E. Pereira | Universidade Federal de Viçosa (original) (raw)

Papers by E. Pereira

PloS one, 2016

Exposure to Bacillus thuringiensis (Bt) toxins in low- and moderate-dose transgenic crops may ind... more Exposure to Bacillus thuringiensis (Bt) toxins in low- and moderate-dose transgenic crops may induce sublethal effects and increase the rate of Bt resistance evolution, potentially compromising control efficacy against target pests. We tested this hypothesis using the fall armyworm Spodoptera frugiperda, a major polyphagous lepidopteran pest relatively tolerant to Bt notorious for evolving field-relevant resistance to single-gene Bt maize. Late-instar larvae were collected from Bt Cry1Ab and non-Bt maize fields in five locations in Brazil, and their offspring was compared for survival, development, and population growth in rearing environment without and with Cry1Ab throughout larval development. Larval survival on Cry1Ab maize leaves varied from 20 to 80% among the populations. Larvae reared on Cry1Ab maize had seven-day delay in development time in relation to control larvae, and such delay was shorter in offspring of armyworms from Cry1Ab maize. Population growth rates were 50-70...

Large-scale adoption of transgenic crops expressing genes from Bacillus thuringiensis (Bt) impose... more Large-scale adoption of transgenic crops expressing genes from Bacillus thuringiensis (Bt) imposes high selection pressure for evolution of field-relevant resistance that can reduce pest control efficacy, such as reported for Cry1F maize (Zea mays L.) in populations of fall armyworm, Spodoptera frugiperda (J. E. Smith) (Lepidoptera: Noctuidae), of Puerto Rico, Brazil, and the United States. As part of our effort to improve fall armyworm resistance management to Bt crops, here we determined the genetic basis of Cry1F resistance in two S. frugiperda strains originated from field collections in different regions of Brazil and further selected in the laboratory for high levels of resistance to Cry1F maize. Continuous exposure to the TC1507 event for 11 generations resulted in more than 183-fold resistance to Cry1F in the two strains studied, and such a high resistance level enabled the insects to complete larval development on the Bt maize plants. Genetic analyses using concentration-response bioassays with progenies from reciprocal crosses between resistant and susceptible insects indicated that the inheritance of the resistance is autosomal, recessive and without maternal effects. Backcross of the F 1 progeny with the parental resistant strains revealed that the resistance in the two selected strains is conferred by a single locus or set of tightly linked loci. These results support some of the assumptions of the strategy in use for fall armyworm resistance management to Bt Cry1F maize, but survival rates of heterozygotes on the Bt plants were higher than 5%, showing that the Cry1F maize does not produce a high dose of the insec-ticidal protein for S. frugiperda. Additionally, we detected a delay in larval development time that may favor assortative mating of individuals carrying resistance alleles. These findings are consistent with the rapid evolution of Cry1F resistance in certain field populations of fall armyworm. Implications for resistance management of S. frugiperda to Bt maize are discussed.

Crop management practices can affect the population of phytophagous pest species and beneficial a... more Crop management practices can affect the population of phytophagous pest species and beneficial arthropods with consequences for integrated pest management. In this study, we determined the effect of no-tillage and crop residue management on the arthropod community associated with the canopy of common beans (Phaseolus vulgaris L.). Abundance and species composition of herbivorous, detritivorous, predaceous and parasitoid arthropods were recorded during the growing seasons of 2003 and 2004 in Coimbra County, Minas Gerais State, Brazil. Arthropod diversity and guild composition were similar among crop management systems, but their abundance was higher under no-tillage relative to conventional cultivation and where residues from the preceding crop were maintained in the field. Thirty-four arthropod species were recorded, and those most representative of the impact of the crop management practices were Hypogastrura springtails, Empoasca kraemeri and Circulifer leafhoppers, and Solenopsis ants. The infestation levels of major insect-pests, especially leafhoppers (Hemiptera: Cicadellidae), was on average seven-fold lower under no-tillage with retention of crop residues relative to the conventional system with removal of residues, whereas the abundance of predatory ants (Hymenoptera: Formicidae) and springtails (Collembola: Hypogastruridae) were, respectively, about seven-and 15-fold higher in that treatment. Importantly, a significant trophic interaction among crop residues, detritivores, predators and herbivores was observed. Plots managed with no-tillage and retention of crop residues had the highest bean yield, while those with conventional cultivation and removal of the crop residues yielded significantly less beans. This research shows that cropping systems that include zero tillage and crop residue retention can reduce infestation by foliar insect-pests and increase abundance of predators and detritivores, thus having direct consequences for insect pest management.

Transgenic maize (Zea mays L., Poaceae) event TC1507, producing the Cry1F protein of Bacillus thu... more Transgenic maize (Zea mays L., Poaceae) event TC1507, producing the Cry1F protein of Bacillus thuringiensis Berliner, has been used for management of the fall armyworm, Spodoptera frugiperda (JE Smith) (Lepidoptera: Noctuidae), in Brazil since 2009. A strain of S. frugiperda, obtained from field collections of larvae in TC1507 maize in Minas Gerais state in 2010, was selected in the laboratory for resistance to Cry1F using leaves of TC1507 maize in two selection regimes. Continuous exposure of larvae to Cry1F was more effective than exposure for 6, 8, and 10 days in the selection of resistant S. frugiperda individuals. With only four generations of laboratory selection, a strain with high levels of resistance to Cry1F was obtained, as indicated by the survival of insects reared on leaves of TC1507 maize plants and by the more than 300-fold resistance level measured in bioassays with the purified Cry1F protein. Importantly, reciprocal crosses between control and the Cry1F-selected strains revealed that the resistance is autosomal and incompletely recessive, and the response obtained in the backcross of the F 1 generation with the resistant strain was consistent with simple monogenic inheritance. Additionally, there were no apparent fitness costs associated with resistance either for survival or larval growth on non-Bt maize leaves. Our findings provide experimental evidence for rapid evolution of Cry1F resistance in S. frugiperda in the laboratory and further reinforce the potential of this species to evolve field resistance to the TC1507 maize as previously reported. The resistant strain isolated in this study provides an opportunity to estimate the resistance allele frequency in the field and to determine the biochemical and molecular basis of the resistance, which should provide further information to assist in the resistance management of S. frugiperda on transgenic maize producing B. thuringiensis proteins.

The green scale, Coccus viridis (Green) (Hemiptera: Coccidae), is an insect pest of coffee and se... more The green scale, Coccus viridis (Green) (Hemiptera: Coccidae), is an insect pest of coffee and several other perennial cultivated plant species. We investigated changes in alkaloid and phenolic contents in coffee plants as a response to herbivory by this insect. Greenhouse-grown, 11-month-old coffee plants were artificially infested with the coccid and compared with control, uninfested plants. Leaf samples were taken at 15, 30, 45, and 60 days after infestation, and high-performance liquid chromatography was used to identify and quantify alkaloid and phenolic compounds induced by the coccids at each sampling date. Of the compounds investigated, caffeine was the main coffee alkaloid detected in fully developed leaves, and its concentration in infested plants was twice as high as in the control plants. The main coffee phenolics were caffeic and chlorogenic acid, and a significant increase in their concentrations occurred only in plants infested by C. viridis. A positive and significant relationship was found between alkaloid and phenolic concentrations and the infestation level by adults and nymphs of C. viridis. Caffeine and chlorogenic acid applied on coffee leaves stimulated the locomo-tory activity of the green scale, thus reducing their feeding compared to untreated leaves. This is the first study to show increased levels of coffee alkaloids and phenolics in response to herbivory by scale insects. The elevation of caffeine and chlorogenic acid levels in coffee leaves because of C. viridis infestation seems to affect this generalist insect by stimulating the locomotion of crawlers.

Acta Scientiarum. Agronomy, 2005

Resistência de berinjela transgênica contendo o gene orizacistatina a Mechanitis polymnia e Mecha... more Resistência de berinjela transgênica contendo o gene orizacistatina a Mechanitis polymnia e Mechanitis lysimnia Acta Scientiarum. Biological Sciences, vol. 27, núm. 4, octubre-diciembre, 2005, p. 647, mail: picanco@ufv.br RESUMO. Esta pesquisa objetivou avaliar a resistência da berinjela (Solanum melongena) transgênica, contendo o gene orizacistatina, a Mechanitis polymnia L. e Mechanitis lysimnia Fabr (Lepidoptera: Nymphalidae). Foram avaliadas a área foliar consumida, a mortalidade e duração das fases larval e pupal de M. polymnia e M. lysimnia. A berinjela transgênica não apresentou resistência aos Lepidoptera M. polymnia e M. lysimnia. Palavras-chave: Solanum melongena, Lepidoptera, proteinase, biotecnologia, resistência de plantas. ABSTRACT. Resistance of transgenic eggplant carrying the oryzacystatin gene to Mechanitis polymnia and Mechanitis lysimnia. This research aimed to evaluate the resistance of transgenic eggplant (Solanum melongena) carrying the oryzacystatin gene to Mechanitis polymnia L. and Mechanitis lysimnia Fabr. (Lepidoptera: Nymphalidae). The leaf area consumed, the mortality and duration of the larval and pupal phases of M. polymnia and M. lysimnia were evaluated. Results showed the transgenic eggplant is not resistant to Lepidoptera M. polymnia and M. lysimnia.

BACKGROUND: The presence of fitness costs of resistance to Bacillus thuringiensis (Bt) insecticid... more BACKGROUND: The presence of fitness costs of resistance to Bacillus thuringiensis (Bt) insecticidal proteins in insect populations may delay or even reverse the local selection of insect resistance to Bt transgenic crops, and deserves rigorous investigation. Here we assessed the fitness costs associated with Cry1Fa resistance in two strains of fall armyworm, Spodoptera frugiperda (Lepidoptera: Noctuidae), derived from field collections in different Brazilian regions and further selected in the laboratory for high levels of resistance to Cry1Fa using leaves of TC1507 corn. RESULTS: Fitness components were compared using paired resistant and susceptible strains with similar genetic backgrounds and F 1 generations from reciprocal crosses, all of them reared on non-transgenic corn leaves. No apparent life history costs in the larval stage were observed in the Bt-resistant strains. Moreover, the resistance remained stable for seven generations in the absence of selection, with no decrease in the proportion of resistant individuals. Larval respiration rates were also similar between resistant and susceptible homozygotes, and heterozygotes displayed respiration rates and demographic performance equal or superior to those of susceptible homozygotes. CONCLUSION: In combination, these results indicate the lack of strong fitness costs associated with resistance to Cry1Fa in the fall armyworm strains studied. These findings suggest that Cry1Fa resistance in S. frugiperda populations is unlikely to be counterselected in Cry1Fa-free environments.

Transgenic crop " pyramids " producing two or more Bacillus thuringiensis (Bt) toxins active agai... more Transgenic crop " pyramids " producing two or more Bacillus thuringiensis (Bt) toxins active against the same pest are used to delay evolution of resistance in insect pest populations. Laboratory and greenhouse experiments were performed with fall armyworm, Spodoptera frugiperda, to characterize resistance to Bt maize producing Cry1A.105 and Cry2Ab and test some assumptions of the " pyramid " resistance management strategy. Selection of a field-derived strain of S. frugiperda already resistant to Cry1F maize with Cry1A.105 + Cry2Ab maize for ten generations produced resistance that allowed the larvae to colonize and complete the life cycle on these Bt maize plants. Greenhouse experiments revealed that the resistance was completely recessive (Dx = 0), incomplete, autosomal, and without maternal effects or cross-resistance to the Vip3Aa20 toxin produced in other Bt maize events. This profile of resistance supports some of the assumptions of the pyramid strategy for resistance management. However, laboratory experiments with purified Bt toxin and plant leaf tissue showed that resistance to Cry1A.105 + Cry2Ab2 maize further increased resistance to Cry1Fa, which indicates that populations of fall armyworm have high potential for developing resistance to some currently available pyramided maize used against this pest, especially where resistance to Cry1Fa was reported in the field. Transgenic crops producing toxins from the soil bacterium Bacillus thuringiensis (Bt) are a major tool for controlling insect pests worldwide 1. Benefits of Bt crops include effective management of target pests, decreased use of conventional insecticides, and reduced risks to non-target organisms, including humans 2–6. However, rapid evolution of resistance to Bt crops in several pest species has compromised some of these benefits 7,8. To increase pest control efficacy and delay resistance evolution, a gene pyramiding strategy has been employed in transgenic plants to produce two or more Bt toxins of dissimilar mode of action effective against the same target pest species 7. As a result, pyramided Bt crops have been adopted rapidly and are expected to become even more prevalent in the future 9. As a strategy for resistance management, pyramided Bt crops are expected to be more effective when insect resistance to each Bt toxin is recessive and associated with reduced fitness of resistant individuals on both non-Bt and Bt plants (i.e., with fitness costs and incomplete resistance), when selection with any one of the pyramided toxins does not cause cross-resistance to the others, and when alleles conferring resistance to each toxin are rare 10–13. However, little is known about that evolutionary response from target species to the selective pressure imposed by two or more toxins with independent mechanisms of action. In this study we characterize the inheritance of resistance to pyramided maize producing the Cry1A.105 and Cry2Ab toxins in the fall armyworm, Spodoptera frugiperda, a major migratory pest of maize in Neotropical America 14 , and recently also in southern United States 15. Previous studies with resistant strains of S. frugiperda to the event TC1507 (i.e., non-pyramided Bt maize producing the Cry1Fa toxin) showed that the genetic basis of resistance did not fit some assumptions of the high dose-refuge strategy, i.e., lack of fitness costs associated with resistance, high frequency of resistance alleles, and heterozygote survival greater than 5% on leaf tissues of Cry1F maize plants 16,17. The result has been the rather quick appearance of Cry1F resistance in Brazilian populations of the fall armyworm 16,18 .

Exposure to Bacillus thuringiensis (Bt) toxins in low-and moderate-dose transgenic crops may indu... more Exposure to Bacillus thuringiensis (Bt) toxins in low-and moderate-dose transgenic crops may induce sublethal effects and increase the rate of Bt resistance evolution, potentially compromising control efficacy against target pests. We tested this hypothesis using the fall armyworm Spodoptera frugiperda, a major polyphagous lepidopteran pest relatively tolerant to Bt notorious for evolving field-relevant resistance to single-gene Bt maize. Late-instar larvae were collected from Bt Cry1Ab and non-Bt maize fields in five locations in Brazil, and their offspring was compared for survival, development, and population growth in rearing environment without and with Cry1Ab throughout larval development. Larval survival on Cry1Ab maize leaves varied from 20 to 80% among the populations. Larvae reared on Cry1Ab maize had seven-day delay in development time in relation to control larvae, and such delay was shorter in offspring of armyworms from Cry1Ab maize. Population growth rates were 50–70% lower for insects continuously exposed to Cry1Ab maize relative to controls , showing the population-level effect of Cry1Ab, which varied among the populations and prior exposure to Cry1Ab maize in the field. In three out of five populations, armyworms derived from Bt maize reared on Cry1Ab maize showed higher larval weight, faster larval development and better reproductive performance than the armyworms derived from non-Bt maize, and one of these populations showed better performance on both Cry1Ab and control diets, indicating no fitness cost of the resistance trait. Altogether, these results indicate that offspring of armyworms that developed on field-grown, single-gene Bt Cry1Ab maize had reduced performance on Cry1Ab maize foliage in two populations studied, but in other three populations, these offspring had better overall performance on the Bt maize foliage than that of the armyworms from non-Bt maize fields, possibly because of Cry1Ab resistance alleles in these populations. Implications of these findings for resistance management of S. frugiperda in Bt crops are discussed.

PloS one, 2016

Exposure to Bacillus thuringiensis (Bt) toxins in low- and moderate-dose transgenic crops may ind... more Exposure to Bacillus thuringiensis (Bt) toxins in low- and moderate-dose transgenic crops may induce sublethal effects and increase the rate of Bt resistance evolution, potentially compromising control efficacy against target pests. We tested this hypothesis using the fall armyworm Spodoptera frugiperda, a major polyphagous lepidopteran pest relatively tolerant to Bt notorious for evolving field-relevant resistance to single-gene Bt maize. Late-instar larvae were collected from Bt Cry1Ab and non-Bt maize fields in five locations in Brazil, and their offspring was compared for survival, development, and population growth in rearing environment without and with Cry1Ab throughout larval development. Larval survival on Cry1Ab maize leaves varied from 20 to 80% among the populations. Larvae reared on Cry1Ab maize had seven-day delay in development time in relation to control larvae, and such delay was shorter in offspring of armyworms from Cry1Ab maize. Population growth rates were 50-70...

Large-scale adoption of transgenic crops expressing genes from Bacillus thuringiensis (Bt) impose... more Large-scale adoption of transgenic crops expressing genes from Bacillus thuringiensis (Bt) imposes high selection pressure for evolution of field-relevant resistance that can reduce pest control efficacy, such as reported for Cry1F maize (Zea mays L.) in populations of fall armyworm, Spodoptera frugiperda (J. E. Smith) (Lepidoptera: Noctuidae), of Puerto Rico, Brazil, and the United States. As part of our effort to improve fall armyworm resistance management to Bt crops, here we determined the genetic basis of Cry1F resistance in two S. frugiperda strains originated from field collections in different regions of Brazil and further selected in the laboratory for high levels of resistance to Cry1F maize. Continuous exposure to the TC1507 event for 11 generations resulted in more than 183-fold resistance to Cry1F in the two strains studied, and such a high resistance level enabled the insects to complete larval development on the Bt maize plants. Genetic analyses using concentration-response bioassays with progenies from reciprocal crosses between resistant and susceptible insects indicated that the inheritance of the resistance is autosomal, recessive and without maternal effects. Backcross of the F 1 progeny with the parental resistant strains revealed that the resistance in the two selected strains is conferred by a single locus or set of tightly linked loci. These results support some of the assumptions of the strategy in use for fall armyworm resistance management to Bt Cry1F maize, but survival rates of heterozygotes on the Bt plants were higher than 5%, showing that the Cry1F maize does not produce a high dose of the insec-ticidal protein for S. frugiperda. Additionally, we detected a delay in larval development time that may favor assortative mating of individuals carrying resistance alleles. These findings are consistent with the rapid evolution of Cry1F resistance in certain field populations of fall armyworm. Implications for resistance management of S. frugiperda to Bt maize are discussed.

Crop management practices can affect the population of phytophagous pest species and beneficial a... more Crop management practices can affect the population of phytophagous pest species and beneficial arthropods with consequences for integrated pest management. In this study, we determined the effect of no-tillage and crop residue management on the arthropod community associated with the canopy of common beans (Phaseolus vulgaris L.). Abundance and species composition of herbivorous, detritivorous, predaceous and parasitoid arthropods were recorded during the growing seasons of 2003 and 2004 in Coimbra County, Minas Gerais State, Brazil. Arthropod diversity and guild composition were similar among crop management systems, but their abundance was higher under no-tillage relative to conventional cultivation and where residues from the preceding crop were maintained in the field. Thirty-four arthropod species were recorded, and those most representative of the impact of the crop management practices were Hypogastrura springtails, Empoasca kraemeri and Circulifer leafhoppers, and Solenopsis ants. The infestation levels of major insect-pests, especially leafhoppers (Hemiptera: Cicadellidae), was on average seven-fold lower under no-tillage with retention of crop residues relative to the conventional system with removal of residues, whereas the abundance of predatory ants (Hymenoptera: Formicidae) and springtails (Collembola: Hypogastruridae) were, respectively, about seven-and 15-fold higher in that treatment. Importantly, a significant trophic interaction among crop residues, detritivores, predators and herbivores was observed. Plots managed with no-tillage and retention of crop residues had the highest bean yield, while those with conventional cultivation and removal of the crop residues yielded significantly less beans. This research shows that cropping systems that include zero tillage and crop residue retention can reduce infestation by foliar insect-pests and increase abundance of predators and detritivores, thus having direct consequences for insect pest management.

Transgenic maize (Zea mays L., Poaceae) event TC1507, producing the Cry1F protein of Bacillus thu... more Transgenic maize (Zea mays L., Poaceae) event TC1507, producing the Cry1F protein of Bacillus thuringiensis Berliner, has been used for management of the fall armyworm, Spodoptera frugiperda (JE Smith) (Lepidoptera: Noctuidae), in Brazil since 2009. A strain of S. frugiperda, obtained from field collections of larvae in TC1507 maize in Minas Gerais state in 2010, was selected in the laboratory for resistance to Cry1F using leaves of TC1507 maize in two selection regimes. Continuous exposure of larvae to Cry1F was more effective than exposure for 6, 8, and 10 days in the selection of resistant S. frugiperda individuals. With only four generations of laboratory selection, a strain with high levels of resistance to Cry1F was obtained, as indicated by the survival of insects reared on leaves of TC1507 maize plants and by the more than 300-fold resistance level measured in bioassays with the purified Cry1F protein. Importantly, reciprocal crosses between control and the Cry1F-selected strains revealed that the resistance is autosomal and incompletely recessive, and the response obtained in the backcross of the F 1 generation with the resistant strain was consistent with simple monogenic inheritance. Additionally, there were no apparent fitness costs associated with resistance either for survival or larval growth on non-Bt maize leaves. Our findings provide experimental evidence for rapid evolution of Cry1F resistance in S. frugiperda in the laboratory and further reinforce the potential of this species to evolve field resistance to the TC1507 maize as previously reported. The resistant strain isolated in this study provides an opportunity to estimate the resistance allele frequency in the field and to determine the biochemical and molecular basis of the resistance, which should provide further information to assist in the resistance management of S. frugiperda on transgenic maize producing B. thuringiensis proteins.

The green scale, Coccus viridis (Green) (Hemiptera: Coccidae), is an insect pest of coffee and se... more The green scale, Coccus viridis (Green) (Hemiptera: Coccidae), is an insect pest of coffee and several other perennial cultivated plant species. We investigated changes in alkaloid and phenolic contents in coffee plants as a response to herbivory by this insect. Greenhouse-grown, 11-month-old coffee plants were artificially infested with the coccid and compared with control, uninfested plants. Leaf samples were taken at 15, 30, 45, and 60 days after infestation, and high-performance liquid chromatography was used to identify and quantify alkaloid and phenolic compounds induced by the coccids at each sampling date. Of the compounds investigated, caffeine was the main coffee alkaloid detected in fully developed leaves, and its concentration in infested plants was twice as high as in the control plants. The main coffee phenolics were caffeic and chlorogenic acid, and a significant increase in their concentrations occurred only in plants infested by C. viridis. A positive and significant relationship was found between alkaloid and phenolic concentrations and the infestation level by adults and nymphs of C. viridis. Caffeine and chlorogenic acid applied on coffee leaves stimulated the locomo-tory activity of the green scale, thus reducing their feeding compared to untreated leaves. This is the first study to show increased levels of coffee alkaloids and phenolics in response to herbivory by scale insects. The elevation of caffeine and chlorogenic acid levels in coffee leaves because of C. viridis infestation seems to affect this generalist insect by stimulating the locomotion of crawlers.

Acta Scientiarum. Agronomy, 2005

Resistência de berinjela transgênica contendo o gene orizacistatina a Mechanitis polymnia e Mecha... more Resistência de berinjela transgênica contendo o gene orizacistatina a Mechanitis polymnia e Mechanitis lysimnia Acta Scientiarum. Biological Sciences, vol. 27, núm. 4, octubre-diciembre, 2005, p. 647, mail: picanco@ufv.br RESUMO. Esta pesquisa objetivou avaliar a resistência da berinjela (Solanum melongena) transgênica, contendo o gene orizacistatina, a Mechanitis polymnia L. e Mechanitis lysimnia Fabr (Lepidoptera: Nymphalidae). Foram avaliadas a área foliar consumida, a mortalidade e duração das fases larval e pupal de M. polymnia e M. lysimnia. A berinjela transgênica não apresentou resistência aos Lepidoptera M. polymnia e M. lysimnia. Palavras-chave: Solanum melongena, Lepidoptera, proteinase, biotecnologia, resistência de plantas. ABSTRACT. Resistance of transgenic eggplant carrying the oryzacystatin gene to Mechanitis polymnia and Mechanitis lysimnia. This research aimed to evaluate the resistance of transgenic eggplant (Solanum melongena) carrying the oryzacystatin gene to Mechanitis polymnia L. and Mechanitis lysimnia Fabr. (Lepidoptera: Nymphalidae). The leaf area consumed, the mortality and duration of the larval and pupal phases of M. polymnia and M. lysimnia were evaluated. Results showed the transgenic eggplant is not resistant to Lepidoptera M. polymnia and M. lysimnia.

BACKGROUND: The presence of fitness costs of resistance to Bacillus thuringiensis (Bt) insecticid... more BACKGROUND: The presence of fitness costs of resistance to Bacillus thuringiensis (Bt) insecticidal proteins in insect populations may delay or even reverse the local selection of insect resistance to Bt transgenic crops, and deserves rigorous investigation. Here we assessed the fitness costs associated with Cry1Fa resistance in two strains of fall armyworm, Spodoptera frugiperda (Lepidoptera: Noctuidae), derived from field collections in different Brazilian regions and further selected in the laboratory for high levels of resistance to Cry1Fa using leaves of TC1507 corn. RESULTS: Fitness components were compared using paired resistant and susceptible strains with similar genetic backgrounds and F 1 generations from reciprocal crosses, all of them reared on non-transgenic corn leaves. No apparent life history costs in the larval stage were observed in the Bt-resistant strains. Moreover, the resistance remained stable for seven generations in the absence of selection, with no decrease in the proportion of resistant individuals. Larval respiration rates were also similar between resistant and susceptible homozygotes, and heterozygotes displayed respiration rates and demographic performance equal or superior to those of susceptible homozygotes. CONCLUSION: In combination, these results indicate the lack of strong fitness costs associated with resistance to Cry1Fa in the fall armyworm strains studied. These findings suggest that Cry1Fa resistance in S. frugiperda populations is unlikely to be counterselected in Cry1Fa-free environments.

Transgenic crop " pyramids " producing two or more Bacillus thuringiensis (Bt) toxins active agai... more Transgenic crop " pyramids " producing two or more Bacillus thuringiensis (Bt) toxins active against the same pest are used to delay evolution of resistance in insect pest populations. Laboratory and greenhouse experiments were performed with fall armyworm, Spodoptera frugiperda, to characterize resistance to Bt maize producing Cry1A.105 and Cry2Ab and test some assumptions of the " pyramid " resistance management strategy. Selection of a field-derived strain of S. frugiperda already resistant to Cry1F maize with Cry1A.105 + Cry2Ab maize for ten generations produced resistance that allowed the larvae to colonize and complete the life cycle on these Bt maize plants. Greenhouse experiments revealed that the resistance was completely recessive (Dx = 0), incomplete, autosomal, and without maternal effects or cross-resistance to the Vip3Aa20 toxin produced in other Bt maize events. This profile of resistance supports some of the assumptions of the pyramid strategy for resistance management. However, laboratory experiments with purified Bt toxin and plant leaf tissue showed that resistance to Cry1A.105 + Cry2Ab2 maize further increased resistance to Cry1Fa, which indicates that populations of fall armyworm have high potential for developing resistance to some currently available pyramided maize used against this pest, especially where resistance to Cry1Fa was reported in the field. Transgenic crops producing toxins from the soil bacterium Bacillus thuringiensis (Bt) are a major tool for controlling insect pests worldwide 1. Benefits of Bt crops include effective management of target pests, decreased use of conventional insecticides, and reduced risks to non-target organisms, including humans 2–6. However, rapid evolution of resistance to Bt crops in several pest species has compromised some of these benefits 7,8. To increase pest control efficacy and delay resistance evolution, a gene pyramiding strategy has been employed in transgenic plants to produce two or more Bt toxins of dissimilar mode of action effective against the same target pest species 7. As a result, pyramided Bt crops have been adopted rapidly and are expected to become even more prevalent in the future 9. As a strategy for resistance management, pyramided Bt crops are expected to be more effective when insect resistance to each Bt toxin is recessive and associated with reduced fitness of resistant individuals on both non-Bt and Bt plants (i.e., with fitness costs and incomplete resistance), when selection with any one of the pyramided toxins does not cause cross-resistance to the others, and when alleles conferring resistance to each toxin are rare 10–13. However, little is known about that evolutionary response from target species to the selective pressure imposed by two or more toxins with independent mechanisms of action. In this study we characterize the inheritance of resistance to pyramided maize producing the Cry1A.105 and Cry2Ab toxins in the fall armyworm, Spodoptera frugiperda, a major migratory pest of maize in Neotropical America 14 , and recently also in southern United States 15. Previous studies with resistant strains of S. frugiperda to the event TC1507 (i.e., non-pyramided Bt maize producing the Cry1Fa toxin) showed that the genetic basis of resistance did not fit some assumptions of the high dose-refuge strategy, i.e., lack of fitness costs associated with resistance, high frequency of resistance alleles, and heterozygote survival greater than 5% on leaf tissues of Cry1F maize plants 16,17. The result has been the rather quick appearance of Cry1F resistance in Brazilian populations of the fall armyworm 16,18 .

Exposure to Bacillus thuringiensis (Bt) toxins in low-and moderate-dose transgenic crops may indu... more Exposure to Bacillus thuringiensis (Bt) toxins in low-and moderate-dose transgenic crops may induce sublethal effects and increase the rate of Bt resistance evolution, potentially compromising control efficacy against target pests. We tested this hypothesis using the fall armyworm Spodoptera frugiperda, a major polyphagous lepidopteran pest relatively tolerant to Bt notorious for evolving field-relevant resistance to single-gene Bt maize. Late-instar larvae were collected from Bt Cry1Ab and non-Bt maize fields in five locations in Brazil, and their offspring was compared for survival, development, and population growth in rearing environment without and with Cry1Ab throughout larval development. Larval survival on Cry1Ab maize leaves varied from 20 to 80% among the populations. Larvae reared on Cry1Ab maize had seven-day delay in development time in relation to control larvae, and such delay was shorter in offspring of armyworms from Cry1Ab maize. Population growth rates were 50–70% lower for insects continuously exposed to Cry1Ab maize relative to controls , showing the population-level effect of Cry1Ab, which varied among the populations and prior exposure to Cry1Ab maize in the field. In three out of five populations, armyworms derived from Bt maize reared on Cry1Ab maize showed higher larval weight, faster larval development and better reproductive performance than the armyworms derived from non-Bt maize, and one of these populations showed better performance on both Cry1Ab and control diets, indicating no fitness cost of the resistance trait. Altogether, these results indicate that offspring of armyworms that developed on field-grown, single-gene Bt Cry1Ab maize had reduced performance on Cry1Ab maize foliage in two populations studied, but in other three populations, these offspring had better overall performance on the Bt maize foliage than that of the armyworms from non-Bt maize fields, possibly because of Cry1Ab resistance alleles in these populations. Implications of these findings for resistance management of S. frugiperda in Bt crops are discussed.