Initial Resistance Allele Frequency Research Articles

Modeling the evolution of resistance in cotton bollworm to concurrently planted Bt cotton and Bt maize in China

Transgenic maize expressing toxins derived from the bacterium Bacillus thuringiensis (Bt) may be commercially planted in northern China where Bt cotton has been planted for more than two decades. While Bt maize brings additional benefits for insect control, it complicates the resistance management of cotton bollworm (CBW), Helicoverpa armigera (Lepidoptera, Noctuidae), a common target of Bt cotton and Bt maize. Here we developed two-locus population genetic models to assess the risk of resistance in CBW in which four cases of Bt cotton and Bt maize and two types of refuges are considered. Model simulations showed that the time to resistance (TTR) is longest in the case of two-toxin Bt cotton & two-toxin Bt maize, followed by the cases of two-toxin Bt cotton & one-toxin Bt maize, one-toxin Bt cotton & two-toxin Bt maize, and one-toxin Bt cotton & one-toxin Bt maize. With 25% of cotton seed mixed refuge and 20% of maize seed mixed refuge, the TTRs in the four cases by order are 54, 13, 9, and 7 generations, respectively. With additional natural refuges, the differences in the TTRs among the four cases are greater. Sensitivity analysis showed that among the parameters examined, the initial frequency of resistance alleles and fitness cost are the ones to which the TTRs are most and least sensitive, respectively. We concluded that when natural refuges are scarce, planting both two-toxin Bt cotton and two-toxin Bt maize instead of one-toxin ones are necessary to combat CBW resistance to concurrently planted Bt cotton and Bt maize in northern China.

An Interactive Teaching Tool Describing Resistance Evolution and Basic Economics of Insecticide-Based Pest Management.

Simple SummaryTo attract bright, creative, and curious students to the academic fields of applied entomology and sustainable food production, instructors of undergraduate and graduate student courses should discuss experiences with lectures and lab sessions and share effective interactive teaching tools. This communication describes how a simple population model in an Excel spreadsheet can be used in teaching both insecticide resistance evolution and basic economics of insecticide-based pest management. A tutorial video and the model as an Excel spreadsheet are freely available. Through hands-on experience with this and similar interactive teaching tools, students will acquire fundamental knowledge about basic structures population models and they will acquire experience with quantitative data interpretation. Teachers can use this tool and accompanying tutorials to demonstrate how models can be used to describe and visualize complex interactions between insect genetics and crop management. Furthermore, data from published studies can be analyzed and discussed using this interactive teaching tool.Effective teaching of complex concepts relies heavily on the ability to establish relevance of topics and to engage students in a constructive dialogue. To connect students with abstract concepts and basic theory, instructors foster and facilitate an engaging teaching environment. Population modeling is a cornerstone in applied entomology. However, it is also a topic and skill set that requires both basic mathematical and biological knowledge, and it may be perceived by students as being abstract and exceedingly theoretical. As a way to introduce entomology students at both that undergraduate and graduate levels to hands-on experience with population modeling, a well-established and widely used deterministic genetic population model is presented as an interactive teaching tool. Moreover, the general model describes three genotypes (SS = homozygous susceptible, SR = heterozygous, and RR = homozygous resistant) during 30 discrete and univoltine generations under a shared population density dependence (carrying capacity). Based on user inputs for each genotype (survival, fitness cost, reproductive rate, emigration, and immigration) and an initial resistance allele frequency, model outputs related to resistance evolution are produced. User inputs related to insecticide-based pest management (pest density action threshold, crop damage rate, insecticide treatment costs, and profit potential) can also be introduced to examine and interpret the basic economic effects of different insect pest management scenarios. The proposed model of resistance evolution and basic economics of pest management relies on a large number of important simplifications, so it may only have limited ability to predict the outcomes of real-world (commercial) scenarios. However, as a teaching tool and to introduce students to a well-known and widely used genetic population model structure, the interactive teaching tool is believed to have considerable utility and relevance.

Pyrethroid Resistance and Fitness Cost Conferred by the super-kdr Mutation M918L in Rhopalosiphum padi (Hemiptera: Aphididae).

Pyrethroid insecticides have been widely utilized for insect pest control. Target-site resistance is one of the major mechanisms explaining pest resistance to pyrethroids. This study quantified pyrethroid resistance and fitness cost conferred by the voltage-gated sodium channel (VGSC) M918L mutation in Rhopalosiphum padi. Six s-kdr-SS and six s-kdr-RS parthenogenetic lineages were established from the same field population and were reared in the laboratory without exposure to pesticides for more than one year. Enzyme activity analysis demonstrated that metabolic resistance had no impact on these lineages. Bioassays showed that the M918L mutation strongly affected pyrethroid efficiency, conferring moderate resistance to bifenthrin (type I) (39.0-fold) and high resistance to lambda-cyhalothrin (type II) (194.7-fold). Compared with the life table of s-kdr-SS lineages, s-kdr-RS lineages exhibited a relative fitness cost with significant decreases in longevity and fecundity. Meanwhile, competitive fitness was measured by blending various ratios of s-kdr-SS and s-kdr-SS aphids. The results indicated that M918L-mediated resistance showed a significant fitness cost in the presence of wild aphids without insecticide pressure. The fitness cost strongly correlated with the initial resistance allele frequency. This work characterized the novel s-kdr M918L mutation in R. padi, defined its function in resistance to different types of pyrethroids, and documented that the M918L-mediated resistance has a significant fitness cost.

An individual-based model of seed- and rhizome-propagated perennial plant species and sustainable management of Sorghum halepense in soybean production systems in Argentina.

Perennial plants which propagate through both seeds and rhizomes are common in agricultural and nonagricultural systems. Due to their multifaceted life cycle, few population models are available for studying such species. We constructed a novel individual‐based model to examine the effects of ecological, evolutionary, and anthropogenic factors on the population dynamics of perennial species. To exemplify the application of the model, we presented a case study of an important weed, Sorghum halepense (L.) Pers. (Johnsongrass), in soybean productions in Argentina. The model encompasses a full perennial weed life cycle with both sexual (seeds) and asexual (rhizomes) propagations. The evolution of herbicide resistance was modeled based on either single genes or quantitative effects. Field experiments were conducted in the species' native environment in Argentina to parameterize the model. Simulation results showed that resistance conferred by single‐gene mutations was predominantly affected by the initial frequency of resistance alleles and the associated fitness cost. Population dynamics were influenced by evolved resistance, soil tillage, and rhizome fecundity. Despite the pivotal role of rhizomes in driving the population dynamics of Johnsongrass, most herbicides target the aboveground biomass, and chemical solutions to control rhizomes are still very limited. To maintain effective (short‐term) and sustainable (long‐term) weed management, it is recommended to combine soil tillage with herbicide applications for suppressing the rhizomes and delaying the evolution of resistance. This novel model of seed‐ and rhizome‐propagated plants will also be a useful tool for studying the evolutionary processes of other perennial weeds, cash crops, and invasive species.

Combining the high-dose/refuge strategy and self-limiting transgenic insects in resistance management-A test in experimental mesocosms.

The high‐dose/refuge strategy has been the primary approach for resistance management in transgenic crops engineered with Bacillus thuringiensis toxins. However, there are continuing pressures from growers to reduce the size of Bt toxin‐free refugia, which typically suffer higher damage from pests. One complementary approach is to release male transgenic insects with a female‐specific self‐limiting gene. This technology can reduce population sizes and slow the evolution of resistance by introgressing susceptible genes through males. Theory predicts that it could be used to facilitate smaller refugia or reverse the evolution of resistance. In this study, we used experimental evolution with caged insect populations to investigate the compatibility of the self‐limiting system and the high‐dose/refuge strategy in mitigating the evolution of resistance in diamondback moth, Plutella xylostella. The benefits of the self‐limiting system were clearer at smaller refuge size, particularly when refugia were inadequate to prevent the evolution of resistance. We found that transgenic males in caged mesocosms could suppress population size and delay resistance development with 10% refugia and 4%–15% initial resistance allele frequency. Fitness costs in hemizygous transgenic insects are particularly important for introgressing susceptible alleles into target populations. Fitness costs of the self‐limiting gene in this study (P. xylostella OX4139 line L) were incompletely dominant, and reduced fecundity and male mating competitiveness. The experimental evolution approach used here illustrates some of the benefits and pitfalls of combining mass release of self‐limiting insects and the high‐dose/refuge strategy, but does indicate that they can be complementary.

Impact of Insect Management on Population Dynamics and Insecticide Resistance of Tarnished Plant Bug (Hemiptera: Miridae).

While transgenic plants targeting lepidopteran and coleopteran insects have been available for almost 20 yr, there are no transgenic crops that target hemipteran insects such as tarnished plant bug, Lygus lineolaris (Palisot de Beauvois), though at least one company lists potential products in advanced stages of development. A resistance management model for the U.S. Mid-South was developed to aid in resistance risk assessments for transgenic crops targeting L. lineolaris, and validated against the prior case of pyrethroid resistance in this insect. The model predicted that resistance to a pyrethroid would evolve in 17.6-21.0 yr (depending on the initial resistance allele frequency), which was close to the 15-20 years observed in the field. Due to uncertainty in female fecundity, we varied fecundity from 3 eggs/female/day to 12 eggs/female/day. Sprays were applied based on action thresholds, and increasing fecundity therefore increased the number of sprays applied per year and decreased durability. We also varied the action threshold in cotton (Gossypium hirsutum L.) fields (the population levels at which sprays were applied) from 17,600/ha to as low as 1,100/ha. Lowering the threshold increased the number of sprays applied, but also increased durability of the pesticide. Removing the noncotton host refuge present at the same time as cotton changed the relationship so that increasing the action threshold increased durability. The impact of insect resistance management on action threshold cost estimates will vary depending on the landscape, and cannot be assumed to always move the economic injury level in the same direction.

High Susceptibility to Cry1Ac and Low Resistance Allele Frequency Reduce the Risk of Resistance of Helicoverpa armigers to Bt Soybean in Brazil.

The Old World bollworm, Helicoverpa armigera (Hübner), was recently introduced into Brazil, where it has caused extensive damage to cotton and soybean crops. MON 87701 × MON 89788 soybean, which expresses the Bt protein Cry1Ac, was recently deployed in Brazil, providing high levels of control against H. armigera. To assess the risk of resistance to the Cry1Ac protein expressed by MON 87701 × MON 89788 soybean in Brazil, we conducted studies to evaluate the baseline susceptibility of H. armigera to Cry1Ac, in planta efficacy including the assessment of the high-dose criterion, and the initial resistance allele frequency based on an F2 screen. The mean Cry1Ac lethal concentration (LC50) ranged from 0.11 to 1.82 μg·mL−1 of diet among all H. armigera field populations collected from crop seasons 2013/14 to 2014/15, which indicated about 16.5-fold variation. MON 87701 × MON 89788 soybean exhibited a high level of efficacy against H. armigera and most likely met the high dose criterion against this target species in leaf tissue dilution bioassays up to 50 times. A total of 212 F2 family lines of H. armigera were established from field collections sampled from seven locations across Brazil and were screened for the presence of MON 87701 × MON 89788 soybean resistance alleles. None of the 212 families survived on MON 87701 × MON 89788 soybean leaf tissue (estimated allele frequency = 0.0011). The responses of H. armigera to Cry1Ac protein, high susceptibility to MON 87701 × MON 89788 soybean, and low frequency of resistance alleles across the main soybean-producing regions support the assumptions of a high-dose/refuge strategy. However, maintenance of reasonable compliance with the refuge recommendation will be essential to delay the evolution of resistance in H. armigera to MON 87701 × MON 89788 soybean in Brazil.

A significant fitness cost associated with ACE1 target site pirimicarb resistance in a field isolate of Aphis gossypii Glover from Australian cotton

The aphid Aphis gossypii Glover is an important pest of Australian cotton and has developed resistance to many chemicals used for its control. Its resistance management is partially based on chemical rotation that relies on insecticide resistance being associated with fitness costs. Therefore, understanding fitness costs associated with insecticide resistance is critical to its sustainable resistance management. We studied the fitness cost of pirimicarb resistance in A. gossypii caused by a single mutation in the acetylcholinesterase gene ACE1 by mixing different ratios of susceptible and resistant aphids. This was achieved by establishing A. gossypii populations of a known starting allele frequency in aphid proof cages and measuring allele frequency change over time via qPCR. Unlike traditional cohort fitness studies, we used competitive fitness as a measure of relative fitness of resistant versus susceptible aphids in the same environment. We demonstrate that competitive fitness measured in this study is an accurate predictor of overall relative fitness. We found that pirimicarb resistance had a significant fitness cost in the presence of susceptible aphids in the absence of insecticide pressure and that the fitness cost was related to the initial resistance allele frequency. By using the competitive fitness measure and knowing the initial allele frequency, it is possible to predict the likely time from resistant to an essentially susceptible population. As resistance was stable in the absence of susceptible competition, we recommend the use of resistance management tactics that do not completely eliminate the susceptible genotype such as complimentary integrated pest management.

Frequency of resistance to Vip3Aa20 toxin from Bacillus thuringiensis in Spodoptera frugiperda (Lepidoptera: Noctuidae) populations in Brazil

High-dose/refuge is the most common resistant management strategy used to delay or prevent pest adaptation to transgenic plants expressing Bacillus thuringiensis Berliner toxins. The success of this strategy depends, among other factors, on a low initial resistance allele frequency. In this study, we used genotypic and phenotypic methods to estimate the frequency of resistance to Vip3Aa20 toxin in Spodoptera frugiperda (J. E. Smith) in Brazil. In genotypic monitoring (F2 screen), 11 populations were screened from 2013 to 2014 in diet-overlay or excised-leaf bioassays. In phenotypic monitoring, 50 populations were screened from 2013 to 2015, by exposition of neonates at a diagnostic concentration of 3600 ng/cm2 of Vip3Aa20 in diet-overlay bioassay. The resistance allele frequency to Vip3Aa20 overlay on diet or expressed on corn leaves was similar 0.0012 (95% CI, 0–0.0033) and 0.0011 (95% CI, 0–0.0031), with an overall frequency of 0.0009 (95% CI, 0–0.0021). The resistance allele frequency does not differ among populations from different Brazilian states. In the phenotypic monitoring, only three populations MS19, MT24 and PR51 from Chapadão do Sul (MS), Lucas do Rio Verde (MT) and Castro (PR) showed larval survival ranging from 0.87 to 0.97%, differing from a susceptible reference population. In summary, the frequency of resistance to Vip3Aa20 toxin in Brazilian populations of S. frugiperda was low. Therefore, if other factors such as recessive inheritance of resistance and high level of refuge compliance are met, the risk of resistance to Vip3Aa20 in S. frugiperda populations in Brazil can be minimized.

Modeling the Evolution of Glyphosate Resistance in Barnyardgrass (Echinochloa crus-galli) in Cotton-Based Production Systems of the Midsouthern United States

Glyphosate-resistant (GR) weeds have been a prime challenge to the sustainability of GR cotton-based production systems of the midsouthern United States. Barnyardgrass is known to be a high-risk species for evolving herbicide resistance, and a simulation model was developed for understanding the likelihood of glyphosate resistance evolution in this species in cotton-based systems. Under a worst-case scenario of five glyphosate applications in monoculture GR cotton, the model predicts resistance evolution in about 9 yr of continuous glyphosate use, with about 47% risk by year 15. A unique insight from this model is that management in response to GR Palmer amaranth in this system (a reactive response) provided a proactive means to greatly reduce the risks of glyphosate resistance evolution in barnyardgrass. Subsequent model analysis revealed that the risk of resistance is high in fields characterized by high barnyardgrass seedbank levels, seedling emergence, and seed production per square meter, whereas the risk is low in fields with high levels of postdispersal seed loss and annual seedbank loss. The initial frequency of resistance alleles was a high determinant of resistance evolution (e.g., 47% risk at year 15 at an initial frequency of 5e−8vs. 4% risk at 5e−10). Monte Carlo simulations were performed to understand the influence of various glyphosate use patterns and production practices in reducing the rate and risk of glyphosate resistance evolution in barnyardgrass. Early planting and interrow cultivation are useful tools. Crop rotation is effective, but the diversity of weed management options practiced in the rotational crop is more important. Diversifying weed management options is the key, yet application timing and the choice of management option is critical. Model analyses illustrate the relative effectiveness of a number of diversified glyphosate use strategies in preventing resistance evolution and preserving the long-term utility of glyphosate in midsouthern U.S. cotton-based production systems.

Insect resistance to Bt crops: lessons from the first billion acres

Evolution of resistance in pests can reduce the effectiveness of insecticidal proteins from Bacillus thuringiensis (Bt) produced by transgenic crops. We analyzed results of 77 studies from five continents reporting field monitoring data for resistance to Bt crops, empirical evaluation of factors affecting resistance or both. Although most pest populations remained susceptible, reduced efficacy of Bt crops caused by field-evolved resistance has been reported now for some populations of 5 of 13 major pest species examined, compared with resistant populations of only one pest species in 2005. Field outcomes support theoretical predictions that factors delaying resistance include recessive inheritance of resistance, low initial frequency of resistance alleles, abundant refuges of non-Bt host plants and two-toxin Bt crops deployed separately from one-toxin Bt crops. The results imply that proactive evaluation of the inheritance and initial frequency of resistance are useful for predicting the risk of resistance and improving strategies to sustain the effectiveness of Bt crops.

Effects of Entomopathogenic Nematodes on Evolution of Pink Bollworm Resistance to <I>Bacillus thuringiensis</I> Toxin Cry1Ac

The evolution of resistance by pests can reduce the efficacy of transgenic crops that produce insecticidal toxins from Bacillus thuringiensis (Bt). However, fitness costs may act to delay pest resistance to Bt toxins. Meta-analysis of results from four previous studies revealed that the entomopathogenic nematode Steinernema riobrave (Rhabditida: Steinernematidae) imposed a 20% fitness cost for larvae of pink bollworm, Pectinophora gossypiella (Saunders) (Lepidoptera: Gelechiidae), that were homozygous for resistance to Bt toxin Cry1Ac, but no significant fitness cost was detected for heterozygotes. We conducted greenhouse and laboratory selection experiments to determine whether S. riobrave would delay the evolution of pink bollworm resistance to Cry1Ac. We mimicked the high dose/refuge scenario in the greenhouse with Bt cotton (Gossypium hirsutum L.) plants and refuges of non-Bt cotton plants, and in the laboratory with diet containing Cry1Ac and refuges of untreated diet. In both experiments, half of the replicates were exposed to S. riobrave and half were not. In the greenhouse, S. riobrave did not delay resistance. In the laboratory, S. riobrave delayed resistance after two generations but not after four generations. Simulation modeling showed that an initial resistance allele frequency > 0.015 and population bottlenecks can diminish or eliminate the resistance-delaying effects of fitness costs. We hypothesize that these factors may have reduced the resistance-delaying effects of S. riobrave in the selection experiments. The experimental and modeling results suggest that entomopathogenic nematodes could slow the evolution of pest resistance to Bt crops, but only under some conditions.

A computer simulation study to evaluate resistance development with a derquantel–abamectin combination on UK sheep farms

UK guidelines for the sustainable control of parasites in sheep (SCOPS) aim to delay further development of anthelmintic drug resistance. This study describes a computer model evaluation of resistance development with a novel oral formulation of derquantel–abamectin, to inform recommendations for use. Two different farm management scenarios, based on UK field data, were modelled to simulate low refugia (non SCOPS) or high refugia (SCOPS) worm populations. The effect on resistance allele frequencies and field efficacy of several treatment scenarios using the novel active derquantel (DQL), a spiroindole (SI), as either a single or multiple active formulation with abamectin (ABA), a macrocyclic lactone (ML), under the two farm management systems was evaluated. The initial resistance allele frequency for DQL was set at 0.0001, assuming that resistance in the UK is low, and for ML at 0.165 or 0.8, assuming that resistant nematode populations exist in the UK. DQL resistance reached a level at which a reduction in field efficacy might be detected (resistance allele frequency 0.25) by year 16 when used sequentially, and by year 31 when used in annual rotation (ABA) with SCOPS management inputs, and by year 5 (sequential) and by year 10 (annual rotation) with non SCOPS management inputs. ML resistance reached a level at which a reduction in ABA field efficacy might be detected (resistance allele frequency 0.25) by year 4 when used sequentially, and by year 8 when used in annual rotation with DQL and SCOPS management inputs, and by year 1 (sequential) and by year 2 (annual rotation) with non SCOPS management inputs. No detectable reduction in field efficacy was observed for DQL–ABA after 40 years of use with SCOPS management inputs for simulations using an initial ML resistance allele frequency of 0.165 and 0.8. A detectable reduction in field efficacy was observed for DQL–ABA by year 32 (initial ML resistance allele frequency=0.165) and by year 6 (initial ML resistance allele frequency=0.8) with non SCOPS management inputs. In summary, the results suggest that formulating DQL in combination with ABA confers a substantial advantage in delaying the development of both DQL and ML resistance, and the provision of adequate refugia further extends this advantage.

Success of the high-dose/refuge resistance management strategy after 15 years of Bt crop use in North America

Transgenic maize and cotton expressing Bacillus thuringiensis (Bt) toxins were first commercialized in 1996. By 2009, Bt crops were planted on ca. 47.6 Mha in 22 countries worldwide, with the USA and Canada accounting for 54% of this area. Resistance (virulence) development in target insect pests is a major threat to the sustainable use of Bt crops. Four major target pests of Bt crops in the USA and Canada – European corn borer, Ostrinia nubilalis (Hubner), southwestern corn borer, Diatraea grandiosella Dyar (both Lepidoptera: Crambidae), tobacco budworm, Heliothis virescens Fabricius (Lepidoptera: Noctuidae), and pink bollworm, Pectinophora gossypiella (Saunders) (Lepidoptera: Gelechiidae) – remain susceptible to Bt toxins after 15 years of intensive use of Bt maize and Bt cotton. The success in sustaining susceptibility in these major pests is associated with successful implementation of the ‘high-dose/refuge’ insecticide resistance management (IRM) strategy: (i) Bt crop cultivars express a ‘high dose’, (ii) initial frequency of resistance alleles is very low, and (iii) a refuge is maintained nearby in the environment. Field resistance (including control failure) to a Bt crop has been clearly documented in three situations: fall armyworm [Spodoptera frugiperda JE Smith] in Puerto Rico, African stem borer [Busseola fusca Fuller (Lepidoptera: Noctuidae)] in South Africa, and P. gossypiella in India. Factors associated with these cases of field resistance include: failure to use high-dose Bt cultivars and lack of sufficient refuge. These observations support the claim that implementation of the ‘high-dose/refuge’ IRM strategy has been successful in substantially delaying field resistance to Bt crops.

Population genetics of concurrent selection with albendazole and ivermectin or diethylcarbamazine on the possible spread of albendazole resistance in Wuchereria bancrofti

The Global Program for the Elimination of Lymphatic Filariasis (GPELF) intends to achieve its aims through yearly mass treatments with albendazole (ABZ) combined with ivermectin (IVM) or diethylcarbamazine (DEC). The use of ABZ and IVM separately to combat parasites of veterinary importance has, on many occasions, resulted in widespread drug resistance. In order to help predict the spread of potential ABZ resistance alleles through a population of Wuchereria bancrofti, we have developed a mathematical model that incorporates population genetics into EPIFIL, a model which examines the transmission dynamics of the parasite. Our model considers the effect of the combined treatments on the frequency of a recessive allele, which confers ABZ resistance. The model predicts that after 10 yearly treatments with ALB and DEC, 85% coverage and an initial resistance allele frequency of 5%, the frequency of the resistance genotype will increase from 0.25 to 12.7%. If non-random mating is assumed, the initial genotype frequency will be 2.34% and will increase to 62.7%. ABZ and IVM combination treatment may lead to weaker selection for this genotype. Treatment coverage, initial allele frequencies and number of treatments also affect the rate of selection.

Assessing Risk of Resistance to Aerial Applications of Methyl-Parathion in Western Corn Rootworm (Coleoptera: Chrysomelidae)

We validated a stochastic model of the evolution of resistance to adulticidal sprays of methyl-parathion in western corn rootworm, Diabrotica virgifera virgifera LeConte, populations in Nebraska. The population dynamics predicted by the model resembled that reported for field populations, and time until control failures occurred closely matched reports by commercial crop consultants. We incorporated uncertainty about the values used for 18 model parameters by replacing default values with random draws taken from a normal distribution. One parameter, the initial resistance allele frequency, was no longer measurable because of the evolution of resistance. We therefore proposed five candidate initial allele frequencies and developed probability distributions for the time to resistance for each by running 1000 simulations with parameters randomly varied. These distributions included variation because of stochastic effects as well as parameter uncertainty. We used Bayesian inference to estimate the candidate frequency most likely, given reported times to field control failures. The initial allele frequency of 10(-4) was most likely (29%), 10(-3) was less likely (28%), whereas 10(-6) was relatively unlikely (5%). Results from sensitivity analysis depended upon how evolution of resistance was measured. When resistance was examined as a genetic phenomenon, the rate of increase of the resistance allele depended almost entirely on genetic factors (LC50 values), the characteristics of the pesticide (residual activity), and the variance associated with emergence of adults. When resistance was measured as failure of methyl-parathion to reduce populations below threshold levels (0.5 gravid females per plant), parameters that contributed to population growth rate (mortality and fecundity) were also important. These data suggest two important phases in resistance evolution in corn rootworms: a genetic phase associated with negative growth rates and rapid changes in resistance allele frequencies and a rebound phase associated with positive growth rates and near fixation of the resistance allele.

Assessing Risk of Resistance to Aerial Applications of Methyl-Parathion in Western Corn Rootworm (Coleoptera: Chrysomelidae)

We validated a stochastic model of the evolution of resistance to adulticidal sprays of methyl-parathion in western corn rootworm, Diabrotica virgifera virgifera LeConte, populations in Nebraska. The population dynamics predicted by the model resembled that reported for field populations, and time until control failures occurred closely matched reports by commercial crop consultants. We incorporated uncertainty about the values used for 18 model parameters by replacing default values with random draws taken from a normal distribution. One parameter, the initial resistance allele frequency, was no longer measurable because of the evolution of resistance. We therefore proposed five candidate initial allele frequencies and developed probability distributions for the time to resistance for each by running 1000 simulations with parameters randomly varied. These distributions included variation because of stochastic effects as well as parameter uncertainty. We used Bayesian inference to estimate the candidate frequency most likely, given reported times to field control failures. The initial allele frequency of 10−4 was most likely (29%), 10−3 was less likely (28%), whereas 10−6 was relatively unlikely (5%). Results from sensitivity analysis depended upon how evolution of resistance was measured. When resistance was examined as a genetic phenomenon, the rate of increase of the resistance allele depended almost entirely on genetic factors (LC50 values), the characteristics of the pesticide (residual activity), and the variance associated with emergence of adults. When resistance was measured as failure of methyl-parathion to reduce populations below threshold levels (0.5 gravid females per plant), parameters that contributed to population growth rate (mortality and fecundity) were also important. These data suggest two important phases in resistance evolution in corn rootworms: a genetic phase associated with negative growth rates and rapid changes in resistance allele frequencies and a rebound phase associated with positive growth rates and near fixation of the resistance allele.

Initial frequency of alleles conferring resistance to Bacillus thuringiensis poplar in a field population of Chrysomela tremulae.

Globally, the estimated total area planted with transgenic plants producing Bacillus thuringiensis (Bt) toxins was 12 million hectares in 2001. The risk of target pests becoming resistant to these toxins has led to the implementation of resistance-management strategies. The efficiency and sustainability of these strategies, including the high-dose plus refuge strategy currently recommended for North American maize, depend on the initial frequency of resistance alleles. In this study, we estimated the initial frequencies of alleles conferring resistance to transgenic Bt poplars producing Cry3A in a natural population of the poplar pest Chrysomela tremulae (Coleoptera: Chrysomelidae). We used the F(2) screen method developed for detecting resistance alleles in natural pest populations. At least three parents of the 270 lines tested were heterozygous for a major Bt resistance allele. We estimated mean resistance-allele frequency for the period 1999-2001 at 0.0037 (95% confidence interval = 0.00045-0.0080) with a detection probability of 90%. These results demonstrate that (i) the F(2) screen method can be used to detect major alleles conferring resistance to Bt-producing plants in insects and (ii) the initial frequency of alleles conferring resistance to Bt toxin can be close to the highest theoretical values that are expected prior to the use of Bt plants if considering fitness costs and typical mutation rates.

Reversing insect adaptation to transgenic insecticidal plants.

The refuge-high-dose strategy for delaying insect adaptation to transgenic plants produces non-transgenic plants that enable survival of susceptible individuals. Previous theoretical work has suggested three requirements for success of the refuge-high-dose strategy: a low initial frequency of the resistance allele, extensive mating between resistant and susceptible adults and recessive inheritance of resistance. In order to understand an observed decrease in resistance frequency and improve the potential for managing resistance better, we used analytical and simulation models for exploring the conditions that prevent or reverse the evolution of resistance, even when resistance is not rare initially. Assuming random mating and recessive or nearly recessive inheritance of resistance, the factors favouring reversal of resistance are non-recessive costs of resistance, low initial resistance allele frequency, large refuges, incomplete resistance and density-independent population growth in refuges.

Selection for and against resistance to insecticides in the absence of insecticide: a case study of malathion resistance in the saw-toothed grain beetle,Oryzaephilus surinamensis(Coleoptera: Silvanidae)

AbstractIn the absence of insecticide there may be intense selection against alleles conferring insecticide resistance, so that these alleles will decline in frequency. On the other hand, selection will act to favour alleles at modifier loci which cause resistance alleles to become coadapted and increase in fitness even in the absence of insecticide. The relative effectiveness of these two processes is of practical importance and theoretical interest. The fate of polymorphism for malathion resistance was studied in the saw-toothed grain beetle,Oryzaephilus surinamensis(Linnaeus). Five to six lines of each of four strains (8401, 8401sel, 8518B and L1040), each polymorphic for resistance phenotypes, were maintained in insecticide-free culture for up to nine generations, over which period the response of individuals to a discriminating dose of 65 mg m-2malathion was determined. There was significant reversion to susceptibility in the related strains 8401, 8401se1 and 8518B. Assuming resistance to be primarily controlled by a single major locus, resistant homozygote and heterozygote fitnesses in the absence of insecticide were estimated to be 0.92, 0.83 and 0.68, respectively. The initial frequency of the resistant allele was inversely related to the time since last exposure to insecticide, and also to the fitness of the resistant allele. This could be explained by the selection of modifier alleles that reduce the fitness cost of resistance in the absence of insecticide. In strain L1040 the proportion of resistant individuals actually increased over time (after the second generation) in the absence of insecticide. The fitness of the susceptible allele in this case was 0.83. In all strains there was significant variation among lines in relative fitness and initial resistance allele frequency. Analysis of knockdown proportions revealed highly significant variation between generations that is not easily interpreted in genetic terms but is better explained by limitations of the knockdown bioassay.