David W. Onstad

Seeds of Change: Corn Seed Mixtures for Resistance Management and Integrated Pest Management

ABSTRACT 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.

Behaviour and ecology of the western corn rootworm ( Diabrotica virgifera virgifera LeConte)

1 The western corn rootworm (WCR) is a historic pest with a legacy of resistance and behavioural plasticity. Its behaviour and nutritional ecology are important to rootworm management. The success of the most effective and environmentally benign rootworm management method, annual crop rotation, was based on an understanding of rootworm behaviour and host–plant relationships. Enthusiastic adoption of crop rotation, provided excellent rootworm management, but also selected for behavioural resistance to this cultural control.

Seed Mixtures as a Resistance Management Strategy for European Corn Borers (Lepidoptera: Crambidae) Infesting Transgenic Corn Expressing Cry1Ab Protein

Abstract Dispersal of neonate European corn borers, Ostrinia nubilalis (Hübner), in seed mixtures of transgenic corn expressing Cry1Ab protein (Bt+) and nontransgenic corn (Bt−) was evaluated in a 2-yr field study. The main objective was to determine if larval dispersal limits the effectiveness of seed mixtures as a resistance management strategy. Mixtures evaluated included (1) all Bt+ plants, (2) every fifth plant Bt− with remaining plants Bt+, (3) every fifth plant Bt+ with remaining plants Bt−, and (4) all Bt− plants. The transformation events MON 802 (B73 BC1F2 × Mo17) and MON 810 (B73 BC1F1 × Mo17), which express the Cry1Ab endotoxin isolated from Bacillus thuringiensis subsp. kurstaki, were used as the sources of Bt+ seed in 1994 and 1995, respectively (YieldGard, Monsanto, St. Louis, MO). At corn growth stage V6-V8, subplots within each mixture (15–20 plants each) were infested so that every fifth plant in mixtures 1 and 4, every Bt− plant in mixture 2, and every Bt+ plant in mixture 3 received two egg masses. Larval sampling over a 21-d period indicated increased neonate dispersal off of Bt+ plants, reduced survival of larvae that dispersed from Bt+ plants to Bt− plants, and a low incidence of late-instar movement from Bt− plants to Bt+ plants. Computer simulations based on mortality and dispersal estimates from this study indicate that seed mixtures will delay the evolution of resistant European corn borer populations compared with uniform planting of transgenic corn. However, resistant European corn borer populations likely will develop faster in seed mixes compared with separate plantings of Bt and non-Bt corn.

Modeling Evolution of Diabrotica virgifera virgifera (Coleoptera: Chrysomelidae) to Transgenic Corn With Two Insecticidal Traits

ABSTRACT 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.

Modeling the Dynamics of Adaptation to Transgenic Corn by Western Corn Rootworm (Coleoptera: Chrysomelidae)

Abstract A simulation model of the population dynamics and genetics of the western corn rootworm, Diabrotica virgifera virgifera LeConte, was created for a landscape of corn, soybean, and other crops. Although the model was created to study a 2-locus problem for beetles having genes for resistance to both crop rotation and transgenic corn, during this first phase of the project, the model was simulated to evaluate only resistance management plans for transgenic corn. Allele expression in the rootworm and toxin dose in the corn plant were the two most important factors affecting resistance development. A dominant resistance allele allowed quick evolution of resistance to transgenic corn, whereas a recessive allele delayed resistance >99 yr. With high dosages of toxin and additive expression, the time required to reach 3% resistance allele frequency ranged from 13 to >99 yr. With additive expression, lower dosages permitted the resistant allele frequency to reach 3% in 2–9 yr with refuges occupying 5–30% of the land. The results were sensitive to delays in emergence by susceptible adults and configuration of the refuge (row strips versus blocks).

Density-Dependent and Density-Independent Mortality of the Western Corn Rootworm: Impact on Dose Calculations of Rootworm-Resistant Bt Corn

ABSTRACT 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, ≈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 ≈800 viable eggs per 30.5 cm. These data are discussed in terms of dose calculations for products targeting the western corn rootworm.

Modeling evolution of behavioral resistance by an insect to crop rotation

Crop rotation has traditionally been a valuable method for managing pests, but now a serious insect pest of maize (Diabrotica virgifera virgifera LeConte [Coleoptera: Chrysomelidae]) has developed behavioral resistance to rotation. A simple model of adult behavior and population genetics can explain how this resistance may have developed. This general model indicates that evolution may be caused by selection on a single gene for adult movement and that behavioral resistance only develops at high levels of rotation (>80% of plant landscape). In less diverse landscapes, crop rotation selects for the expansion of host preferences (polyphagy) by adults. More diverse landscapes may delay the evolution of resistance to crop rotation depending on the fitness costs and the nature of the genetic system.

Does Landscape Diversity Slow the Spread of Rotation-Resistant Western Corn Rootworm (Coleoptera: Chrysomelidae)?

Abstract A behavioral change in some western corn rootworm (Diabrotica virgifera virgifera LeConte) populations is threatening the effectiveness of crop rotation, a successful management strategy for controlling this pest. We created a set of simple meteorologic and behavioral models that can be used to predict the spread of the beetle infesting soybean (Glycine max (L.)) throughout the midwestern United States. We used data collected in Illinois, IN, MI, and Ohio to create maps of observations to evaluate the model. We displayed data on the maps using detection thresholds for western corn rootworm in soybean fields of 10 or 20 beetles per 100 sweeps and one or two beetles per yellow sticky trap per day. Counts greater than a detection threshold represent populations with a lack of fidelity to corn (Zea mays L.) and adapted to circumvent corn-soybean rotation. Some of the models invoked a landscape-diversity function that included the proportion of noncorn, nonrotated soybean vegetation on farmland in each county (i.e., extra vegetation). The best model for the period from 1997 to 2001 is based on heavy-storm data, with distance that beetles spread each year reduced by the proportion of extra vegetation in a county. This version is superior to a previously published model and to two new models that do not consider landscape diversity. Most of the models predicted spread at too high a rate between 1997 and 2001, compared with observations, but a few new models with rates of spread reduced by a landscape-diversity function matched the observations relatively well. Results suggest that the conclusions based on a linear model using proportion of extra vegetation as the key parameter are likely to be robust. Thus, we hypothesize that as the landscape diversity represented by the proportion of noncorn and nonrotated soybean vegetation in a geographic region increases, the rate of regional spread of the rotation-resistant western corn rootworm decreases over several years.

Western Corn Rootworm (Coleoptera: Chrysomelidae) Dispersal and Adaptation to Single-Toxin Transgenic Corn Deployed With Block or Blended Refuge

ABSTRACT 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.

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 Darwins 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.