Preference of Bt‐resistant and susceptible Busseola fusca moths and larvae for Bt and non‐Bt maize

The sustainability of genetically engineered insecticidal Bacillus thuringiensis Berliner (Bt) maize, Zea mays L. (Poaceae), is threatened by the evolution of resistance by target pest species. Several Lepidoptera species have evolved resistance to Cry proteins expressed by Bt maize over the last decade, including the African maize stem borer, Busseola fusca (Fuller) (Lepidoptera: Noctuidae). The insect resistance management (IRM) strategy (i.e., the high‐dose/refuge strategy) deployed to delay resistance evolution is grounded on certain assumptions about the biology and ecology of a pest species, for example, the interactions between the insect pest and crop plants. Should these assumptions be violated, the evolution of resistance within pest populations will be rapid. This study evaluated the assumption that B. fusca adults and larvae select and colonize maize plants at random, and do not show any preference for either Bt or non‐Bt maize. Gravid female B. fusca moths of a resistant and susceptible population were subjected to two‐choice oviposition preference tests using stems of Bt and non‐Bt maize plants. Both the number of egg batches as well as the total number of eggs laid on each stem were recorded. The feeding preference of Bt‐resistant and susceptible neonate B. fusca larvae were evaluated in choice test bioassays with whorl leaf samples of specific maize cultivars. Although no differential oviposition preference was observed for either resistant or susceptible female moths, leaf damage ratings indicated that neonate larvae were able to detect Bt toxins and that they displayed feeding avoidance behaviour on Bt maize leaf samples.     

Dispersal capacity in the Mediterranean corn borer, Sesamia nonagrioides

Corn (Zea mays L.) borers are the primary target of Bacillus thuringiensis Berliner (Bt) transgenic maize. Management of corn borer resistance to Bt requires information on larval and adult dispersal capacities, a feature that is particularly unknown in Sesamia nonagrioides Lefèbvre (Lepidoptera: Noctuidae), the most damaging corn borer in Spain. Larval dispersal was studied over a 3 year period by infesting plants with egg masses and dissecting the neighbouring plants 7, 14, and 32 days later to measure larval dispersal at several ages. The number and age of larvae were recorded in the dissected plants. Only mature larvae dispersed in significant numbers; they moved at least to rows adjacent to those containing the infested plant, and down the row five plants. The percentage of larvae that dispersed from the infested plant was density‐dependent. Adult dispersal was studied with directional light and pheromone uni‐traps over 5 and 3 year periods, respectively. Directional light traps were placed in the margins between Bt and non‐Bt maize fields, half oriented towards each of the two kinds of maize field. Pheromone traps were placed in the Bt and non‐Bt fields at increasing distances (0–100 m) from the border. The numbers of males and females caught in directional light traps were not different in traps oriented towards Bt or non‐Bt fields, but the number of males caught in the third flight in Bt fields was lower than in non‐Bt fields. These results suggest that males from adjacent Bt and non‐Bt fields mate indiscriminately with females emerging in any of the two kinds of maize fields. However, male movement in the third flight may not be sufficient to randomly distribute males between the two fields.     

Oviposition and feeding avoidance in Helicoverpa armigera (Hübner) against transgenic Bt cotton

Helicoverpa armigera (Hübner), the major target pest of transgenic Bacillus thuringiensis (Bt) cotton, remains susceptible to Bt cotton in China at present. Behavioural avoidance by ovipositing females might lead to reduced exposure to Bt cotton and minimize selection for physiological resistance. We examined the behavioural responses of H. armigera to Bt and non‐Bt cottons to determine whether behavioural avoidance to Bt cotton may be present. In oviposition choice tests, the number of eggs on non‐Bt cotton plants was significantly higher than on Bt cotton plants. Similarly, in no‐choice tests, Bt cotton plants attracted significantly fewer eggs compared with non‐Bt cotton plants. H. armigera neonates showed higher dispersal and lower establishment on Bt cotton than on non‐Bt cotton. First instars were found to feed consistently on non‐Bt cotton leaves, creating large feeding holes, but only produced tiny feeding holes on Bt cotton leaves. The H. armigera population used in this study showed avoidance of oviposition and feeding on Bt cotton. Our results provide important insights into one possible mechanism underlying the durability of Bt cotton resistance and may be useful for improving strategies to sustain the effectiveness of Bt crops.     

Effects of Bt maize on Frankliniella tenuicornis and exposure of thrips predators to prey-mediated Bt toxin

As a part of a risk assessment procedure, the impact of Bt maize expressing Cry1Ab toxin on the thrips Frankliniella tenuicornis (Uzel) (Thysanoptera: Thripidae) was investigated, and the potential risks for predators feeding on thrips on Bt maize were evaluated. The effects of Bt maize on F. tenuicornis were assessed by measuring life‐table parameters when reared on Bt and non‐Bt maize. The content of Cry1Ab toxin in different stages of F. tenuicornis reared on Bt maize and the persistence of the toxin in adults where determined in order to evaluate the possible exposure of predators when feeding on thrips. In addition, Chrysoperla carnea (Stephens) (Neuroptera: Chrysopidae) was used as a model predator to assess how the behaviour of prey and predator may influence the exposure of a natural enemy to the Bt toxin. Life‐table parameter results showed that F. tenuicornis was not affected when it was reared on Bt maize. This indicates that the potential for prey quality‐mediated effects on predators is low. Bt content was highest in thrips larvae and adults, and negligible in the non‐feeding prepupal and pupal stages. The persistence of the Cry1Ab toxin in adult F. tenuicornis was short, resulting in a decrease of 97% within the first 24 h. Predation success by young C. carnea larvae varied among the thrips stages, indicating that exposure of predators to Bt toxin can additionally depend on the prey stage. When combining the current knowledge of the susceptibility of major thrips predators with our findings showing no potential for prey quality‐mediated effects, relatively low toxin content in thrips as well as short persistence, it can be concluded that the risks for predators when feeding on thrips in or next to Bt maize fields are negligible.     

The impact of Bt maize as a natal host on adult head capsule width in field populations of western corn rootworm

The success of the current resistance management plan for transgenic maize, Zea mays L. (Poaceae), targeting the rootworm complex hinges upon high rates of mating between resistant and susceptible beetles. However, differences in the fitness of adult beetles could result in assortative mating, which could, in turn, change the rate of resistance evolution. Adult head capsule widths of naturally occurring populations of western corn rootworm, Diabrotica virgifera virgifera LeConte (Coleoptera: Chrysomelidae), were examined in a variety of refuge configurations. Beetles were classified into treatments based on the hybrid – non‐Bt refuge or Bt maize targeting larval rootworms (hereafter Bt‐RW maize) – and location – proximity to other Bt‐RW or refuge plants – of the natal host plant. Treatments included the following: a refuge plant surrounded by other refuge plants, a refuge plant located near a Bt‐RW plant, a Bt‐RW plant surrounded by Bt‐RW plants, and a Bt‐RW plant located near a refuge plant. The mean head capsule width of males emerging from Bt‐RW plants was significantly smaller than the mean head capsule width of males emerging from refuge plants. These results indicate that males emerging from Bt‐RW maize plants may be exposed to sublethal doses of the Bt toxin as larvae. No differences were detected between females emerging from refuge plants compared with Bt‐RW plants. Overall mean head capsule width decreased as the season progressed, regardless of treatment. The diminished head capsule width of western corn rootworm males emerging from Bt‐RW maize may act to enhance resistance management, particularly in a seed mix refuge system.     

Growth and development of Bacillus thuringiensis Cry1Ab-susceptible and Cry1Ab-resistant sugarcane borer on diet and conventional maize plants

The sugarcane borer, Diatraea saccharalis (F.) (Lepidoptera: Crambidae), is a dominant maize borer pest and a major target of Bacillus thuringiensis (Bt)‐maize in Louisiana and the Gulf Coast area of Texas (USA). Growth and development of D. saccharalis on non‐toxic diet, diet treated with three low concentrations (0.01, 0.05, and 0.1 μg g^?1) of Cry1Ab toxin, and on non‐Bt maize plants were compared for five insect genotypes: a Bt‐susceptible strain (BT‐SS), a Cry1Ab‐resistant strain (BT‐RR), a back‐crossed and re‐selected resistant strain (BT‐R’R’), and two F₁ progeny of the BT‐SS and BT‐R’R’ strains. Fitness of the five genotypes was examined by infesting neonates on diet with/without Cry1Ab toxin in the laboratory and on intact non‐Bt maize plants in the greenhouse. Biological parameters measured were neonate‐to‐pupa development time and pupation rate, larval survival, larval and pupal weight, and sex ratio. Larvae of BT‐SS and BT‐R’R’ on non‐toxic diet and non‐Bt maize plants grew normally and there were no significant differences between the two strains in all measured parameters, suggesting a lack‐of‐fitness cost of the Cry1Ab resistance in D. saccharalis. Except for the development time on non‐Bt diet, all other parameters on both non‐Bt diet and non‐Bt maize plants were similar among the five genotypes. Larval development of BT‐SS was significantly affected on diet treated with Cry1Ab toxin at 0.05 and 0.1 μg g^?1, whereas the effect to BT‐RR and BT‐R’R’ was not significant. Pupal weight and sex ratio reared on Cry1Ab‐diet were similar and there were no significant differences among the five genotypes. Neonate‐to‐pupation rate decreased as Cry1Ab concentrations increased but the decrease was more significant for BT‐SS than for the other four genotypes. The lack‐of‐fitness costs of Bt resistance in D. saccharalis imply a greater challenge in managing Bt resistance for this maize borer species.     

Fitness costs associated with Cry1F resistance in the European corn borer

Crops producing insecticidal toxins derived from the bacterium Bacillus thuringiensis (Bt) are widely planted to manage insect pests. Bt crops can provide an effective tool for pest management; however, the evolution of Bt resistance can diminish this benefit. The European corn borer, Ostrinia nubilalis Hübner, is a significant pest of maize and is widely managed with Bt maize in the Midwest of the United States. When Bt crops are grown in conjunction with non‐Bt refuges, fitness costs of Bt resistance can delay the evolution of resistance. Importantly, fitness costs often vary with ecological factors, including host‐plant genotype and diapause. In this study, we examined fitness costs associated with Cry1F resistance in O. nubilalis when insects were reared on three maize lines. Fitness costs were tested in two experiments. One experiment assessed the fitness costs when Cry1F‐resistant and Cry1F‐susceptible insects were reared on plants as larvae and experienced diapause. The second experiment tested resistant, susceptible and F1 heterozygotes that were reared on plants but did not experience diapause. Despite some evidence of greater adult longevity for Cry1F‐resistant insects, these insects produced fewer fertile eggs than Cry1F‐susceptible insects, and this occurred independent of diapause. Reduced fecundity was not detected among heterozygous individuals, which indicated that this fitness cost was recessive. Additionally, maize lines did not affect the magnitude of this fitness cost. The lower fitness of Cry1F‐resistant O. nubilalis may contribute to the maintenance of Cry1F susceptibility in field populations more than a decade after Cry1F maize was commercialized.     

Stay or move: how Bt‐susceptible Helicoverpa armigera neonates behave on Bt cotton plants

Helicoverpa armigera (Hübner) (Lepidoptera: Noctuidae) larvae occasionally have been reported to survive at management threshold levels in fields of Bollgard II^® cotton, Gossypium hirsutum L. (Malvaceae). The pattern and degree of larval survival is not easily predicted but depends on the ability of first instars to establish on host plants. Experiments were conducted with Bacillus thuringiensis Berliner (Bt)‐susceptible and Bt‐resistant larvae of H. armigera to understand how physiologically Bt‐susceptible H. armigera survive on Bt cotton plants, and examine how their first meal influences survival rates. In assays using cotton plant parts, both strains of larvae displayed similar tendencies to drop‐off specific plant parts of Bt and non‐Bt cotton. However, significantly more Bt‐susceptible larvae dropped off young leaves, mature leaves, and squares of Bt cotton compared to non‐Bt cotton plants. Egg cannibalism significantly improved the survival of Bt‐susceptible H. armigera larvae on Bt cotton plants. Larvae were more likely to eat live aged eggs, than newly laid or dead eggs. Survival significantly improved when larvae cannibalized eggs before feeding on Bt leaves. The behavior of Bt‐susceptible larvae with respect to drop‐off and egg cannibalism may help enhance their survival on Bt cotton plants.     

Six Years after the Commercial Introduction of Bt Maize in Spain: Field Evaluation, Impact and Future Prospects

We carried out a 6-year-field evaluation to assess potential hazards of growing Compa, a transgenic Bt maize variety based on the transformation event CG 00256-176. Two categories of hazards were investigated: the potential of the target corn borer Sesamia nonagrioides to evolve resistance to Bt maize and effects on non-target organisms. In order to address the first hazard, dispersal capacity of the corn borer was measured and our results indicated that larvae move to plants other than those onto which the female oviposited - even to plants in adjacent rows - in remarkable numbers and they do so mostly at a mature age, suggesting that mixing Bt and non-Bt seeds in the same field would not be a very useful deployment strategy to delay/prevent resistance. In addition, adults move among fields to mate and males may do so for up to 400 m. Three different aspects of potential non-target effects were investigated: sub-lethal effects on the target S. nonagrioides, effects on non-target maize pests, and effects on maize-dwelling predators. Larvae collected in Bt fields at later growth stages, in which event 176 Bt maize expresses Bt toxin at sub-lethal concentrations, had longer diapause and post-diapause development than larvae collected in non-Bt fields, a feature that might lead to a certain isolation between populations in both type of fields and accelerate Bt resistance evolution. Transgenic maize did not have a negative impact on non-target pests in the field; more aphids and leafhoppers but similar numbers of cutworms and wireworms were counted in Bt versus non-Bt fields; in any case differences in damage or yield were recorded. We observed no difference in the numbers of the most relevant predators in fields containing transgenic or no transgenic maize.     

The effect of Bt maize on Sesamia calamistis in South Africa

Bt maize, Zea mays L. (Poaceae) expressing Cry 1Ab insecticidal proteins was introduced for control of Busseola fusca (Fuller) (Lepidoptera: Noctuidae) and Chilo partellus (Swinhoe) (Lepidoptera: Crambidae) in South Africa after its development for control of crambid borers in North America. In the light of the reportedly lower toxicity of Bt maize to certain Noctuidae borers, the effect of Bt maize was evaluated on Sesamia calamistis (Hampson) (Lepidoptera: Noctuidae). The characteristic larval behaviour of S. calamistis may result in reduced exposure to Bt toxin and subsequent high levels of survival. Larvae do not feed on plant whorls like other borer species but penetrate stems directly from behind leaf sheaths where eggs are laid. Greenhouse and laboratory bioassays were done with three Bt maize hybrids and their iso‐hybrids. ‘Whole plant methods’ were used and potted plants artificially infested with eggs or larvae and survival recorded over time. Larval survival was also determined on different plant parts (whorls, stems, tillers, and ears) over time. Bt maize was shown to be highly toxic to S. calamistis. No larvae survived longer than 12–18 days on Bt maize plants in any of the experiments. Adults did not differentiate between Bt and non‐Bt plants in oviposition choice experiments. Sesamia calamistis is polyphagous and occurs in mixed populations with other borer species with which it shares many parasitoid species in Africa. The ecological impact of local extinction of S. calamistis caused by this highly effective transgenic event is therefore not expected to be great.     

Effect of water-deficit stress on cotton plants expressing the Bacillus thuringiensis toxin

Bacillus thuringiensis (Bt) crops require a high dosage of Bt toxin to delay development of insect resistance, in particular, when the refuge strategy is applied. This strategy is threatened by plant developmental and environmental factors that might reduce Bt toxin concentration and Bt efficacy in Bt crops. Growth of Bt (Cry1Ac) cotton under prolonged, moderate water deficit as a single stress factor was evaluated. Bt cotton plants were analysed for physiological performance, Bt toxin concentration and Bt efficacy. For performance analysis, leaf and total plant dry weight and leaf area were measured. Bt toxin concentration was determined by an immuno‐assay. Effects of Bt toxin on growth and mortality of African cotton bollworm, Helicoverpa armigera, larvae were measured in different plant organs. Leaves from young plants exposed for 30 days to moderate water deficit had both higher Bt toxin concentrations and were more effective against larvae than leaves, flowers or bolls from mature flowering plants exposed to 60 days of moderate water deficit. Although growth of Bt cotton plants under moderate water‐deficit conditions decreased Bt concentrations in leaves, flowers and bolls, this had no effect on efficacy against first‐instar cotton bollworm larvae. No significant evidence was found that moderate water deficit, as a single stress factor, decreases Bt efficacy in Bt cotton.     

Assessment of fitness costs in Cry3Bb1‐resistant and susceptible western corn rootworm (Coleoptera: Chrysomelidae) laboratory colonies

Abstract Maize production in the United States is dominated by plants genetically modified with transgenes from Bacillus thuringiensis (Bt). Cry3Bb delta endotoxins expressed by Bt maize specifically target corn rootworms (genus Diabrotica) and have proven highly efficacious. However, development of resistance to Bt maize, especially among western corn rootworm (Diabrotica virgifera virgifera) populations, poses a significant threat to the future viability of this pest control biotechnology. The structured refuge insect resistance management (IRM) strategy implemented in the United States for Bt maize adopts a conservative approach to managing resistance by assuming no fitness costs of Bt resistance, even though these trade‐offs strongly influence the dynamics of Bt resistance within numerous agricultural pest species. To investigate the effects of Bt resistance on fitness components of western corn rootworm, we compared survivorship, fecundity and viability of five Bt‐resistant laboratory lines reared on MON863 (YieldGard Rootworm), a Bt maize product that expresses Cry3Bb1 delta endotoxin, and on its non‐transgenic isoline. Analysis of performance on the isoline maize demonstrated no fitness costs associated with Bt resistance. In fact, resistant lines emerged approximately 2–3 days earlier than control lines when reared on both MON863 and the isoline, indicating that selection for Bt resistance resulted in a general increase in the rate of larval development. In addition, resistant lines reared on Bt maize displayed higher fecundity than those reared on the isoline, which may have significant management implications. These data will be valuable for formulating improved IRM strategies for a principal agricultural pest of maize.     

Feeding behaviour of Helicoverpa armigera larvae on insect-resistant transgenic cotton and non-transgenic cotton

Abstract: Feeding behaviour of Helicoverpa armigera Hübner (Lep.; Noctuidae) larvae on non‐transgenic Bacillus thuringiensis (Bt) cotton (Gossypium hirsutum L.), Zhong 30, and transgenic cowpea trypsin inhibitor (CpTI)‐Bt cotton, SGK 321, and non‐transgenic cotton, Shiyuan 321, was investigated in both choice tests and no‐choice tests. The results of choice tests suggested that neonates have the ability to detect and avoid transgenic cotton. In the choice tests of neonates with both transgenic and non‐transgenic cotton leaves, a significantly greater proportion of larvae and higher consumption were observed on non‐transgenic cotton than on the transgenic Bt or CpTI‐Bt cotton. In the choice tests with leaves of two transgenic cotton lines, the proportion of neonates on leaf discs of the two lines was not significantly different, but there was significantly higher consumption on CpTI‐Bt transgenic cotton than that on Bt transgenic cotton. In addition, significantly more neonates were found away from the leaf discs, lower consumption and higher mortality were achieved in the choice test with two transgenic cotton leaves than in the choice tests containing non‐transgenic cotton leaves. Leaves and buds were examined in choice tests of fourth instars. It appeared that fourth instars were found in equal numbers on transgenic and non‐transgenic cotton, except when larvae were exposed to leaves for 3 h. However, the total consumption on transgenic cotton was lower than that of the non‐transgenic cotton, so fourth instars may still have the capacity to detect transgenic cotton and reduce feeding on it, although they showed no preference on either transgenic or non‐transgenic cotton. More larvae were found off diet in the treatments with leaves than that of buds, and the number of injured leaf discs by per fourth instar was significantly higher than that of buds in choice tests, suggesting that leaf is a less preferred organ for H. armigera larvae, elicited more larval movements. Similarly, in no‐choice tests of fifth instars, significantly fewer feeding time and more moving time occurred on leaf than that of bud, boll and petal. When cotton line was considered, compared with non‐transgenic cotton, significantly lower feeding time and higher resting time occurred on the two transgenic cottons. Overall, H. armigera larvae have the ability to detect the transgenic Bt and CpTI‐Bt cottons or the less preferred organs and selectively feed more on the non‐transgenic cotton or the preferred organs, especially the neonates, which have a high capacity for avoiding transgenic cotton.     

Video-tracking and On-plant Tests Show Cry1Ab Resistance Influences Behavior and Survival of Neonate <Emphasis Type="Italic">Ostrinia nubilalis</Emphasis> Following Exposure to Bt Maize

To examine how resistance to Bacillus thuringiensis (Bt) toxins influences movement and survival of European corn borer (Ostrinia nubilalis [Hübner]) neonates, the responses of Cry1Ab-resistant , -susceptible, and hybrid (F1) larvae were examined using two different techniques. First, using an automated video-tracking system, aspects of O. nubilalis movement were quantified in the presence of artificial diet incorporating 50% non-Bt or insect-resistant Cry1Ab maize tissue. Second, O. nubilalis dispersal and survival were measured 48–72 h after hatching on a Cry1Ab maize plant surrounded by two non-Bt maize plants. Video tracking indicated the presence of Cry1Ab tissue increased the total distance moved (m), time moving (%), and time away from the diet (%) for O. nubilalis while decreasing meander (degrees/cm). However, resistant larvae showed reduced movement and increased meander (≈localized searching) relative to susceptible or hybrid larvae on diet incorporating Cry1Ab tissue. Conversely, when placed onto Cry1Ab maize plants, resistant larvae were more likely than susceptible O. nubilalis to disperse onto adjacent non-Bt plants. The difference in on-plant dispersal seems to reflect greater survival after toxin exposure for resistant larvae rather than increased activity. These results suggest that simplified ‘Petri dish’ tests may not be predictive of larval movement among non-Bt and insect-resistant Bt maize plants. Because models of O. nubilalis resistance evolution incorporate various movement and survival parameters, improved data for on-plant behavior and survival of Bt- resistant , -susceptible, and hybrid larvae should help preserve the efficacy of transgenic insect-resistant maize.     

Feeding of fall armyworm,Spodoptera frugiperda,on Bt transgenic cotton and its isoline

Studies on insect food intake and utilization are important for determining the degree of insect/plant association and host species’ resistance, and also for helping design pest management programs by providing estimates of potential economic losses, techniques for mass breeding of insects, and identifying physiological differences between species. We studied the feeding and development of fall armyworm, Spodoptera frugiperda (JE Smith) (Lepidoptera: Noctuidae), on transgenic (Bt) and non‐transgenic (non‐Bt) cotton. The larvae of S. frugiperda fed on Bt cotton had a longer development period (23.0 days) than those fed on non‐Bt cotton (20.2 days). Survivorship of S. frugiperda larvae fed on Bt cotton (74.1%) was lower than that of larvae fed on non‐Bt cotton (96.7%). Pupal weight of larvae fed on Bt cotton (0.042 g) was lower than that of larvae fed on non‐Bt cotton (0.061 g). The cotton cultivar significantly affected food intake, feces production, metabolization, and food assimilation by S. frugiperda larvae. However, it did not affect their weight gain. Intake of Bt‐cotton leaf (0.53 g dry weight) per S. frugiperda larva was lower than the intake of non‐Bt‐cotton leaf (0.61 g dry weight). Larvae fed on Bt‐cotton leaves produced less feces (0.25 g dry weight) than those fed on non‐Bt‐cotton leaves (0.37 g dry weight). Weight gain per S. frugiperda larva fed on Bt‐cotton leaves (0.058 g dry weight) was similar to the weight gain for larvae fed on non‐Bt‐cotton leaves (0.056 g dry weight). The cotton cultivar significantly affected the relative growth, consumption, and metabolic rates, as well as other nutritional indices: the figures were lower for larvae fed on Bt‐cotton leaves than for larvae fed on non‐transgenic cotton leaves.     

Effects of Bt maize expressing Cry1Ab and Bt spray on Spodoptera littoralis

Various studies have been conducted to assess the damage caused by secondary lepidopteran pests to transgenic Bt maize expressing Cry1Ab. However, to date little is known on the effects of transgenic maize on Spodoptera littoralis (Boisduval) (Lepidoptera: Noctuidae), a polyphagous herbivore which is considered a pest in Mediterranean maize growing areas. Here we present results on the effects of Bt maize (Bt‐11) and Bt spray (Dipel) on the various life stage parameters of this herbivore. We further assess the expression of Cry1Ab in different leaves and leaf parts in maize at a given plant growth stage, and determine whether the feeding damage of 3rd instar S. littoralis is influenced by Bt toxin expression. Contrary to previous literature reporting that S. littoralis is not sensitive to Bt Cry1Ab toxin, our results show that insects fed on either transgenic or Bt sprayed plants were negatively affected. Young S. littoralis larvae (1st and 2nd instars) were found to be the most sensitive to the Bt toxin. This was represented by a higher mortality and a slower developmental time of larvae maintained on transgenic or sprayed plants when compared to insects maintained on control plants. Moreover, Bt maize had a stronger and prolonged detrimental effect on insects when compared to Bt spray in maize. This was revealed by the fact that insects maintained on transgenic plants from 3rd instar to pupation took longer to reach adult emergence compared to insects that were maintained on sprayed plants. This was likely due to the continuous exposure of insects to the toxin when kept on transgenic maize. ELISA results showed a variation in the amount of Bt toxin among different leaf sections in transgenic maize at a given plant growth stage. These differences in Bt toxin were primarily found in the youngest leaf of growing plants. Although the lowest amounts of Bt toxin were detected in the growing leaf section of young leaves, this difference did not appear to influence the feeding behavior of 3rd instar S. littoralis.     

Susceptibility of field populations of sugarcane borer from non‐Bt and Bt maize plants to five individual Cry toxins

Abstract Sugarcane borer, Diatraea saccharalis (F.), is a major target of transgenic maize expressing Bacillus thuringiensis (Bt) proteins in South America and the US mid‐south region. Resistance development in target pest populations is a major threat to the sustainable use of Bt crops. In our field trials in 2009, a significant number of live borers and plant injury from D. saccharalis were observed in an experimental SmartStax? maize line. The objective of this study was to assess the relative susceptibility of two field populations of D. saccharalis collected from non‐Bt and Bt maize plants containing SmartStax? traits to five individual Cry proteins. The five Bt proteins included two proteins (Cry1A.105 and Cry2Ab2) that were expressed in SmartStax? maize plants and three other common Bt proteins (Cry1Aa, Cry1Ab and Cry1Ac) that were not produced in SmartStax?. Larval mortality and growth inhibition on Bt diet of the fourth generation after field collections were evaluated 7 days after release of neonates on the diet surface. The laboratory bioassays showed that 50% lethal concentration (LC₅₀) values for Cry1A.105 and Cry2Ab2 for the population originated from Bt plants were 3.55‐ and 1.34‐fold greater, respectively, than those of the population collected from non‐Bt plants. In contrast, relative to the population from non‐Bt plants, the LC₅₀ of the population sampled from Bt plants were 3.85‐, 2.5‐ and 1.64‐fold more sensitive to Cry1Aa, Cry1Ab and Cry1Ac, respectively. The results did not provide clear evidence to conclude that the observed field survival of D. saccharalis on Bt plants was associated with increased levels of resistance.     

Regional assessment of Helicoverpa zea populations on cotton and non-cotton crop hosts

Selection pressure on bollworm, Helicoverpa zea (Boddie) (Lepidoptera: Noctuidae), by cotton, Gossypium hirsutum (L.) (Malvaceae), that produces one or more Bacillus thuringiensis Berliner (Bt) proteins is reduced by plantings of non‐Bt refuge cotton that produce non‐selected individuals. However, the contributions of non‐Bt, non‐cotton crop hosts to the overall effective refuge for H. zea on Bt cotton have not been estimated. A 2‐year, season‐long study was conducted in five US cotton‐producing states to assess the spatial and temporal population dynamics and host use of H. zea. Helicoverpa zea larval estimates in commercial crop fields demonstrated that non‐cotton crop hosts, such as maize, Zea mays L. (Poaceae), grain sorghum, Sorghum bicolor (L.) Moench (Poaceae), peanut, Arachis hypogaea L. (Fabaceae), and soybean, Glycine max (L.) Merrill (Fabaceae), collectively support much larger larval populations than cotton throughout the season. Larval populations were almost entirely restricted to maize in the middle part of the season (June and portions of July), and were observed in non‐cotton crop hosts more frequently and typically in larger numbers than in cotton during the period when production would be expected in cotton (July and August). Numbers of H. zea larvae produced in replicated strip trials containing various crop hosts paralleled production estimates from commercial fields. In contrast, the number of H. zea adults captured in pheromone traps at interfaces of fields of Bt cotton and various crop hosts rarely varied among interfaces, except in instances where maize was highly attractive. With the exception of this early season influence of maize, moth numbers were not related to local larval production. These data demonstrate that H. zea adults move extensively from their natal host origins. Therefore, non‐cotton crop hosts, and even relatively distant hosts, contribute significantly to effective refuge for H. zea on Bt cotton. The results presented here demonstrate that substantial natural refuge is present for Bt‐resistance management of H. zea throughout the mid‐South and Southeast portions of the US cotton belt.     

Contamination and management of resistance evolution to high-dose transgenic insecticidal crops

For maize and cotton, transgenic varieties that express toxins derived from Bacillus thuriengensis (Bt) are now planted in several countries. To slow resistance evolution, the “high-dose/refuge” strategy is broadly implemented in which resistance is recessive and some fields (or areas within fields) are planted exclusively with Bt crops and other fields planted exclusively with non-transgenic refuge crops for susceptible insects. This strategy, however, could potentially be undermined by contamination. Here, we investigate general models of resistance evolution for high-dose events in which fields are contaminated due to the inadvertent mixing of seeds, volunteer plants, or pollen flow between Bt and non-Bt varieties coupled with seed-saving by farmers. Contamination of the refuge by Bt plants increases selection for resistance, thereby speeding resistance evolution. Nonetheless, in most situations this effect is small. Contamination of Bt fields by non-transgenic plants might be expected to have the opposite effect and always reduce the rate of resistance evolution. While this is often the case, it is not always so. If larvae move among plants within a field, then high movement rates may reverse the effect of contamination of Bt fields to slow resistance evolution. Furthermore, if the dispersal rates of adult females between Bt and refuge fields are low, then contamination of Bt fields may speed resistance. These results suggest that contamination has the potential to undermine the efficacy of the high-dose/refuge strategy, yet depending upon the particular pest and situation, contamination may not be a concern.     

Environmental risk assessment for the small tortoiseshell Aglais urticae and a stacked Bt-maize with combined resistances against Lepidoptera and Chrysomelidae in central European agrarian landscapes

The cultivation of Lepidoptera‐resistant Bt‐maize may affect nontarget butterflies. We assessed the risk posed by event MON89034 × MON88017 (expressing Cry1A.105 and Cry2Ab2 against corn borers) to nontarget Lepidoptera. Using the small tortoiseshell Aglais urticae, a butterfly species common in central Europe, as a test organism we (i) assessed the toxicity of Bt‐maize pollen on butterfly larvae; (ii) measured pollen deposition on leaves of the host plant Urtica dioica; (iii) mapped the occurrence and distribution of host plants and larvae in two arable landscapes in Germany during maize anthesis; and (iv) described the temporal occurrence of a 1‐year population of A. urticae. (i) Larvae‐fed 200 Bt‐maize pollen grains/cm² had a reduced feeding activity. Significant differences in developmental time existed at pollen densities of 300 Bt‐maize pollen grains/cm² and in survival at 400 grains/cm². (ii) The highest pollen amount found was 212 grains/cm² at the field margin. Mean densities were much lower. (iii) In one region, over 50% of A. urticae nests were located within 5 m of a maize field, while in the other, all nests were found in more than 25 m distance to a maize field. (iv) The percentage of larvae developing during maize anthesis was 19% in the study area. The amount of pollen from maize MON89034 × MON88017 found on host plants is unlikely to adversely affect a significant proportion of larvae of A. urticae. This paper concludes that the risk of event MON89034 × MON88017 to populations of this species is negligible.