Efficacy of insecticides of different chemistries against Helicoverpa zea (Lepidoptera: Noctuidae) in transgenic Bacillus thuringiensis and conventional cotton

Six insecticides of different chemistries were evaluated against the cotton bollworm, Helicoverpa zea (Boddie), in non-B.t. (Deltapine 'DP 5415', Deltapine 'DP 5415RR') and transgenic Bacillus thuringiensis (Berliner) (B.t.) (Deltapine 'NuCOTN 33B', Deltapine 'DP 458 B/RR') cotton. In 1998, treatments consisted of three rates each of a pyrethroid (lambda-cyhalothrin), spinosyn (spinosad), carbamate (thiodicarb), pyrrole (chlorfenapyr), oxadiazine (indoxacarb), and avermectin (emamectin benzoate) in a nonirrigated field. In 1999, treatments consisted of three rates each of lambda-cyhalothrin, spinosad, thiodicarb, and indoxacarb in an irrigated and a nonirrigated (dryland) field. The highest rate of each insecticide corresponded to normal grower-use rates. Spinosad and thiodicarb controlled H. zea in non-B.t. cotton, whereas other materials were less effective. Even though H. zea is becoming increasingly resistant to pyrethroid insecticides, lambda-cyhalothrin was highly effective in dryland B. thuringiensis cotton. Spinosad and thiodicarb were equally effective. Data indicated that reduced rates of lambda-cyhalothrin, spinosad, and thiodicarb could be used for control of H. zea in dryland B.t. cotton systems. However, reduced rates of these insecticides in a heavily irrigated B.t. cotton system did not provide adequate control.     

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.     

Toxicity of Bacillus thuringiensis insecticidal proteins for Helicoverpa armigera and Helicoverpa punctigera (Lepidoptera: Noctuidae), major pests of cotton

The susceptibilities of the major pests of cotton in Australia, Helicoverpa armigera and Helicoverpa punctigera, to some insecticidal proteins from Bacillus thuringiensis were tested by bioassay. A commercial formulation, DiPel, and individual purified insecticidal proteins were tested. H. armigera was consistently more tolerant to B. thuringiensis insecticidal proteins than was H. punctigera, although both were susceptible to only a limited range of these proteins. Only Cry1Ab, Cry1Ac, Cry2Aa, Cry2Ab, and Vip3A killed H. armigera at dosages that could be considered acceptable. There was no significant difference in the toxicities of Cry1Fa and Cry1Ac for H. punctigera but Cry1Fa had little toxicity for H. armigera. The five instars of H. armigera did not differ significantly in their susceptibility to DiPel on the basis of LC(50). However, there were significant differences in the susceptibility to Cry1Ac and Cry2Aa of three strains of H. armigera. Bioassays conducted with Cry1Ac and Cry2Aa showed that there was a small but significant negative interaction between these delta-endotoxins.     

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.     

Efficacy of transgenic cotton containing a cry1Ac gene from Bacillus thuringiensis against Helicoverpa armigera (Lepidoptera: Noctuidae) in northern China

NuCOTN 33B, a Bt transgenic variety of upland cotton (Gossypium hirsutum L.) expressing the insecticidal protein Cry1Ac from Bacillus thuringiensis Berliner sp. kurstaki, was evaluated for resistance to Helicoverpa armigera (Hübner) during 1998-2000 in northern China. The results indicated that there was no significant difference in egg densities between NuCOTN 33B and three nontransgenic varieties (DP5415, Zhongmian12, and Shiyuan321) during the season, although the survival of larvae on NuCOTN 33B seemed significantly reduced. High larval densities observed on non-Bt cotton appeared in great contrast to the low larval populations observed on NuCOTN 33B plants during the seasons. In an environment without insecticide sprays, the annual ginned cotton yields in NuCOTN 33B plots, ranging from 1391.17 to 1511.35 kg/ha, were significantly higher than those in non-Bt cotton (340.34-359.58 kg/ha). These high levels of field efficacy for NuCOTN 33B against H. armigera in northern China may pave the way for reduced pesticide applications and an expansion of alternative pest-control strategies.     

Genetic variation for resistance to Bacillus thuringiensis toxins in Helicoverpa zea (Lepidoptera: Noctuidae) in eastern North Carolina

To evaluate resistance to Bacillus thuringiensis Berliner (Bt) toxins, adult female bollworms, Helicoverpa zea (Boddie) (Lepidoptera: Noctuidae), were collected from four light trap locations in two eastern North Carolina counties from August to October during 2001 and 2002. Females were allowed to oviposit, and upon hatching, 24 neonates from each female (F1 lines) were screened for survival and growth rate on each of three diets: non-Bt diet, diet containing 5.0 microg/ml Cry1Ac toxin, or diet containing 5.0 microg/ml Cry2Ab toxin. These screens were designed to identify nonrecessive Bt resistance alleles present in field populations of bollworm. Of 561 and 691 families screened with both Cry1Ac- and Cry2Ab-containing diets in 2001 and 2002, respectively, no F1 lines were identified that seemed to carry a gene conferring substantial resistance to either Cry1Ac or Cry2Ab. Adults from F1 lines with growth scores in the highest (R) and lowest (S) quartiles were mated in four combinations, RxR, SxR, RxS, and SxS. Differences in growth rates of larvae from these crosses demonstrated that there is substantial quantitative genetic variation in eastern North Carolina populations for resistance to both Cry1Ac and Cry2Ab toxins. These findings, in addition to results suggesting partially dominant inheritance of resistance to Cry1Ac and Cry2Ab, are critically important for determining appropriate resistance management strategies that impact the sustainability of transgenic cotton, Gossypium hirsutum (L.).     

Toxicity of Bacillus thuringiensis Cry proteins to Helicoverpa armigera (Lepidoptera: Noctuidae) in South Africa

The susceptibility of one of the most important pests in southern Africa, Helicoverpa armigera (Lepidoptera: Noctuidae), to Bacillus thuringiensis Cry proteins was evaluated by bioassay. Cry proteins were produced in Escherichia coli BL21 cells that were transformed with plasmids containing one of six cry genes. The toxicity of each Cry protein to H. armigera larvae was determined by the diet contamination method for second instar larvae and the droplet feeding method for neonate larvae. For each of the proteins, dose-mortality and dose-growth inhibition responses were analyzed and the median lethal dose (LD(50)) and median inhibitory dose (ID(50)) determined. Second instar larvae were consistently less susceptible to the evaluated Cry proteins than neonate larvae. The relative toxicity of Cry proteins ranked differently between neonate larvae and second instar larvae. On the basis of the LD(50) and ID(50) values, Cry1Ab, Cry1Ac, and Cry2Aa were the most toxic of the evaluated proteins to H. armigera larvae. The study provides an initial benchmark of the toxicity of individual Cry proteins to H. armigera in South Africa.     

Cold hardiness of Helicoverpa zea (Lepidoptera: Noctuidae) pupae

An insect's cold hardiness affects its potential to overwinter and outbreak in different geographic regions. In this study, we characterized the response of Helicoverpa zea (Boddie) pupae to low temperatures by using controlled laboratory measurements of supercooling point (SCP), lower lethal temperature (LT(50)), and lower lethal time (LLTime). The impact of diapause, acclimation, and sex on the cold hardiness of the pupae also were evaluated. Sex did not significantly affect the SCP, LT(50), or LLTime. However, the mean SCP of diapausing pupae (-19.3°C) was significantly lower than nondiapausing pupae (-16.4°C). Acclimation of nondiapausing pupae to constant temperatures from 10 to 20°C before supercooling also produced a significantly lower SCP than nondiapausing pupae held at 25°C. The LT(50)s of nondiapausing and diapausing were not significantly different, but confirmed that H. zea pupae are chill-intolerant because these lethal temperatures are warmer than the corresponding mean SCPs. Diapausing pupae survived longer than nondiapausing pupae at the same, constant, cold temperatures, a finding consistent with the SCP results. Both of these results suggest enhanced cold hardiness in diapausing pupae. When laboratory results were compared with field temperatures and observed distributions of H. zea in the contiguous United States, the laboratory results corroborated what is currently perceived to be the northern overwintering limit of H. zea; approximately the 40(th) parallel. Moreover, our research showed that areas north of this limit are lethal to overwintering pupae not because of low temperature extremes, but rather the length of time spent at near-zero temperatures.     

Field and laboratory evaluations of transgenic cottons expressing one or two Bacillus thuringiensis var. kurstaki Berliner proteins for management of noctuid (Lepidoptera) pests

Field studies were conducted from 1999 to 2001 to evaluate the efficacy of the transgenic cotton, Gossypium hirsutum (L.), genotype, Bollgard II (Monsanto 15985), which expresses two Bacillus thuringiensis Berliner (Bt) proteins (Cry1Ac + Cry2Ab) that are active against lepidopterous pests. Bollgard II was compared with Bollgard (DP50B), which expresses only one Bt protein (Cry1Ac), and, in all tests, the conventional variety, DP50, was used as a non-Bt control. Larval populations of the bollworm, Helicoverpa zea (Boddie), and the soybean looper, Pseudoplusia includens (Walker), were significantly lower in Bollgard II than in Bollgard and conventional cotton, and the proportion of fruit damaged by H. zea was also lower. Fall armyworm, Spodoptera frugiperda (J. E. Smith), populations were lower in Bollgard II than in Bollgard, although not significantly. Field tests were supplemented with laboratory bioassays in 2001 to compare mortality of S. frugiperda, and beet armyworms, Spodoptera exigua (Hübner), feeding on these genotypes. Mortality of both species was significantly greater on Bollgard II plant material than on either Bollgard or conventional cotton. This study demonstrated that the dual-toxin Bollgard II genotype is highly effective against lepidopterous pests that are not adequately controlled by the current single-toxin Bollgard varieties. If toxin expression in future Bollgard II varieties remains consistent with that of Monsanto 15985, supplemental insecticides will be reduced, and may be eliminated for lepidopterous pests in South Carolina.     

Evaluation of the natural refuge function for Helicoverpa arnigera (Lepidoptera: Noctuidae) within Bacillus thuringiensis transgenic cotton growing areas in north China

The density of Helicoverpa armigera (Hübner) populations on Bacillus thuringiensis Berliner (Bt) transgenic cotton, corn, peanut, and soybean; differences in its development on Bt cotton and common (nontransgenic) cotton; and the potential for mating among populations from Bt cotton fields and other crop fields were investigated in the suburbs of Xinxiang City (Henan Province) and Langfang City (Hebei Province) in the southern and northern parts of north China, respectively. Although development of H. armigera on Bt cotton was much slower than on common cotton, there was a still high probability of mating between populations from Bt cotton and other sources due to the scattered emergence pattern of H. armigera adults, and overlap of the second and third generations. In a cotton and corn growing region, early and late planted corn provided suitable refugia for the third and fourth generations of H. armigera, but not for the second generation. In a cotton and soybean/ peanut mix system, noncotton crops provided a natural refugia from the second- to fourth-generation H. armigera, but function of the refuge would closely depend on the proportion of Bt cotton. Consequently, it may be necessary to compensate the original mixed cropping patterns in different areas for delaying resistance development of H. armigera to Bt cotton.     

Toxicity of several delta-endotoxins of Bacillus thuringiensis against Helicoverpa armigera (Lepidoptera: Noctuidae) from Spain

Toxicity and larval growth inhibition of 11 insecticidal proteins of Bacillus thuringiensis were evaluated against neonate larvae of Helicoverpa armigera, a major pest of important crops in Spain and other countries, by a whole-diet contamination method. The most active toxins were Cry1Ac4 and Cry2Aa1, with LC50 values of 3.5 and 6.3 microg/ml, respectively. At the concentrations tested, Cry1Ac4, Cry2Aa1, Cry9Ca, Cry1Fa1, Cry1Ab3, Cry2Ab2, Cry1Da, and Cry1Ja1, produced a significant growth inhibition, whereas Cry1Aa3, Cry1Ca2, and Cry1Ea had no effect.     

Interaction of Bacillus thuringiensis toxins with larval midgut binding sites of Helicoverpa armigera (Lepidoptera: Noctuidae)

In 1996, Bt-cotton (cotton expressing a Bacillus thuringiensis toxin gene) expressing the Cry1Ac protein was commercially introduced to control cotton pests. A threat to this first generation of transgenic cotton is the evolution of resistance by the insects. Second-generation Bt-cotton has been developed with either new B. thuringiensis genes or with a combination of cry genes. However, one requirement for the "stacked" gene strategy to work is that the stacked toxins bind to different binding sites. In the present study, the binding of (125)I-labeled Cry1Ab protein ((125)I-Cry1Ab) and (125)I-Cry1Ac to brush border membrane vesicles (BBMV) of Helicoverpa armigera was analyzed in competition experiments with 11 nonlabeled Cry proteins. The results indicate that Cry1Aa, Cry1Ab, and Cry1Ac competed for common binding sites. No other Cry proteins tested competed for either (125)I-Cry1Ab or (125)I-Cry1Ac binding, except Cry1Ja, which competed only at the highest concentrations used. Furthermore, BBMV from four H. armigera populations were also tested with (125)I-Cry1Ac and Cry1Ab to check the influence of the insect population on the binding results. Finally, the inhibitory effect of selected sugars and lectins was also determined. (125)I-Cry1Ac binding was strongly inhibited by N-acetylgalactosamine, sialic acid, and concanavalin A and moderately inhibited by soybean agglutinin. In contrast, (125)I-Cry1Ab binding was only significantly inhibited by concanavalin A. These results show that Cry1Ac and Cry1Ab use different epitopes for binding to BBMV.     

Survivorship of Helicoverpa zea and Heliothis virescens on cotton plant structures expressing a Bacillus thuringiensis vegetative insecticidal protein

A series of tests quantified bollworm, Helicoverpa zea (Boddie), and tobacco budworm, Heliothis virescens (F.), larval survival on plant structures of a nontransgenic cotton (Gossypium hirsutum L.), 'Coker 312', and two transgenic cottons expressing Vip3A protein or both Vip3A + CrylAb proteins (VipCot). Vegetative and reproductive structures including terminal leaves, flower bud (square) bracts, whole debracted squares, flower petals, flower anthers, and intact capsules (bolls) were harvested from plants in field plots. Each structure was infested with 2-d-old larvae from one of the two heliothine species. Larvae were allowed to feed for 96 h on fresh tissue. Survivorship at 96 h after infestation was significantly lower on all structures of Vip3A and VipCot cotton lines compared with similar structures of Coker 312. VipCot plant structures generally resulted in lower larval survivorship compared with similar structures of the Vip3A cotton line. H. zea survivorship ranged from 4 to 28% and from 1 to 18% on Vip3A and VipCot plant structures, respectively. H. virescens survivorship ranged from 10 to 43% and from 2 to 12% on Vip3A and VipCot plant structures, respectively. H. virescens survivorship was higher on VIP3A plant structures compared with that for H. zea on similar structures. These differences between species were not observed on plants from the cotton line expressing VipCot proteins. The results for these plant structures demonstrate that the combination of proteins expressed in VipCot cotton lines are more effective than Vip3A cotton lines against this heliothine complex.     

Interaction of Bacillus thuringiensis Toxins with Larval Midgut Binding Sites of Helicoverpa armigera (Lepidoptera: Noctuidae)

In 1996, Bt-cotton (cotton expressing a Bacillus thuringiensis toxin gene) expressing the Cry1Ac protein was commercially introduced to control cotton pests. A threat to this first generation of transgenic cotton is the evolution of resistance by the insects. Second-generation Bt-cotton has been developed with either new B. thuringiensis genes or with a combination of cry genes. However, one requirement for the “stacked” gene strategy to work is that the stacked toxins bind to different binding sites. In the present study, the binding of ¹²⁵I-labeled Cry1Ab protein (¹²⁵I-Cry1Ab) and ¹²⁵I-Cry1Ac to brush border membrane vesicles (BBMV) of Helicoverpa armigera was analyzed in competition experiments with 11 nonlabeled Cry proteins. The results indicate that Cry1Aa, Cry1Ab, and Cry1Ac competed for common binding sites. No other Cry proteins tested competed for either ¹²⁵I-Cry1Ab or ¹²⁵I-Cry1Ac binding, except Cry1Ja, which competed only at the highest concentrations used. Furthermore, BBMV from four H. armigera populations were also tested with ¹²⁵I-Cry1Ac and Cry1Ab to check the influence of the insect population on the binding results. Finally, the inhibitory effect of selected sugars and lectins was also determined. ¹²⁵I-Cry1Ac binding was strongly inhibited by N-acetylgalactosamine, sialic acid, and concanavalin A and moderately inhibited by soybean agglutinin. In contrast, ¹²⁵I-Cry1Ab binding was only significantly inhibited by concanavalin A. These results show that Cry1Ac and Cry1Ab use different epitopes for binding to BBMV.     

Resistance to the Cry1Ac delta-endotoxin of Bacillus thuringiensis in the cotton bollworm, Helicoverpa armigera (Lepidoptera: Noctuidae)

Three laboratory strains of Helicoverpa armigera (Hübner) were established by mating of field-collected insects with an existing insecticide-susceptible laboratory strain. These strains were cultured on artificial diet containing the Cry1Ac protoxin of Bacillus thuringiensis using three different protocols. When no response to selection was detected after 7-11 generations of selection, the three strains were combined by controlled mating to preserve genetic diversity. The composite strain (BX) was selected on the basis of growth rate on artificial diet containing Cry1Ac crystals. Resistance to Cry1Ac was first detected after 16 generations of continuous selection. The resistance ratio (RR) peaked approximately 300-fold at generation 21, after which it declined to oscillate between 57- and 111-fold. First-instar H. armigera from generation 25 (RR = 63) were able to complete their larval development on transgenic cotton expressing Cry1Ac and produce fertile adults. There appeared to be a fitness cost associated with resistance on cotton and on artificial diet. The BX strain was not resistant to the commercial Bt spray formulations DiPel and XenTari, which contain multiple insecticidal crystal proteins, but was resistant to the MVP formulation, which only contains Cry1Ac. The strain was also resistant to Cry1Ab but not to Cry2Aa or Cry2Ab. Toxin binding assays showed that the resistant insects lacked the high affinity binding site that was detected in early generations of the strain. Genetic analysis confirmed that resistance in the BX strain of H. armigera is incompletely recessive.     

Cross-resistance to alpha-cypermethrin after xanthotoxin ingestion in Helicoverpa zea (Lepidoptera: Noctuidae)

Cytochrome P450 monooxygenases (P450) are membrane-bound hemoproteins that play important roles in conferring protection against both naturally occurring phytochemicals and synthetic organic insecticides. Despite the potential for common modes of detoxification, cross-resistance between phytochemicals and synthetic organic insecticides has rarely been documented. In this study, we examined the responses of a susceptible strain of corn earworm, Helicoverpa zea (Boddie), a polyphagous noctuid, to exposure by an allelochemical infrequently encountered in its host plants and by an insecticide widely used for control purposes. Within a single generation, survivors of xanthotoxin exposure displayed higher levels of tolerance to alpha-cypermethrin than did unexposed control larvae. The F1 offspring of xanthotoxin-exposed survivors also displayed higher alpha-cypermethrin tolerance than did offspring of unexposed control larvae, suggesting that increased alpha-cypermethrin tolerance after xanthotoxin exposure represents, at least in part, heritable resistance. Administration of piperonyl butoxide, a P450 synergist, demonstrated that resistance to both xanthotoxin and alpha-cypermethrin is P450-mediated. Alpha-cypermethrin-exposed survivors, however, failed to show superior growth on xanthotoxin diets. Assays with control larvae, larvae induced by both xanthotoxin and alpha-cypermethrin, and survivors of LD50 doses of both compounds indicated that H. zea midgut P450s are capable of metabolizing both xanthotoxin and alpha-cypermethrin. Metabolism of each compound is significantly inhibited by the presence of the other compound, suggesting that at least one form of P450 in H. zea midguts degrades both compounds and may constitute the biochemical basis for possible cross-resistance. Compared with control larvae, xanthotoxin- and alpha-cypermethrin-induced larvae displayed 2- to 4-fold higher P450-mediated metabolism of both compounds. However, xanthotoxin- and alpha-cypermethrin-exposed survivors exhibited much higher (2.5- to 11-fold) metabolism of both compounds than did the induced larvae. The metabolism results, like the bioassay results, are consistent with the interpretation that increased alpha-cypermethrin tolerance after xanthotoxin exposure is attributable mainly to heritable resistance.     

Resistance monitoring of Helicoverpa armigera (Lepidoptera Noctuidae) to Bacillus thuringiensis insecticidal protein in China

Sensitivities of Helicoverpa armigera (Hübner) field populations to Bacillus thuringiensis (Bt) insecticidal protein CrylAc were monitored during 1998-2000 in China. A total of 41 strains was sampled, and most of them were collected from Bt cotton planting regions. The range of IC50 values (concentration producing 50% inhibition of larval development to third instar) among different populations in 1998,1999, and 2000 were 0.020-0.105 microg/ml, 0.016-0.099 microg/ml, and 0.016-0.080 microg/ml, respectively. Diagnostic concentration studies (IC99) showed that the percentage of individuals reaching third instar ranged from 0 to 4.35%, with only eight of the 41 tested populations showing values above 0%. Also interesting was a trend over successive years in which fewer populations contained individuals that survived the diagnostic concentration by reaching third instar. Considering these data, it was determined that the field populations sampled during the 3-yr study were susceptible to CrylAc protein, and that movement toward resistance among H. armigera populations was not apparent.     

Species diagnosis and Bacillus thuringiensis resistance monitoring of Heliothis virescens and Helicoverpa zea (Lepidoptera: Noctuidae) field strains from the southern United States using feeding disruption bioassays

Validation of a feeding disruption bioassay for the detection of resistance to Bacillus thuringiensis toxin and species identification is reported using field strains of Heliothis virescens and Helicoverpa zea collected from the southern United States in 1998. Feeding disruption is measured by a lack of fecal production from larvae exposed to a diagnostic concentration of CryIAc in a blue indicator diet. The bioassay provided rapid (24 h) diagnosis of the species composition of larvae tested and also monitored for the presence of resistance in H. virescens. An additional diagnostic concentration was established for monitoring resistance in H. zea. A probit model was used to compare the fecal production responses of insect strains over a range of CryIAc doses. Probability calculations, derived from our assay results, are also presented to aid in the interpretation of future results from field trials. Integration of the feeding disruption bioassay into integrated pest management programs is discussed.     

Overwintering of Heliothis virescens (F.) and Helicoverpa zea (Boddie) (Lepidoptera: Noctuidae) in cotton fields of northeast Mississippi

Following the 1995-2001 crop seasons, population densities of pupae of tobacco budworm, Heliothis virescens (F.), and corn earworm, Helicoverpa zea (Boddie), overwintering in cotton fields, Gossypium hirsutum L., in Monroe County, MS, were estimated by digging a total of 43,542 m-row. Densities of pupae varied greatly among years with the highest densities occurring in years with low rainfall in the late summer and early fall. The sex ratio of pupae was variable and significantly male-biased in some years. The total area-wide, tobacco budworm population emerging from all overwintering sites was estimated from catches in pheromone traps whose absolute efficiency is known. After the introduction of Bacillus thuringiensis Berliner (Bt)-transgenic cotton in 1996 but before a several-fold increase in use of herbicide resistant (HR)-transgenic cotton cultivars in 2001, local cotton fields accounted for <2% of the total overwintered tobacco budworm population. In the pretransgenic cotton production era, it is estimated cotton fields typically accounted for <10% of the overwintered tobacco budworm population. Driving surveys of the tillage status of cotton fields and pheromone trapping to determine the timing of adult emergence in the spring indicate that--until the 2000 crop season--postseason tillage destroyed most tobacco budworm and corn earworm pupae overwintering in cotton fields. Adoption of HR-transgenic cultivars has quintupled cotton field area planted no-till, but the contribution of cotton fields to overwintered tobacco budworm populations has remained low. Preemptive tillage of cotton fields to kill overwintering tobacco budworm pupae would not appreciably suppress in-season population densities of this pest. However, the absence of such tillage might increase the rate at which the tobacco budworm develops resistance to insecticides and counteradaptation to antibiotic host plant resistance traits including the Bt-transgenic trait.