Insecticide‐induced hormesis in an insecticide‐resistant strain of the maize weevil,Sitophilus zeamais

Sublethal responses to insecticides are frequently neglected in studies of insecticide resistance, although stimulatory effects associated with low doses of compounds toxic at higher doses, such as insecticides, have been recognized as a general toxicological phenomenon. Evidence for this biphasic dose–response relationship, or hormesis, was recognized as one of the potential causes underlying pest resurgence and secondary pest outbreaks. Hormesis has also potentially important implications for managing insecticide‐resistant populations of insect‐pest species, but evidence of its occurrence in such context is lacking and fitness parameters are seldom considered in these studies. Here, we reported the stimulatory effect of sublethal doses of the pyrethroid insecticide deltamethrin sprayed on maize grains infested with a pyrethroid‐resistant strain of the maize weevil (Sitophilus zeamais) (Coleoptera: Curculionidae). The parameters estimated from the fertility tables of resistant insects exposed to deltamethrin indicated a peak in the net reproductive rate at 0.05 ppm consequently leading to a peak in the intrinsic rate of population growth at this dose. The phenomenon is consistent with insecticide‐induced hormesis and its potential management implications are discussed.     

High dose refuge strategies and genetically modified crops – reply to Tabashnik et al.

Cultivating non‐toxic conventional crops (refuges) in the proximity to transgenic crops that produce Bacillus thuringienesis (Bt) toxins is widely recommended to delay pest adaptation to these toxins. Using a spatially structured model of resistance evolution, Vacher and co‐workers (Vacher, C., Bourguet, D., Rousset, F., Chevillon, C. & Hochberg, M.E. 2003. J. Evol. Biol. 16 : 378–387.) show that the percentage of refuge fields required for the sustainable control of pests can be reduced through intermediate levels of refuge field aggregation and by lowering the toxin dose produced by Bt plants. Tabashnik, B.E., Gould, F. & Carrière, Y. (2004 J. Evol. Biol doi: 10.1111/j1420–9101.2004.00695.x) call into question the results of Vacher et al. (2003) concerning the effect of toxin dose. They argue that these results arise from invalid assumptions about larval concentration–mortality responses for the insect considered, the cotton pest Heliothis virescens. We show here that the models presented by Vacher et al. (2003) and Tabashnik et al. (2004) both show inaccuracies in their definitions of genotypic fitness. The level of dominance estimated by Tabashnik et al. (2004) from larval mortality rates data is irrelevant to resistance evolution, and the fitness cost of resistance evolution, and the fitness cost of resistance is inaccurately integrated into their framework. Neverthless, the comments of Tabashnik et al. (2004) are very helpful in elucidating the definitions of genotypic fitness used in Vacher et al. (2003) and in pointing out the essential factors in predicting the evolution of insect resistance to Bt transgenic crops, namely, accurate estimations of the fitness cost of resistance, of the dominance level of this cost, and of the variations in the dominance level of the advantage conferred by the resistance with Bt toxin dose.     

棉铃虫对Bt杀虫剂的抗性遗传方式

随着转基因棉花种植面积的日益增加,棉铃虫Helicoverpa armigera（Hübner)对Bt的抗性已经成为一个不容忽视的问题。发展转多价基因作物是当前缓解害虫对Bt抗性的最有效措施。本研究以经室内多年筛选的、抗性倍数达2 000多倍的Bt杀虫剂（含多种蛋白）抗性品系为材料,通过生物测定和不同的杂交试验,测定棉铃虫对Bt杀虫剂的抗性遗传方式,以期为Bt生物农药的抗性治理提供一定的依据,同时为制定棉铃虫对转多基因作物的抗性治理策略提供一定的参考。对敏感亲本和抗性亲本杂交产生的F1代的研究结果表明,杂交品系的抗性倍数分别为22.2倍和24.6倍;抗性显性度D值均小于0,分别为-0.20和-0.17,抗性为常染色体不完全隐性遗传。对4种回交后代和2种自交后代F2的研究结果表明,实际死亡率与期望死亡率差异较大,说明抗性是由单基因多个位点或多基因控制。     

Comparing the refuge strategy for managing the evolution of insect resistance under different reproductive strategies

Genetically modified (GM) crops are used extensively worldwide to control diploid agricultural insect pests that reproduce sexually. However, future GM crops will likely soon target haplodiploid and parthenogenetic insects. As rapid pest adaptation could compromise these novel crops, strategies to manage resistance in haplodiploid and parthenogenetic pests are urgently needed. Here, we developed models to characterize factors that could delay or prevent the evolution of resistance to GM crops in diploid, haplodiploid, and parthenogenetic insect pests. The standard strategy for managing resistance in diploid pests relies on refuges of non-GM host plants and GM crops that produce high toxin concentrations. Although the tenets of the standard refuge strategy apply to all pests, this strategy does not greatly delay the evolution of resistance in haplodiploid or parthenogenetic pests. Two additional factors are needed to effectively delay or prevent the evolution of resistance in such pests, large recessive or smaller non-recessive fitness costs must reduce the fitness of resistance individuals in refuges (and ideally also on GM crops), and resistant individuals must have lower fitness on GM compared to non-GM crops (incomplete resistance). Recent research indicates that the magnitude and dominance of fitness costs could be increased by using specific host–plants, natural enemies, or pathogens. Furthermore, incomplete resistance could be enhanced by engineering desirable traits into novel GM crops. Thus, the sustainability of GM crops that target haplodiploid or parthenogenetic pests will require careful consideration of the effects of reproductive mode, fitness costs, and incomplete resistance.     

The importance of modelling the spread of insecticide resistance in a heterogeneous environment: the example of adding synergists to bed nets

ABSTRACT: BACKGROUND: Insecticides are an effective and practical tool for reducing malaria transmission but the development of resistance to the insecticides can potentially compromise controls efforts. In this study a mathematical model was developed to explore the effects on mosquito populations of spatial heterogeneous deployment of insecticides. This model was used to identify important parameters in the evolution of insecticide resistance and to examine the contribution of new generation long-lasting insecticidal bed nets, that incorporate a chemical synergist on the roof panel, in delaying insecticide resistance. METHODS: A genetic model was developed to predict changes in mosquito fitness and resistance allele frequency. Parameters describing insecticide selection, fitness cost and the additional use of synergist were incorporated. Uncertainty and sensitivity analysis were performed followed by investigation of the evolution of resistance under scenarios of fully effective or ineffective synergists. RESULTS: The spread of resistance was most sensitive to selection coefficients, fitness cost and dominance coefficients while mean fitness was most affected by baseline fitness levels. Using a synergist delayed the spread of resistance but could, in specific circumstances that were thoroughly investigated, actually increase the rate of spread. Different spread dynamics were observed, with simulations leading to fixation, loss and most interestingly, equilibrium (without explicit overdominance) of the resistance allele. CONCLUSIONS: This strategy has the potential to delay the spread of resistance but note that in an heterogeneous environment it can also lead to the opposite effect, i.e., increasing the rate of spread. This clearly emphasizes that selection pressure acting inside the house cannot be treated in isolation but must be placed in context of overall insecticide use in an heterogeneous environment.     

Inheritance of Cry1F resistance in laboratory-selected European corn borer and its survival on transgenic corn expressing the Cry1F toxin

A major assumption of the high-dose/refuge strategy proposed for insect resistance management strategies for transgenic crop plants that express toxins from Bacillus thuringiensis is that resistance traits that evolve in pest species will be recessive. The inheritance of Cry1F resistance and larval survival on commercially available Cry1F corn hybrids were determined in a laboratory-selected strain of European corn borer, Ostrinia nubilalis (Hübner), displaying more than 3000-fold resistance to Cry1F. Concentration-response bioassays of reciprocal parental crosses indicated that the resistance is autosomal and recessive. Bioassays of the backcross of the F1 generation with the selected strain were consistent with the hypothesis that a single locus, or a set of tightly linked loci, is responsible for the resistance. Greenhouse experiments with Cry1F-expressing corn hybrids indicated that some resistant larvae survived the high dose of toxin delivered by Cry1F-expressing plants although F1 progeny of susceptible by resistant crosses had fitness close to zero. These results provide the first direct evidence that the high dose/refuge strategy currently in place to manage resistance in Cry1F-expressing corn is appropriate.     

Frequency and fitness cost of resistance to Bacillus thuringiensis in Chrysomela tremulae (Coleoptera: Chrysomelidae)

The "high dose-refuge" (HDR) strategy is commonly recommended and currently used for delaying or preventing pest adaptation to transgenic plants producing Bacillus thuringiensis (Bt) toxins. The efficiency of this strategy depends, among other factors, on the initial frequency of Bt resistance alleles and on the fitness costs associated with these alleles. Two years ago, an allele conferring resistance to Bt poplar was detected in a French population of the poplar pest Chrysomela tremulae F. Although this pest had never been subjected to Bt selection pressure due to human activities, the frequency of this allele was estimated at 0.0037, with a 95% credible (CI) interval of 0.00045-0.0080. We investigated the frequency of this allele in a second sample of C. tremulae collected more than 500 km from the site of the initial population. The estimated frequency in this sample was 0.0113 (95% CI 0.0031-0.0247), reinforcing the conclusion that resistance to Bt plants may be present at detectable frequencies in pest populations before selection resulting from pest management by humans. The frequency of the Bt resistance allele over the two samples was 0.0049 (95% CI 0.0020-0.0091). We also followed five laboratory lines in which the frequency of this allele was initially fixed at 0.500. After five generations maintained on non-Bt poplar leaves, the frequency of this allele decreased in all lines, whereas allelic frequencies at a neutral locus were unaffected. Thus, the Bt resistance allele detected in French populations of C. tremulae is probably associated with a fitness cost.     

The biology of insecticidal activity and resistance

Identifying insecticide resistance mechanisms is paramount for pest insect control, as the understandings that underpin insect control strategies must provide ways of detecting and managing resistance. Insecticide resistance studies rely heavily on detailed biochemical and genetic analyses. Although there have been many successes, there are also many examples of resistance that still challenge us. As a precursor to rational pest insect control, the biology of the insect, within the contexts of insecticide modes of action and insecticide metabolism, must be well understood. It makes sense to initiate this research in the best model insect system, Drosophila melanogaster, and translate these findings and methodologies to other insects. Here we explore the usefulness of the D. melanogaster model in studying metabolic-based insecticide resistances, target-site mediated resistances and identifying novel insecticide targets, whilst highlighting the importance of having a more complete understanding of insect biology for insecticide studies.     

Mating behaviour and reproductive output in insecticide‐resistant and ‐susceptible strains of the maize weevil (Sitophilus zeamais)

Insecticide resistance is a broadly recognised and well‐studied management problem resulting from intensive insecticide use, which also provides useful evolutionary models of newly adapted phenotypes to changing environments. Two common assumptions in such models are the existence of fitness costs associated with insecticide resistance, which will place the resistant individuals at a disadvantage in insecticide‐free environments, and the prevalence of random mating among insecticide‐resistant and ‐susceptible individuals. However, cases of insecticide resistance lacking apparent fitness disadvantages do exist impacting the evolution and management of insecticide resistance. Assortative mating, although rarely considered, may also favour the evolution and spread of insecticide resistance. Thus, the possible existence of both conditions in the maize weevil (Sitophilus zeamais), a key pest of stored cereals, led to the assessment of the mating behaviour and reproductive fitness of insecticide‐resistant and ‐susceptible weevil strains and their reciprocal crosses. The patterns of female and male mating choice also were assessed. Although mating behaviour within and between weevil strains was similar without mate choice, mating within the resistant strain led to higher reproductive output than within the susceptible strain; inter‐strain matings led to even higher fertility. Thus, no apparent fitness cost associated with resistance seems to exist in these weevils, favouring the evolution of this phenotype that is further aided by the higher fertility of inter‐strain matings. Mate choice reduced latency to mate and no inter‐strain preference was detected, but female weevils were consistent in their mate selection between 1st and 2nd matings indicating existence of female mating preference among maize weevils. Therefore, if female mate selection comes to favour trait(s) associated with insecticide resistance, higher reproductive fitness will be the outcome of such matings favouring the evolution and spread of insecticide resistance among maize weevil populations reverting into a management concern.     

Effect of entomopathogenic nematodes on the fitness cost of resistance to Bt toxin crylac in pink bollworm (Lepidoptera: Gelechiidae)

The widespread use of crop plants genetically engineered to produce toxins from the bacterium Bacillus thuringiensis (Bt) imposes selection on insect populations to evolve resistance. The pink bollworm, Pectinophora gossypiella (Saunders) (Lepidoptera: Gelechiidae), is a major pest of cotton in the southwestern United States that is currently controlled with transgenic cotton that produces Bt toxin Cry1Ac. Previously reported theoretical work suggests that, in conjunction with a high dose/refuge strategy, fitness costs of Bt resistance can slow or prevent the evolution of resistance. We report here that the entomopathogenic nematode Steinernema riobrave (Rhabditida: Steinernematidae) increased the fitness cost of resistance to Cry1Ac in P. gossypiella. Mortality of P. gossypiella from fourth instar to adult eclosion was significantly higher for a Bt-resistant strain than a susceptible strain in tests with two to 14 infective juveniles of S. riobrave per larva, but it did not differ between strains when nematodes were absent. Nematodes established in P. gossypiella larvae at all concentrations tested, and nematode reproduction in infected P. gossypiella larvae occurred at nematode concentrations of four to 14 infective juveniles per larva. Our results suggest that incorporation of entomopathogenic nematodes into an integrated resistance management strategy could help to delay pest resistance to Bt toxins.     

Assessment of fitness costs in Cry3Bb1‐resistant and susceptible western corn rootworm (Coleoptera: Chrysomelidae) laboratory colonies

Abstract Maize production in the United States is dominated by plants genetically modified with transgenes from Bacillus thuringiensis (Bt). Cry3Bb delta endotoxins expressed by Bt maize specifically target corn rootworms (genus Diabrotica) and have proven highly efficacious. However, development of resistance to Bt maize, especially among western corn rootworm (Diabrotica virgifera virgifera) populations, poses a significant threat to the future viability of this pest control biotechnology. The structured refuge insect resistance management (IRM) strategy implemented in the United States for Bt maize adopts a conservative approach to managing resistance by assuming no fitness costs of Bt resistance, even though these trade‐offs strongly influence the dynamics of Bt resistance within numerous agricultural pest species. To investigate the effects of Bt resistance on fitness components of western corn rootworm, we compared survivorship, fecundity and viability of five Bt‐resistant laboratory lines reared on MON863 (YieldGard Rootworm), a Bt maize product that expresses Cry3Bb1 delta endotoxin, and on its non‐transgenic isoline. Analysis of performance on the isoline maize demonstrated no fitness costs associated with Bt resistance. In fact, resistant lines emerged approximately 2–3 days earlier than control lines when reared on both MON863 and the isoline, indicating that selection for Bt resistance resulted in a general increase in the rate of larval development. In addition, resistant lines reared on Bt maize displayed higher fecundity than those reared on the isoline, which may have significant management implications. These data will be valuable for formulating improved IRM strategies for a principal agricultural pest of maize.     

Resistance management: the stable zone strategy

The different strategies of insecticide resistance management that have been formulated so far consist of delaying the appearance and spread of resistance genes. In this paper, we propose a strategy that can be used even if resistance genes are already present. This strategy consists of applying insecticides in an area smaller than a certain critical size, so that gene flow from the untreated area, combined with the fitness cost of the resistance genes, prevents its frequency reaching high equilibrium value. A two-locus model was analysed numerically to determine population densities at equilibrium as a function of selection coefficients (insecticide selection, fitness costs of resistance genes and dominances), gene flow and size of the treated area. This model indicates that there is an optimal size for the treated area where a minimal and stable density reach equilibrium, and where resistance genes cannot invade. This resistance management strategy seems applicable to a large variety of field situations, but eventually it may encounter obstacles due to a modifier which reduces the fitness costs of resistance genes.     

Evolution of resistance to Bt crops: directional selection in structured environments

Abstract Widespread use of transgenic crops that express an insecticidal endotoxin from Bacillus thuringiensis increases the risk of evolution of resistance by the European corn borer and other insect pests. To delay resistance evolution, the high‐dose/refuge strategy is being implemented for Bt maize and Bt cotton. We develop a general modelling framework to understand the invasion and spread of alleles conferring resistance. We show that at least three processes are involved in explaining the effectiveness of the high‐dose/refuge strategy: the intensity of selection, assortative (non‐random) mating due to spatial subdivision, and variation in male mating success also due to spatial subdivision. Understanding these processes leads to a greater range of possible resistance management tactics. For example, efforts to encourage adults to leave their natal fields may have the unwanted effect of speeding rather than slowing resistance evolution. Furthermore, when Bt maize causes high mortality to susceptible target pests, spraying insecticides in refuges to reduce pest populations may not greatly disrupt resistance management.     

Mortality and knockdown effects of imidacloprid and methomyl in house fly (Musca domestica L., Diptera: Muscidae) populations

In this study, the knockdown and mortality effects of imidacloprid and methomyl were investigated. The residual surface applications were carried out to determine the knockdown effects (KDt₅₀ and KDt₉₅) and mortality (LD₅₀ and LD₉₅) induced by each insecticide. For mortality comparisons, the susceptible house fly (Musca domestica L., Diptera: Muscidae) of a WHO population and three natural field‐collected M. domestica populations from Turkey were used. In conclusion, it was found that the resistance to imidacloprid and methomyl was significantly higher in the field populations when compared to the susceptible population from WHO. The results showed that applicators and pest management decision‐makers should control and conduct an integrated pest management strategy by including biological agents to prevent the development of high levels of resistance in the field populations.     

Suppressing resistance to Bt cotton with sterile insect releases

Genetically engineered crops that produce insecticidal toxins from Bacillus thuringiensis (Bt) are grown widely for pest control. However, insect adaptation can reduce the toxins' efficacy. The predominant strategy for delaying pest resistance to Bt crops requires refuges of non-Bt host plants to provide susceptible insects to mate with resistant insects. Variable farmer compliance is one of the limitations of this approach. Here we report the benefits of an alternative strategy where sterile insects are released to mate with resistant insects and refuges are scarce or absent. Computer simulations show that this approach works in principle against pests with recessive or dominant inheritance of resistance. During a large-scale, four-year field deployment of this strategy in Arizona, resistance of pink bollworm (Pectinophora gossypiella) to Bt cotton did not increase. A multitactic eradication program that included the release of sterile moths reduced pink bollworm abundance by >99%, while eliminating insecticide sprays against this key invasive pest.     

Monitoring western corn rootworm (Coleoptera: Chrysomelidae) susceptibility to carbaryl and cucurbitacin baits in the areawide management pilot program

Areawide pest management involves the uniform application of a pest control strategy over wide geographic areas. Therefore, these programs are likely to impose intense selective pressures, and the risk for resistance development among pest species for which areawide management programs are implemented is likely to be high. Pilot studies for areawide management of western corn rootworm, Diabrotica virgifera virgifera LeConte, were conducted from 1996 to 2002 at four different sites across the Corn Belt. This program used cucurbitacin baits to deliver high doses of a traditional neurotoxic insecticide (carbaryl) to individual insects while reducing the overall rate of insecticide use. Because of the concern and potential for resistance evolution, annual assessments of susceptibility to the active ingredient carbaryl were conducted both within the managed area as well as from untreated control areas. Significantly reduced susceptibility to carbaryl based on survival at a diagnostic concentration was detected in three of the four management sites (Kansas, Iowa, and Illinois/Indiana), whereas susceptibility of beetles collected outside the managed areas remained unchanged. Additionally, significantly reduced responsiveness to cucurbitacin baits was observed in beetles collected from the managed area relative to the control area at the same three sites. These results suggest strongly that areawide management has the potential to select for resistance and that a strategy for managing resistance and reducing selective pressure should be proactively implemented.     

Sublethal effects of deltamethrin exposure of parental generations on physiological traits and overwintering in Leptinotarsa decemlineata

Although the evolution of insecticide resistance has received a lot of attention, less is known about sublethal effects of insecticide stress experienced by the preceding generations on the performance of pest populations. We investigated whether three generations of parental exposure to a deltamethrin insecticide influences physiological traits and overwintering success of offspring in Leptinotarsa decemlineata (Say) (Coleoptera: Chrysomelidae). Beetles descending from insecticide‐exposed parents had lower adult body mass but higher relative lipid content and resting metabolic rate than those descending from non‐insecticide‐exposed parents. Also, a higher proportion of beetles descending from insecticide‐exposed parents than from control parents overwintered on the soil surface, which was associated with very low overwintering survival. When burrowed into the soil for overwintering, both groups had similar probability to survive the overwintering period. Parental insecticide exposure can reduce overall overwintering survival in the next generation by disturbing overwintering behaviour. Although beetles descending from insecticide‐exposed parents were small, the overwintering conditions seem to select for high lipid content, which could override the negative effects of small weight, and possibly improve fitness in the following growing season. Thus, insecticide application may have unintended consequences, which should be considered in pest management.     

Inheritance of lambda-cyhalothrin resistance in the predator lady beetle Eriopis connexa (Germar) (Coleoptera: Coccinellidae)

Lambda-cyhalothrin is a pyrethroid insecticide widely used to control pests in various crop ecosystems, in which the lady beetle occurs naturally. Therefore, lady beetle populations are exposed to lambda-cyhalothrin sprays that may foster tolerance to this insecticide. This study was conducted to confirm the occurrence of resistance in the lady beetle Eriopis connexa (Germar) (Coleoptera: Coccinellidae) to lambda-cyhalothrin and to characterize the inheritance of resistance after eight progressive selection with insecticide dosages based on the LD₅₀ determined for the F1 generation. Dose–mortality curves were determined for parental populations, F1 hybrids and backcross progenies. Parameters regarding the heritability of resistance to lambda-cyhalothrin in E. connexa allow estimating 10-fold increase in the initial LD₅₀ after 54.5 generations of successive selections. The resistance of E. connexa to lambda-cyhalothrin was characterized as autosomally inherited and incompletely dominant, and influenced by a major gene with possible influence of secondary genes. Additionally, the resistance in E. connexa varies from functionally dominant to functionally recessive depending on the dose used. These findings indicate that insecticide resistance in E. connexa can be selected in the field as determined for field-collected individuals, and subsequently enhanced under laboratory conditions. Its characterization presented here is an important step toward linking biological and chemical control within pest management regarding the lady beetle and lambda-cyhalothrin targeting different pest groups.     

Combining genetic engineering and traditional breeding to provide elevated resistance in potatoes to Colorado potato beetle

The sustainable deployment of resistant crop varieties is a critical issue for the implementation of biotechnology in crop pest management. Feeding, biomass accumulation, and mortality were evaluated for susceptible, insecticide‐resistant, and Bacillus thuringiensis (Bt) Cry 3A‐selected Colorado potato beetle (Leptinotarsa decemlineata Say) (Coleoptera, Chrysomelidae) larvae fed on: cultivated potato, a Solanum chacoense line expressing leptine glycoalkaloids, a transformed line expressing Bt toxin, or the leptine line transformed to express Bt toxin. Larvae selected for resistance to Bt‐Cry3A performed better on Bt foliage, but not as well on the leptine foliage, compared to susceptible or insecticide‐resistant larvae. Neither leptine nor Bt toxin completely inhibited the feeding and growth of 3rd and 4th instars of all three strains of Colorado potato beetle. However, for all three strains of Colorado potato beetle on leptine + Bt foliage, feeding was almost zero, growth was zero or negative, and mortality was near 100%.