Models of Diabrotica Populations: Demography, Population Genetics, Geographic Spread, and Management.

Both Diabrotica virgifera virgifera LeConte and D. barberi Smith and Lawrence are among the most damaging insects impacting corn in North America. D. virgifera virgifera has also invaded Europe and has become an important pest in that region. Computer models have become an important tool for understanding the impact and spread of these important pests. Over the past 30 years, over 40 models have been published related to these pests. The focus of these models range from occupancy models (particularly for Europe), impact of climate change, range expansion, economics of pest management, phenology, to the evolution of resistance to toxins and crop rotation. All of these models share characteristics. We elaborate on the methods in which modelers have incorporated the biology of these pests, including density-dependence, movement, fecundity and overwintering mortality. We discuss the utility of both spatially-explicit, complex models and spatially-implicit, generational models and where each might be appropriate. We review resistance models that either explain past evolution to crop rotation, insecticides or insecticidal traits or attempt to predict the consequences of resistance management strategies.

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.

The macroecology of infectious diseases: a new perspective on global-scale drivers of pathogen distributions and impacts.

Identifying drivers of infectious disease patterns and impacts at the broadest scales of organisation is one of the most crucial challenges for modern science, yet answers to many fundamental questions remain elusive. These include what factors commonly facilitate transmission of pathogens to novel host species, what drives variation in immune investment among host species, and more generally what drives global patterns of parasite diversity and distribution? Here we consider how the perspectives and tools of macroecology, a field that investigates patterns and processes at broad spatial, temporal and taxonomic scales, are expanding scientific understanding of global infectious disease ecology. In particular, emerging approaches are providing new insights about scaling properties across all living taxa, and new strategies for mapping pathogen biodiversity and infection risk. Ultimately, macroecology is establishing a framework to more accurately predict global patterns of infectious disease distribution and emergence.

Evolution of Resistance by Helicoverpa zea (Lepidoptera: Noctuidae) Infesting Insecticidal Crops in the Southern United States.

We created a deterministic, frequency-based model of the evolution of resistance by corn earworm, Helicoverpa zea (Boddie) (Lepidoptera: Noctuidae), to insecticidal traits expressed in crops planted in the heterogeneous landscapes of the southern United States. The model accounts for four generations of selection by insecticidal traits each year. We used the model results to investigate the influence of three factors on insect resistance management (IRM): 1) how does adding a third insecticidal trait to both corn and cotton affect durability of the products, 2) how does unstructured corn refuge influence IRM, and 3) how do block refuges (50% compliance) and blended refuges compare with regard to IRM? When Bt cotton expresses the same number of insecticidal traits, Bt corn with three insecticidal traits provides longer durability than Bt corn with two pyramided traits. Blended refuge provides similar durability for corn products compared with the same level of required block refuge when the rate of refuge compliance by farmers is 50%. Results for Mississippi and Texas are similar, but durabilities for corn traits are surprisingly lower in Georgia, where unstructured corn refuge is the highest of the three states, but refuge for Bt cotton is the lowest of the three states. Thus, unstructured corn refuge can be valuable for IRM but its influence is determined by selection for resistance by Bt cotton.

Natural enemies delay insect resistance to Bt crops.

We investigated whether development of resistance to a Bt crop in the presence of a natural enemy would be slower than without the natural enemy and whether biological control, in conjunction with a Bt crop, could effectively suppress the pest population. Additionally, we investigated whether insecticide-sprayed refuges of non-Bt crops would delay or accelerate resistance to the Bt crop. We used a system of Bt broccoli expressing Cry1Ac, a population of the pest Plutella xylostella with a low frequency of individuals resistant to Cry1Ac and the insecticide spinosad, and a natural enemy, Coleomegilla maculata, to conduct experiments over multiple generations. The results demonstrated that after 6 generations P. xylostella populations were very low in the treatment containing C. maculata and unsprayed non-Bt refuge plants. Furthermore, resistance to Bt plants evolved significantly slower in this treatment. In contrast, Bt plants with no refuge were completely defoliated in treatments without C. maculata after 4-5 generations. In the treatment containing sprayed non-Bt refuge plants and C. maculata, the P. xylostella population was low, although the speed of resistance selection to Cry1Ac was significantly increased. These data demonstrate that natural enemies can delay resistance to Bt plants and have significant implications for integrated pest management (IPM) with Bt crops.

Modeling a western corn rootworm, Diabrotica virgifera virgifera (Coleoptera: Chrysomelidae), maturation delay and resistance evolution in Bt corn.

BACKGROUND: Emergence delay and female-skewed sex ratios among adults of Diabrotica virgifera virgifera LeConte (Coleoptera: Chrysomelidae) from Bt corn have been reported in field studies. The authors used a simulation model to study the effect of a maturation delay and a female-skewed sex ratio for D. v. virgifera emerging from Bt corn on the evolution of Bt resistance. RESULTS: The effect of skewed toxin mortality in one sex on evolution of Bt resistance was insignificant. An emergence delay among resistant beetles from Bt corn slowed resistance evolution. A shift in the time of emergence for homozygous susceptible beetles from Bt corn did not have a significant effect on the evolution of Bt resistance in D. v. virgifera. CONCLUSION: This simulation study suggested that skewed toxin mortality in one sex and an emergence delay for beetles in Bt corn are not major concerns for managing resistance by D. v. virgifera to single-toxin or pyramided Bt corn.

Modeling the integration of parasitoid, insecticide, and transgenic insecticidal crop for the long-term control of an insect pest.

The tools of insect pest management include host plant resistance, biological control, and insecticides and how they are integrated will influence the durability of each. We created a detailed model of the population dynamics and population genetics of the diamondback moth, Plutella xylostella L., and its parasitoid, Diadegma insulare (Cresson), to study long-term pest management in broccoli Brassica oleracea L. Given this pest's history of evolving resistance to various toxins, we also evaluated the evolution of resistance to transgenic insecticidal Bt broccoli (expressing Cry1Ac) and two types of insecticides. Simulations demonstrated that parasitism provided the most reliable, long-term control of P. xylostella populations. Use of Bt broccoli with a 10% insecticide-free refuge did not reduce the long-term contribution of parasitism to pest control. Small refuges within Bt broccoli fields can delay evolution of resistance > 30 generations if resistance alleles are rare in the pest population. However, the effectiveness of these refuges can be compromised by insecticide use. Rainfall mortality during the pest's egg and neonate stages significantly influences pest control but especially resistance management. Our model results support the idea that Bt crops and biological control can be integrated in integrated pest management and actually synergistically support each other. However, the planting and maintenance of toxin-free refuges are critical to this integration.

Insect resistance management for stored product pests: a case study of cowpea weevil (Coleoptera: Bruchidae).

The cowpea weevil, Callosobruchus maculatus F. (Coleoptera: Bruchidae), can cause up to 100% yield loss of stored cowpea seeds in a few months in West Africa. Genes expressing toxins delaying insect maturation (MDTs) are available for genetic engineering. A simulation model was used to investigate the possible use of MDTs for managing C. maculatus. Specifically, we studied the effect of transgenic cowpea expressing an MDT, an insecticide, or both, on the evolution of resistance by C. maculatus at constant temperature. Transgenic cowpea expressing only a nonlethal MDT causing 50-100% maturation delay did not control C. maculatus well. Mortality caused by a maturation delay improved the efficacy of transgenic cowpea expressing only a lethal MDT, but significantly reduced the durability of transgenic cowpea Transgenic cowpea expressing only a lethal MDT causing 50% maturation delay and 90% mortality controlled C. maculatus better than one expressing only a nonlethal MDT, but its durability was only 2 yr. We concluded that transgenic cowpea expressing only an MDT has little value for managing C. maculatus. The resistance by C. maculatus to transgenic cowpea expressing only an insecticide rapidly evolved. Stacking a gene expressing a nonlethal MDT and a gene expressing an insecticide in transgenic cowpea did not significantly improve the durability of an insecticide, but stacking a gene expressing a lethal MDT and a gene expressing an insecticide in transgenic cowpea significantly improved the durability of an insecticide and an MDT. We also discussed this approach within the idea of using transgenic RNAi in pest control strategies.

Effect of insecticides and Plutella xylostella (Lepidoptera: Plutellidae) genotype on a predator and parasitoid and implications for the evolution of insecticide resistance.

In the laboratory and in cages in the greenhouse, we evaluated the toxicity of two insecticides (lambda-cyhalothrin and spinosad) on the parasitoid, Diadegma insulare (Cresson), and the predator, Coleomegilla maculate (DeGeer), both natural enemies of the diamondback moth, Plutella xylostella (L.). Lambda-cyhalothrin was very toxic to both natural enemies. Spinosad was less toxic to C. maculata adults and larvae, and slightly toxic to D. insulare. Both natural enemies suppressed P. xylostella populations in cages with 80% spinosad-treated and 20% nontreated plants; such suppression was not seen when lambda-cyhalothrin was used. Using broccoli, Brassica oleracea L. variety italica, a common host for P. xylostella, we also studied direct and indirect effects of both natural enemies in the presence and absence of the two insecticides and to different P. xylostella genotypes: resistant to the insecticide, susceptible, or heterozygous. Neither natural enemy could distinguish host genotype if P. xylostella were feeding on nontreated plants. They could also not distinguish between larvae feeding on spinosad-treated plants and nontreated plants, but D. insulare could distinguish between larvae feeding on lambda-cyhalothrin treated and nontreated plants. Our studies suggest that lambda-cyhalothrin has direct toxicity to these two natural enemies, can affect their host foraging and acceptance of P. xylostella and consequently would not be compatible in conserving these natural enemies in a program for suppression of P. xylostella. In contrast, our studies suggest that treatment with spinosad has much less effect on these natural enemies and would allow them to help suppress populations of P. xylostella. These findings are discussed in relation to the evolution of insecticide resistance and suppression of the pest populations.

Putting the brakes on a cycle: bottom-up effects damp cycle amplitude.

Pest population density oscillations have a profound effect on agroecosystem functioning, particularly when pests cycle with epidemic persistence. Here, we ask whether landscape-level manipulations can be used to restrict the cycle amplitude of the European corn borer moth [Ostrinia nubilalis (Hübner)], an economically important maize pest. We analysed time series from Minnesota (1963-2009) and Wisconsin (1964-2009) to quantify the extent of regime change in the US Corn Belt where rates of transgenic Bt maize adoption varied. The introduction of Bt maize explained cycle damping when the adoption of the crop was high (Minnesota); oscillations were damped but continued to persist when Bt maize was used less intensely (Wisconsin). We conclude that host plant quality is key to understanding both epidemic persistence and the success of intervention strategies. In particular, the dichotomy in maize management between states is thought to limit the spatial autocorrelation of O. nubilalis.

Seeds of change: corn seed mixtures for resistance management and integrated pest management.

The use of mixtures of transgenic insecticidal seed and nontransgenic seed to provide an in-field refuge for susceptible insects in insect-resistance-management (IRM) plans has been considered for at least two decades. However, the U.S. Environmental Protection Agency has only recently authorized the practice. This commentary explores issues that regulators, industry, and other stakeholders should consider as the use of biotechnology increases and seed mixtures are implemented as a major tactic for IRM. We discuss how block refuges and seed mixtures in transgenic insecticidal corn, Zea mays L., production will influence integrated pest management (IPM) and the evolution of pest resistance. We conclude that seed mixtures will make pest monitoring more difficult and that seed mixtures may make IRM riskier because of larval behavior and greater adoption of insecticidal corn. Conversely, block refuges present a different suite of risks because of adult pest behavior and the lower compliance with IRM rules expected from farmers. It is likely that secondary pests not targeted by the insecticidal corn as well as natural enemies will respond differently to block refuges and seed mixtures.

Effect of Bt broccoli and resistant genotype of Plutella xylostella (Lepidoptera: Plutellidae) on development and host acceptance of the parasitoid Diadegma insulare (Hymenoptera: Ichneumonidae).

The ecological implications on biological control of insecticidal transgenic plants, which produce crystal (Cry) proteins from the soil bacterium Bacillus thuringiensis (Bt), remain a contentious issue and affect risk assessment decisions. In this study, we used a unique system of resistant insects, Bt plants and a parasitoid to critically evaluate this issue. The effects of broccoli type (normal or expressing Cry1Ac protein) and insect genotype (susceptible or Cry1Ac-resistant) of Plutella xylostella L. (Lepidoptera: Plutellidae) were examined for their effects on the development and host foraging behavior of the parasitoid, Diadegma insulare (Cresson) (Hymenoptera: Ichneumonidae) over two generations. Parasitism rate and development of D. insulare were not significantly different when different genotypes (Bt-resistant or susceptible) of insect host larvae fed on non-Bt broccoli plants. D. insulare could not discriminate between resistant and susceptible genotypes of P. xylostella, nor between Bt and normal broccoli plants with different genotypes of P. xylostella feeding on them. No D. insulare could emerge from Bt broccoli-fed susceptible and heterozygous P. xylostella larvae because these larvae were unable to survive on Bt broccoli. The parasitism rate, developmental period, pupal and adult weights of D. insulare that had developed on Bt broccoli-fed Cry1Ac-resistant P. xylostella larvae were not significantly different from those that developed on non-Bt broccoli-fed larvae. Female D. insulare emerged from Cry1Ac-resistant P. xylostella that fed on Bt plants could successfully parasitize P. xylostella larvae. The life parameters of the subsequent generation of D. insulare from P. xylostella reared on Bt broccoli were not significantly different from those from non-Bt broccoli. The Cry1Ac protein was detected in P. xylostella and in D. insulare when hosts fed on Bt broccoli. These results are the first to indicate that Cry1Ac did not harm the development or host acceptance of an important endoparasitoid after two generations of exposure. We suggest that using other Bt crops and resistant insect species would likely lead to similar conclusions about the safety of the presently used Bt proteins on parasitoids.

Western corn rootworm (Coleoptera: Chrysomelidae) dispersal and adaptation to single-toxin transgenic corn deployed with block or blended refuge.

A simulation model of the temporal and spatial dynamics and population genetics of western corn rootworm, Diabrotica virgifera virgifera LeConte, was created to evaluate the use of block refuges and seed blends in the management of resistance to transgenic insecticidal corn (Zea mays L.). This Bt corn expresses one transgenic corn event, DAS-59122-7, that produces a binary insecticidal protein toxin (Cry34Ab1/Cry35Ab1) and provides host-plant resistance. The model incorporates the latest information about larval and adult behavior. Results of this modeling effort indicate that the seed-blend scenarios in many cases produced equal or greater durability than block refuges that were relocated each year. Resistance evolved in the most likely scenarios in 10-16 yr. Our standard analysis presumed complete adoption of 59122 corn by all farmers in our hypothetical region, no crop rotation, and 100% compliance with Insect Resistant Management (IRM) regulations. As compliance levels declined, resistance evolved faster when block refuges were deployed. Seed treatments that killed the pest when applied to all seeds in a seed blend or just to seeds in Bt corn blocks delayed evolution of resistance. Greater control of the pest population by the seed treatment facilitated longer durability of the transgenic trait. Therefore, data support the concept that pyramiding a transgenic insecticidal trait with a highly efficacious insecticidal seed treatment can delay evolution of resistance.

Modeling evolution of Diabrotica virgifera virgifera (Coleoptera: Chrysomelidae) to transgenic corn with two insecticidal traits.

A simulation model of the population dynamics and genetics of western corn rootworm, Diabrotica virgifera virgifera LeConte (Coleoptera: Chrysomelidae), was created to evaluate the use of refuges in the management of resistance to transgenic insecticidal corn, Zea mays L., expressing one or two toxin traits. Hypothetical scenarios and a case study of a corn hybrid pyramided with existing toxins are simulated. In the hypothetical situations, results demonstrated that evolution is generally delayed by pyramids compared with deployment of a single-toxin corn hybrid. However, soil insecticide use in the refuge reduced this delay and quickened the evolution of resistance. Results were sensitive to the degree of male beetle dispersal before mating and to the effectiveness of both toxins in the pyramid. Resistance evolved faster as fecundity increased for survivors of insecticidal corn. Thus, effects on fecundity must be measured to predict which resistance management plans will work well. Evolution of resistance also occurred faster if the survival rate due to exposure to the two toxins was not calculated by multiplication of two independent survival rates (one for each insect gene) but was equivalent to the minimum of the two. Furthermore, when single-trait and pyramided corn hybrids were planted within rootworm-dispersal distance of each other, the toxin traits lost efficacy more quickly than they did in scenarios without single-trait corn. For the case study involving transgenic corn expressing Cry34/35Ab1 and Cry3Bb1, the pyramid delayed evolution longer than a single trait corn hybrid and longer than a sequence of toxins based on at least one resistance-allele frequency remaining below 50%. Results are discussed within the context of a changing transgenic corn marketplace and the landscape dynamics of resistance management.

Density-dependent and density-independent mortality of the western corn rootworm: impact on dose calculations of rootworm-resistant Bt corn.

The percentage of viable eggs of the western corn rootworm, Diabrotica virgifera virgifera LeConte, which survived to the adult stage was evaluated for the effect of egg density in 2005 and 2007 in central Missouri. In 2005, each plot was 2.44 by 3.05 m and contained 64 maize (corn), Zea mays L., plants. In 2007, plots were 3.05 by 3.05 m and again contained 64 corn plants. Seven egg densities (2,400, 1,200, 600, 300, 100, 50, and 25 viable eggs per 30.5 cm) were evaluated with four to six replications in each year in a completely randomized design. In 2007 only, an additional row was infested near each plot to evaluate plant damage. In both years, there was no correlation of infestation level and percentage of emergence between infestation levels of 25-600 viable eggs per 30.5 cm, indicating that density-dependent mortality did not occur at these egg densities. In 2005, 8.04% of the viable eggs established on a corn plant and produced an adult at these lower infestation rates. In 2007, this value was 2.9%. Regardless of egg density, approximately 92-97% failed to establish and produce adults (density-independent mortality). In 2005 and in the combined analysis, as viable egg densities increased from 600 to 2400 per 30.5 cm there was a significant decrease in percentage of emergence. In a broken line analysis of the 2005 data, the point where density-dependent mortality began in the combined analysis was 851 eggs per 30.5 cm with a 95% confidence interval from 678 to 1024. That year density-dependent mortality was important at high infestations and killed 54.4% of those larvae that successfully established on a plant at the highest egg density. However, little or no density-dependent mortality occurred at infestation levels <850 viable eggs per 30.5 cm in either year of the study. Combining data from both years with all previously published data in a broken line analysis indicated that density-dependent mortality began at approximately 800 viable eggs per 30.5 cm. These data are discussed in terms of dose calculations for products targeting the western corn rootworm.

Microarray analysis yields candidate markers for rotation resistance in the western corn rootworm beetle, Diabrotica virgifera virgifera.

As pest species may evolve resistance to chemical controls, they may also evolve resistance to cultural control methods. Yearly rotation of corn (Zea mays) with another crop interrupts the life cycle of the western corn rootworm beetle (Diabrotica virgifera virgifera, Coleoptera: Chrysomelidae), but behavioral resistance to crop rotation is now a major problem in the Midwest of the USA. Resistant adult females exhibit reduced fidelity to corn as a host and lay their eggs in the soil of both corn and soybean (Glycine max) fields. Behavioral assays suggest that the adaptation is related to increased locomotor activity, but finding molecular markers has been difficult. We used microarray analysis to search for gene expression differences between resistant and wild-type beetles. Candidates validated with real-time polymerase chain reaction exhibit predicted patterns from the microarray in independent samples across time and space. Many genes more highly expressed in the rotation-resistant females have no matches to known proteins, and most genes that were more lowly expressed are involved in antimicrobial defense.

Evolutionary analysis of herbivorous insects in natural and agricultural environments.

Herbivorous insects offer a remarkable example of the biological diversity that formed the foundation for Darwin's theory of evolution by natural selection. The ability of insects to evolve resistance rapidly to insecticides and host-plant resistance present a continual challenge for pest management. This paper considers the manner in which genetic constraints, host-plant availability and trade-offs affect the evolution of herbivorous insects in natural and agricultural environments, and the extent to which lessons learned from studying natural systems may be applied to improve insect resistance management in agricultural systems. Studies on the genetic architecture of adaptation by herbivores to host plants and to insecticides are reviewed. The genetic basis of resistance is an important component of simulation models that predict the evolution of resistance. These models often assume monogenic resistance, but available data suggest that this assumption may be overly narrow and that modeling of resistance as oligogenic or polygenic may be more appropriate. As omics (e.g. genomics and proteomics) technologies become more accessible, a better understanding of the genetic basis of resistance will be possible. Trade-offs often accompany adaptations by herbivores. Trade-offs arise when the benefit of a trait, such as the ability to feed on a novel host plant or to survive in the presence of an insecticide, is counterbalanced by fitness costs that decrease fitness in the absence of the selective agent. For resistance to insecticides, and resistance to insecticidal transgenic crops in particular, fitness costs may act as an evolutionary constraint and delay or prevent the evolution of resistance. An important observation is that certain ecological factors such as host plants and entomopathogens can magnify fitness costs, which is termed ecological negative cross-resistance. The application of omics technologies may allow for more efficient identification of factors that will impose ecological negative cross-resistance, thereby bolstering insect resistance management.

Finding the economics in economic entomology.

To recommend new pest management tactics and strategies to farmers and policy makers, economic entomologists must evaluate the economics of biologically reasonable approaches. We collected data to determine how frequently these economic evaluations occur. We discovered from our survey of entomological journals representing the discipline of economic entomology that < 1% of research papers published since 1972 include economic evaluations of pest management tactics. At least 85% of these analyses were performed by entomologists and not economists. Much of the research on economic evaluations is performed without special funds granted by agencies separate from the authors' institutions. In the United States, USDA competitive grants supported 20% of the economic evaluations published since 2000. However, only approximately 12% of the projects funded since 2000 by three sections of the USDA (Crops at Risk, Risk Avoidance and Mitigation Program, and Pest Management Alternatives Program) resulted in publications concerning economic evaluations. If the purpose of economic entomology is to ultimately determine the value of different kinds of tactics, the discipline may need to take steps to enhance the research that supports these evaluations.

Planting transgenic insecticidal corn based on economic thresholds: consequences for integrated pest management and insect resistance management.

A simulation model of the western corn rootworm, Diabrotica virgifera virgifera LeConte, was used to investigate whether sampling and economic thresholds can improve integrated pest management (IPM) and insect resistance management (IRM) when transgenic insecticidal crops are used for insect pest management. When transgenic corn killed at least 80% of susceptible larvae, the calculated economic threshold increased linearly as the proportion of susceptible beetles surviving the toxin increased. The use of economic thresholds slightly slowed the evolution of resistance to transgenic insecticidal crops. In areas with or without rotation-resistant western corn rootworm phenotypes, the use of sampling and economic thresholds generated similar returns compared with strategies of planting transgenic corn, Zea mays L., every season. Because transgenic crops are extremely effective, farmers may be inclined to plant transgenic crops every season rather than implementing costly and time-consuming sampling protocols.