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.     

Field efficacy of sweet corn hybrids expressing a Bacillus thuringiensis toxin for management of Ostrinia nubilalis (Lepidoptera: Crambidae) and Helicoverpa zea (Lepidoptera: Noctuidae)

Field studies were done in 1995-1996 to assess the efficacy of three sweet corn hybrids that express the Bacillus thuringiensis (Bt) toxin, CrylAb, against two lepidopteran pests, Ostrinia nubilalis (Hubner) and Helicoverpa zea (Boddie). The Bt hybrids tested were developed by Novartis Seeds, using the event BT-11, which expresses Bt toxin in green tissue as well as reproductive tissues including the tassel, silk, and kernel. Bt hybrids were compared with a standard non-Bt control or the non-Bt isoline for each hybrid; none of the hybrids were treated with insecticides during the study. Hybrid efficacy was based on larval control of each pest, as well as plant or ear damage associated with each pest. In both years, control of O. nubilalis larvae in primary ears of all Bt hybrids was 99-100% compared with the appropriate non-Bt check. Plant damage was also significantly reduced in all Bt hybrids. In 1996, control of H. zea in Bt hybrids ranged from 85 to 88% when compared with the appropriate non-Bt control. In 1996, a University of Minnesota experimental non-Bt hybrid (MN2 x MN3) performed as well as the Bt hybrids for control of O. nubilalis. Also, in 1996, two additional University of Minnesota experimental non-Bt hybrids (A684su X MN94 and MN2 X MN3) performed as well as Bt hybrids for percent marketable ears (ears with no damage or larvae). In addition, compared with the non-Bt hybrids, percent marketable ears were significantly higher for all Bt hybrids and in most cases ranged from 98 to 100%. By comparison, percent marketable ears for the non-Bt hybrids averaged 45.5 and 37.4% in 1995 and 1996, respectively. Results from the 2-yr study strongly suggest that Bt sweet corn hybrids will provide high levels of larval control for growers in both fresh and processing markets. Specifically, Bt sweet corn hybrids, in the absence of conventional insecticide use, provided excellent control of O. nubilalis, and very good control of H. zea. However, depending on location of specific production regions, and the associated insect pests of sweet corn in each area, some insecticide applications may still be necessary.     

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.     

Feeding behavior of neonate Ostrinia nubilalis (Lepidoptera: Crambidae) on Cry1Ab Bt corn: implications for resistance management

The European corn borer, Ostrinia nubilalis (Hübner) (Lepidoptera: Crambidae), is an economically important insect pest of corn, Zea mays L., in the United States and Canada. The development of genetically modified corn expressing genes derived from Bacillus thuringiensis (Bt) that encodes insecticidal crystalline (Cry) proteins has proven to be effective in controlling this insect. To assess the feeding behavior of neonate O. nubilalis on Bt corn, we examined differences in feeding behavior, based on presence of plant material in the gut, between Cry1Ab Bt corn and non-Bt near isoline corn for four intervals over a 48-h period. Feeding experiments revealed that there was significantly less feeding on Bt corn compared with non-Bt near isoline corn. The behavior of neonates on the plant corresponded with the differences in feeding on the two corn lines. The findings also showed that > 50% of the larvae initially left the plant before there was evidence in the gut of feeding regardless of whether the source was Bt or non-Bt corn. A higher quantity of plant material was found in the gut of larvae recovered from leaves of non-Bt compared with Bt corn. At the end of 48 h among the larvae that had left the plant, a greater proportion from Bt corn had plant material in the gut than did those from non-Bt corn.     

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.     

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.     

DiPel-selected Ostrinia nubilalis larvae are not resistant to transgenic corn expressing Bacillus thuringiensis Cry1Ab

The survival of KS-SC DiPel-resistant and -susceptible European corn borer, Ostrinia nubilalis (Hübner), was evaluated on different tissues from corn, Zea mays L., hybrids, including a nontransgenic and two transgenic corn plants (events MON810 and Bt11) expressing high doses of Bacillus thuringiensis (Bt) Cry1Ab. The survival of Bt-resistant and -susceptible third instars was similar after a 5-d exposure to transgenic plant tissues. Survivors eventually died when returned to Bt corn tissues, but many were able to continue development when transferred to non-Bt corn tissues. Survival of resistant and susceptible larvae also was evaluated in bioassays with dilutions of leaf extracts from the three corn hybrids incorporated in an artificial diet. In these assays, survival was significantly higher for resistant O. nubilalis neonates at three of the five dilutions compared with the susceptible strain, but the resistance ratio was only 2.2- and 2.4-fold for MON810 and Bt11, respectively. The data demonstrate that Bt-resistant and unselected control O. nubilalis larvae were similar in susceptibility to MON810 and Bt11 event corn hybrids. Although we were unable to evaluate the Cry1Ab protein that larvae were exposed to in the transgenic tissue because of company restrictions, Cry1Ab protoxin produced in Escherichia coli was incubated with extracts from non-Bt corn leaves to simulate the in planta effect on the transgenic protein. Cry1Ab protoxin was hydrolyzed rapidly by enzymes in the corn extract into peptide fragments with molecular masses ranging from 132 to 74 kDa, and eventually 58 kDa. Overall, these data suggest that plant enzymes hydrolyze transgenic toxin to one that is functionally activated. Therefore, resistant insect populations with reduced proteinase activity do not seem to pose a threat to the efficacy of commercial MON810 and Bt11 corn hybrids.     

Cannibalism of Helicoverpa zea (Lepidoptera: Noctuidae) from Bacillus thuringiensis (Bt) transgenic corn versus non-Bt corn

Because of the importance of cannibalism in population regulation of Helicoverpa zea (Boddie) (Lepidoptera: Noctuidae) in corn, Zea mays L., it is useful to understand the interactions between Bacillus thuringiensis (Bt) transgenic corn and cannibalism. To determine the effects of Bt corn on cannibalism in H. zea, pairs of the same or different instars were taken from Bt or non-Bt corn and placed on artificial diet in proximity. Cannibalism occurred in 91% of pairs and was approximately 7% greater for pairs of larvae reared from Bt transgenic corn (95%) than from non-Bt corn (88%). Also, first instar by first instar pairs had a lower rate of cannibalism than other pairs. Time until cannibalism was not different for larvae from Bt corn versus non-Bt corn. Pupation rate of cannibals and surviving victims was not different for pairs from Bt corn versus non-Bt corn. Finally, cannibalism increased pupation rate of cannibals from both Bt and non-Bt corn by approximately 23 and 12%, respectively, although the increases were not significant. Thus, negative effects of Bt on larvae were compensated by increased cannibalism in comparison with larvae reared on non-Bt corn, which increased larval survival to levels comparable with larvae reared on non-Bt plants.     

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.     

Performance of transgenic corn hybrids in Missouri for insect control and yield

The efficacy of Bacillus thuringiensis-transformed corn (Zea mays L.) hybrids compared with comparable nontransformed corn hybrids for controlling first- and second-generation European corn borer, Ostrinia nubilalis (Hübner), and second-generation southwestern corn borer, Diatraea grandiosella Dyar, was determined. Yield comparisons were obtained from the same plots of corn hybrids. Both generations of European and the second-generation of southwestern corn borer were effectively controlled, but the Bt hybrids varied in degree of control. Hybrids from Ciba Seeds, DEKALB, and Mycogen had more European corn borer tunneling than those from Novartis or Cargill, and this was generally ascribed to different transgenic events. The Bt-transformed hybrids had virtually no leaf-feeding damage and less tunneling than the non-Bt corn hybrids. Some Bt corn hybrids had no tunneling, whereas other Bt hybrids had a small amount of tunneling. All of the non-Bt hybrids had significant leaf-feeding damage and stalk tunneling from both insects. Only three live European corn borer larvae (stunted) were found in the Bt corn hybrids while splitting stalks to assess tunnel length. When insect damage was significant, and in some evaluations where damage was not significant, differences in yields among hybrids were observed. No significant insect population differences were observed for five genera of beneficial insects for Bt versus non-Bt corn hybrids. Corn hybrids that have been transformed with the Bt gene provide an effective means of control for corn borers and efforts to reduce the likelihood of development of borer resistance are warranted.     

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.     

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.     

Performance of Bt-susceptible and -heterozygous dual-gene resistant genotypes of Spodoptera frugiperda (J.E. Smith) (Lepidoptera: Noctuidae) in seed blends of non-Bt and pyramided Bt maize

A seed blend refuge has been implemented in the U.S. Corn Belt for Bt maize resistance management. The fall armyworm, Spodoptera frugiperda (J.E. Smith), is a target pest of Bt maize in the Americas. The larvae of this pest are mobile, which may affect the efficacy of seed blend refuges. In this study, field and greenhouse trials were conducted to determine the performance of Bt-susceptible (aabb) and -heterozygous dual-gene-resistant (AaBb) genotypes of S. frugiperda in seed blends of non-Bt and pyramided Bt maize. Three field trials evaluated larval survival, larval growth, and plant injury with aabb in seed blends of Bt maize expressing Cry1A.105/Cry2Ab2/Vip3A with 0–30% non-Bt seeds. Greenhouse tests investigated the performance of aabb and AaBb in seed blends of Cry1A.105/Cry2Ab2 with 0–30% non-Bt seeds. In pure non-Bt maize plots, after 9–13 d of neonates being released on the plants, 0.39 and 0.65 larvae/plant survived with leaf injury ratings of 4.7 and 5.9 (Davis's 1–9 scale) in the field and greenhouse, respectively. In contrast, live larvae and plant injury were virtually not observed on Bt plants across all planting patterns. Larval occurrence and plant injury by aabb on non-Bt plants were similar between seed blends and pure non-Bt plantings, suggesting that the blended refuges could provide an equivalent susceptible population as structured refuge under the test conditions. In the greenhouse, the two insect genotypes in seed blends performed similarly, indicating that the seed blends did not provide more favorable conditions for AaBb over aabb. The information generated from this study should be useful in managing S. frugiperda and evaluating if send blends could be suitable refuge options for Bt resistance management in the regions where the insect is a primary target pest.     

Evaluating popcorn as a potential refuge of Ostrinia nubilalis (Lepidoptera: Crambidae)

Popcorn was evaluated in a series of experiments conducted over four growing seasons for its potential as a refuge for European corn borer, Ostrinia nubilalis (Hübner). Objectives of these studies were to determine whether more larvae were produced in popcorn than in field corn and to determine how popcorn influenced female oviposition and larval distribution in neighboring field corn. Two varieties of popcorn (M140, 105d and M3374Y, 118d), one mixture of popcorn (50% 105d and 50% 118d), and field corn (DK580, 108d) were evaluated. Number of egg masses, eggs per egg mass, and larvae were significantly higher in popcorn compared with field corn. Moth oviposition and larval distribution were evaluated using 105d popcorn embedded in several cornfields across Iowa. The row of field corn adjacent to popcorn had significantly more larvae compared with background field corn. In larger field experiments, O. nubilalis larval survival after overwintering was significantly different, with 2.2-18.7 times more O. nubilalis larvae surviving in popcorn than field corn. The potential use of popcorn as an O. nubilalis refuge for genetically engineered corn is considered.     

Evaluation of transgenic Bacillus thuringiensis corn hybrids against Cry1Ab-susceptible and -resistant sugarcane borer (Lepidoptera: Crambidae)

A Louisiana strain of the sugarcane borer, Diatraea saccharalis (F.) (Lepidoptera: Crambidae), was selected for resistance to the CrylAb protein of Bacillus thuringiensis (Bt) by using an F2 screening procedure. Survival of Bt-resistant, -susceptible, and -heterozygous genotypes of sugarcane borer was evaluated on vegetative and reproductive stages of five non-Bt and seven Bt field corn, Zea mays L., hybrids in a greenhouse study. Larval survival was recorded 21 d after infestation of neonates on potted plants. Larval survival across the three sugarcane borer genotypes and five non-Bt corn hybrids after 21 d ranged from 23.6 +/- 5.2% (mean +/- SEM) to 57.5 +/- 5.2%. Mean survival of Cry1Ab-resistant larvae on vegetative and reproductive plant stages was 12 and 21%, respectively. During the vegetative stages, all seven Bt corn hybrids were highly efficacious against Cry1Ab-susceptible and -heterozygous genotypes of sugarcane borer, with a larval survival rate of <2% for the Bt-susceptible genotype and < or =5% for the heterozygotes. However, 8-18% of the heterozygous genotype survived on reproductive stage plants for four of the seven Bt corn hybrids tested. The variation in performance of Bt corn cultivars at vegetative and reproductive growth stages against Cry1Ab resistant sugarcane borer suggests differential seasonal expression that may hasten resistance in the field. Bt corn hybrids expressing a "high dose" for European corn borer, Ostrinia nubilalis (Hübner), may not produce a sufficient high dose for the sugarcane borer.     

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.     

转Bt基因玉米对甜菜夜蛾幼虫存活和发育的影响

在室内测定了2种转Cry1Ab基因的Bt玉米MON810和Bt11不同组织对甜菜夜蛾 Spodoptera exigua （Hübner）初孵幼虫以及心叶对4龄幼虫存活和发育的影响,在田间比较了甜菜夜蛾幼虫取食Bt 和非Bt玉米雌穗的存活和为害情况。结果表明,转Cry1Ab基因的Bt玉米的不同组织对甜菜夜蛾初孵幼虫都具有明显的杀虫活性,取食Bt玉米心叶、苞叶、籽粒时甜菜夜蛾均在幼虫期死亡; 取食MON810和Bt11雄穗的初孵幼虫化蛹率分别为5.2%和2.1%,羽化率为2.1%和1.0%;取食MON810和Bt11花丝的初孵幼虫化蛹率分别为1.0%和2.1%,但不能羽化。4龄幼虫取食MON810玉米心叶的化蛹率与对照差异不显著,而取食Bt11的化蛹率与对照差异显著; 取食两种Bt玉米心叶的４龄幼虫化蛹后的雌、雄蛹重和羽化率与对照组差异显著,但蛹期和平均单雌产卵量差异不显著,虽然对照组羽化的成虫平均产卵量高于Bt玉米组。田间接种初孵幼虫10 天后的调查结果表明,在MON810和Bt11玉米花丝上幼虫存活率分别为1.3%和0.3%, 而对照组分别为12.9%和16.2%;MON810和Bt11玉米雌穗被害率分别为18.3%和5.0%,而对照组分别为93.3%和95.0%,均显著低于对照组。     

An in-field screen for early detection and monitoring of insect resistance to Bacillus thuringiensis in transgenic crops

We present a field-based approach to detect and monitor insects with resistance to insecticidal toxins produced by transgenic plants. Our objective is to estimate the phenotypic frequency of resistance in a population by relating the densities of insects on genetically transformed plants to densities on nontransformed plants. We focus on European corn borer, Ostrinia nubilalis (Hübner), in sweet corn, Zea mays L., expressing Cry1Ab from Bacillus thuringiensis subsp. kurstaki Berliner to illustrate principles underlying the method. The probability of detecting one or more rare, resistant larvae depends on sample size, the density of larvae on nontransformed plants, and an assumed frequency of resistant phenotypes in a given population. Probability of detection increases with increases in sample size, background density, or the frequency of resistant individuals. Following binomial probability theory, if a frequency of 10(-4) is expected, 10(3)-10(4) samples must be collected from a B. thuringiensis (Bt) crop to have at least a 95% probability of locating one or more resistant larvae. In-field screens using transgenic crops have several advantages over traditional laboratory-based methods, including exposure to a large number of feral insects, discrimination of resistant individuals based on Bt dosages expressed in the field, incorporation of natural and Bt-induced mortality factors, simultaneous monitoring for more than one insect species, and ease of use. The approach is amenable to field survey crews working in research, extension, and within the seed corn industry. Estimates of the phenotypic frequency of resistance from the in-field screen can be useful for estimating initial frequency of resistant alleles. Bayesian statistical methods are outlined to estimate phenotype frequencies, allele frequencies, and associated confidence intervals from field data. Results of the approach are discussed relative to existing complementary methods currently available for O. nubilalis and corn earworm, Helicoverpa zea (Boddie).     

Effects of Cry1Ab-expressing corn anthers on the movement of monarch butterfly larvae

Decreased larval feeding and weight of the monarch butterfly, Danaus plexippus L., have been detected after 4 d of exposure in the laboratory to a high density of Bacillus thuringiensis (Bt)-expressing anthers. One hypothesis is that larvae exposed to Bt anthers exhibit increased wandering, resulting in less feeding and lower weight gain. To test this hypothesis, 2-d-old monarch butterfly larvae exposed to milkweed leaf disks with no anthers, anthers that express Bt (Cry1Ab, event MON810), or other non-Bt anthers were observed using a video-tracking system. As had been shown in previous studies, larvae exposed to Bt anthers fed less and gained less weight than larvae exposed to non-Bt or no anthers, yet there was no evidence of feeding on anthers. Total distance moved, maximum displacement from release point, percentage of time spent moving or near anthers, or mean turn angle did not differ across treatments. However, larvae exposed to Bt anthers spent more time off milkweed leaf disks than those exposed to no anthers and were more likely to move off the leaf than larvae exposed to non-Bt anthers. Results suggest that larvae exposed to Bt anthers behave differently and that ingestion may not be the only way Bt can affect nontarget insects like the monarch butterfly.     

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.