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Transgenic corn and cotton that express Cry and Vip insecticidal proteins from the bacterium, Bacillus thuringiensis (Bt), have been valuable tools for the management of lepidopteran pests. In 2019, a Vip3Aa-resistant strain of Helicoverpa zea (CEW-Vip-RR) was isolated from F2 screens of field populations in Texas. Characterizing the resistance mechanism in this strain is important for predicting the sustained efficacy of current commercial Bt traits and guiding the development of future transgenic traits. Resistance to insecticidal proteins in Bt traits is commonly associated with reduced toxin binding, with the exception of Vip3Aa resistance being associated to altered proteolytic processing in the insect host gut. Therefore, Vip3Aa protoxin processing was tested by incubation with midgut fluids from CEW-Vip-RR relative to a susceptible strain (CEW-SS). Finding no significant processing differences, alterations in Vip3Aa binding were tested by comparing binding of radiolabeled and biotinylated Vip3Aa toxin to midgut brush border membrane vesicles (BBMV) from CEW-Vip-RR and CEW-SS larvae. Specific Vip3Aa binding to CEW-Vip-RR BBMV in these experiments was consistently reduced when compared with CEW-SS BBMV. These results support that an altered Vip3Aa-receptor is associated with resistance in CEW-Vip-RR. Understanding this resistance mechanism could have important implications for resistance management decisions, considering widespread Cry1 and Cry2 resistance in H. zea populations. IMPORTANCE Helicoverpa zea is a major crop pest in the United States that is managed with transgenic corn and cotton that produce insecticidal proteins from the bacterium, Bacillus thuringiensis (Bt). However, H. zea has evolved widespread resistance to the Cry proteins produced in Bt corn and cotton, leaving Vip3Aa as the only plant-incorporated protectant in Bt crops that consistently provides excellent control of H. zea. The benefits provided by Bt crops will be substantially reduced if widespread Vip3Aa resistance develops in H. zea field populations. Therefore, it is important to identify resistance alleles and mechanisms that contribute to Vip3Aa resistance to ensure that informed resistance management strategies are implemented. This study is the first report of reduced binding of Vip3Aa to midgut receptors associated with resistance. [ABSTRACT FROM AUTHOR]

The corn earworm, Helicoverpa zea, is a major target pest of the insecticidal Vip3Aa protein used in pyramided transgenic Bt corn and cotton with Cry1 and Cry2 proteins in the U.S. The widespread resistance to Cry1 and Cry2 proteins in H. zea will challenge the long-term efficacy of Vip3Aa technology. Determining the frequency of resistant alleles to Vip3Aa in field populations of H. zea is critically important for resistance management. Here, we provided the first F2 screen study to estimate the resistance allele frequency for Vip3Aa in H. zea populations in Texas, U.S. In 2019, 128 H. zea neonates per isofamily for a total of 114 F2 families were screened with a diagnostic concentration of 3.0 μg/cm2 of Vip3Aa39 protein in diet-overlay bioassays. The F2 screen detected two families carrying a major Vip3Aa resistance allele. The estimated frequency of major resistance alleles against Vip3Aa39 in H. zea in Texas from this study was 0.0065 with a 95% CI of 0.0014–0.0157. A Vip3Aa-resistant strain (RR) derived from the F2 screen showed a high level of resistance to Vip3Aa39 protein, with a resistance ratio of >588.0-fold relative to a susceptible population (SS) based on diet-overlay bioassays. We provide the first documentation of a major resistance allele conferring high levels of Vip3Aa resistance in a field-derived strain of H. zea in the U.S. Data generated from this study contribute to development of management strategies for the sustainable use of the Vip3Aa technology to control H. zea in the U.S. [ABSTRACT FROM AUTHOR]

In Texas, the cotton fleahopper (Pseudatomoscelis seriatus (Reuter)) is considered a highly economically damaging pest of cotton (Gossypium hirsutum L.). Current control methods rely heavily on foliar chemical insecticides throughout the growing season. Considering the cost of insecticides and the critical timeliness of their application, chemical control methods are often not optimized to reduce potential yield losses. The Mpp51Aa2.834_16 gene in cotton (ThryvOn) has shown effectiveness against thrips and several piercing and sucking mirid insect pests, suggesting it has the potential to mitigate yield losses caused by the cotton fleahopper. Choice and no-choice caged feeding assays were conducted to assess the impact of cotton fleahoppers on ThryvOn cotton square retention under controlled laboratory conditions. In the choice assay, feeding by cotton fleahoppers significantly reduced square retention in the gene-lacking cotton to 46%, while the ThryvOn cotton retained 60% of the squares. In the no-choice assay, cotton fleahopper nymph feeding significantly reduced square retention in the cotton not expressing Mpp51Aa2 to 61%, whereas the ThryvOn cotton was unaffected. Based on the differences in square retention observed in both the choice and no-choice feeding assays, our findings indicate that the Mpp51Aa2 protein influences cotton fleahopper feeding preferences and the susceptibility of cotton plants to damage caused by cotton fleahoppers. Our study offers confirmation of the activity of ThryvOn on cotton fleahoppers observed in the field. The ThryvOn trait's activity towards cotton fleahoppers is consistent with that found for other mirid pests in cotton. [ABSTRACT FROM AUTHOR]

The cotton fleahopper (Pseudatomoscelis seriatus Reuter) is considered a highly economically damaging pest of cotton (Gossypium hirsutum L.) in Texas and Oklahoma. Current control methods rely heavily on the use of foliar-applied chemical insecticides, but considering the cost of insecticides and the critical timeliness of applications, chemical control methods are often not optimized to reduce potential yield losses from this pest. The Bacillus thuringiensis (Bt) Mpp51Aa2 (formerly Cry51Aa2.834_16) protein has proven effective against thrips and plant bugs with piercing and sucking feeding behaviors, but the impact of this toxin on cotton fleahoppers has not been investigated. To evaluate the Mpp51Aa2 trait effectiveness towards the cotton fleahopper, field trials were conducted in 2019, 2020, and 2021, comparing a cotton cultivar containing the Mpp51Aa2 trait to a non-traited isoline cultivar under insecticide-treated and untreated conditions. Populations of cotton fleahopper nymphs and adults were estimated weekly by visually inspecting cotton terminals. Square retention was also assessed during the first week of bloom to provide some insight on how the Bt trait may influence yield. While cotton fleahopper population differences between the traited and non-traited plants were not consistently noted during the pre-bloom squaring period, there was a consistent increase in square retention in cotton expressing Mpp51Aa2 relative to non-traited cotton. Additionally, cotton expressing Mpp51Aa2 offered similar square protection relative to non-traited cotton treated with insecticides for the cotton fleahopper. These findings indicate that the Mpp51Aa2 protein should provide benefits of delayed nymphal growth, population suppression, and increased square retention. [ABSTRACT FROM AUTHOR]