0000000000262895
AUTHOR
Joel González-cabrera
Toxicity of several d-endotoxins of Bacillus thuringiensis against Helicoverpa armigera (Lepidoptera: Noctuidae) from Spain
Abstract Toxicity and larval growth inhibition of 11 insecticidal proteins of Bacillus thuringiensis were evaluated against neonate larvae of Helicoverpa armigera, a major pest of important crops in Spain and other countries, by a whole-diet contamination method. The most active toxins were Cry1Ac4 and Cry2Aa1, with LC50 values of 3.5 and 6.3 μg/ml, respectively. At the concentrations tested, Cry1Ac4, Cry2Aa1, Cry9Ca, Cry1Fa1, Cry1Ab3, Cry2Ab2, Cry1Da, and Cry1Ja1, produced a significant growth inhibition, whereas Cry1Aa3, Cry1Ca2, and Cry1Ea had no effect.
Supplementary figures and tables from Plant guttation provides nutrient-rich food for insects
Plant guttation is a fluid from xylem and phloem sap secreted at the margins of leaves from many plant species. All previous studies have considered guttation as a water source for insects. Here, we hypothesized that plant guttation serves as a reliable and nutrient-rich food source for insects with effects on their communities. Using highbush blueberries as a study system, we demonstrate that guttation droplets contain carbohydrates and proteins. Insects from three feeding lifestyles, an herbivore, a parasitic wasp and a predator, increased their longevity and fecundity when fed on these guttation droplets compared to those fed on control water. Our results also show that guttation droplet…
Mutations associated with pyrethroid resistance in Varroa mite, a parasite of honey bees, are widespread across the United States.
BACKGROUND Managed honey bees are key pollinators of many crops and play an essential role in the United States food production. For more than ten years, beekeepers in the United States have been reporting high rates of colony losses. One of the drivers of these losses is the parasitic mite Varroa destructor. Maintaining healthy honey bee colonies in the United States is dependent on a successful control of this mite. The pyrethroid tau-fluvalinate (Apistan®) was among the first synthetic varroacides registered in the United States. With over 20 years of use, mites resistant to Apistan® have emerged, and so it is unsurprising that treatment failures have been reported. Resistance to tau-flu…
Mutations in the Bacillus thuringiensis Cry1Ca toxin demonstrate the role of domains II and III in specificity towards Spodoptera exigua larvae
Several mutants of the Bacillus thuringiensis Cry1Ca toxin affected with regard to specific activity towards Spodoptera exigua were studied. Alanine was used to replace single residues in loops 2 and 3 of domain II (mutant pPB19) and to replace residues 541– 544 in domain III (mutant pPB20). Additionally, a Cry1Ca mutant combining all mutations was constructed (mutant pPB21). Toxicity assays showed a marked decrease in toxicity against S. exigua for all mutants, while they retained their activity against Manduca sexta, confirming the importance of these residues in determining insect specificity. Parameters for binding to the specific receptors in BBMV (brush border membrane vesicles) of S.…
Mutations associated with pyrethroid resistance in Varroa mites, a parasite of honey bees, are widespread across the USA
ABSTRACTBACKGROUNDManaged honey bees are key pollinators of many crops and play an essential role in the United States food production. For more than 10 years, beekeepers in the US have been reporting high rate of colony losses. One of the drivers of this colony loss is the parasitic mite Varroa destructor. Preserving healthy honey bee colonies in the US is dependent on a successful control of this mite. The pyrethroid tau-fluvalinate (Apistan®) was among the first synthetic varroacide registered in the US. With over 20 years of use, population of mites resistant to Apistan® have emerged, and so it is unsurprising that treatment failures have been reported. Resistance in US mite populations…
Toxicity and mode of action of Bacillus thuringiensis Cry proteins in the Mediterranean corn borer, Sesamia nonagrioides (Lefebvre)
ABSTRACT Sesamia nonagrioides is one of the most damaging pests of corn in Spain and other Mediterranean countries. Bt corn expressing the Bacillus thuringiensis Cry1Ab toxin is being grown on about 58,000 ha in Spain. Here we studied the mode of action of this Cry protein on S. nonagrioides (binding to specific receptors, stability of binding, and pore formation) and the modes of action of other Cry proteins that were found to be active in this work (Cry1Ac, Cry1Ca, and Cry1Fa). Binding assays were performed with 125 I- or biotin-labeled toxins and larval brush border membrane vesicles (BBMV). Competition experiments indicated that these toxins bind specifically and that Cry1Aa, Cry1Ab, an…
Resistance to amitraz in the parasitic honey bee mite Varroa destructor is associated with mutations in the β−adrenergic-like octopamine receptor
AbstractVarroa destructor is considered a major reason for high loss rate of Western honey bee (Apis mellifera) colonies. To prevent colony losses caused by V. destructor it is necessary to actively manage the mite population. Beekeepers, particularly commercial beekeepers, have few alternative treatments other than synthetic acaricides to control the parasite, resulting in intensive treatment regimens that led to the evolution of resistance in mite populations.To investigate the mechanism of the resistance to amitraz detected in V. destructor mites from French and U.S. apiaries, we identified and characterized octopamine and tyramine receptors (the known targets of amitraz) in this species…
Variation in Susceptibility to Bacillus thuringiensis Toxins among Unselected Strains of Plutella xylostella
ABSTRACT So far, the only insect that has evolved resistance in the field to Bacillus thuringiensis toxins is the diamondback moth ( Plutella xylostella ). Documentation and analysis of resistant strains rely on comparisons with laboratory strains that have not been exposed to B. thuringiensis toxins. Previously published reports show considerable variation among laboratories in responses of unselected laboratory strains to B. thuringiensis toxins. Because different laboratories have used different unselected strains, such variation could be caused by differences in bioassay methods among laboratories, genetic differences among unselected strains, or both. Here we tested three unselected st…
Analyses of Cry1Ab binding in resistant and susceptible strains of the European corn borer, Ostrinia nubilalis (Hubner) (Lepidoptera: Crambidae).
ABSTRACT Cry1Ab toxin binding analysis was performed to determine whether resistance in laboratory-selected Ostrinia nubilalis strains is associated with target site alteration. Brush border membrane vesicles were prepared using dissected midguts from late instars of susceptible and resistant strains (Europe-R and RSTT) of O. nubilalis . Immunoblot analysis indicated that three different proteins bound to Cry1Ab toxin and were recognized by an anticadherin serum. In a comparison of resistant and susceptible strains, reduced Cry1Ab binding was apparent for all three bands corresponding to cadherin-like proteins in the Europe-R strain, while reduced binding was apparent in only one band for t…
Tomato trichomes are deadly hurdles limiting the establishment of Amblyseius swirskii Athias-Henriot (Acari: Phytoseiidae)
[EN] Amblyseius swirskii is a predatory mite widely used for the control of very important pest species, such as whiteflies and thrips, in organic farming and conventional agriculture. However, this species cannot establish on tomato crops, probably due to the toxic effects of plant trichomes and their exudates. We evaluated tomato plants for effects on: a) A. swirskii preference mediated by plant volatiles, b) A. swirskii development, predation capacity and reproductive performance, c) the dispersal and survival of mites as affected by stem trichomes, and d) mite survival as a function of secondary metabolites secreted by tomato trichomes. The results showed that A. swirskii mites which ga…
Identification of new viruses specific to the honey bee mite Varroa destructor
AbstractLarge-scale colony losses among managed Western honey bees have become a serious threat to the beekeeping industry in the last decade. There are multiple factors contributing to these losses but the impact of Varroa destructor parasitism is by far the most important, along with the contribution of some pathogenic viruses vectored by the mite. So far, more than 20 viruses have been identified infecting the honey bee, most of them RNA viruses. They may be maintained either as covert infections or causing severe symptomatic infections, compromising the viability of the colony. In silico analysis of available transcriptomic data obtained from mites collected in the USA and Europe as wel…
Mutations in the voltage-gated sodium channel gene associated with deltamethrin resistance in commercially sourced Phytoseiulus persimilis
The implementation of Integrated Pest Management (IPM) in current agricultural practice is a convenient and very effective strategy to maintain pest populations under control. The use of Biological Control Agents, like Phytoseiulus persimilis, is key for the success of such approach. This predatory mite is widely used since it is very effective for controlling the two spotted spider mite (Tetranychus urticae), one of the most devastating pests worldwide. Here we show the identification of mutations located in the Voltage Gated Sodium Channel (VGSC) of P. persimilis, that correlate with the reduced susceptibility to deltamethrin observed in commercially sourced colonies of this predatory mit…
High Genetic Variability for Resistance to Bacillus thuringiensis Toxins in a Single Population of Diamondback Moth
ABSTRACT The long-term benefit of insecticidal products based on Cry toxins, either in sprays or as transgenic crops, is threatened by the development of resistance by target pests. The models used to predict evolution of resistance to Cry toxins most often are monogenic models in which two alleles are used. Moreover, the high-dose/refuge strategy recommended for implementation with transgenic crops relies on the assumption that the resistance allele is recessive. Using selection experiments, we demonstrated the occurrence in a laboratory colony of diamondback moth of two different genes (either allelic or nonallelic) that confer resistance to Cry1Ab. At the concentration tested, resistance…
Effect of mass rearing on the genetic diversity of the predatory mite Amblyseius swirskii
Amblyseius swirskii Athias-Henriot (Acari: Phytoseiidae) is a predatory mite used to control whiteflies and thrips in protected crops. This biocontrol agent, originating from the Eastern Mediterranean region, has been mass-reared for commercial use since 2005 and is widely used in augmentative biocontrol programs. As a polyphagous predator, it has to cope with different biotic and abiotic factors. However, possible adaptation to mass rearing for production might be hindering its resilience and capacity for optimum performance in the field. In this study, we investigated the effect of long-term mass rearing on the genetic diversity of A. swirskii. We identified six microsatellite loci from w…
Neonicotinoids from coated seeds toxic for honeydew-feeding biological control agents
Seed coating (‘seed treatment’) is the leading delivery method of neonicotinoid insecticides in major crops such as soybean, wheat, cotton and maize. However, this prophylactic use of neonicotinoids is widely discussed from the standpoint of environmental costs. Growing soybean plants from neonicotinoid-coated seeds in field, we demonstrate that soybean aphids (Aphis glycines) survived the treatment, and excreted honeydew containing neonicotinoids. Biochemical analyses demonstrated that honeydew excreted by the soybean aphid contained substantial concentrations of neonicotinoids even one month after sowing of the crop. Consuming this honeydew reduced the longevity of two biological control …
Binding analyses of Cry1Ab and Cry1Ac with membrane vesicles from Bacillus thuringiensis-resistant and -susceptible Ostrinia nubilalis.
The binding properties of Bacillus thuringiensis toxins to brush border membrane vesicles of Dipel-resistant and -susceptible Ostrinia nubilalis larvae were compared using ligand-toxin immunoblot analysis, surface plasmon resonance (SPR), and radiolabeled toxin binding assays. In ligand-toxin immunoblot analysis, the number of Cry1Ab or Cry1Ac toxin binding proteins and the relative toxin binding intensity were similar in vesicles from resistant and susceptible larvae. Surface plasmon resonance with immobilized activated Cry1Ab toxin indicated that there were no significant differences in binding with fluid-phase vesicles from resistant and susceptible larvae. Homologous competition assays …
IPM-recommended insecticides harm beneficial insects through contaminated honeydew
The use of some systemic insecticides has been banned in Europe because they are toxic to beneficial insects when these feed on nectar. A recent study shows that systemic insecticides can also kill beneficial insects when they feed on honeydew. Honeydew is the sugar-rich excretion of hemipterans and is the most abundant carbohydrate source for beneficial insects such as pollinators and biological control agents in agroecosystems. Here, we investigated whether the toxicity of contaminated honeydew depends on i) the hemipteran species that excretes the honeydew; ii) the active ingredient, and iii) the beneficial insect that feeds on it. HPLC-MS/MS analyses demonstrated that the systemic insec…
Next-generation biological control
Biological control is widely successful at controlling pests, but effective biocontrol agents are now more difficult to import from countries of origin due to more restrictive international trade laws (the Nagoya Protocol). Coupled with increasing demand, the efficacy of existing and new biocontrol agents needs to be improved with genetic and genomic approaches. Although they have been underutilised in the past, application of genetic and genomic techniques is becoming more feasible from both technological and economic perspectives. We review current methods and provide a framework for using them. First, it is necessary to identify which biocontrol trait to select and in what direction. Nex…
Plant guttation provides nutrient-rich food for insects
Plant guttation is a fluid from xylem and phloem sap secreted at the margins of leaves from many plant species. All previous studies have considered guttation as a water source for insects. Here, we hypothesized that plant guttation serves as a reliable and nutrient-rich food source for insects with effects on their communities. Using highbush blueberries as a study system, we demonstrate that guttation droplets contain carbohydrates and proteins. Insects from three feeding lifestyles, a herbivore, a parasitic wasp and a predator, increased their longevity and fecundity when fed on these guttation droplets compared to those fed on control water. Our results also show that guttation droplets…
Large-Scale Monitoring of Resistance to Coumaphos, Amitraz, and Pyrethroids in Varroa destructor
ABSTRACTVarroa destructor is an ectoparasitic mite causing devastating damages to honey bee colonies around the world. Its impact is considered a major factor contributing to the significant seasonal losses of colonies recorded every year. Beekeepers are usually relying on a reduced set of acaricides to manage the parasite, usually the pyrethroids tau-fluvalinate or flumethrin, the organophosphate coumaphos and the formamidine amitraz. However, the evolution of resistance in the populations is leading to an unsustainable scenario with almost no alternatives to reach an adequate control of the mite.Here we present the results from the first, large-scale and extensive monitoring of the suscep…
Binding of Bacillus thuringiensis toxins in resistant and susceptible strains of pink bollworm (Pectinophora gossypiella)
Abstract Evolution of resistance by pests could cut short the success of transgenic plants producing toxins from Bacillus thuringiensis, such as Bt cotton. The most common mechanism of insect resistance to B. thuringiensis is reduced binding of toxins to target sites in the brush border membrane of the larval midgut. We compared toxin binding in resistant and susceptible strains of Pectinophora gossypiella, a major pest of cotton worldwide. Using Cry1Ab and Cry1Ac labeled with 125I and brush border membrane vesicles (BBMV), competition experiments were performed with unlabeled Cry1Aa, Cry1Ab, Cry1Ac, Cry1Ba, Cry1Ca, Cry1Ja, Cry2Aa, and Cry9Ca. In the susceptible strain, Cry1Aa, Cry1Ab, Cry1…
Shared Binding Sites in Lepidoptera for Bacillus thuringiensis Cry1Ja and Cry1A Toxins
ABSTRACT Bacillus thuringiensis toxins act by binding to specific target sites in the insect midgut epithelial membrane. The best-known mechanism of resistance to B. thuringiensis toxins is reduced binding to target sites. Because alteration of a binding site shared by several toxins may cause resistance to all of them, knowledge of which toxins share binding sites is useful for predicting cross-resistance. Conversely, cross-resistance among toxins suggests that the toxins share a binding site. At least two strains of diamondback moth ( Plutella xylostella ) with resistance to Cry1A toxins and reduced binding of Cry1A toxins have strong cross-resistance to Cry1Ja. Thus, we hypothesized that…
Neonicotinoids in excretion product of phloem-feeding insects kill beneficial insects
Significance The use of insecticides in agriculture is one of the suggested causes of the decline in insect populations. Neonicotinoids are among the most widely used insecticides. However, they have important negative side effects, especially for pollinators and other beneficial insects feeding on floral nectar and pollen. We identified an exposure route: Neonicotinoids reach and kill beneficial insects when they feed on the most abundant carbohydrate source for insects in agroecosystems, honeydew. Honeydew is the excretion product of phloem-feeding hemipteran insects such as aphids, mealybugs, whiteflies, or psyllids. This route of exposure is likely to affect a much wider range of benefi…