The coleopteran insect western corn rootworm (WCR) (Diabrotica virgifera virgifera LeConte) is a devastating crop pest in North America and Europe. Although crop plants that produce Bacillus thuringiensis (Bt) proteins can limit insect infestation, some insect populations have evolved resistance to Bt proteins. Here we describe an insecticidal protein, designated IPD072Aa, that is isolated from Pseudomonas chlororaphis. Transgenic corn plants expressing IPD072Aa show protection from WCR insect injury under field conditions. IPD072Aa leaves several lepidopteran and hemipteran insect species unaffected but is effective in killing WCR larvae that are resistant to Bt proteins produced by currently available transgenic corn. IPD072Aa can be used to protect corn crops against WCRs.

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.

Several Bt maize events expressing various insecticidal Cry protein genes have been commercialized for management of western corn rootworm,Diabrotica virgifera virgiferaLeConte (Coleoptera: Chrysomelidae). We used high efficacy (>99.7%) experimental maize events that express mCry3A for selections under laboratory conditions to develop a western corn rootworm colony resistant to mCry3A at higher levels than published results. The resistance ratio (RR) to mCry3A was >97-fold based on LCvalues in diet-based bioassays after six generations of selections when compared to that of an unselected Control colony. Using a sublethal seedling assay (SSA) method, we confirmed that the colony had no cross-resistance to maize event DAS-59122-7, which expresses Cry34/35Ab. Reciprocal crosses between the mCry3A-resistant colony and the susceptible colony were performed to test the inheritance of resistance. Larval survival and development evaluated by the SSA method indicated that resistance to mCry3A was inherited autosomally and was incompletely recessive (h = 0.23-0.25). Specific binding of mCry3A to brush border membrane vesicles of midgut tissue revealed reduced binding in the resistant colony when compared to binding in the susceptible colony. This is the first report where resistance in western corn rootworm has been shown to involve reduced binding of a Cry3-class protein in midgut tissue.

To delay evolution of insect resistance to insecticidal Bacillus thuringiensis Berliner (Bt) transgenic crops, USA and Canadian maize growers commonly set aside a portion of each field as a refuge, where susceptible pests can develop without exposure to the toxin(s) expressed in the insecticidal crop. Abundant mate-seeking refuge adults are expected to move into insecticidal crop areas and mate with rare, resistant insects. Production of heterozygous offspring, rather than homozygous offspring reduces the rate of resistance evolution. Expectations about the refuge strategy are predicated on assumptions about when and where pest insects move and mate. Accumulating evidence indicates that the behavior of western corn rootworm beetles, Diabrotica virgifera virgifera LeConte (Coleoptera: Chrysomelidae) (WCR), in refuge and corn rootworm (CRW)-active Bt-transgenic maize does not always conform to assumptions about random mating and individual movement. Contrary to expectations, up to 23.6% of unmated refuge females left refuge and were non-teneral by the time they mated in CRW-active maize. Consequences of protandry, limited male mating capacity, premating male and female movement patterns, skewed sex ratios, and delayed adult emergence from CRW-active maize all may contribute to unexpected or undesirable patterns of WCR reproductive behavior. Modeling suggests that temporal and spatial components of WCR mating that limit interactions in block refuges can be reduced by deploying blended refuges.

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.

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.

Zea mays L. (maize) hybrids producing the Cry1F protein from Bacillus thuringiensis were first commercialized in the United States in 2003. These products demonstrated varying levels of moderate control, but not immunity to Striacosta albicosta (Smith) (Lepidoptera: Noctuidae) (western bean cutworm). Susceptibility of western bean cutworm to Cry1F protein was assessed in field populations collected in the mid- and western United States in 2003, 2004, 2013, and 2014 using diet bioassay. A meta-analysis of 32 western bean cutworm field collections assessed for susceptibility to Cry1F was conducted to investigate changes in susceptibility over time. Based on meta-analysis results, these data suggest a 5.2-fold increase in median lethal concentration (LC50) response to Cry1F in the 2013–2014 populations compared with collections that were assessed 10 yr earlier. Widespread use of Cry1F-producing maize hybrids over the past 10 yr may have contributed to favoring western bean cutworm populations with tolerance to the Cry1F protein.

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