This work has implications for resistance management of O. nubilalis and demonstrates that it is important to determine whether N. pyrausta is present when testing susceptibility of larvae to transgenic corn hybrids.     

Effect of Inoculative Releases of Trichogramma ostriniae on Populations of Ostrinia nubilalis and Damage to Sweet Corn and Field Corn

European corn borer (Ostrinia nubilalis) is one of the most injurious pests of sweet and field corn in the United States. We report here on controlled experiments in which an egg parasitoid (Trichogramma ostriniae) was released inoculatively (75,000 females ha⁻¹) early in the growing season (when corn plants were at the early to mid-whorl stage) to test its efficacy as a biological control agent of O. nubilalis. Releases were made in fields of sweet corn and field corn. Numbers of eggs laid in experimental plots, larval tunnels, and larvae and proportion of damaged ears were determined. Mass of ears was determined for field corn plots. In sweet corn, despite greater oviposition by O. nubilalis in T. ostriniae release plots, the number of borer larvae, stalk tunnels, and damaged ears was reduced by ∼50% compared with those in nonrelease plots. This reduction in damage was consistent for early and late-planted sweet corn. In the field corn plots, larger numbers of O. nubilalis eggs were again laid in some release plots than in control plots. However, O. nubilalis damage appeared to be suppressed in T. ostriniae release plots, although no significant differences were found in most. These results were promising, but further work is required in field corn. The results for sweet corn demonstrated that inoculative releases of T. ostriniae provide suppression of O. nubilalis populations adequate to reduce damage significantly.     

Binomial sequential sampling plans for late instars of European corn borer (Lepidoptera: Crambidae), corn earworm (Lepidoptera: Noctuidae), and damaged kernels in sweet corn ears

Late-season infestations of European corn borer, Ostrinia nubilalis (Hübner), and corn earworm, Helicoverpa zea (Boddie), were sampled to develop binomial sequential sampling plans for larval infestations and damaged kernels in sweet corn, Zea mays L., ears, near harvest. Fields were sampled to obtain a range of larval densities likely to be encountered over a range of infestation levels and field conditions. Binomial sampling plans were developed for O. nubilalis larvae, H. zea larvae, O. nubilalis, and H. zea larvae combined, and for damaged sweet corn kernels. Observed densities ranged from 0.01 to 4.40 larvae per ear for O. nubilalis, 0.005-1.62 larvae per ear for H. zea, and 0.004-36.12 damaged kernels per ear. Results of resampling analyses, based on the proportion of ears infested with one or more larvae, or damaged kernels, indicated an average sample size of 34-37 ears was necessary to classify whether larval infestations, or the incidence of damaged kernels, exceeded 5%. Two operating characteristic curves are presented for each of the four sampling plans. Initial results, with upper bounds of 0.10, and alpha (type I) and beta (type II) error rates at 0.10 and 0.05, respectively, resulted in a 90% probability of making the correct management decision at infestation levels >10%. To improve performance of the sampling plans, we modified the binomial plans by reducing the upper bound to 0.075, while maintaining the same error rates. This plan resulted in a higher probability (>95%) of making the correct management decision to reject a sweet corn load when infestation levels are >10%.     

Potential of Trichogramma ostriniae (Hymenoptera: Trichogrammatidae) for biological control of European corn borer (Lepidoptera: Crambidae) in solanaceous crops

We assessed the ability of Trichogramma ostriniae (Peng & Chen) to locate and parasitize Ostrinia nubilalis (Hübner) eggs in crops other than corn, and we evaluated the efficacy of inundative releases of the parasitoid in two solanaceous crops, pepper and potato. Despite a greater plant surface area to search, parasitism of O. nubilalis eggs was consistently higher in sweet corn than dicotyledonous crops such as pepper, snap bean, broccoli, potato, and melon, in choice and no-choice experiments. Nonetheless, in 2002 and 2003, we made four to five separate inundative releases of approximately 30,000-50,000 T. ostriniae per 0.02 ha in nine pepper fields in Virginia, Pennsylvania, and Massachusetts and compared O. nubilalis egg parasitization and fruit damage in those plots with spatially isolated nonrelease plots. Egg parasitization averaged 48.7% in T. ostriniae release plots, which was significantly higher than in nonrelease plots (1.9%). Also, cumulative pepper fruit damage averaged 8.7% in release plots, which was significantly less than in nonrelease plots (27.3%). In potatoes in 2002 and 2003, we made two releases of approximately 75,000 T. ostriniae per 0.2 ha in nine fields in Maine and Virginia and compared O. nubilalis damage in those plots with that in nonrelease plots. T. ostriniae releases significantly reduced the number of tunnel holes and number of O. nubilalis larvae in potato stems. We conclude that this parasitoid has great potential as a biocontrol agent for O. nubilalis in solanaceous crops.     

Effect of planting dates and Bacillus thuringiensis corn on the population dynamics of European corn borer (Lepidoptera: Crambidae).

Field studies were conducted to determine how field corn, Zea mays L., phenologies in combination with transgenic Bacillus thuringiensis Berliner (Bt) corn and non-Bt (near isogenic) corn could affect egg laying by female European corn borer, Ostrinia nubilalis (Hubner), and subsequent larval injury. Transgenic Bt (events 176 and Bt11) and non-Bt corn was planted at three different times to assess the use of early- and late- planted Bt corn as a means for egg recruitment to these targeted planting dates. Plant growth stages, egg densities, and stalk tunneling was recorded at four locations in southwestern, central, and northern Iowa for three summers (1996-1998). No significant differences in egg densities were observed between Bt and non-Bt corn during the first and second generation for all three years. Significant differences did occur among planting dates. Between 50 and 100% of the eggs were laid in the early planting during the first generation. In addition, between 40 and 65% of the eggs were laid in the late planting for the second generation. Correlations between egg density and larval tunneling were inconsistent from year to year. Additional inconsistencies stemming from yearly phenological differences among sequential plantings and variable O. nubilalis populations increases the difficulty in recommending planting date adjustments as a practical management tool for European corn borer and Bt corn.     

Insect pest densities across site-specific management zones of irrigated corn in northeastern Colorado

The ability to manage insect pests in a site-specific manner is hindered by the costs and time required to describe pest densities and distributions. The purpose of this study was to determine whether insect pest distributions are related to site-specific management zones (SSMZs). Site-specific management zones, as described in this study, delineate fields into three zones of similar yield potential: high, medium, and low productivity. If insect densities vary across SSMZs, it is possible that management decisions could be made at the SSMZ level instead of treating the whole field. This research was conducted during summers 2001 and 2002 on cooperators' farms in northeastern Colorado. Surveys were conducted within corn, Zea mays L., fields, so that densities of three common insect pests of Colorado corn could be compared across SSMZ. The three insect pests were western corn rootworm, Diabrotica virgifera virgifera LeConte; European corn borer, Ostrinia nubilalis (HiAbner); and western bean cutworm, Richia albicosta (Smith). D. v. virgifera larvae and adults were most common in the high-productivity SSMZ. O. nubilalis larval abundance was similar at three fields, whereas in a fourth field the larvae were most common in the high-productivity SSMZ. In one field that contained substantial numbers of R. albicosta, egg abundance was similar across SSMZs, whereas larvae were most common in the high-productivity SSMZ. Site-specific management zones seemed to correlate well with the abundance of some insect pests and might prove useful for managing insects in a site-specific manner.     

Frequency and severity of western bean cutworm (Lepidoptera: Noctuidae) ear damage in transgenic corn hybrids expressing different Bacillus thuringiensis cry toxins

The frequency and severity of corn ear damage caused by western bean cutworm, Striacosta albicosta (Smith), were measured on transgenic corn, Zea mays L., hybrids expressing two different insecticidal Bacillus thuringiensis (Bt) (Berliner) Cry toxins (Bt) selected to protect against damage caused by larval European corn borer, Ostrinia nubilalis (Hübner). A field cage experiment deliberately infested with western bean cutworm egg masses resulted in less damage in the hybrid expressing the Cry1F protein and supported fewer western bean cutworm larvae than its non-Bt isoline. Corn hybrids expressing Cry1F, grown in small plot field experiments at three locations over two separate years and exposed to natural western bean cutworm infestations suffered less damage than non-Bt or Bt-hybrids expressing a Cry1Ab protein. Later maturing hybrids suffered more damage than shorter-season hybrids. Finally, corn ears observed in strip trials for several years in diverse agronomic conditions in farmer-cooperator fields corroborated the in-plant protection conferred by corn hybrids expressing the Cry1F protein in small plot field trials.     

Understanding successful resistance management: the European corn borer and Bt corn in the United States

The European corn borer, Ostrinia nubilalis Hübner (Lepidoptera: Crambidae) has been a major pest of corn and other crops in North America since its accidental introduction nearly a hundred years ago. Wide adoption of transgenic corn hybrids that express toxins from Bacillus thuringiensis, referred to as Bt corn, has suppressed corn borer populations and reduced the pest status of this insect in parts of the Corn Belt. Continued suppression of this pest, however, will depend on managing potential resistance to Bt corn, currently through the high-dose refuge (HDR) strategy. In this review, we describe what has been learned with regard to O. nubilalis resistance to Bt toxins either through laboratory selection experiments or isolation of resistance from field populations. We also describe the essential components of the HDR strategy as they relate to O. nubilalis biology and ecology. Additionally, recent developments in insect resistance management (IRM) specific to O. nubilalis that may affect the continued sustainability of this technology are considered.     

Effect of corn hybrids expressing the coleopteran-specific cry3Bb1 protein for corn rootworm control on aboveground insect predators

Field and laboratory studies were conducted to determine the effect of transgenic Bacillus thuringiensis (Bt) corn, Zea mays L. (YieldGard Rootworm), expressing the Cry3Bb1 protein on aboveground nontarget insect predators (minute pirate bug, ladybird beetles, and carabids). Visual counts of adult and immature Orius insidiosus (Say), Coleomegilla maculata (DeGeer), Hippodamia convergens Gurin-Meneville, and Scymnus spp. occurring in Bt corn and its non-Bt isoline were made at Manhattan, KS, in 2002 and at Manhattan and Scandia, KS, in 2003. No significant differences were found between the Bt corn and non-Bt isoline plots in the abundance (number per plant) of O. insidiosus, C. maculata, H. convergens, and Scymnus spp. Field predation on Ostrinia nubilalis (Hübner) (Lepidoptera: Crambidae) egg masses was also observed during the silking stage of corn at Manhattan and Scandia in 2003. No significant differences were observed among treatments in predation rate for predators with chewing versus sucking mouthparts. Two laboratory studies determined the effect of Cry3Bb1 protein expressed in Bt corn pollen on C. maculata and carabids. The larvae of C. maculata were reared on Bt pollen, non-Bt pollen, or greenbugs, Schizaphis graminum (Rondani). The duration of larval and pupal stages, developmental time from egg hatch to adult emergence, percentage of survival, and elytra length were compared among treatments. There were no significant differences in developmental time of larvae fed pollen or greenbugs during their first two instars. However, significantly prolonged development of the third (1 d) and fourth instars (2 d) was observed for larvae fed greenbugs only. Total time for larval development was significantly longer for larvae that fed on greenbugs versus larvae fed on pollen. No significant differences were observed among treatments in the percentage of larvae that pupated or pupal stage duration. Larvae that fed on greenbugs had higher pupal and adult weights compared with pollen-fed larvae. However, pupal and adult weights did not vary between the Bt and non-Bt pollen treatments. No significant differences occurred in longevity and elytra length of beetles among all treatments. Two carabid species, Harpalus caliginosus F. and Harpalus pensylvanicus DeGeer, were reared on moistened dog food sprinkled with Bt or non-Bt corn pollen. No significant differences in mortality of H. caliginosus and H. pensylvanicus were detected among any of the treatments. There was no significant effect of Bt pollen on fecundity and egg viability of H. caliginosus. Our studies showed that YieldGard Rootworm had no effect on the selected coleopteran predators; therefore, this Bt corn hybrid could be used in an integrated pest management system.     

Large-scale releases of Trichogramma ostriniae to suppress Ostrinia nubilalis in commercially grown processing and fresh market sweet corn

Abstract:  We evaluated the egg parasitoid Trichogramma ostriniae (Pang et Chen) (Hym.: Trichogrammatidae) for control of European corn borer [Lep.:Crambidae: Ostrinia nubilalis (Hübner)] in fields of commercial processing sweet corn in New York in 2002 and 2003. We made inoculative releases of 75 000 T. ostriniae /ha when corn was at mid-whorl and regional O. nubilalis activity had begun. Each release field was paired with a similar non-release control. The incidence of egg mass parasitism, number of stalk tunnels, incidence of ear damage and effect on insecticide spray decisions were evaluated. Parasitism of cumulative number of O. nubilalis egg masses was 51.3 ± 7.2% and 43.5 ± 5.7% (mean ± SEM) in release fields in 2002 and 2003 respectively. The incidence of ear damage by O. nubilalis was not affected by T. ostriniae in either year of the study. There was no difference in stalk damage between release fields and control fields in either 2002 or 2003. However, individual comparisons between paired release and control fields showed reduced stalk tunnelling in 10 of 19 fields, but ear damage was reduced in only two of 19 fields. Neither the number of actual insecticide sprays nor the number of decisions to spray based on sequential sampling were affected significantly by releasing T. ostriniae . In a separate but related study, where releases were conducted in commercial fresh market sweet corn and used higher rates of release, T. ostriniae had no effect on the number of insecticidal sprays. In an additional experiment using small plots of sweet corn, ear damage was similar whether plots were treated with T. Ostriniae or with insecticide. Additional research should focus on improving the timing, frequency and rate of releases.     

Inheritance of Cry1F resistance in laboratory-selected European corn borer and its survival on transgenic corn expressing the Cry1F toxin

A major assumption of the high-dose/refuge strategy proposed for insect resistance management strategies for transgenic crop plants that express toxins from Bacillus thuringiensis is that resistance traits that evolve in pest species will be recessive. The inheritance of Cry1F resistance and larval survival on commercially available Cry1F corn hybrids were determined in a laboratory-selected strain of European corn borer, Ostrinia nubilalis (Hübner), displaying more than 3000-fold resistance to Cry1F. Concentration-response bioassays of reciprocal parental crosses indicated that the resistance is autosomal and recessive. Bioassays of the backcross of the F1 generation with the selected strain were consistent with the hypothesis that a single locus, or a set of tightly linked loci, is responsible for the resistance. Greenhouse experiments with Cry1F-expressing corn hybrids indicated that some resistant larvae survived the high dose of toxin delivered by Cry1F-expressing plants although F1 progeny of susceptible by resistant crosses had fitness close to zero. These results provide the first direct evidence that the high dose/refuge strategy currently in place to manage resistance in Cry1F-expressing corn is appropriate.     

Comparative susceptibility of European corn borer, southwestern corn borer, and sugarcane borer (Lepidoptera: Crambidae) to Cry1Ab protein in a commercial Bacillus thuringiensis corn hybrid

One field strain each of the European corn borer, Ostrinia nubilalis (Hübner); southwestern corn borer, Diatraea grandiosella Dyar; and sugarcane borer, Diatraea saccharalis (F.); were collected from cornfields in northeastern Louisiana. Susceptibilities of the field strain and a corresponding laboratory strain of the three borer species to Cry1Ab protein in DK69-70 Bacillus thuringiensis (Bt) corn hybrid were determined by exposing neonates to intact leaf tissues from whorl stage plants or by feeding neonates or third instars on a meridic diet treated with different concentrations of Cry1lAb protein extracted from Bt corn leaves. Mortality and growth of larvae were evaluated after 2 and 4 d posttreatment in the bioassays by using intact leaf tissues or after 7 d in the bioassays by using diet incorporating Cry1Ab protein. D. saccharalis was the least susceptible species to Cry1Ab protein among the three species, followed by D. grandiosella, whereas O. nubilalis was most susceptible. The 2-d mortality of D. saccharalis neonates on intact Bt leaf tissues was lower than that of O. nubilalis and D. grandiosella. All neonates of O. nubilalis were killed on the diet treated with Cry1Ab protein at 0.5 and 1 mg/kg. The mortality of D. grandiosella was > 75% at 1 mg/kg, but it was < 6% for D. saccharalis at 1 mg/kg. The LC50 values of D. saccharalis were 3- and 11-fold higher than those of D. grandiosella and O. nubilalis, respectively. The LC90 values of D. saccharalis were 8- and 32-fold higher than those of D. grandiosella and O. nubilalis, respectively. Larval growth of the three species on Cry1Ab-treated diet was inhibited, but the inhibition was greater for O. nubilalis and D. grandiosella than for D. saccharalis. The lower susceptibility of D. saccharalis to Cry1Ab protein suggests that it is necessary to verify if a high-dose Bt corn for O. nubilalis and D. grandiosella is also a high dose for D. saccharalis.     

甘蔗、玉米气味对亚洲玉米螟成虫产卵和幼虫取食的影响

【目的】农作物间套作对害虫的影响很大程度上取决于害虫的行为反应,通过研究玉米/甘蔗套作系统中植物气味对亚洲玉米螟Ostrinia furnacalis(Guenee)行为的影响,将为利用化学生态手段防治套作田中的害虫提供依据。【方法】通过昆虫触角电位实验、成虫产卵和幼虫取食实验,测定了甘蔗、玉米植株及二氯甲烷漂洗物对亚洲玉米螟成虫产卵、幼虫取食的影响。【结果】亚洲玉米螟成虫在甘蔗植株上的落卵量(卵粒数、卵块数)与玉米植株差异不显著,取食甘蔗植株的幼虫数量显著低于玉米植株。亚洲玉米螟对甘蔗、玉米叶片漂洗物均可产生EAG反应,且在同一浓度下的反应值之间无显著差异。成虫产卵量(卵粒数、卵块数)在0.1 gE/mL浓度的甘蔗和玉米漂洗物间差异不显著;幼虫对经甘蔗叶片漂洗物处理过的饲料的相对取食率均低于65.00%,显著低于玉米漂洗物处理的饲料。【结论】甘蔗和玉米气味对亚洲玉米螟产卵的影响没有差异,但甘蔗气味会导致初孵幼虫表现出很强的逃逸行为。     

Evaluation of thiamethoxam and imidacloprid as seed treatments to control European corn borer and Indianmeal moth (Lepidoptera: Pyralidae) larvae

Efficacy of thiamethoxam (Cruiser) and imidacloprid (Gaucho) were evaluated as seed treatments for controlling European corn borer, Ostrinia nubilalis (Hübner) and Indianmeal moth, Plodia interpunctella (Hübner) larvae in stored grain. At approximately 22-26 degrees C, all fifth instar European corn borers died after two or 4 d of exposure to corn treated with 250 and 500 ppm thiamethoxam, respectively, while mortality of larvae exposed for two and 4 d on corn treated with 6.3-937.5 ppm imidacloprid did not exceed 48% at any concentration. At 29 degrees C, all nondiapausing fifth instars were killed after 3, 4, and 6-d exposure to 400, 300 and 200-ppm thiamethoxam, respectively, while survival increased at successively lower concentrations of 100, 50, 25, and 12.5 ppm. At 29 degrees C, the LC50 decreased from 85.9 to 7.2 ppm as the duration of exposure on treated corn increased from 2 to 6 d. All second and third instar Indianmeal moth larvae died after a 5 d exposure period to corn grain treated with thiamethoxam at 50 ppm or higher, but as the larvae aged, higher concentrations and longer exposure periods were required to give 100% mortality of each larval instar. Similar results were obtained when larval Indianmeal moths were exposed on corn treated with imidacloprid, or on sorghum treated with thiamethoxam. Mature wandering phase fifth instars were the most tolerant larval stage of the Indianmeal moth.     

Role of egg density on establishment and plant-to-plant movement by western corn rootworm larvae (Coleoptera: Chrysomelidae)

The effect of egg density on establishment and dispersal of larvae of the western corn rootworm, Diabrotica virgifera virgifera LeConte, was evaluated in a 3-yr field study. Implications of these data for resistance management plans for Bt crops are discussed. Viable egg levels of 100, 200, 400, 800, and 1600 eggs per infested plant were evaluated in 2000, 2001, and 2002. A 3200 viable egg level was also tested in 2001 and 2002. All eggs were infested on one plant per subplot in a field that was planted to soybean, Glycine max (L.), in the previous year. For each subplot, the infested plant, three plants down the row, the closest plant in the adjacent row of the plot, and a control plant at least 1.5 m from any infested plant (six plants total) were sampled. In 2000, there were five sample dates between egg hatch and pupation, and in 2001 and 2002, there were six sample dates. On each sample date, four replications of each egg density were sampled for both larval recovery and plant damage. Initial establishment on a corn plant seemed to not be density-dependent because a similar percentage of larvae was recovered from all infestation rates. Plant damage and, secondarily, subsequent postestablishment larval movement were density-dependent. Very little damage and postestablishment movement occurred at lower infestation levels, but significant damage and movement occurred at higher infestation rates. Movement generally occurred at a similar time as significant plant damage and not at initial establishment, so timing of movement seemed to be motivated by available food resources rather than crowding. At the highest infestation level in 2001, significant movement three plants down the row and across the 0.76 m row was detected, perhaps impacting refuge strategies for transgenic corn.     

Seed mixtures as a resistance management strategy for European corn borers (Lepidoptera: Crambidae) infesting transgenic corn expressing Cry1Ab protein

Dispersal of neonate European corn borers, Ostrinia nubilalis (Hübner), in seed mixtures of transgenic corn expressing Cry1Ab protein (Bt+) and nontransgenic corn (Bt-) was evaluated in a 2-yr field study. The main objective was to determine if larval dispersal limits the effectiveness of seed mixtures as a resistance management strategy. Mixtures evaluated included (1) all Bt+ plants, (2) every fifth plant Bt- with remaining plants Bt+, (3) every fifth plant Bt+ with remaining plants Bt-, and (4) all Bt- plants. The transformation events MON 802 (B73 BC1F2 x Mol7) and MON 810 (B73 BC1F1 x Mo17), which express the Cry1Ab endotoxin isolated from Bacillus thuringiensis subsp. kurstaki, were used as the sources of Bt+ seed in 1994 and 1995, respectively (YieldGard, Monsanto, St. Louis, MO). At corn growth stage V6-V8, subplots within each mixture (15-20 plants each) were infested so that every fifth plant in mixtures 1 and 4, every Bt- plant in mixture 2, and every Bt+ plant in mixture 3 received two egg masses. Larval sampling over a 21-d period indicated increased neonate dispersal off of Bt+ plants, reduced survival of larvae that dispersed from Bt+ plants to Bt- plants, and a low incidence of late-instar movement from Bt- plants to Bt+ plants. Computer simulations based on mortality and dispersal estimates from this study indicate that seed mixtures will delay the evolution of resistant European corn borer populations compared with uniform planting of transgenic corn. However, resistant European corn borer populations likely will develop faster in seed mixes compared with separate plantings of Bt and non-Bt corn.