Author Correction: A transgenic approach for controlling Lygus in cotton.

An amendment to this paper has been published and can be accessed via a link at the top of the paper.

Quantitative Differences in Feeding Behavior of Lygus lineolaris (Hemiptera: Miridae) on Transgenic and Nontransgenic Cotton.

Lygus lineolaris (Palisot de Beauvois) is one of the most important pests on cotton in the United States. Previous research showed that transgenic cotton plants expressing the Bacillus thurigiensis (Bt) crystalline protein Cry51Aa2.834_16 (designated MON 88702) have insecticidal effects on nymphal L. lineolaris. The present study is the first to examine effects of a Bt-expressing cotton on feeding by a heteropteran like L. lineolaris. We compared stylet probing behaviors of third-instar nymphs on pin-head squares (i.e., buds <3 mm wide) of MON 88702 cotton versus nontransgenic (control) DP393 plants using AC-DC electropenetrography. Waveforms were quantified based on appearances previously characterized and correlated with adult L. lineolaris feeding behaviors; nymphal and adult waveforms had the same appearance. Generalized third-instar feeding included maceration of tissues during cell rupturing (waveform CR), tasting/testing during a waveform called transition (T), and ingestion (I); all were similar between MON 88702 and DP393 plants. However, the number of events and duration of each waveform were different between treatments. Relative to nymphs on DP393, those on MON 88702 spent more time overall in stylet probing, due to increased number of maceration events per probe and longer durations of tasting/testing, per waveform event, per probe, and per insect; yet, ingestion events were shorter and more frequent. These findings support that MON 88702 cotton plants were less palatable and/or preorally digestible to L. lineolaris nymphs than DP393, suggesting antixenosis for MON 88702. Transgenic cotton antixenosis could positively affect cotton pest management by reducing feeding of L. lineolaris nymphs and protecting crop yield.

First transgenic trait for control of plant bugs and thrips in cotton.

BACKGROUND: Plant bugs (Lygus spp.) and thrips (Thrips spp.) are two of the most economically important insect pest groups impacting cotton production in the USA today, but are not controlled by current transgenic cotton varieties. Thus, seed or foliar-applied chemical insecticides are typically required to protect cotton from these pest groups. Currently, these pests are resistant to several insecticides, resulting in fewer options for economically viable management. Previous publications documented the efficacy of transgenic cotton event MON 88702 against plant bugs and thrips in limited laboratory and field studies. Here, we report results from multi-location and multi-year field studies demonstrating efficacy provided by MON 88702 against various levels of these pests. RESULTS: MON 88702 provided a significant reduction in numbers of Lygus nymphs and subsequent yield advantage. MON 88702 also had fewer thrips and minimal injury. The level of control demonstrated by this transgenic trait was significantly better compared with its non-transgenic near-isoline, DP393, receiving insecticides at current commercial rates. CONCLUSION: The level of efficacy demonstrated here suggests that MON 88702, when incorporated into existing IPM programs, could become a valuable additional tool for management of Lygus and thrips in cotton agroecosystems experiencing challenges of resistance to existing chemical control strategies. © 2018 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Certain applied electrical signals during EPG cause negative effects on stylet probing behaviors by adult Lygus lineolaris (Hemiptera: Miridae).

This study is the first to fully evaluate whether electrical signals applied to large insects during electropenetrography (EPG; also called electrical penetration graph) negatively affect insect behavior. During EPG, electrical signals are applied to plants, and thus to the gold-wire-tethered insects feeding on them. The insect completes an electrical circuit whose changes in voltage reflect the insect's stylet probing/penetration behaviors, recorded as waveform output. For nearly 50 years of EPG science, evidence has supported that there are no or negligible effects on tiny insects from applied electricity during EPG. Recently however, EPG studies of large-bodied hemipterans such as heteropterans and sharpshooter leafhoppers have been published. The wider stylet diameters of such large insects cause them to have lower inherent resistances to applied signals compared with smaller insects, conveying more electrical current. The present study asked whether such increased currents would affect insect stylet probing, by comparing Lygus lineolaris behaviors on pin-head cotton squares using an AC-DC electropenetrograph. Effects of AC or DC applied signals were separately examined in two factorial studies, each comparing four input resistor (Ri) levels (106, 107, 108 and 109 Ω) and four applied voltage levels (2, 60, 150 and 250 mV). Results showed that changes in both probing and non-probing behaviors were indeed caused by changing signal type, Ri level, or applied voltage. Negative effects on feeding were numerically greater overall for DC than AC applied signals, perhaps due to muscular tetany from DC; however, AC versus DC could not be statistically tested. Results strongly support the need for flexible Ri and applied voltage levels and types, to tailor instrument settings to the size and special needs of each insect subject. Our findings will facilitate further EPG studies of Lygus spp., such as host plant resistance or insecticidal assays/bioassays to assess mode of action and appropriate dosage. It is hoped that this study will also inform EPG studies of similar, large heteropterans in the future.

Correlation of Electropenetrography Waveforms From Lygus lineolaris (Hemiptera: Miridae) Feeding on Cotton Squares With Chemical Evidence of Inducible Tannins.

Probing behavior of Lygus lineolaris (Palisot de Beauvois) has previously been characterized with electropenetrography (EPG). Cell rupturing (CR) and ingestion (I) EPG waveforms were identified as the two main stylet-probing behaviors by adult L. lineolaris. However, characterization and identification of EPG waveforms are not complete until specific events of a particular waveform are correlated to insect probing. With the use of EPG, histology, microscopy, and chemical analysis, probing behavior of L. lineolaris on pin-head cotton squares was studied. Occurrences of waveforms CR and I were artificially terminated during the EPG recording. Histological samples of probed cotton squares were prepared and analyzed to correlate specific types and occurrences of feeding damage location and plant responses to insect feeding. Both CR and I occurred in the staminal column of the cotton square. Cell rupturing events elicited the production of dark-red deposits seen in histological staining that were demonstrated via chemical analysis to contain condensed tannins. We hypothesize that wounding and saliva secreted during CR triggered release of tannins, because tannin production was positively correlated with the number of probes with single CR events performed by L. lineolaris. Degraded plant tissue and tannins were removed from the staminal column during occurrence of waveform I. These results conclude the process of defining CR and I as probing waveforms performed by L. lineolaris on pin-head cotton squares. These biological definitions will now allow EPG to be used to quantitatively compare L. lineolaris feeding among different plant treatments, with the goal of improving pest management tactics against this pest.

Evaluation of SmartStax and SmartStax PRO maize against western corn rootworm and northern corn rootworm: efficacy and resistance management.

BACKGROUND: Cases of western corn rootworm (WCR) field-evolved resistance to Cry3Bb1 and other corn rootworm (CRW) control traits have been reported. Pyramid products expressing multiple CRW traits can delay resistance compared to single trait products. We used field studies to assess the pyramid CRW corn products, SmartStax (expressing Cry3Bb1 and Cry34Ab1/Cry35Ab1) and SmartStax PRO (expressing Cry3Bb1, Cry34Ab1/Cry35Ab1 and DvSnf7), at locations with high WCR densities and possible Cry3Bb1 resistance, and to assess the reduction in adult emergence attributable to DvSnf7 and other traits. Insect resistance models were used to assess durability of SmartStax and SmartStax PRO to WCR resistance. RESULTS: SmartStax significantly reduced root injury compared to non-CRW-trait controls at all but one location with measurable WCR pressure, while SmartStax PRO significantly reduced root injury at all locations, despite evidence of Cry3Bb1 resistance at some locations. The advantage of SmartStax PRO over SmartStax in reducing root damage was positively correlated with root damage on non-CRW-trait controls. DvSnf7 was estimated to reduce WCR emergence by approximately 80-95%, which modeling indicated will improve durability of Cry3Bb1 and Cry34Ab1/Cry35Ab1 compared to SmartStax. CONCLUSION: The addition of DvSnf7 in SmartStax PRO can reduce root damage under high WCR densities and prolong Cry3Bb1 and Cry34Ab1/Cry35Ab1 durability. © 2017 Society of Chemical Industry.

Characterization of the Activity Spectrum of MON 88702 and the Plant-Incorporated Protectant Cry51Aa2.834_16.

The spectrum of insecticidal activity of Cry51Aa2.834_16 protein targeting hemipteran and thysanopteran insect pests in cotton was characterized by selecting and screening multiple pest and non-pest species, based on representation of ecological functional groups, taxonomic relatedness (e.g. relationship to species where activity was observed), and availability for effective testing. Seven invertebrate orders, comprising 12 families and 17 representative species were screened for susceptibility to Cry51Aa2.834_16 protein and/or the ability of the protein to protect against feeding damage in laboratory, controlled environments (e.g. greenhouse/growth chamber), and/or field studies when present in cotton plants. The screening results presented for Cry51Aa2.834_16 demonstrate selective and limited activity within three insect orders. Other than Orius insidiosus, no activity was observed for Cry51Aa2.834_16 against several groups of arthropods that perform key ecological roles in some agricultural ecosystems (e.g. pollinators, decomposers, and natural enemies).

A transgenic approach for controlling Lygus in cotton.

Lygus species of plant-feeding insects have emerged as economically important pests of cotton in the United States. These species are not controlled by commercial Bacillus thuringiensis (Bt) cotton varieties resulting in economic losses and increased application of insecticide. Previously, a Bt crystal protein (Cry51Aa2) was reported with insecticidal activity against Lygus spp. However, transgenic cotton plants expressing this protein did not exhibit effective protection from Lygus feeding damage. Here we employ various optimization strategies, informed in part by protein crystallography and modelling, to identify limited amino-acid substitutions in Cry51Aa2 that increase insecticidal activity towards Lygus spp. by >200-fold. Transgenic cotton expressing the variant protein, Cry51Aa2.834_16, reduce populations of Lygus spp. up to 30-fold in whole-plant caged field trials. One transgenic event, designated MON88702, has been selected for further development of cotton varieties that could potentially reduce or eliminate insecticide application for control of Lygus and the associated environmental impacts.

Identification, inheritance, and fitness costs of Cry2Ab2 resistance in a field-derived population of sugarcane borer, Diatraea saccharalis (F.) (Lepidoptera: Crambidae).

The sugarcane borer, Diatraea saccharalis (F.), is one of the major target pests of transgenic maize, Zea mays, expressing Bacillus thuringiensis (Bt) proteins in South America and mid-southern region of the U.S. The MON89034 maize expresses Cry2Ab2 and Cry1A.105 Bt proteins and it was developed to provide better control of key lepidopteran pests of maize including D. saccharalis. The objectives of this study were to select and characterize the resistance of D. saccharalis to Cry2Ab2 using a non-commercial Cry2Ab2 single gene Bt maize line. A Cry2Ab2-resistant strain (Cry2Ab2-RR) of D. saccharalis was established from 28 two-parent families collected from fields in northeast Louisiana, U.S. The Cry2Ab2-RR showed a high level of resistance to Cry2Ab2 in both diet-incorporated and whole maize plant bioassays. The Cry2Ab2 resistance in D. saccharalis was likely inherited as a single or a few tightly linked autosomal genes. The resistance was non-recessive and not associated with fitness costs. The results should provide valuable information in resistance monitoring, assessing resistance risk, and developing effective management strategies for the sustainable use of Bt maize technology for managing maize stalk borers.

Genetic markers for western corn rootworm resistance to Bt toxin.

Western corn rootworm (WCR) is a major maize (Zea mays L.) pest leading to annual economic losses of more than 1 billion dollars in the United States. Transgenic maize expressing insecticidal toxins derived from the bacterium Bacillus thuringiensis (Bt) are widely used for the management of WCR. However, cultivation of Bt-expressing maize places intense selection pressure on pest populations to evolve resistance. Instances of resistance to Bt toxins have been reported in WCR. Developing genetic markers for resistance will help in characterizing the extent of existing issues, predicting where future field failures may occur, improving insect resistance management strategies, and in designing and sustainably implementing forthcoming WCR control products. Here, we discover and validate genetic markers in WCR that are associated with resistance to the Cry3Bb1 Bt toxin. A field-derived WCR population known to be resistant to the Cry3Bb1 Bt toxin was used to generate a genetic map and to identify a genomic region associated with Cry3Bb1 resistance. Our results indicate that resistance is inherited in a nearly recessive manner and associated with a single autosomal linkage group. Markers tightly linked with resistance were validated using WCR populations collected from Cry3Bb1 maize fields showing significant WCR damage from across the US Corn Belt. Two markers were found to be correlated with both diet (R2 = 0.14) and plant (R2 = 0.23) bioassays for resistance. These results will assist in assessing resistance risk for different WCR populations, and can be used to improve insect resistance management strategies.

Carbon isotope ratios document that the elytra of western corn rootworm (Coleoptera: Chrysomelidae) reflects adult versus larval feeding and later instar larvae prefer Bt corn to alternate hosts.

In much of the Corn Belt and parts of Europe, the western corn rootworm, Diabrotica virgifera virgifera LeConte, is the most important insect pest of maize. The need for additional basic knowledge of this pest has been highlighted while developing resistance management plans for insecticidal genetically modified crops. This study evaluated the possibility of tracking feeding habits of western corn rootworm larvae using stable carbon isotope signatures. Plants accumulate different ratios of (13)C:(12)C isotopes, usually expressed as δ(13)C, according to whether they use the C3 or C4 photosynthetic pathway. Herbivore biomass is expected to reflect the δ(13)C of the food they eat. For the current experiment, western corn rootworm larvae were grown on different species of plants exhibiting different δ(13)C values. The δ(13)C values were then measured in elytra of emerged beetles. When beetles were unfed, biomass reflected larval feeding. When beetles were fed for 31 d postemergence, δ(13)C values of elytra almost exclusively reflected adult feeding. These results suggest the use of caution in the interpretation of δ(13)C data aiming to document larval diet history when adult feeding history is unknown. The technique was also used to evaluate western corn rootworm larval choice between alternate hosts and maize with and without genetically modified (Bt) traits aimed at their control. Propensity for feeding on alternate hosts versus maize was biased toward feeding on maize regardless whether the maize had Bt or not, suggesting western corn rootworm larvae were not repelled by Bt. These data will be helpful for regulators in interpreting western corn rootworm feeding data on Bt maize.

Cotton plants expressing a hemipteran-active Bacillus thuringiensis crystal protein impact the development and survival of Lygus hesperus (Hemiptera: Miridae) nymphs.

The plant bugs Lygus hesperus Knight (Hemiptera: Miridae) and L. lineolaris (Palisot de Beauvois) have emerged as economic pests of cotton in the United States. These hemipteran species are refractory to the insect control traits found in genetically modified commercial varieties of cotton. In this article, we report the isolation and characterization of a 35 kDa crystal protein from Bacillus thuringiensis, designated TIC807, which causes reduced mass gain and mortality of L. hesperus and L. lineolaris nymphs when presented in an artificial diet feeding assay. Cotton plants expressing the TIC807 protein were observed to impact the survival and development of L. hesperus nymphs in a concentration-dependent manner. These results, demonstrating in planta activity of a Lygus insecticidal protein, represent an important milestone in the development of cotton varieties protected from Lygus feeding damage.

Population growth of soybean aphid, Aphis glycines, under varying levels of predator exclusion.

Although soybean aphid, Aphis glycines Matsumura (Hemiptera: Aphididae), has caused economic damage in several Midwestern states, growers in Missouri have experienced relatively minor damage. To evaluate whether existing predatory insect populations are capable of suppressing or preventing soybean aphid population growth or establishment in Missouri, a predator exclusion study was conducted to gauge the efficacy of predator populations. Three levels of predator exclusion were used; one that excluded all insects (small mesh), one that excluded insects larger than thrips (medium mesh), and one that excluded insects larger than Orius insidiosus (Say) (Hemiptera: Anthocoridae), a principal predator (large mesh). Along with manipulating predator exposure, timing of aphid arrival (infestation) was manipulated. Three infestation times were studied; vegetative (V5), beginning bloom (R1), and beginning pod set (R3). Timing of aphid and predator arrival in a soybean field may affect the soybean aphid's ability to establish and begin reproducing. Cages infested at V5 and with complete predator exclusion reached economic threshold within two weeks, while cages with predators reached economic threshold in four and a half weeks. Cages infested at R1 with complete predator exclusion reached economic threshold within five weeks; cages with predators reached economic threshold within six weeks. Cages infested at R3 never reached threshold (with or without predators). The predator population in Missouri seems robust, capable of depressing the growth of soybean aphid populations once established, and even preventing establishment when the aphid arrived late in the field.

Microbial symbionts in insects influence down-regulation of defense genes in maize.

Diabrotica virgifera virgifera larvae are root-feeding insects and significant pests to maize in North America and Europe. Little is known regarding how plants respond to insect attack of roots, thus complicating the selection for plant defense targets. Diabrotica virgifera virgifera is the most successful species in its genus and is the only Diabrotica beetle harboring an almost species-wide Wolbachia infection. Diabrotica virgifera virgifera are infected with Wolbachia and the typical gut flora found in soil-living, phytophagous insects. Diabrotica virgifera virgifera larvae cannot be reared aseptically and thus, it is not possible to observe the response of maize to effects of insect gut flora or other transient microbes. Because Wolbachia are heritable, it is possible to investigate whether Wolbachia infection affects the regulation of maize defenses. To answer if the success of Diabrotica virgifera virgifera is the result of microbial infection, Diabrotica virgifera virgifera were treated with antibiotics to eliminate Wolbachia and a microarray experiment was performed. Direct comparisons made between the response of maize root tissue to the feeding of antibiotic treated and untreated Diabrotica virgifera virgifera show down-regulation of plant defenses in the untreated insects compared to the antibiotic treated and control treatments. Results were confirmed via QRT-PCR. Biological and behavioral assays indicate that microbes have integrated into Diabrotica virgifera virgifera physiology without inducing negative effects and that antibiotic treatment did not affect the behavior or biology of the insect. The expression data and suggest that the pressure of microbes, which are most likely Wolbachia, mediate the down-regulation of many maize defenses via their insect hosts. This is the first report of a potential link between a microbial symbiont of an insect and a silencing effect in the insect host plant. This is also the first expression profile for a plant attacked by a root-feeding insect.

Mortality of western corn rootworm larvae on MIR604 transgenic maize roots: field survivorship has no significant impact on survivorship of F1 progeny on MIR604.

Mortality of western corn rootworm, Diabrotica virgifera virgifera LeConte, larvae due to MIR604 transgenic corn, Zea mays L., expressing the modified Cry3A (mCry3A) protein relative to survivorship on corn with the same genetic background without the gene (isoline corn) was evaluated at three Missouri sites in both 2005 and 2006. We made these comparisons by using wild-type western corn rootworm at three different egg densities (6,000, 3,000, and 1,500 eggs per m) so that the role of density-dependent mortality would be known. The mortality due to the mCry3A protein was 94.88% when averaged across all environments and both years. Fifty percent emergence of beetles was delayed approximately 5.5 d. Beetles were kept alive and their progeny evaluated on MIR604 and isoline corn in the greenhouse to determine whether survivorship on MIR604 in the field for one generation increased survivorship on MIR604 in the greenhouse in the subsequent generation. There was no significant difference in survivorship on MIR604 in greenhouse assays between larvae whose parents survived isoline and larvae whose parents survived MIR604 in the field the previous generation, indicating that many susceptible beetles survived MIR604 in the field the previous season along with any potentially resistant beetles. The data are discussed in terms of rootworm insect resistance management.

Phylogenetic analysis of an economically important species complex of wireworms (Coleoptera: Elateridae) in the midwest.

Wireworms are a common soil-dwelling pest of maize, Zea mays L., in the midwestern United States. Wireworms are a problematic group to control and study due to the difficulty involved in identification. The objectives of this study are to identify this species complex of wireworms by using molecular diagnostic techniques and to reconstruct a phylogeny of economically important wireworm species. The cytochrome oxidase I gene of mitochondrial DNA was sequenced from > 300 individuals. The species analyzed include all economically important members of the genus Melanotus Eschscholtz as well as Conoderus lividus (De Geer). The species that are indistinguishable in the larval stage were successfully separated using nucleotide p-distances, and sequence data were then used in phylogenetic analyses. The data presented here represent an initial phylogenetic hypothesis concerning economically important wireworms. Our results indicate that the molecular phylogeny of the mitochondrial cytochrome oxidase subunit I gene provides a fast and accurate method of separating wireworm species. By increasing the ease and accuracy of identification, we hope to facilitate further investigations into their biology and control.

Prairie grasses as hosts of the northern corn rootworm (Coleoptera: Chrysomelidae).

We evaluated 27 prairie grass species thought to be among those dominant 200 yr ago in the northern midwest as larval hosts of the northern corn rootworm, Diabrotica barberi Smith and Lawrence. Maize (Zea mays L.), spring wheat (Triticum aestivum L.), and grain sorghum (Sorghum bicolor L.) were included as controls for a total of 30 species. Twenty pots of each species were planted in a randomized complete block design. Each pot was infested 5 wk later with 20 neonate northern corn rootworm larvae. Two pots within each species and block were assigned an extraction date of 7 or 14 d after infestation. The remaining two pots from each block were used to monitor adult emergence. The percentage of larvae recovered, change in larval head capsule width, and change in average dry weights varied significantly among the grass species. The highest percentage of larvae was recovered from slender wheatgrass, Elymus trachycaulus (Link), and this was significantly greater than the percentage recovered from all other species including maize for the 14-d sample date. Several additional species were also relatively good hosts, in that the percentage of larvae recovered from these species was not significantly different from maize. The average dry weight of larvae recovered was significantly greater for larvae recovered from maize than for larvae recovered from all other species except slender wheatgrass, when the two samples dates were combined. Overall, adults were produced from only 6 of the 28 species evaluated, and no analysis was performed because of the low numbers. The results of this study are discussed in relation to the potential of alternate hosts of northern corn rootworm to serve as a bridge to survival on transgenic maize.

Interactions of alternate hosts, postemergence grass control, and rootworm-resistant transgenic corn on western corn rootworm (Coleoptera: Chrysomelidae) damage and adult emergence.

Field studies were conducted in 2003 and 2004 to determine the effects of grassy weeds, postemergence grass control, transgenic rootworm-resistant corn, Zea mays L., expressing the Cry3Bb1 endotoxin and glyphosate herbicide tolerance (Bt corn), and the interactions of these factors on western corn rootworm, Diabrotica virgifera virgifera LeConte, damage and adult emergence. Three insect management tactics (Bt corn, its nontransgenic isoline, and isoline plus tefluthrin) were evaluated with two weed species (giant foxtail, Setaria faberi Herrm, and large crabgrass, Digitaria sanquinalis L. Scop), and four weed management regimes (weed free, no weed management, early [V3-4] weed management and late [V5-6] weed management) in a factorial arrangement of a randomized split split-plot design. In each case, the isoline was also tolerant to glyphosate. Root damage was significantly affected by insect management tactics in both years, but weed species and weed management did not significantly affect damage to Bt corn in either year. Adult emergence was significantly affected by insect management tactics in both years and by weed species in 2003, but weed management and the interaction of all three factors was not significant in either year. The sex ratio of female beetles produced on Bt corn without weeds was generally greater than when weeds were present and this difference was significant for several treatments each year. Average dry weight of male and female beetles emerging from Bt corn was greater than the weights of beetles emerging from isoline or isoline plus tefluthrin in 2003, but there was no difference for females in 2004 and males weighed significantly less than other treatments in 2004. The implications of these results in insect resistance management are discussed.

Western corn rootworm (Coleoptera: Chrysomelidae) beetle emergence from weedy Cry3Bbl rootworm-resistant transgenic corn.

Three greenhouse experiments were conducted to evaluate western corn rootworm, Diabrotica virgifera virgifera LeConte, beetle emergence from individual pots containing glyphosate-tolerant transgenic corn, Zea mays L., expressing the Cry3Bbl endotoxin from the soil bacterium Bacillus thuringiensis Berliner (MON863), nontransgenic glyphosate-tolerant isoline corn, grassy weeds (giant foxtail, Setariafaberi R.A.W. Herrm; and large crabgrass, Digitaria sanguinalis (L.) Scop.), and combinations thereof infested with 40 neonate larvae. In the first two experiments, pots with corn and weed combinations were treated with glyphosate 5 d after larval infestation to kill the weeds. The third experiment was similar to the first two except that untreated corn-weed combinations were added. In all three experiments beetle emergence varied significantly. Beetle recovery generally did not vary between the nontransgenic, nontransgenic + weeds, and MON863 + weeds. Significantly more beetles were recovered from MON863 + weeds than MON863 alone or weeds alone. Beetle emergence from MON863 + weeds was likely enhanced by larvae that initially survived on weeds before application of glyphosate. Preliminary data indicated that fecundity was highest from beetles reared on nontransgenic isoline corn and fewer eggs were laid by beetles reared on MON863 alone. Egg viability was generally lowest from beetles reared on MON863. The implications of these results in relation to insect resistant management are discussed.