Cross-resistance and mechanism of resistance to Cry1Ab toxin from Bacillus thuringiensis in a field-derived strain of European corn borer, Ostrinia nubilalis.

The cross-resistance spectrum and biochemical mechanism of resistance to the Bacillus thuringiensis Cry1Ab toxin was studied in a field-derived strain of Ostrinia nubilalis (Hübner) (Lepidoptera: Crambidae) that was further selected in the laboratory for high levels (>1000-fold) of resistance to Cry1Ab. The resistant strain exhibited high levels of cross-resistance to Cry1Ac and Cry1Aa but only low levels of cross-resistance (<4-fold) to Cry1F. In addition, there was no significant difference between the levels of resistance to full-length and trypsin-activated Cry1Ab protein. No differences in activity of luminal gut proteases or altered proteolytic processing of the toxin were observed in the resistant strain. Significantly reduced binding of radiolabeled Cry1Aa was observed in the resistant strain whereas binding of Cry1Ab and Cry1Ac was practically the same in both resistant and susceptible strains. The interpretation of the overall data seems to suggest the involvement of an alteration in the binding of Cry1A toxins to a common receptor, which is more clearly revealed by the binding assays using radiolabeled Cry1Aa.

Fitness costs of Cry1Ab resistance in a field-derived strain of Ostrinia nubilalis (Lepidoptera: Crambidae).

The study of fitness costs associated with resistance to toxins from Bacillus thuringiensis Berliner (Bt) in Ostrinia nubilalis (Hübner) (Lepidoptera: Crambidae) is important for understanding resistance evolution and for evaluating resistance management practices that prevent or mitigate resistance to transgenic corn, Zea mays L. Resistant individuals identified from a field collection in Kandiyohi, MN, were used to generate a Cry1Ab-resistant strain. We used susceptible and resistant strains with similar genetic background to establish crosses and estimate dominance of fitness costs by measuring fitness components and population parameters determined by fertility life tables. Spermatophore volume and mating frequency also were compared to identify potential effects of resistance on fertility. Inheritance of fitness costs in O. nubilalis varied from recessive to incompletely recessive among the parameters evaluated. Selection for resistance to Cry1Ab significantly reduced the fitness of O. nubilalis. Resistant insects exhibited reduced pupal weight and increased developmental time compared with susceptible and F1 larvae derived from reciprocal crosses of resistant and susceptible parents. In addition, it was observed that resistant insects exhibited a higher proportion of unsuccessful matings and lower fertility than the susceptible strain. Despite the differences observed in resistant insects, our results did not indicate strong evidence of fitness costs in the F1 progeny.

Conventional Sampling Plan for Scouting Neoleucinodes elegantalis (Lepidoptera: Crambidae) Eggs on Tomato Fruits.

The small tomato borer, Neoleucinodes elegantalis (Guenée), is an important pest of tomato (Solanum lycopersicum L.) in South and Central America. This pest is a potential threat for many tomato-producing areas and was listed in 2014 as an A1 quarantine pest by the European and Mediterranean Plant Protection Organization. Soon after hatching, the neonate N. elegantalis larvae penetrate the fruits where they feed until pupation. Therefore, effective N. elegantalis management relies on the timely scouting of egg densities to allow decision-making prior to penetration of neonates into fruits. This study aimed to develop a conventional sampling plan for scouting N. elegantalis eggs on tomato plants. The most representative and precise sampling unit was the most basal fruit cluster for plants bearing up to three clusters (S1 plants). For plants with more than three fruit clusters (S2 plants), the most representative and precise sampling unit was the combination of the second and third fruit clusters. Among the four variables evaluated (eggs/fruit, egg masses/fruit, percentage fruit with eggs, and percentage clusters with eggs), the percentage of clusters with eggs was the most economical for N. elegantalis sampling based on the number of samples and cost required. For this variable, the number of samples determined at the 25% error level was 42 and 36 samples for S1 and S2 plants, respectively. The sampling plan developed for scouting N. elegantalis is fast, reflects pest infestation in tomato fields, and costs less than US$1.50 per field scouted.

Comparison and validation of methods to quantify Cry1Ab toxin from Bacillus thuringiensis for standardization of insect bioassays.

Standardization of toxin preparations derived from Bacillus thuringiensis (Berliner) used in laboratory bioassays is critical for accurately assessing possible changes in the susceptibility of field populations of target pests. Different methods were evaluated to quantify Cry1Ab, the toxin expressed by 80% of the commercially available transgenic maize that targets the European corn borer, Ostrinia nubilalis (Hübner). We compared three methods of quantification on three different toxin preparations from independent sources: enzyme-linked immunosorbent assay (ELISA), sodium dodecyl sulfate-polyacrylamide gel electrophoresis and densitometry (SDS-PAGE/densitometry), and the Bradford assay for total protein. The results were compared to those obtained by immunoblot analysis and with the results of toxin bioassays against susceptible laboratory colonies of O. nubilalis. The Bradford method resulted in statistically higher estimates than either ELISA or SDS-PAGE/densitometry but also provided the lowest coefficients of variation (CVs) for estimates of the Cry1Ab concentration (from 2.4 to 5.4%). The CV of estimates obtained by ELISA ranged from 12.8 to 26.5%, whereas the CV of estimates obtained by SDS-PAGE/densitometry ranged from 0.2 to 15.4%. We standardized toxin concentration by using SDS-PAGE/densitometry, which is the only method specific for the 65-kDa Cry1Ab protein and is not confounded by impurities detected by ELISA and Bradford assay for total protein. Bioassays with standardized Cry1Ab preparations based on SDS-PAGE/densitometry showed no significant differences in LC(50) values, although there were significant differences in growth inhibition for two of the three Cry1Ab preparations. However, the variation in larval weight caused by toxin source was only 4% of the total variation, and we conclude that standardization of Cry1Ab production and quantification by SDS-PAGE/densitometry may improve data consistency in monitoring efforts to identify changes in insect susceptibility to Cry1Ab.

Blended Refuge and Insect Resistance Management for Insecticidal Corn.

In this review, we evaluate the intentional mixing or blending of insecticidal seed with refuge seed for managing resistance by insects to insecticidal corn (Zea mays). We first describe the pest biology and farming practices that will contribute to weighing trade-offs between using block refuges and blended refuges. Case studies are presented to demonstrate how the trade-offs will differ in different systems. We compare biological aspects of several abstract models to guide the reader through the history of modeling, which has played a key role in the promotion or denigration of blending in various scientific debates about insect resistance management for insecticidal crops. We conclude that the use of blended refuge should be considered on a case-by-case basis after evaluation of insect biology, environment, and farmer behavior. For Diabrotica virgifera virgifera, Ostrinia nubilalis, and Helicoverpa zea in the United States, blended refuge provides similar, if not longer, delays in the evolution of resistance compared to separate block refuges.

Toxicity of insecticides to the sweetpotato whitefly (Hemiptera: Aleyrodidae) and its natural enemies.

Efficient chemical control is achieved when insecticides are active against insect pests and safe to natural enemies. In this study, the toxicity of 17 insecticides to the sweetpotato whitefly, Bemisia tabaci (Gennadius), and the selectivity of seven insecticides to natural enemies of this insect pest were evaluated. To determine the insecticide toxicity, B. tabaci adults were exposed to abamectin, acephate, acetamiprid, cartap, imidacloprid, malathion, methamidophos, bifenthrin, cypermethrin, deltamethrin, esfenvalerate, fenitrothion, fenpropathrin, fenthion, phenthoate, permethrin and trichlorphon at 50 and 100% of the field rate (FR), and to water (untreated control). To determine the insecticide selectivity, adults of Encarsia sp., Acanthinus sp., Discodon sp. and Lasiochilus sp. were exposed to abamectin, acephate, acetamiprid, cartap, imidacloprid, malathion and methamidophos at 50 and 100% FR, and to water. Groups of each insect species were exposed to kale leaves preimmersed in each treatment under laboratory conditions. Mortality of exposed individuals was recorded 24 h after treatment. Cartap and imidacloprid at 50 and 100% FR and abamectin and acetamiprid at 100% FR showed insecticidal activity to B. tabaci adults. Abamectin at 50 and 100% FR was the least insecticidal compound to the natural enemies Acanthinus sp., Discodon sp. and Lasiochilus sp. The present results suggest that abamectin at 100% FR may decrease B. tabaci field populations but can still be harmless to predators. Implications of these results within an integrated pest management context are discussed.

Seletividade de Inseticidas a Brevicoryne brassicae (L. ) (Hemiptera: Aphididae) e ao Predador Doru luteipes (Scudder) (Dermaptera: Forficulidae) / Insecticide Selectivity to Brevicoryne brassicae (L. ) (Hemiptera: Aphididae) and the Predator Doru luteipes (Scudder) (Dermaptera: Forficulidae)

The objective of this work was to study the selectivity of the insecticides acephate, deltamethrin, dimethoate, methamidophos, methyl parathion, and pirimicarb to first, second, and third instar nymphs and adults of the predator Doru luteipes (Scudder) (Dermaptera: Forficulidae) in comparison with last instar nymphs of the prey Brevicoryne brassicae (L.) (Hemiptera: Aphididae). Pirimicarb and deltamethrin were highly selective to all developmental stages of D. luteipes; the same was observed with acephate except for first instar nymphs to whom it showed median selectivity. Dimethoate showed median selectivity, but methamidophos and methyl parathion were low selective to the predator. Second and third instar nymphs and adults of D. luteipes were more tolerant to acephate, methyl parathion and pirimicarb than first instar nymphs. First and second instar nymphs showed similar tolerance to deltamethrin, dimethoate and methamidophos. Concentration-response curves for dimethoate (to adults and first and second instar nymphs) and methamidophos (to third instar nymphs) showed steeper slopes indicating an homogeneous response of these developmental stages to these insecticides.

Este trabalho teve por objetivo estudar a seletividade dos inseticidas acefato, deltametrina, dimetoato, metamidofós, paratiom metílico e pirimicarbe a ninfas de último estádio de Brevicoryne brassicae (L.) (Hemiptera: Aphididae), em relação a adultos e ninfas de primeiro, segundo e terceiro estádios do predador Doru luteipes (Scudder) (Dermaptera: Forficulidae). Pirimicarbe e deltametrina foram altamente seletivos em favor de todos os estádios de desenvolvimento de D. luteipes, sendo que o mesmo ocorreu para o acefato, com exceção de ninfas de primeiro estádio do predador às quais ele é medianamente seletivo. Dimetoato foi, em geral, medianamente seletivo enquanto metamidofós e paratiom metílico foram pouco seletivos em favor do predador. Adultos e ninfas de segundo e terceiro estádios de D. luteipes foram mais tolerantes ao acefato, paratiom metílico e pirimicarbe do que ninfas de primeiro estádio do predador. Ninfas de primeiro e segundo estádios apresentaram tolerância semelhante a deltametrina, dimetoato e metamidofós. As curvas de concentração-mortalidade do dimetoato (para adultos, ninfas de primeiro e segundo estádios do predador) e metamidofós (para ninfas de terceiro estádio) apresentaram as maiores inclinações, indicando que os estádios estudados apresentam resposta homogênea a esses inseticidas.