Economic Injury Levels for Bt-resistant Helicoverpa zea (Lepidoptera: Noctuidae) in Cotton.

Thresholds for Helicoverpa zea (Boddie) in cotton Gossypium hirsutum L. have been understudied since the widespread adoption of Bt cotton in the United States. Our study was possible due to the widespread presence of H. zea populations with Cry-toxin resistance. We initiated progressive spray timing experiments using three Bt cotton brands (Deltapine, Stoneville, and Phytogen) widely planted across the U.S. Cotton Belt expressing pyramided toxins in the Cry1A, Cry2, and Vip3Aa19 families. We timed foliar insecticide treatments based on week of bloom to manipulate H. zea populations in tandem with crop development during 2017 and 2018. We hypothesized that non-Bt cotton, cotton expressing Cry toxins alone, and cotton expressing Cry and Vip3Aa19 toxins would respond differently to H. zea feeding. We calculated economic injury levels to support the development of economic thresholds from significant responses. Pressure from H. zea was high during both years. Squares and bolls damaged by H. zea had the strongest negative yield associations, followed by larval number on squares. There were fewer yield associations with larval number on bolls and with number of H. zea eggs on the plant. Larval population levels were very low on varieties expressing Vip3Aa19. Yield response varied across experiments and varieties, suggesting that it is difficult to pinpoint precise economic injury levels. Nonetheless, our results generally suggest that current economic thresholds for H. zea in cotton are too high. Economic injury levels from comparisons between non-Bt varieties and those expressing only Cry toxins could inform future thresholds once H. zea evolves resistance to Vip3Aa19.

Helicoverpa zea (Lepidoptera: Noctuidae) Preference for Plant Structures, and Their Location, Within Bt Cotton Under Different Nitrogen and Irrigation Regimes.

Helicoverpa zea Boddie is a common economic pest of cotton (Gossypium hirsutum L.), including transgenic cotton varieties that express Bacillus thuringiensis (Bt). Helicoverpa zea oviposition is similar in Bt and non-Bt cotton, but behavior of H. zea larvae can be different in the presence of Bt, with neonates moving away from terminals faster in single-toxin Bt than non-Bt cotton or avoiding Bt-treated diet in the lab. We quantified H. zea oviposition and larval distribution on structures within cotton plants in small plot experiments of Cry1Ac + Cry1F cotton for 2 yr under different irrigation and nitrogen treatments. More eggs were oviposited on plants receiving nitrogen application during 2016 and on leaves in the top section of irrigated plants during 2017, but other treatment effects on eggs or larvae were minimal. Helicoverpa zea eggs were most common on leaves in the top third of plants at position zero and middle section of cotton plants throughout the season, but some oviposition occurred on fruiting structures as well. First and second instars were more common on squares in the top section of plants during 2016 and bolls in the middle and lower sections during 2017 due to oviposition lower in the canopy during 2017. During both years, third through fifth instars were more common on bolls in the middle and lower section of plants closer to the main stem. These findings have resistance management implications as extended larval feeding on bolls could optimize nutrition, decrease Bt susceptibility, and potentially influence behavioral resistance.

Helicoverpa zea (Lepidoptera: Noctuidae) Oviposition and Larval Vertical Distribution in Bt Cotton Under Different Levels of Nitrogen and Irrigation.

In some Bt cotton (Gossypium hirsutum L.) varieties, bollworm (Helicoverpa zea Boddie) larval behavior differs from non-Bt varieties. Laboratory assays indicate bollworm larvae can detect Bt proteins, which may cause behavioral differences. Plant stress from factors including fertility and water availability causes changes in plant physiology and Bt expression. Our objective was to determine whether nitrogen and irrigation influenced bollworm behavior in Bt cotton by recording the vertical distribution of eggs and larvae over time. We conducted small plot experiments with Cry1Ac + Cry1F cotton in 2016 and 2017 with three nitrogen rates, along with irrigated and nonirrigated treatments during 2017. Bollworm locations were determined by in-field examination of 10-20 cotton plants per plot over 6-8 wk. The location of each egg and larva was recorded by node, with instar estimation of each larva. Oviposition was higher in in plots receiving nitrogen; first and second instars were also more common in plots receiving nitrogen or irrigation, whereas older instars had similar numbers among treatments. Oviposition was more evenly distributed throughout the canopy earlier in the sampling period than during later weeks, with more eggs in the top third of the canopy in only three of 14-wk. Early instars were also evenly distributed throughout the canopy. Later, instars moved to the middle portions of the canopy, away from bottom nodes, and did not move toward the terminal. Understanding bollworm behavior can inform both crop scouting and resistance management decisions.

Development and Dispersal of Helicoverpa zea (Lepidoptera: Noctuidae) on Non-Bt and Bt Pyramided Cotton.

Bollworm (Helicoverpa zea Boddie) (Lepidoptera: Noctuidae) can cause economic losses in both non-Bt and Bt cotton. Larvae modify their behavior in the presence of Bt by moving away from terminals faster in Bt cotton compared to non-Bt cotton and avoiding Bt-treated diets. Our objectives were to understand differences in bollworm egg and larvae populations within, and dispersal away from, non-Bt and Bt pyramided-toxin cotton. We conducted small plot experiments in 2016 and 2017 to monitor on-plant egg and larval numbers, and off-plant dispersal of larvae, from non-Bt and different Bt toxin pyramided cotton. Bollworm adults preferred to oviposit in most Bt toxin pyramids compared to non-Bt; this was likely unrelated to detection of Bt by adults, but rather density-dependent aversion from high larval populations. First instar numbers were similar in all non-Bt/Bt toxin pyramids and dispersed at a similar rate. Second through fifth instar numbers were higher in non-Bt than Bt toxin pyramids but dispersed equally from all non-Bt/Bt toxin pyramids, regardless of Bt pyramid type. Development times of larvae were often slower in Bt toxin pyramids compared to non-Bt. Fifth instars were found in, and dispersing from, Bt toxin pyramids containing Vip3A, raising concerns of resistance development. Furthermore, differences in oviposition rate among non-Bt/Bt toxin pyramids and slowed development rate of larvae on Bt varieties could create inconsistencies in generation times emerging from Bt and non-Bt hosts, which could contribute to resistance development.

Long-Term Empirical and Observational Evidence of Practical Helicoverpa zea Resistance to Cotton With Pyramided Bt Toxins.

Evidence of practical resistance of Helicoverpa zea (Boddie) (Lepidoptera: Noctuidae) to Bt cotton in the United States is debatable, supported with occasional reports of boll damage in the field. Our objective was to provide both empirical and long-term observational evidence of practical resistance by linking both in-season and end-of-season measurements of H. zea damage to pyramided Bt cotton bolls and to provide Cry1Ac diet-based bioassay data in support of these damage estimates. In-season boll damage from H. zea was highly correlated to end-of-season damaged bolls. Across North Carolina, Bt cotton fields with end-of-season bolls damaged by H. zea increased during 2016 compared to previous years. Elevated damage was coupled with an increase in field sprays targeting H. zea during 2016, but not related to an increase in H. zea abundance. Bioassay data indicated that there was a range of Cry1Ac susceptibility across the southeastern United States. Given the range of susceptibility to Cry1Ac across the southeastern United States, it is probable that resistant populations are common. Since H. zea is resistant to cotton expressing pyramided Cry toxins, the adoption of new cotton varieties expressing Vip3Aa will be rapid. Efforts should be made to delay resistance of H. zea to the Vip3Aa toxin to avoid foliar insecticide use.