Transgenic maize, Zea mays L., modified to express insecticidal proteins from the bacterium Bacillus thuringiensis Berliner, was introduced in 1996 to control Ostrinia nubilalis Hübner (Lepidoptera: Crambidae), a key maize pest in North America. The high-dose/refuge concept, developed to delay or prevent resistance evolution to this technology, has been exemplified by O. nubilalis as no cases of practical resistance were identified in >20 years. This study documents the first case of practical resistance to Cry1F Bt maize by O. nubilalis in North America. Four collections of O. nubilalis were made from Cry1F maize in Nova Scotia, Canada with unexpected injury (UXI) ranging from 30–70%. Greater survival of UXI collections was observed when larvae were exposed to the highest concentration of 200 ng Cry1F cm−2 in diet-overlay bioassays compared to susceptible laboratory colonies. Larvae also fed and survived on Cry1F leaf tissue in 7 d bioassays. A collection from non-Bt maize, 120 km west of the UXI region, also survived 200 ng Cry1F cm−2, but was susceptible to Cry1F leaf tissue. Detection of Cry1F-resistant O. nubilalis in what might be considered an insignificant maize-growing region indicates that a number of preventable causal factors may have been related to inadequate stewardship of Bt maize technology.

Western bean cutworm, Striacosta albicosta (Smith) (Lepidoptera: Noctuidae), is a pest of corn (Zea mays L.) that has recently expanded its range into Ontario, Canada. Control of S. albicosta damage to corn hybrids containing event TC1507-expressing Cry1F Bacillus thuringiensis protein alone or pyramided with event MON 89034 expressing Cry1A.105 and Cry2Ab2 Bt proteins was tested in 2011-2015 in Ontario in small- and large-scale field plots with natural infestation. In 2011, significantly lower incidence and severity of kernel damage was sustained by Cry1F × Cry1A.105 + Cry2Ab2 corn compared with a non-Bt near-isogenic hybrid. However, from 2012 to 2015, there was no difference in incidence or severity of damage comparing non-Bt hybrids with Cry1F hybrids alone or pyramided with Cry1A.105 and Cry2Ab2 planted as a pure stand or with an integrated refuge (95% Bt: 5% non-Bt seeds). In 2015, neonate larvae derived from Ontario field-collections were tested in concentration-response diet-overlay bioassays with lyophilized Cry1F protein at concentrations up to 75 µg cm-2. The concentrations at which mortality of 50% (LC50) of the collections occurred ranged from approximately 10 µg cm-2 (F0) to >28 µg cm-2 (F1) in a 7-d bioassay, indicating relative insensitivity to Cry1F. Results from field experiments, laboratory bioassays, and the history of exposure to Cry1F in corn show that S. albicosta in Ontario are not controlled by Cry1F-expressing corn hybrids and provide evidence for the conclusion that the evolution of resistance to Cry1F has occurred.

Canadian and United States (US) insect resistance management (IRM) programs for lepidopteran pests in Bacillus thuriengiensis (Bt)-expressing crops are optimally designed for Ostrinia nubilalis Hübner in corn (Zea mays L.) and Chloridea virescens Fabricius in cotton (Gossypium hirsutum L.). Both Bt corn and cotton express a high dose for these pests; however, there are many other target pests for which Bt crops do not express high doses (commonly referred to as nonhigh dose pests). Two important lepidopteran nonhigh dose (low susceptibility) pests are Helicoverpa zea Boddie (Lepidoptera: Noctuidae) and Striacosta albicosta Smith (Lepidoptera: Noctuidae). We highlight both pests as cautionary examples of exposure to nonhigh dose levels of Bt toxins when the IRM plan was not followed. Moreover, IRM practices to delay Bt resistance that are designed for these two ecologically challenging and important pests should apply to species that are more susceptible to Bt toxins. The purpose of this article is to propose five best management practices to delay the evolution of Bt resistance in lepidopteran pests with low susceptibility to Bt toxins in Canada and the US: 1) better understand resistance potential before commercialization, 2) strengthen IRM based on regional pest pressure by restricting Bt usage where it is of little benefit, 3) require and incentivize planting of structured corn refuge everywhere for single toxin cultivars and in the southern US for pyramids, 4) integrate field and laboratory resistance monitoring programs, and 5) effectively use unexpected injury thresholds.

The first case of field-evolved resistance in European corn borer, Ostrinia nubilalis (Hübner) (Lepidoptera: Crambidae) to transgenic corn (Zea mays L.) producing a Bacillus thuringiensis (Bt) Berliner toxin was discovered in Nova Scotia, Canada in 2018. This case involved resistance to Bt corn producing Cry1Fa toxin. As a mitigation response, Bt corn hybrids producing only Cry1Fa were replaced in that region with hybrids producing two or three Bt toxins targeting O. nubilalis. In this study, we collected O. nubilalis in several corn-growing regions of Canada during 2018 to 2020 and tested their progeny for susceptibility to four Bt toxins produced by currently available Bt corn that targets O. nubilalis: Cry1Fa, Cry1Ab, Cry1A.105, and Cry2Ab. Based on toxin concentrations killing 50% of larvae from 23 field-derived strains relative to two susceptible laboratory strains, the resistance ratio was at least 10 for Cry1Fa for 12 strains (52% of strains) consisting of 10 strains from Nova Scotia, as well as strains from near Montreal, Quebec and Roseisle, Manitoba. We found low but statistically significant resistance relative to at least one of two susceptible strains for Cry1Ab (23% of strains), Cry1A.105 (45% of strains), and Cry2Ab (14% of strains), with maximum resistance ratios of 3.9, 5.8, and 2.0, respectively. These results provide key information for addressing O. nubilalis resistance to Bt corn in Canada.

Fusarium graminearum Schwabe (Hypocreales: Nectriaceae) and Fusarium verticillioides (Saccardo) (Hypocreales: Nectriaceae) Nirenberg infection results in accumulation of deoxynivalenol (DON), zearalenone (ZON), and fumonisin (FBs) mycotoxins in infected corn, Zea mays L. Lepidopteran insect feeding may exacerbate fungal infection by providing entry points on the ear resulting in increased mycotoxin contamination of grain. The objective of the current study was to simulate different types and severity levels (extent of injury) of lepidopteran injury to corn ears at different stages of ear development and its effect on mycotoxin accumulation in grain corn. Field experiments were conducted under conditions favorable for F. graminearum development where insect injury was simulated to corn ears and inoculated with F. graminearum. All simulated injury treatments resulted in elevated mycotoxin concentration compared with ears without simulated injury; however, the severity of injury within a treatment had little effect. Injury to kernels on the side of the ear resulted in greater DON and ZON concentration than injury to tip kernels, grazing injury applied at physiological maturity, or when no injury was simulated. Greater FBs was measured when tip kernel injury was simulated at the blister stage or when side kernel injury was simulated at milk and dent stages compared with noninjured ears, silk clipping, tip injury at milk and dent stages, or grazing injury at physiological maturity. The current study confirms that the risk of mycotoxin accumulation in the Great Lakes region is greater in the presence of ear-feeding insect pests and may differ depending on the feeding behavior of pest species.