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Effect of husk characters on resistance to corn earworm (Lepidoptera: Noctuidae) in high-maysin maize populations.

Two maize (Zea mays L.) breeding populations with very high concentrations of maysin, a silk-expressed flavone glycoside, were tested for their ability to resist ear damage by the corn earworm, Helicoverpa zea Boddie, under field conditions. Tests were conducted in 2000 and 2001 at multiple locations in Georgia. The high maysin populations, EPM6 and SIM6, as well as resistant and susceptible checks, were scored for silk-maysin content, H. zea damage, and husk characters. In 2000, there was a negative correlation between husk tightness and earworm damage at three of five locations, while there was no significant correlation between damage and maysin content at any location. In 2001, EPM6 and SIM6 had approximately ten times the maysin content of the low-maysin control genotypes; nevertheless, earworm damage to EPM6 and SIM6 was either greater than or not significantly different from the low-maysin genotypes at all locations. The resistant control genotype, Zapalote Chico, had significantly less earworm damage than EPM6 and SIM6 for both years at all locations. The results of this study highlight the importance of identifying and quantifying husk and ear traits that are essential to H. zea resistance in maize.

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Effect of maysin on wild-type, deltamethrin-resistant, and Bt-resistant Helicoverpa armigera (Lepidoptera: Noctuidae).

Larvae of the Old World corn earworm, Helicoverpa armigera (Hübner), were fed diets containing lyophilized silks from maize genotypes expressing varying levels of maysin, a flavone glycoside known to be toxic to the New World corn earworm, Helicoverpa zea Boddie. Three different H. armigera colonies were tested: a wild-type colony (96-S), a colony selected for resistance to deltamethrin (Del-R), and a colony selected for resistance to the Cry1Ac protoxin of Bacillus thuringiensis (Bt-R). A colony of H. zea was also tested as a control. High-maysin silk diets significantly slowed the growth and arrested the development of larvae from all H. armigera colonies compared with low-maysin silk diets, maysin-lacking silk diets, and no-silk control diets. The effects on the H. armigera and H. zea colonies were similar across maysin levels, although H. zea is a larger insect than H. armigera and this overall size difference was observed. Among the H. armigera colonies, maysin effects were generally similar, although 7-d-old Del-R larvae were significantly smaller than 7-d-old Bt-R and 96-S larvae for one no-silk control and two maysin-containing silk treatments. The toxic effect of maysin on the Bt-R and Del-R colonies suggests that physiological mechanisms of H. armigera resistance to Cry1Ac and deltamethrin do not confer cross-resistance to maysin.

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Development, survival and fitness performance of Helicoverpa zea (Lepidoptera: Noctuidae) in MON810 Bt field corn.

Helicoverpa zea (Boddie) development, survival, and feeding injury in MON810 transgenic ears of field corn (Zea mays L.) expressing Bacillus thuringiensis variety kurstaki (Bt) Cry1Ab endotoxins were compared with non-Bt ears at four geographic locations over two growing seasons. Expression of Cry1Ab endotoxin resulted in overall reductions in the percentage of damaged ears by 33% and in the amount of kernels consumed by 60%. Bt-induced effects varied significantly among locations, partly because of the overall level and timing of H. zea infestations, condition of silk tissue at the time of egg hatch, and the possible effects of plant stress. Larvae feeding on Bt ears produced scattered, discontinuous patches of partially consumed kernels, which were arranged more linearly than the compact feeding patterns in non-Bt ears. The feeding patterns suggest that larvae in Bt ears are moving about sampling kernels more frequently than larvae in non-Bt ears. Because not all kernels express the same level of endotoxin, the spatial heterogeneity of toxin distribution within Bt ears may provide an opportunity for development of behavioral responses in H. zea to avoid toxin. MON810 corn suppressed the establishment and development of H. zea to late instars by at least 75%. This level of control is considered a moderate dose, which may increase the risk of resistance development in areas where MON810 corn is widely adopted and H. zea overwinters successfully. Sublethal effects of MON810 corn resulted in prolonged larval and prepupal development, smaller pupae, and reduced fecundity of H. zea. The moderate dose effects and the spatial heterogeneity of toxin distribution among kernels could increase the additive genetic variance for both physiological and behavioral resistance in H. zea populations. Implications of localized population suppression are discussed.

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Effects of MON810 Bt field corn on adult emergence of Helicoverpa zea (Lepidoptera: Noctuidae).

A 3-yr study (1996-1998) was conducted to evaluate the effects of MON810 Bt corn on Helicoverpa zea (Boddie) emergence and to determine whether delayed larval development as a result of Bt intoxication results in higher levels of diapause induction and pupal mortality. In the 1997 study, there was no difference in prepupal mortality between corn types, although significantly more prepupae from Bt plots than from non-Bt plots died in emergence buckets before constructing pupal chambers in 1998. In all years, significantly fewer moths emerged from prepupae collected from Bt plots, suggesting that effects of the expressed Cry1Ab extended to the prepupal and pupal stages. Late plantings of corn showed the greatest reductions in moth emergence from Bt corn because environmental conditions were more conducive to trigger diapause at the time H. zea was developing in these plantings. This was supported by a significantly greater proportion of diapausing pupae remaining in the ground in the late plantings of both Bt and non-Bt corn. For April and early May plantings, larval feeding on Bt corn delayed the time to pupation, although there was no significant difference in moth emergence between corn types for those larvae that successfully pupated. Although Bt expression had less impact on the proportion of moths emerging, the actual number of moths emerging from Bt corn was significantly reduced because fewer larvae reached pupation. Delays in adult emergence, along with significant reductions in adult emergence from MON810 Bt corn, should reduce the rates of colonization in soybean and other late host crops but may also result in asynchrony of mating between individuals emerging from Bt and non-Bt corn. This, in turn, may contribute to the evolution of resistance to Bt corn.

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Effect of MON810 Bt field corn on Helicoverpa zea (Lepidoptera: Noctuidae) cannibalism and its implications to resistance development.

Pairs of Helicoverpa zea (Boddie) larvae reared on diet-incorporated MON810 transgenic leaf tissue of field corn (Zea mays L.) were observed in the laboratory to characterize effects of sublethal levels of Bacillus thuringiensis variety kurstaki (Bt) Cry1Ab endotoxins on cannibalistic behavior and mortality. Feeding on sublethal levels of Bt corn reduced the frequency of cannibalistic behaviors exhibited by H. zea when uneven instars were paired together. Exposure to the Bt endotoxin had no significant effect on when cannibalistic mortality occurred or the level of mortality as a result of cannibalism. Assuming that H. zea larvae reared on nonBt corn tissue behaved in a similar way that resistant larvae would if feeding on Bt tissue, sublethal effects of Cry1Ab intoxication may reduce the chances of successful cannibalism by susceptible larvae and thus play a disproportionate role in the survival of multiple ear infestations. Furthermore, cannibalistic encounters could result in partially resistant larvae feeding on nontoxic food, thus temporarily providing an escape from exposure to the Bt endotoxin. These behavior alterations could increase the selective differential between susceptible individuals and those carrying resistance genes.

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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.

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Plant-incorporated Bacillus thuringiensis resistance for control of fall armyworm and corn earworm (Lepidoptera: Noctuidae) in corn.

Fall armyworm, Spodoptera frugiperda (J.E. Smith), and corn earworm, Helicoverpa zea (Boddie), perennially cause leaf and ear damage to corn, Zea mays L., in the southeastern United States. Transgenic Bacillus thuringiensis (Bt) hybrids with the Bt11, MON810, or 176 events expressing the Cry1Ab insecticidal endotoxin from were evaluated for control fall armyworm and corn earworm at seven locations in Georgia during 1999 and 2000. Corn was planted at the recommended time for each location and 1 and 2 mo later in the southern locations. All Bt events consistently reduced whorl infestation and damage, although event 176 did not prevent whorl damage in the later plantings in the southern locations in both years. All events also reduced seedling damage by the lesser cornstalk borer, Elasmopalpus lignosellus (Zeller), in one trial and stalk infestations and tunnel length by southwestern corn borers, Diatraea grandiosella Dyar, in another trial. Hybrids containing Bt11 and MON810 events reduced ear infestations in all trials, although reductions were small in later plantings. Nevertheless, both events reduced grain damage from earworms and armyworms by an average +/- SE of 52.5 +/- 5.1% in all trials. The hybrid containing event 176 did not reduce ear infestations and damage. Total grain aflatoxin concentrations were not significantly affected by Bt resistance in any trial (N = 17). Yield responses were variable with the prevention of yield loss being proportional to the severity of insect damage. Although plantings made after the recommended time did not consistently benefit from Bt resistance, Bt11 and MON810 events were effective in reducing damage to field corn when large infestations occurred. The Bt11 and MON810 events mitigated the risk of severe lepidopteran damage to corn, thereby making later plantings of corn feasible in double-cropping systems.

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Relative fitness of Cry1A-resistant and -susceptible Helicoverpa armigera (Lepidoptera: Noctuidae) on conventional and transgenic cotton.

Glasshouse and laboratory experiments were conducted to evaluate the relative fitness of Cry1A-susceptible and laboratory-selected resistant strains of Helicoverpa armigera (Hübner). Life history parameters of H. armigera larvae feeding on young cotton plants showed a significant developmental delay of up to 7 d for the resistant strain compared with the susceptible strain on non-Bacillus thuringiensis (Bt) cotton. This fitness cost was not evident on artificial diet. There was no developmental delay in the F1 hybrid progeny from the reciprocal backcross of the resistant and susceptible strains, indicating that the fitness cost is recessive. In two cohorts tested, survival to pupation of resistant larvae on Bt cotton expressing Cry1Ac was 54 and 51% lower than on non-Bt cotton, whereas all susceptible and F1 larvae tested on Cry1Ac cotton were killed. Mortality of susceptible larvae occurred in the first or second instar, whereas the F1 larvae were able to develop to later instars before dying, demonstrating that resistance is incompletely recessive. The intrinsic rate of increase was reduced by >50% in the resistant strain on Cry1Ac cotton compared with the susceptible strain on non-Bt cotton. There was a significant reduction in the survival of postdiapausal adults from the resistant strain and the F1 strains, indicating that there is a nonrecessive overwintering cost associated with Cry1A resistance in H. armigera.

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Comparative production of Helicoverpa zea (Lepidoptera: Noctuidae) from transgenic cotton expressing either one or two Bacillus thuringiensis proteins with and without insecticide oversprays.

Transgenic cotton, Gossypium hirsutum (L.), expressing either one or two Bacillus thuringiensis ssp. kurstaki Berliner (Bt) proteins was compared with the conventional sister line in field experiments with regard to production of bollworm, Helicoverpa zea (Boddie), and bolls damaged by bollworm. The relative numbers of bollworms that developed on Bollgard (Monsanto Co., St. Louis, MO), Bollgard II (Monsanto Co.), and conventional cotton were estimated under nontreated conditions in 2000 and both insecticide-treated and nontreated conditions in 2001-2002 in North Carolina tests. Averaged across seven field studies under nontreated conditions, Bollgard cotton generated statistically similar numbers of large (L4-L5) bollworm larvae compared with the conventional variety; however, Bollgard cotton produced significantly fewer damaged bolls and bollworm adults than the conventional variety. Production of large larvae, damaged bolls, and adults was decreased dramatically by Bollgard II cotton as compared with Bollgard and conventional varieties. When comparing insecticide-treated and nontreated cotton genotypes, both Bt cotton sustained less boll damage than the conventional variety averaged across insecticide regimes; furthermore, Bollgard II cotton had fewer damaged bolls than the Bollgard variety. When averaged across cotton genotypes, pyrethroid oversprays reduced the numbers of damaged bolls compared with the nontreated cotton. Insecticide-treated Bollgard cotton, along with insecticide-treated and nontreated Bollgard II cotton reduced production of bollworm larvae, pupae, and adults. However, the addition of pyrethroid oversprays to Bollgard II cotton seemed to be the best resistance management strategy available for bollworm because no bollworms were capable of completing development under these conditions.

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Relative concentration of Cry1A in maize leaves and cotton bolls with diverse chlorophyll content and corresponding larval development of fall armyworm (Lepidoptera: Noctuidae) and southwestern corn borer (Lepidoptera: Crambidae) on maize whorl leaf profiles.

To manage insect resistance to transgenic crops that express insecticidal proteins from Bacillus thuringiensis (Bt) Berliner, the U.S. Environmental Protection Agency recommends a refuge-based insect resistance management strategy where a percentage of non-Bt (refuge) crop is grown in proximity to a Bt-expressing crop. An important requirement for this strategy is that the toxin exists at a high effective dose for control of the target pest(s), so that heterozygous individuals in the population do not reach adulthood. Factors that cause reduced levels of toxin in the plant are a threat to this strategy. We quantified Cry1Ab from different areas of the maize, Zea mays L., leaf. In general, the distal tip of the V7 maize leaf had a higher concentration of Cry1Ab compared with the middle section of the V7 leaf, and the middle section of the developing V9 leaf had the lowest concentration of Cry1Ab. When these sections of maize tissue were fed to fall armyworm, Spodoptera frugiperda (J.E. Smith), and southwestern corn borer, Diatraea grandiosella Dyar, there was not a reduction in development or an increase in mortality with tissue that had higher concentrations of toxin. Another study tested the relative concentration of Cry1Ab between the white-yellow, yellow-green, and green portions of the developing ninth leaf within the maize whorl. There were differences in Cry1Ab concentration among these leaf areas. The green tissue had the highest concentration of toxin followed by the yellow-green and white-yellow tissues. Correlations between concentration of Cry1Ab and 5-d fall armyworm larval weights among the three leaf color profiles were all significant and negative, i.e., decreased concentration of Cry1Ab in the leaf tissue resulted in increased 5-d larval weights. There was 100% mortality to the southwestern corn borer larvae fed Cry1Ab maize leaf tissue. Differences in the amount of Cry1Ab in the developing V9 leaf profiles did not alter the absolute susceptibility of the southwestern corn borer to the toxin. In cotton, Gossypium hirsutum L., the amount of Cry1Ac was significantly lower in boll tips where flowers had remained attached compared with normal boll tips. Boll tips where the flowers remained attached are often the site where corn earworms, Helicoverpa zea (Boddie), penetrate Bt cotton bolls. This study demonstrated that, in two diverse plant species, tissue that has low chlorophyll content does not fully express Cry1A. Photosynthesis regulating factors related to mRNA transcription and translation should be studied for their effect on Cry1A production and insect control.

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Selective feeding of tobacco budworm and bollworm (Lepidoptera: Noctuidae) on meridic diet with different concentrations of Bacillus thuringiensis proteins.

Laboratory experiments were conducted to evaluate the behavior of bollworm, Helicoverpa zea (Boddie), and tobacco budworm, Heliothis virescens (F.), larvae on meridic diet with different concentrations of the Cry1Ac and Cry2Ab proteins from Bacillus thuringiensis subsp. kurstaki Berliner. The proteins used in these experiments are the ones in commercially available Bollgard and Bollgard II cotton. Both bollworms and tobacco budworms selectively fed on nontreated diet compared with diet treated with Cry1Ac. In addition, bollworms exhibited a concentration response with Cry1Ac. In general, bollworms selected diet with low concentrations of Cry1Ac compared with diet with higher concentrations of Cry1Ac. For Cry2Ab, the avoidance was not as prominent as that observed for Cry1Ac. Based on results from no-choice assays, the Cry1Ac and Cry2Ab concentrations used in choice assays represented a wide range of biological activity on both species. The lower concentrations provided low levels of mortality, whereas the higher concentrations provided high levels of mortality. Also, the developmental times of larvae were longer at higher concentrations of both proteins. These data provide important information about the behavioral response of key cotton pests to the B. thuringiensis proteins found in commercially available transgenic cotton. This information will be important to develop accurate scouting and management procedures for Bollgard and Bollgard II cotton.

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Effect of aging on pheromone emission from a commercial beet armyworm (Lepidoptera: Noctuidae) lure and trap efficiency.

The effect of aging on a commercial pheromone-based lure for attractiveness to male adult beet armyworms, Spodoptera exigua (Hübner), was assessed in terms of trapping efficiency and volatile emissions of two key components: (Z,E)-9,12-tetradecadienyl acetate and (Z)-9-tetradecen-1-ol. In field assays conducted in the Lower Rio Grande Valley, Texas, for 9 wk (June-August 2000) and for 8 wk (March-May) in Tamaulipas, Mexico, 0-1-wk-old lures collected greater than four-fold more male beet armyworm adults than lures that had been aged for 3-4 wk. Using solid phase microextraction and gas chromatographic analysis of the volatiles, mean (Z,E)-9,12-tetradecadienyl acetate emission declined by 32% after 5 wk of aging in a ventilated environmental chamber at 29.4 degrees C, 3.5% RH, to simulate subtropical summers. Mean (Z)-9-tetradecen-1-ol emission was reduced by 62% after 4 wk of aging. Under the tropical and subtropical conditions of this study, the capacity of the lure to attract moths into the trap declined after aging for 3 wk. The blend ratio (Z,E) -9,12-tetradecadienyl acetate: (Z)-9-tetradecen-1-ol declined 29% when the lure was aged for 5 wk. This study will assist in the development of a standard trapping system for assessing adult beet armyworm populations in the tropical and subtropical conditions to which the species is endemic.

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Season-long variation in expression of Cry1Ac gene and efficacy of Bacillus thuringiensis toxin in transgenic cotton against Helicoverpa armigera (Lepidoptera: Noctuidae).

The insertion of genes encoding insecticidal Cry1A delta-endotoxins from the bacterium Bacillus thuringiensis Berliner variety kurstaki (Bt) into varieties of cotton (Gossypium spp.) was undertaken to assist in the control of a range of lepidopteran pests. In Australia, where Helicoverpa armigera (Hübner) and Helicoverpa punctigera (Wallengren) are major pests, the level of control is useful, but not complete, because efficacy declines as the crop matures. Fluctuations in the efficacy of Bt cotton, to the extent that some insects survive, provide opportunities for H. armigera to develop resistance to the Bt toxin. Therefore, variations in the efficacy of Bt cotton need to be understood if we are to plan rational resistance management strategies to retard the rate of the development of resistance. We measured the changes in efficacy associated with plant development over the growing season, in the field and glasshouse. In addition, the levels of Cry1Ac protein toxin and cry1Ac RNA were determined. In this first demonstration of the relationship between these three factors, we found that the developmental decline in bioefficacy in field-grown plants was associated with reduced cry1Ac transcript levels and Bt toxin levels in postsquaring cotton. In addition, changes in plant chemistry associated with the maturation of the cotton plant were observed to contribute to changes in the efficacy of Bt toxin. Results from the field and glasshouse suggested that variations in efficacy within the growing season and between seasons also may be influenced by environmental factors.

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Correlation between fenvalerate resistance and cytochrome P450-mediated O-demethylation activity in Helicoverpa armigera (Lepidoptera: Noctuidae).

Cytochrome P450 monooxygenases are a major metabolic mechanism responsible for pyrethroid resistance in Helicoverpa armigera (Hübner) from Asia. Cytochrome P450-mediated O-demethylation activity toward p-nitroanisole (PNOD) of individual fourth instars was determined in five strains of H. armigera by using a microplate reader. The four resistant strains of YS, HD, YGF, and YG59 had 6-, 71-, 2540-, and 11,800-fold resistance, respectively, to fenvalerate in comparison with the susceptible BK77 strain. Their mean PNOD activity was 4-, 10-, 24-, and 60-fold, respectively, compared with the BK77 strain. A strong positive correlation (correlation coefficient r = 0.98) between PNOD activity and fenvalerate resistance was found. Of 48 larvae from each strain, only 4% larvae of the susceptible BK77 strain had detectable PNOD activity, whereas 25, 33, 79, and 96% of larvae from the resistant strains YS, HD, YGF, and YG59 exhibited PNOD activity, respectively. There was a clear discrimination of patterns of PNOD frequency distribution between H. armigera strains and their magnitudes of fenvalerate resistance. The PNOD activity can be used as a biochemical marker for monooxygenase-mediated pyrethroid resistance in field populations of H. armigera.

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Effects of selection for the timing of vegetative phase transition on corn borer (Lepidoptera: Noctuidae and Crambidae) damage.

In maize, Zea mays L., the timing of vegetative phase transition from juvenile to adult vegetative phases can be modified through selection. A reduction in the juvenile vegetative phase has been associated with resistance to diseases and pests. The major maize pest in temperate areas is Ostrinia nubilalis (Hübner) and in Europe Sesamia nonagrioides Lefebvre. The objective of our study was to determine the effects of divergent selection for the timing of vegetative phase transition in maize on resistance to corn borers. Three cycles of divergent selection for early and late phase transition in a field corn synthetic and in a sweet corn population were evaluated separately under S. nonagrioides and O. nubilalis artificial infestation. For the field corn experiment, yield and moisture improved with selection for phase transition in both directions, but improvement was due to artifacts of selection, rather than to the change in phase transition. There were no correlated responses for corn borer damage, yield, or grain moisture due to selection for the timing of vegetative phase transition. In the sweet corn experiment, selection for the timing of vegetative phase transition had no significant effects on corn borer damage in sweet corn harvested at the fresh stage. Our results do not support the use of phase transition as an indirect criterion for improving resistance to corn borers in maize. The relationship between phase transition and pest resistance reported by other studies could depend on the genotypes or could be too weak to be detected in a selection program with wild-type maize.

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Fitness of Cry1A-resistant and -susceptible Helicoverpa armigera (Lepidoptera: Noctuidae) on transgenic cotton with reduced levels of Cry1Ac.

The performance of Helicoverpa armigera (Hübner) on 15-wk-old cotton plants was compared for a susceptible strain, a near-isogenic laboratory-selected strain, and F1 progeny of the two strains. Glasshouse experiments were conducted to test the three insect types on conventional plants and transgenic plants that produced the Bacillus thuringiensis (Bt) toxin Cry1Ac. At the time of testing (15 wk), the Cry1Ac concentration in cotton leaves was 75% lower than at 4 wk. On these plants, < 10% of susceptible larvae reached the fifth instar, and none survived to pupation. In contrast, survival to adulthood on Cry1Ac cotton was 62% for resistant larvae and 39% for F1 larvae. These results show that inheritance of resistance to 15-wk-old Cry1Ac cotton is partially dominant, in contrast to results previously obtained on 4-wk-old Cry1Ac cotton. Growth and survival of resistant insects were similar on Cry1Ac cotton and on non-Bt cotton, but F1 insects developed more slowly on Cry1Ac cotton than on non-Bt cotton. Survival was lower and development was slower for resistant larvae than for susceptible and F1 larvae on non-Bt cotton. These results show recessive fitness costs are associated with resistance to Cry1Ac.

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Free phenols in maize pith and their relationship with resistance to Sesamia nonagrioides (Lepidoptera: Noctuidae) attack.

The stem borer Sesamia nonagrioides (Lefèbvre) is the most important insect pest of maize, Zea mays L., in northwestern Spain. Among the metabolites present in maize, phenolic compounds could play an important role in resistance. The objective of this work was to determine whether a relationship between phenols and the amount of resistance exists. Amounts of free phenolic compounds in the pith of 13 inbred maize lines that differ in resistance were measured. The phenolic compounds identified were p-coumaric acid, cafeic acid, ferulic acid, vanillic acid, syringic acid, chorogenic acid, sinapic acid, p-hydroxybenzoic acid, and vanillin. The amount of free p-coumaric acid was correlated with the resistance level. Higher quantities of p-coumaric in the pith could contribute to general resistance to stem borer attack. Jointly with ferulic acid, p-coumaric could provide resistance mechanisms through cell wall fortification and lignification. The other compounds showed no or an unclear relationship with resistance. The vanillic acid showed a decreased tendency after silking, when maize is most attractive for S. nonagrioides, suggesting this acid could act as a chemoattractant for S. nonagrioides larvae or adults. Future studies that focus on these phenolic compounds could be useful in understanding S. nonagrioides resistance.

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Genetic basis of resistance to Cry1Ac and Cry2Aa in Heliothis virescens (Lepidoptera: Noctuidae).

The development of pest resistance to transgenic crop plants producing insecticidal toxins from Bacillus thuringiensis Berliner (Bt) poses a major threat to their sustainable use in agriculture. "Pyramiding" two toxins with different modes of actions in the same plant is now being used to delay the evolution of resistance in the insects, but this strategy could fail if a single gene in a pest confers resistance to both toxins. The CP73 strain of the cotton pest Heliothis virescens (F.) is resistant to both Cry1Ac and Cry2Aa toxins from Bt. We explored the genetic basis of resistance in this strain with a backcross, split-family design. The gene with the largest effect on Cry1Ac resistance in CP73 (BtR-5) maps to linkage group 10 of H. virescens and thus differs from the previously described linkage group 9 BtR-4 resistance found in the YHD2 strain, involving mutation of the gene encoding a 12-domain cadherin-like binding target of the Cry1A toxins. Neither BtR-4 nor BtR-5 seems to confer significant resistance to Cry2Aa. A majority of the linkage groups studied in one backcross family made a small positive contribution to resistance for both toxins. Thus, the Cry2Aa resistance in CP73 is not caused by either of the two major Cry1Ac resistance-conferring genes but instead probably has a quantitative genetic basis.

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