Susceptibility of Helicoverpa armigera from different host plants in northern China to Bacillus thuringiensis toxin Cry1Ac

Helicoverpa armigera (Hübner) (Lepidoptera: Noctuidae) is a serious pest of cotton and many other crops in northern China. To evaluate the contribution of alternative hosts as an effective refuge for transgenic cotton expressing the Bacillus thuringiensis (Bt) Cry1Ac toxin, the susceptibility to this toxin was measured in progeny derived from field-collected H. armigera larvae and pupae from different hosts in the Xiajin’s region of the Shandong Province in northern China. During 2008-2010, progeny from a total of 258,56,184 and 160 single-pair crosses derived from wheat (first-generation), Bt cotton (second-generation), Bt cotton (third-generation), and corn (third-generation) were screened on Cry1Ac diets, respectively. Based on relative average development rates (RADR) of H. armigera larvae in these F₁ tests, the second and third-generation moths emerging from Bt cotton fields were more tolerant to the Bt toxin than the first and third-generation moths emerging from wheat and corn each year. These results suggest that there is significant variation in susceptibility to Bt toxins among H. armigera populations derived from different host crops. Alternate crops, such as corn, that maintain Bt susceptible populations of H. armigera could be used as refugia to minimize the evolution of resistance to Bt cotton.     

Manual topping decreases bollworm infestations in cotton cultivation in Mali

In sub-Saharan areas of Africa, cotton growers no longer cut the shoot tips from plants (topping), although manual topping was promoted at the start of the 20th century to improve yield and, surprisingly, to reduce pest incidence. In these areas, the bollworms Helicoverpa armigera Hübner, Earias spp., and Diparopsis watersi Rothschild are responsible for the majority of cotton yield losses, and the use of pyrethroids has resulted in resistance in field populations of H. armigera. In the face of these problems and given the scarcity of literature on the effects of topping on pest control, we assessed bollworm infestation levels in 12 trials comparing manual topping and non-topping cotton plots in Mali over a six-year period (2002, 2003, and 2005 to 2008). Topping was performed at the emergence of the 15th sympodial branch or at 10 days after the first flower opening. Our results showed no significant difference in seed cotton yields between topped and non-topped cotton. Bollworm infestations (all species) were always lower on topped cotton and 7 out of 12 trials showed significantly lower infestations on topped cotton. In plots of topped cotton, we recorded an average of 56% fewer H. armigera larvae, 68% fewer Earias spp. larvae, and 71% fewer D. watersi larvae with respectively 5, 4, and 3 out of 12 trials with significant differences in favor of topping and no significant difference in favor of non-topping. To our knowledge, our study is the first to report decreases in D. watersi larval infestation with cotton topping. Further studies are required to understand the mechanisms involved in these effects and to ensure that topping is economically attractive for farmers.     

Frequency of Bt resistance alleles in Helicoverpa armigera in 2007-2009 in the Henan cotton growing region of China

Helicoverpa armigera (Hübner) (Lepidoptera: Noctuidae) is a key insect pest of cotton in the Henan cotton growing region of China. In this region, cotton is grown on small acreages in rich agricultural landscapes, contrary to cropping systems in the United States or Australia. Under such cropping regimes, naturally occurring refuges (with non-Bt plants) may be sufficient to prevent H. armigera resistance development to Bt toxins. In order to gain a timely understanding of the evolution of resistance of H. armigera to Bt toxin after continuous cultivation of Bt cotton for c. 10 years, we assessed the frequency of alleles conferring resistance to Cry1Ac toxin in field populations of H. armigera sampled from Xinxiang County in Henan province during 2007-2009. Screening F₁ and F₂ generations from isofemale lines, derived from female moths trapped in the field, were used with a discriminating dose of Cry1Ac diet to estimate the frequency of resistance alleles. Totals of 625, 516 and 488 isofemale lines were screened for the F₁ generation in 2007, 2008 and 2009, respectively. Resistance gene frequency in Xinxiang fluctuated between 0.0000 and 0.0005, and it did not increase significantly from 2007 to 2009. Based on the relative average development rates (RADR) of H. armigera larvae in F₁ tests, no substantial increase in Cry1Ac tolerance was found in the Xinxiang region over the 3-yr period.     

Season-long expression of Cry1Ac and Cry2Ab proteins in Bollgard II cotton in Australia

Bollgard II cotton has been grown commercially in Australia since 2003 for control of the primary target species Helicoverpa armigera (Hübner) and Helicoverpa punctigera (Wallengren) Larvae of both species have been reported to survive at low frequencies on Bollgard II with larvae >8 mm recorded in between 7 and 18% of the area planted to Bollgard II cotton between 2005/06 and 2007/08. F₁ and F₂ tests have shown that this is not due to the presence of resistance genes in the surviving larvae. To understand if fluctuations in the expression of the Cry proteins in Bollgard II allow some larvae to survive, plant tissue samples were taken from five Bollgard II cultivars throughout the growing season within fields and from different farms within a production region between 2007 and 2010. The data indicate that the expression of both Cry proteins is similar to the known resistance-monitoring diagnostic concentrations and relatively uniform between fields within a farm and between farms within a region, with less than one-third of the tests at this level of variation being significant. However, there were intra-seasonal changes in expression of both Cry proteins and differences in expression between plant structures and between cultivars for both Cry proteins. Further work is needed to establish if this variation in Cry protein content in Bollgard II cotton affects the control of Helicoverpa spp. in the field or whether plant-physiological and pest-behavioural factors underlie the occasional occurrence of Helicoverpa larval survival on Bollgard II cotton.     

Impacts of transgenic Bt cotton on a non-target pest, Apolygus lucorum (Meyer-Dür) (Hemiptera: Miridae), in northern China

Transgenic Bt (Bacillus thuringiensis) cotton has been effectively used to control the cotton bollworm, Helicoverpa armigera (Hübner) in China. However, in recent years, following the wide commercialization of Bt cotton in northern China there have been frequent outbreaks of the non-target pest Apolygus lucorum (Meyer-Dür). To clarify how transgenic cotton contributes to these outbreaks, a four-year field investigation of population dynamics and laboratory life table studies were carried out from 2007 to 2010 to evaluate the impact of two transgenic cotton cultivars (SGK321 expressing Cry1Ac + CpTI and GK12 expressing Cry1Ac) and their corresponding parental non-transgenic lines (Shiyuan321 and Simian3) on A. lucorum. There were no significant differences in the population densities of A. lucorum found in Bt cotton and non-Bt cotton plots, whether one compared those that had received insecticide treatments or those that had not. However, population densities of A. lucorum were significantly lower in pesticide treated plots than in controls. Furthermore, there were no significant differences in the net reproductive rates, generation times or intrinsic rates of increase of A. lucorum when reared on either Bt or non-Bt cotton cultivars. These results suggest that Bt cotton has no direct positive or negative effects on the biology of A. lucorum, so the most logical explanation for the observed outbreaks is the decrease in pesticide applications following the commercial release of Bt cotton.     

Damage and survivorship of fall armyworm (Lepidoptera: Noctuidae) on transgenic field corn expressing Bacillus thuringiensis Cry proteins

Field corn, Zea mays L., plants expressing Cry1Ab and Cry1F insecticidal crystal (Cry) proteins of Bacillus thuringiensis (Bt) Berliner are planted on considerable acreage across the Southern region of the United States. The fall armyworm, Spodoptera frugiperda (J.E. Smith), is an economically important pest during the mid-to-late season on non-Bt and some commercial Bt corn hybrids. The objective of this study was to quantify foliar injury and survivorship of fall armyworm on transgenic corn lines expressing Cry1Ab or Cry1F Bt proteins. Corn lines/hybrids expressing Cry1Ab, Cry1F, and a conventional non-Bt cultivar were evaluated against artificial infestations of fall armyworm in field trials. Larvae (second instars) of fall armyworm were placed on corn plants (V8-V10 stages). Leaf injury ratings were recorded 14 d after infestation. Hybrids expressing Cry1F had significantly lower feeding injury ratings than non-Bt corn plants. Development and survivorship of fall armyworm on Bt corn lines/hybrids were also evaluated in no-choice laboratory assays by offering freshly harvested corn leaf tissue to third instars. Transgenic corn hybrids expressing Cry1Ab or Cry1F significantly reduced growth, development, and survivorship of fall armyworm compared to those offered non-Bt corn tissue. However, 25-76% of third instars offered Bt corn leaf tissues successfully pupated and emerged as adults. These results suggest Cry1Ab has limited effects on fall armyworm; whereas Cry1F demonstrated significant reductions in foliar injury and lower survivorship compared to that on non-Bt corn tissues. Although fall armyworm is not considered a primary target for insect resistance management by the U.S. Environmental Protection Agency, these levels of survivorship could impact selection pressures across the farmscape, especially when considering that transgenic Bt cotton cultivars express similar Cry (Cry1Ac or Cry1F) proteins.     

Relative Fitness of Helicoverpa armigera (Lepidoptera: Noctuidae) on Seven Host Plants: A Perspective for IPM in Brazil

The cotton bollworm Helicoverpa armigera (Hübner) (Lepidoptera: Noctuidae) is a widespread pest of many cultivated and wild plants in Europe, Africa, Asia, and Australia. In 2013, this species was reported in Brazil, attacking various host crops in the midwestern and northeastern regions of the country and is now found countrywide. Aiming to understand the effects of different host plants on the life cycle of H. armigera, we selected seven species of host plants that mature in different seasons and are commonly grown in these regions: cotton (Gossypium hirsutum, “FM993”), corn (Zea mays, “2B587”), soybean (Glycine max, “99R01”), rattlepods (Crotalaria spectabilis), millet (Pennisetum glaucum, “ADR300”), sorghum (Sorghum bicolor, “AGROMEN70G35”), and cowpea (Vigna unguiculata, “SEMPRE VERDE”). The development time of immatures, body weight, survivorship, and fecundity of H. armigera were evaluated on each host plant under laboratory conditions. The bollworms did not survive on corn, millet, or sorghum and showed very low survival rates on rattlepods. Survival rates were highest on soybean, followed by cotton and cowpea. The values for relative fitness found on soybean, cotton, cowpea, and rattlepods were 1, 0.5, 0.43, and 0.03, respectively. Survivorship, faster development time, and fecundity on soybean, cotton, and cowpea were positively correlated. Larger pupae and greater fecundity were found on soybean and cotton. The results indicated that soybean, cotton, and cowpea are the most suitable plants to support the reproduction of H. armigera in the field.     

Crop hosts and genotypic resistance influence the biological activity of Bacillus thuringiensis towards Helicoverpa armigera

Studies on the influence of genotypic resistance on biological activity of a commercial formulation of Bacillus thuringiensis (Bt) and pure Bt toxin Cry1Ac were carried out to develop appropriate strategies for pod borer, Helicoverpa armigera management in chickpea, sorghum, pigeonpea and cotton. The interaction effects of host plant resistance and biological activity of commercial Bt/Cry1Ac were studied by incorporating the lyophilized tissues of chickpea leaves, milk stage sorghum grain, pigeonpea pods and cotton squares into the artificial diet with and without Bt formulation or Cry1Ac. The H. armigera larval weights were significantly lower in insects reared on diets with square powder of the insect - resistant Bt-cotton RCH 2 + Bt/Cry1Ac and pod powder of insect - resistant pigeonpea genotype, ICPL 332WR + Bt/Cry1Ac as compared to the larvae reared on diets with leaf powder of H. armigera susceptible chickpea genotype, ICCC 37 and the standard artificial diet. Pupation and adult emergence were significantly lower in insects reared on diets with tissues of pod borer-resistant genotypes + Bt/Cry1Ac as compared to insects reared on diets with tissues of the insect susceptible genotypes + Bt/Cry1Ac. Insects reared on diets containing insect-resistant and -susceptible genotypes of sorghum, pigeonpea and cotton and pod borer-resistant genotype of chickpea (ICC 506EB) + Bt/Cry1Ac did not lay any eggs. However, eggs were laid by the insects reared on diets containing pod borer-susceptible genotype of chickpea, ICCC 37 and on the standard artificial diet + Bt/Cry1Ac. The insects reared on diets with sorghum genotype, ICSV 745, and Bt-cotton, RCH 2 without Bt/Cry1Ac also did not lay eggs. The results suggested that Bt/Cry1Ac is more effective for management of H. armigera when deployed in combination with insect-resistant genotypes of cotton, chickpea, pigeonpea and sorghum.     

Relationships between Cry1Ac and Cry2Ab protein expression in field-grown Bollgard II® cotton and efficacy against Helicoverpa armigera and Helicoverpa punctigera (Lepidoptera: Noctuidae)

Seven-day laboratory bioassays with first-instar Helicoverpa armigera and Helicoverpa punctigera larvae were conducted using leaves from field-grown Bollgard II^® cotton during the 2009/10 season and larval mortality and development recorded. Leaves were from three farms in the St George region and two fields on one farm at Emerald in eastern Australia. The Cry1Ac and Cry2Ab content of leaves from the same samples was determined (see Knight et al., 2013) and the relationships between Cry protein content and larval mortality and development were examined. The Cry1Ac protein content was between 3.98 and 12.08 μg/g during the growing season, while Cry2Ab content ranged between 300.6 and 953.3 μg/g. Cry1Ac and Cry2Ab content of leaves were highly correlated (r = 0.8276, P < 0.001). Seven-day mortality of H. punctigera larvae was close to 100% throughout the season. H. armigera mortality was close to 100% early in the season, but fell to ∼65% by mid-to late February in the laboratory bioassays. Fitting three-dimensional non-linear models associating Cry1Ac and Cry2Ab content with H. armigera and H. punctigera mortality elucidated the relative importance of the two proteins in determining larval mortality; for this analysis, data were pooled with data from an isoline study to provide better sampling of the three-dimensional surface being modelled. For both Helicoverpa species, the fitted mortality response to the Cry1Ac protein was close to its maximum at protein concentrations above ∼3 μg/g Cry1Ac. For H. punctigera, response to the Cry2Ab protein was close to maximal once Cry2Ab was greater than ∼200 μg/g. In contrast, the fitted H. armigera mortality response to Cry2Ab increased steadily with concentration up to ∼1200 μg/g Cry2Ab. These responses led to markedly different response surfaces for the two species; H. punctigera mortality was close to 100% at most places on the response surface, while for H. armigera the response surface showed stronger increases in mortality with concentration for Cry2Ab than for Cry1Ac. These results can be interpreted as meaning that at the plant-expressed range of concentrations in Bollgard II cotton the two proteins are approximately equally important for H. punctigera but that changes in Cry2Ab content more strongly influences changes in larval mortality in H. armigera than does Cry1Ac, with Cry1Ac contributing a consistent 40–45% mortality for concentrations above 3 μg/g. For H. armigera, there was no evidence of either synergism or antagonism between Cry1Ac and Cry2Ab proteins (P > 0.05) but this aspect was not testable for H. punctigera because mortality was mostly close to 100%.     

Comparative susceptibility of Ostrinia furnacalis, Ostrinia nubilalis and Diatraea saccharalis (Lepidoptera: Crambidae) to Bacillus thuringiensis Cry1 toxins

Transgenic corn hybrids that express toxins from Bacillus thuringiensis (Bt) are highly effective against the European corn borer, Ostrinia nubilalis (Hübner), and the closely related Asian corn borer, Ostrinia furnacalis (Guenée). Since the registration of Bt corn hybrids in the U.S. in 1996, there has been a great deal of information generated on O. nubilalis. However, relatively little information exists for O. furnacalis. To help determine whether the information generated for O. nubilalis can be leveraged for decisions regarding the use of transgenic Bt corn against O. furnacalis, experiments were designed to determine whether the pattern of sensitivity to various Bt Cry1 toxins is similar between the two species. Test insects included laboratory-reared O. furnacalis originating from Malaysia, a Bt-susceptible laboratory colony of O. nubilalis maintained at the University of Nebraska-Lincoln (UNL) and an out-group consisting of the sugarcane borer, Diatraea saccharalis (F.), from Louisiana which represents a different genus from the same family. O. furnacalis and O. nubilalis exhibited a similar pattern of susceptibility to all the Cry1 toxins and were highly susceptible to the range of Bt toxins tested including Cry1Aa, Cry1Ab, Cry1Ac and Cry1F. Both of the Ostrinia species were more tolerant to Cry1Ba compared with D. saccharalis, although sensitivity of O. furnacalis was intermediate and did not differ significantly from that of O. nubilalis and D. saccharalis. D. saccharalis was also susceptible to the range of toxins tested but unlike the two Ostrinia species, was more tolerant to Cry1F and more susceptible to Cry1Ba. These results indicate that both of the Ostrinia corn borer species are similar in sensitivity to the Cry1Aa, Cry1Ab, Cry1Ac, Cry1Ba and Cry1F toxins, thus suggesting shared toxin receptors and mechanisms of toxicity for the two species.     

Influences of Cry1Ac Broccoli on Larval Survival and Oviposition of Diamondback Moth

Larval survival and oviposition behavior of three genotypes of diamondback moth, Plutella xylostella L. (Lepidoptera: Plutellidae), (homozygous Cry1Ac-susceptibile, Cry1Ac-resistant, and their F₁ hybrids), on transgenic Bacillus thuringiensis (Bt) broccoli expressing different levels of Cry1Ac protein were evaluated in laboratory. These Bt broccoli lines were designated as relative low, medium, and high, respectively, according to the Cry1Ac content. Untransformed brocccoli plants were used as control. Larval survival of diamondback moth on non-Bt leaves was not significantly different among the three genotypes. The Cry1Ac-resistant larvae could survive on the low level of Bt broccoli plants, while Cry1Ac-susceptible and F₁ larvae could not survive on them. The three genotypes of P. xylostella larvae could not survive on medium and high levels of Bt broccoli. In oviposition choice tests, there was no significant difference in the number of eggs laid by the three P. xylostella genotypes among different Bt broccoli plants. The development of Cry1Ac-susceptible and Cry1Ac-resistant P. xylostella on intact Bt plants was also tested in greenhouse. All susceptible P. xylostella larvae died on all Bt plants, while resistant larvae could survive on broccoli, which expresses low Cry1Ac protein under greenhouse conditions. The results of the greenhouse trials were similar to that of laboratory tests. This study indicated that high dose of Bt toxins in broccoli cultivars or germplasm lines is required for effective resistance management.     

Herbivory tolerance of cotton expressing insecticidal proteins from Bacillus thuringiensis: responses to damage caused by Helicoverpa spp. and to manual bud removal

Transgenic cotton varieties expressing Cry IA(c) insecticidal proteins from Bacillus thuringiensis (`Bt cotton') remain vulnerable to non-lepidopteran insects. In addition, they are susceptible to lepidopteran pests when the efficacy of Bt toxins falls because of ontogenetic and/or environmental factors. Hence the importance of knowing to what extent Bt cotton is able to tolerate damage. The degree of tolerance of Bt cotton to actual and simulated insect damage was assessed in three field experiments. Exp. 1 compared the effects of Helicoverpa spp. (Lepidoptera: Noctuidae) on the growth, development and yield of Bt cotton with those of its near isogenic non-Bt counterpart in two genetic backgrounds (Siokra V15, Sicala V2) under two regimes of chemical control of insects (S1: nine insecticide applications during the growing cycle, S2: six insecticide applications). Exp. 2 compared insecticide-protected Bt crops with crops manually damaged to simulate (a) early-season loss of vegetative buds, (b) loss of flowerbuds, and (c) loss of both vegetative and reproductive buds. Also using manual damage, Exp. 3 evaluated the effect of timing of flowerbud loss on the yield and maturity time of insecticide-protected Bt crops. In Exp. 1, well-protected Bt crops (S1) yielded 24% more than their less-protected counterparts (S2). The less protected crops had, however, substantially more immature fruit at the end of the season highlighting a considerable potential for recovery. Poor soil conditions, interacting with season length accounted for the difference between potential and actual compensation in crops that were exposed to almost continuous damage by Helicoverpa spp. Under more favourable growing conditions, maturity was delayed but yield of damaged Bt crops was unaffected by discrete episodes of simulated herbivory (Exps. 2 and 3). The introduction of B. thuringiensis genes into cotton does not seem to have reduced the considerable capacity of the crop to tolerate insect damage, and this attribute should be considered in the development of pest management strategies for Bt crops.     

Sudanese cotton and the whitefly: a case study of the emergence of a new primary pest

In the late 1970s the whitefly Bemisia tabaci (Gennadius) became the primary cotton pest in the Sudan, superseding the American bollworm Heliothis armigera (Hübner). DDT and a DDT/dimethoate combination were used to control the bollworm and, simultaneously, jassids and whiteflies. B. tabaci, a secondary pest at first, became resistant to dimethoate by frequent selections from 1964 onwards. At the same time, fertility stimulation occurred due to DDT residues on cotton plants. Finally, resistance reached a level so that the whitefly were not controlled by dimethoate, monocrotophos or other organophosphorus insecticides, and stimulation by DDT could exert its full effect. The consequence of this was a tremendous flare-up of the whitefly by 1980/81. This train of events was concluded from laboratory and field studies of the resistance patterns, as well as the acceleration effects from DDT residues on plants to the whitefly. A current hypothesis claiming that the problems arose from the elimination of beneficial insects through insecticide applications is reviewed in the light of experimental evidence and practical experience.     

Feeding disruption tests for monitoring the frequency of larval lepidopteran resistance to Cry1Ac, Cry1F and Cry1Ab

An alternative to traditional larval lepidopteran resistance-monitoring bioassays was developed. Feeding disruption tests were developed for detecting insects resistant to three Bacillus thuringiensis (Bt) proteins: Cry1Ac, Cry1F and Cry1Ab. The assays rely on a diagnostic dose of Bt toxin in 100-μl hydratable meal pads of artificial diet containing blue indicator dye. The assay was formatted as a portable (palm-sized) plastic plate containing an array of 16 test wells, each containing a single hydratable meal pad with one insect added per well. The diagnostic dose was the concentration of Bt in meal pad rehydration solution that reduced 24 h dyed fecal production of Bt-susceptible neonates to ≤2 fecal pellets per larva. Bt-resistant neonates were able to consume the diagnostic dose of the insecticidal protein and produce >2 blue fecal pellets. The feces were distinctly visible on the white background of the feeding disruption test plate. Diagnostic doses were determined with lab-strain Bt-susceptible Heliothis virescens and Helicoverpa zea. For H. virescens, the diagnostic doses were 10, 20 and 15 μg/ml for Cry1Ac, Cry1F and Cry1Ab, respectively. For H. zea, the diagnostic doses were 40, 200 and 500 μg/ml, respectively. The assays were validated against a lab-strain of Bt-resistant H. virescens and with susceptible larval H. virescens collected as eggs from field-grown tobacco in North Carolina.     

Threshold-based interventions for cotton pest control in West Africa: What's up 10 years later?

In the late 1980s, after a long period during which insecticides were sprayed at preset dates to control cotton (Gossypium hirsutum L.) pests and their damage, some French-speaking countries in sub-Saharan areas decided to disseminate a special form of crop protection approach among smallholders, i.e. targeted staggered control (LEC, for Lutte étagée ciblée). According to this approach, decisions on some insecticide applications were made on the basis of infestation levels or the extent of crop injury caused by major pests: Aphis gossypii Glover aphids, Haritalodes (=Syllepte) derogata F., leaf-eating caterpillars, and more generally Helicoverpa armigera (Hübner), Diparopsis watersi Rothschild, Earias insulana (Boisduval) and Earias biplaga (Walker) bollworms. Polyphagotarsonemus latus (Banks) mites were sometimes included on this list. Today, the calendar-based or conventional program is still widely implemented with some changes in insecticides applied due to the development of pyrethroid resistance in H. armigera. Depending on the country, protection programs based on pest monitoring have been preserved or replaced by programs still using thresholds (staggered or true). In Benin, there are two forms of LEC tailored to two regions delineated according to the extent of damage caused by of bollworms that live inside cotton bolls, i.e. Thaumatotibia (=Cryptophlebia) leucotreta (Meyrick). The logical follow-up to LEC, involving true thresholds, was developed in Mali, Cameroon and, recently, Senegal. Cameroon opted for a sequential plan for individual decision program or LOIC (for Lutte après observation individuelle des chenilles), based on control after sequential sampling of bollworms. A calendar program with additional applications of insecticides based on a particular scouting of H. armigera was developed in Togo. In Ivory Coast, the use of true thresholds is limited to the beginning of the cotton crop cycle whereas in Burkina Faso true thresholds are used after the first two calendar sprayings. The present article describes the diversity of these cotton protection programs, sampling methods and associated intervention thresholds based on pests or injury levels in addition to the advantages and constraints associated with their adoption.     

Effects of pigment glands and gossypol on growth,development and insecticide-resistance of cotton bollworm (Heliothis armigera (Hübner))

Cotton bollworm (Heliothis armigera) is a major pest of cotton and other crops. It is important to understand the mechanisms of insecticide tolerance of cotton bollworm on cotton cultivars with host plant resistance to this insect pest. The objectives of this study were to investigate the effects of cotton pigment glands and their gossypol on the growth, development and insecticide tolerance of cotton bollworm. Three pairs of cotton isogenic lines with glanded versus glandless leaves, as well as artificial diets with 5 levels of gossypol, were used to raise cotton bollworm larvae for five generations. The growth, development and insecticide tolerance of larvae were studied. The results indicated that the cotton pigment glands and higher levels of gossypol resulted in a significant decrease in larval weights and moth eclosion rates and delayed the development of larvae and pupae. Larvae that fed on glanded cotton leaves were significantly more tolerant to two insecticides, cyhalothin and monocrotophos, than those fed on glandless cotton leaves. LD₅₀ values were only increased where they were in amount per unit body weight, and not where were calculated in amount per individual. Also the insecticide tolerance of cotton bollworm larvae increased as the gossypol content was raised from 0 to 0.225% in artificial diets. Meanwhile, the activities of two detoxifying enzymes, carboxylesterase and glutathione s-transferase, were significantly higher in the larvae fed on glanded cotton leaves than those fed on glandless cotton leaves. The activities of two enzymes also increased greatly with the increase of gossypol content from 0 to 0.225% in artificial diets. Across 5 generations of feeding and investigation, the significant inhibition effect on larval growth and larval tolerance to two pesticides were found to be only associated with the feeding by current generation, but were not related to previous diets. The activities of two detoxifying enzymes in larvae were also not enhanced significantly when they were fed continuously on glanded cotton leaves or artificial diet with high gossypol. These results indicated that pigment glands and higher levels of gossypol not only inhibited the growth of cotton bollworm larvae but also enhanced their insecticide tolerance. However, the inhibition effect and enhanced insecticide tolerance were inducible but could not be accumulated or inherited. These results will help us better understand the interaction and co-evolution of insecticide tolerance in larvae of cotton bollworm and host chemical components, and also has provided useful information on cotton bollworm management in cotton production, especially in glandless cotton.     

Comparative assessment of the field-susceptibility of Sesamia nonagrioides to the Cry1Ab toxin in areas with different adoption rates of Bt maize and in Bt-free areas

A three year (2003-2005) field study compared the susceptibility to the Cry1Ab toxin, expressed in Bt maize, of Mediterranean corn borer (MCB) Sesamia nonagrioides populations collected from areas with different adoption rates of Bt maize in Spain with Bt-free areas in Greece. Spain is the only European country where the cultivar Compa CB derived from the event Bt176 was commercially grown, from 1998 to 2005. The large decrease of the titer of the toxin in this cultivar at later growth stages represented the worst-case scenario for resistance development of MCB, since larvae of the second and third generations were exposed to sublethal concentrations of Cry1Ab toxin. Our data revealed that the variation in susceptibility to Cry1Ab for the MCB Spanish field populations analyzed in the three years was very low, with LC₅₀ values fluctuating between 12 and 30 ng Cry1Ab/cm², regardless of the region of origin, the type of maize (Bt or non-Bt) and the year. Furthermore, no significant differences were found when comparisons were made with a laboratory population (LC₅₀ values: 18-26 ng Cry1Ab/cm²) or with field populations from Greece (Bt-free areas), which displayed LC₅₀ values ranging between 22 and 27 ng Cry1Ab/cm². Standardizing bioassay protocols proved to be essential for obtaining comparable results. These findings suggest that resistant MCB populations did not evolve in those Spanish maize areas where Compa CB was largely cultivated for eight years, contradicting the expected rapid development of resistance under these unfavourable conditions. Additionally, our results can be used as baseline indices in post-market resistance monitoring programs if Bt maize is introduced in Greece. Further studies should continue, since the insights gained from a resistance monitoring program may help to enhance the durability of Bt maize.     

Susceptibility of Cry1Ab-susceptible and -resistant sugarcane borer to transgenic corn plants containing single or pyramided Bacillus thuringiensis genes

Multiple independent trials were conducted to evaluate the performance of Cry1Ab-susceptible (Cry1Ab-SS), -heterozygous (Cry1Ab-RS), and -resistant (Cry1Ab-RR) genotypes of the sugarcane borer, Diatraea saccharalis (F.), on eight commercial hybrids and six experimental corn lines. The commercial varieties included two non-Bt and six Bt corn hybrids that expressed a single Bt protein (either Cry1Ab or Cry1F) targeting above-ground lepidopteran pests. The six experimental lines consisted of two non-Bt and four Bt corn lines, two expressing just the Cry1Ab protein and two containing the pyramided-genes Cry1A.105 and Cry2Ab2 (event MON 89034). Larval mortality on non-Bt corn leaf tissue ranged from 6 to 45% after 12 d across insect genotypes. The 12 d mortality of Cry1Ab-SS on leaf tissue of commercial Cry1Ab or Cry1F corn was 96-100%, whereas it was 80-96% for Cry1Ab-RS and 68-78% for Cry1Ab-RR. On intact plants, 39-64% of larvae survived on non-Bt corn plants after 21-25 d. Larval survivorship on intact plants of commercial Cry1Ab or Cry1F corn was 0-8.1% for Cry1Ab-SS, 1.3-34% for Cry1Ab-RS, and 19-51% for Cry1Ab-RR. Larvae of Cry1Ab-RR and -RS also caused significant plant injury to most of the commercial Bt corn hybrids, especially to the Cry1Ab corn. Cry1Ab resistance in D. saccharalis was incompletely dominant on commercial Bt corn hybrids. However, both experimental lines with pyramided genes of Cry1A.105 and Cry2Ab2 provided complete control of all three insect genotypes in both leaf tissue and intact plant tests. Results of this study suggest that MON 89034 should offer a means for Bt resistance management in D. saccharalis.     

Monitoring Cry1Ab susceptibility in Asian corn borer (Lepidoptera: Crambidae) on Bt corn in the Philippines

Between 2002 and 2004, collections of egg masses of Asian corn borer (ACB), Ostrinia furnacalis (Guenée) were made from corn-planting sites on the major Philippine islands of Luzon (Laguna, Pangasinan, Camarines Sur and Isabela provinces) and Mindanao (Bukidnon and South Cotabato provinces). The resulting neonates were bioassayed for susceptibility to Bacillus thuringiensis (Bt) Cry1Ab protein. The median lethal concentrations (LC₅₀s) for the different collections ranged from 0.42 to 2.37 ng/cm². The bioassay results suggest that Philippine corn borer populations were highly susceptible to Cry1Ab protein prior to the widespread deployment of Bt corn. The upper limit of the estimated LC₉₉ (104 ng/cm²) from the pooled bioassay data was selected as a candidate diagnostic concentration and subsequently tested on eleven ACB populations. Results of the validation assays showed that the mortality response of all the tested ACB populations was higher than the expected mortality (99%). Therefore, the concentration of 104 ng/cm² was used to monitor susceptibility in ACB populations in the Philippines. Monitoring of field populations during 2009 in areas where Bt corn had been grown for 3 years found some enhanced survival of neonates at the diagnostic concentration but progeny of the diagnostic-concentration survivors did not survive on Bt corn, indicating that ACB populations in the Philippines remain susceptible to Cry1Ab-containing Bt corn hybrids.