The corn earworm, Helicoverpa zea (Boddie), is a major pest of Bacillus thuringiensis (Bt) maize and cotton in the U.S. Reduced efficacy of Bt plants expressing Cry1 and Cry2 against H. zea has been reported in some areas of the U.S. In this study, we evaluated the occurrence and ear damage of H. zea on transgenic Bt maize expressing Cry proteins or a combination of Vip3A and Cry proteins in the field in Texas in 2018. We found that the occurrence of H. zea larvae and the viable kernel damage area on the ear were not different between non-Bt maize and Bt maize expressing Cry1A.105+Cry2Ab2 and Cry1Ab+Cry1F proteins. A total of 67.5% of the pyramided Bt maize expressing Cry1Ab+Cry1F+Vip3A was damaged by 2nd–4th instar larvae of H. zea. Diet bioassays showed that the resistance ratio against Vip3Aa51 for H. zea obtained from Cry1Ab+Cry1F+Vip3A maize was 20.4 compared to a field population collected from Cry1F+Cry1A.105+Cry2Ab2 maize. Leaf tissue bioassays showed that 7-day survivorship on WideStrike3 (Cry1F+Cry1Ac+Vip3A) cotton leaves was significantly higher for the H. zea population collected from Cry1Ab+Cry1F+Vip3A maize than for a Bt-susceptible laboratory population. The results generated from this study suggest that H. zea has evolved practical resistance to Cry1 and Cry2 proteins. Therefore, it is crucial to ensure the sustainable use of the Vip3A technology in Bt maize and cotton.

Evolution of resistance by pests can reduce the benefits of crops genetically engineered to produce insecticidal proteins from Bacillus thuringiensis (Bt). Because of the widespread resistance of Helicoverpa zea to crystalline (Cry) Bt toxins in the United States, the vegetative insecticidal protein Vip3Aa is the only Bt toxin produced by Bt corn and cotton that remains effective against some populations of this polyphagous lepidopteran pest. Here we evaluated H. zea resistance to Vip3Aa using diet bioassays to test 42,218 larvae from three lab strains and 71 strains derived from the field during 2016 to 2020 in Arkansas, Louisiana, Mississippi, Tennessee, and Texas. Relative to the least susceptible of the three lab strains tested (BZ), susceptibility to Vip3Aa of the field-derived strains decreased significantly from 2016 to 2020. Relative to another lab strain (TM), 7 of 16 strains derived from the field in 2019 were significantly resistant to Vip3Aa, with up to 13-fold resistance. Susceptibility to Vip3Aa was significantly lower for strains derived from Vip3Aa plants than non-Vip3Aa plants, providing direct evidence of resistance evolving in response to selection by Vip3Aa plants in the field. Together with previously reported data, the results here convey an early warning of field-evolved resistance to Vip3Aa in H. zea that supports calls for urgent action to preserve the efficacy of this toxin.

Helicoverpa zea is a major target pest of Bt crops expressing Cry and/or Vip3Aa proteins in the U.S.A. Widespread practical resistance of H. zea to the Cry1 and Cry2 proteins makes Vip3Aa the only effective Bt protein against this pest. Understanding the frequency of resistance alleles against Vip3Aa in field populations of H. zea is crucial for resistance management and the sustainability of Vip3Aa technology. Using a modified F₂ screen method by crossing susceptible laboratory female moth with feral male moth of H. zea, we successfully screened a total of 24,576 neonates from 192 F₂ families of H. zea collected from Arkansas, Louisiana, Mississippi, and Tennessee during 2019–2020. We found five F₂ families containing ≥3rd instar survivors on the diagnostic concentration of 3.0 µg/cm² Vip3Aa39. Dose-response bioassays confirmed the high levels of Vip3Aa resistance in these F₂ families, with an estimated resistance ratio of >909.1-fold relative to the susceptible strain. The estimated resistance allele frequency against Vip3Aa in H. zea for these four southern states is 0.0155 with a 95% CI of 0.0057–0.0297. These data should provide critical information for understanding the risks of Vip3Aa resistance in H. zea and help design appropriate resistance management strategies for the sustainability of the Vip3Aa technology.

Evaluating the frequency of resistance alleles is important for resistance management and sustainable use of transgenic crops that produce insecticidal proteins from Bacillus thuringiensis. Helicoverpa zea (Boddie) (Lepidoptera: Noctuidae) is a major crop pest in the United States that has evolved practical resistance to the crystalline (Cry) proteins in Bt corn and cotton. The standard F₂ screen for estimating resistance allele frequency does not work well for H. zea because successful single-pair matings are rare. In this study, we developed and implemented a modified F₂ screen for H. zea that generates F₁ progeny by crossing three laboratory susceptible female moths with one feral male moth instead of single-pair crosses. During 2019–2020, we used this modified method to establish 192 F₂ families from 623 matings between susceptible females and feral males from Arkansas, Louisiana, Mississippi, and Tennessee. From each F₂ family, we screened 128 neonates against discriminating concentrations of Cry1Ac and Cry2Ab in diet overlay bioassays. Based on these discriminating concentration bioassays, families were considered positive for resistance if at least five larvae survived to second instar, including at least one to third instar. The percentage of positive families was 92.7% for Cry1Ac and 38.5% for Cry2Ab, which yields an estimated resistance allele frequency (with 95% confidence interval) of 0.722 (0.688–0.764) for Cry1Ac and 0.217 (0.179–0.261) for Cry2Ab. The modified F₂ screen developed and implemented here may be useful for future resistance monitoring studies of H. zea and other pests.

Foliar symptoms resembling those associated with late-season decline (LSD) disease of corn, caused by the bacteria Pantoea ananatis, were observed in commercial fields of sorghum plants in 2023 in the Texas panhandle. Diagnosis of symptomatic tissues consistently recovered bacterial isolates that were subsequently identified, based on partial sequences of their respective 16S rRNA gene, as P. ananatis. The pathogenicity of a representative isolate on sorghum was investigated using the method of Koch's postulates and resulted in the successful reproduction of LSD-like foliar symptoms identical to those described for corn, as well as those observed in affected sorghum fields.

Pantoea ananatis was recently described as the causative agent of late-season decline, a new bacterial disease first observed affecting field corn plants, in the Texas Panhandle. The rapid spread of the disease throughout the region and the patchy distribution of symptomatic plants in affected fields, as well as routine observations of edge effects, in which plants with severe symptoms are observed on the edges of affected fields, led us to hypothesize that vectors might be involved in the dissemination of the disease pathogen. In this study, we investigated the western corn rootworm (Diabrotica virgifera virgifera LeConte) and southern corn rootworm (Diabrotica undecimpunctata howardi Barber) for any naturally occurring association with P. ananatis and potential to acquire and transmit the bacterial pathogen. Additionally, we investigated the transgenic corn encoding insecticidal Bacillus thuringiensis proteins (Bt) pyramided with RNAi interference anti-rootworm technology for its potential to protect against any larval role in the transmission of the pathogen through their feeding activities on corn roots. We successfully recovered naturally occurring P. ananatis from samples of both rootworm species collected from corn plants in the field. Following acquisition assays, the acquired pathogen was successfully recovered from previously P. ananatis-free adult rootworms, their eggs, as well as first-instar larvae, suggesting an affinity of the bacteria to establish an endosymbiotic and transovarial association with both rootworm species. Additionally, the transgenic Bt corn with RNAi anti-rootworm technology was ineffective in preventing the transmission of the pathogen by the infected larvae. Findings from this study confirm a vector role in the transmission of the disease pathogen.