Rapid Adaptation and Interspecific Introgression in the North American Crop Pest Helicoverpa zea.

Insect crop pests threaten global food security. This threat is amplified through the spread of nonnative species and through adaptation of native pests to control measures. Adaptations such as pesticide resistance can result from selection on variation within a population, or through gene flow from another population. We investigate these processes in an economically important noctuid crop pest, Helicoverpa zea, which has evolved resistance to a wide range of pesticides. Its sister species Helicoverpa armigera, first detected as an invasive species in Brazil in 2013, introduced the pyrethroid-resistance gene CYP337B3 to South American H. zea via adaptive introgression. To understand whether this could contribute to pesticide resistance in North America, we sequenced 237 H. zea genomes across 10 sample sites. We report H. armigera introgression into the North American H. zea population. Two individuals sampled in Texas in 2019 carry H. armigera haplotypes in a 4 Mbp region containing CYP337B3. Next, we identify signatures of selection in the panmictic population of nonadmixed H. zea, identifying a selective sweep at a second cytochrome P450 gene: CYP333B3. We estimate that its derived allele conferred a ∼5% fitness advantage and show that this estimate explains independently observed rare nonsynonymous CYP333B3 mutations approaching fixation over a ∼20-year period. We also detect putative signatures of selection at a kinesin gene associated with Bt resistance. Overall, we document two mechanisms of rapid adaptation: the introduction of fitness-enhancing alleles through interspecific introgression, and selection on intraspecific variation.

Pest population dynamics are related to a continental overwintering gradient.

Overwintering success is an important determinant of arthropod populations that must be considered as climate change continues to influence the spatiotemporal population dynamics of agricultural pests. Using a long-term monitoring database and biologically relevant overwintering zones, we modeled the annual and seasonal population dynamics of a common pest, Helicoverpa zea (Boddie), based on three overwintering suitability zones throughout North America using four decades of soil temperatures: the southern range (able to persist through winter), transitional zone (uncertain overwintering survivorship), and northern limits (unable to survive winter). Our model indicates H. zea population dynamics are hierarchically structured with continental-level effects that are partitioned into three geographic zones. Seasonal populations were initially detected in the southern range, where they experienced multiple large population peaks. All three zones experienced a final peak between late July (southern range) and mid-August to mid-September (transitional zone and northern limits). The southern range expanded by 3% since 1981 and is projected to increase by twofold by 2099 but the areas of other zones are expected to decrease in the future. These changes suggest larger populations may persist at higher latitudes in the future due to reduced low-temperature lethal events during winter. Because H. zea is a highly migratory pest, predicting when populations accumulate in one region can inform synchronous or lagged population development in other regions. We show the value of combining long-term datasets, remotely sensed data, and laboratory findings to inform forecasting of insect pests.

Reduced toxin binding associated with resistance to Vip3Aa in the corn earworm (Helicoverpa zea).

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.

Bacillus thuringiensis Cry1Da_7 and Cry1B.868 Protein Interactions with Novel Receptors Allow Control of Resistant Fall Armyworms, Spodoptera frugiperda (J.E. Smith).

Two new modified Bacillus thuringiensis (Bt) proteins, Cry1Da_7 and Cry1B.868, with activity against fall armyworms (FAW), Spodoptera frugiperda (J.E. Smith), were evaluated for their potential to bind new insect receptors compared to proteins currently deployed as plant-incorporated protectants (PIPs) in row crops. Results from resistant insect bioassays, disabled insecticidal protein (DIP) bioassays, and cell-based assays using insect cells expressing individual receptors demonstrate that receptor utilizations of the newly modified Cry1Da_7 and Cry1B.868 proteins are distinct from each other and from those of commercially available Bt proteins such as Cry1F, Cry1A.105, Cry2Ab, and Vip3A. Accordingly, these two proteins target different insect proteins in FAW midgut cells and when pyramided together should provide durability in the field against this economically important pest.IMPORTANCE There is increased concern with the development of resistance to insecticidal proteins currently expressed in crop plants, especially against high-resistance-risk pests such as fall armyworm (FAW), Spodoptera frugiperda, a maize pest that already has developed resistance to Bacillus thuringiensis (Bt) proteins such as Cry1F. Lepidopteran-specific proteins that bind new insect receptors will be critical in managing current Cry1F-resistant FAW and delaying future resistance development. Results from resistant insect assays, disabled insecticidal protein (DIP) bioassays, and cell-based assays using insect cells expressing individual receptors demonstrate that target receptors of the Cry1Da_7 and Cry1B.868 proteins are different from each other and from those of commercially available Bt proteins such as Cry1F, Cry1A.105, Cry2Ab, and Vip3A. Therefore, pyramiding these two new proteins in maize will provide durable control of this economically important pest in production agriculture.

F2 screen for resistance to Bacillus thuringiensis Cry2Ab2-maize in field populations of Spodoptera frugiperda (Lepidoptera: Noctuidae) from the southern United States.

The fall armyworm, Spodoptera frugiperda (J. E. Smith) (Lepidoptera: Noctuidae), is a target pest of transgenic maize and cotton expressing Bacillus thuringiensis (Bt) proteins in both North and South America. In 2013 and 2014, a total of 215 F2 two-parent families of S. frugiperda were established using single-pair mating of field individuals collected from seven locations in four states of the southern U.S.: Texas, Louisiana, Georgia, and Florida. The objective of the investigation was to detect resistance alleles in field populations to Cry2Ab2, a common Bt protein produced in transgenic maize and cotton. For each F2 family, 128 F2 neonates were screened on leaf tissue of Cry2Ab2 maize plants in the laboratory. A conservative estimate of the frequency of major Cry2Ab2 resistance alleles in S. frugiperda from the four states was 0.0023 with a 95% credibility interval of 0.0003-0.0064. In addition, six families were considered to likely possess minor resistance alleles at a frequency of 0.0082 with a 95% credibility interval of 0.0033-0.0152. One F2 family from Georgia (GA-15) was confirmed to possess a major resistance allele to the Cry2Ab2 protein. Larvae from this family survived well on whole maize plants expressing Cry2Ab2 protein and demonstrated a significant level (>15-fold) of resistance when fed with the same protein incorporated in a meridic diet. The detection of the major resistance allele along with the relatively abundant minor resistance alleles revealed in this study may have important implications for resistance management.

Susceptibility of Field and Laboratory Bt-Susceptible and Resistant Strains of Helicoverpa zea (Boddie) to HearNPV.

During 2021 and 2022, eight field-collected and five laboratory Helicoverpa zea strains with varying susceptibility to different Bt proteins were evaluated for their responses against HearNPV using diet-overlay bioassays. The five laboratory strains included SS (susceptible to all Bt proteins), CRY-RR (resistant to Cry1 and Cry2), VIP-RR-70 (resistant to Vip3Aa), VIP-RR-15 (resistant to Vip3Aa), and TRE-RR (resistant to Cry1, Cry2, and Vip3Aa). Our findings showed that the susceptibility of TRE-RR, VIP-RR-70, and VIP-RR-15 strains to HearNPV was similar to that of the SS strain. However, the field and Cry-RR strains were more resistant to HearNPV compared to the SS strain. Because most feral H. zea strains in the southern U.S. have developed practical resistance to Cry Bt proteins but remain susceptible to Vip3Aa, the results suggest that the reduced susceptibility to HearNPV in H. zea may be associated with the resistance to Cry Bt proteins but not with the resistance to Vip3Aa. Correlation analysis confirmed that there was a significant positive relationship between Cry resistance and HearNPV resistance, but not between the Vip3Aa resistance and HearNPV resistance in H. zea. Our findings provide valuable insights into the relationship between susceptibility to HearNPV and resistance to Bt proteins in H. zea.

Oebalus pugnax (Hemiptera: Pentatomidae) resistance to lambda-cyhalothrin in Texas and efficacy of 2 alternative insecticides in grain sorghum.

Along the Coastal Bend of Texas, the rice stink bug, Oebalus pugnax (F.), is a major pest of grain sorghum and rice that is primarily managed by insecticide applications. Reports of rice stink bug resistance to pyrethroids in Texas first surfaced in 2015 and continued to spread. To determine the status of pyrethroid resistance, rice stink bug populations across Texas and Louisiana were evaluated from 2021 to 2023. Mortality was assessed through glass vial exposures to eight concentrations (0, 0.03, 0.1, 0.3, 1, 3, 10, and 30 µg/vial) of a pyrethroid, lambda-cyhalothrin. The concentration of lambda-cyhalothrin required to kill 50% (LC50) of each population was estimated by probit analysis. Furthermore, the efficacy of insecticides, including lambda-cyhalothrin, dimethoate, and dinotefuran, were evaluated in field experiments conducted in 2021. Our results indicated that 14 of the 21 rice stink bug populations sampled were resistant to lambda-cyhalothrin, with LC50 values ranging from 42 to 1,600 times higher than a susceptible population. In the field trial, lambda-cyhalothrin did not control rice stink bugs. Dinotefuran provided excellent control of nymphs, but dimethoate provided greater control of adult rice stink bugs. To our knowledge, this is the first study to thoroughly evaluate the extent or geographic range of pyrethroid resistance in Texas for rice stink bugs.

Diverse genetic basis of Vip3Aa resistance in five independent field-derived strains of Helicoverpa zea in the US.

BACKGROUND: Practical resistance of Helicoverpa zea to Cry proteins has become widespread in the US, making Vip3Aa the only effective Bacillus thuringiensis (Bt) protein for controlling this pest. Understanding the genetic basis of Vip3Aa resistance in H. zea is essential in sustaining the long-term efficacy of Vip3Aa. The objectives of this study were to characterize the inheritance of Vip3Aa resistance in four distinct field-derived H. zea strains (M1-RR, AC4-RR, R2-RR and R15-RR), and to test for shared genetic basis among these strains and a previously characterized Texas resistant strain (LT#70-RR). RESULTS: Maternal effects and sex linkage were absent, and the effective dominance level (DML) was 0.0 across Vip3Aa39 concentrations ranging from 1.0 to 31.6 µg cm-2, in all H. zea resistant strains. Mendelian monogenic model tests indicated that Vip3Aa resistance in each of the four strains was controlled by a single gene. However, interstrain complementation tests indicated that three distinct genetic loci are involved in Vip3Aa resistance in the five resistant H. zea strains: one shared by M1-RR and LT#70-RR; another shared by R2-RR and R15-RR; and a distinct one for AC4-RR. CONCLUSION: Results of this study indicate that Vip3Aa resistance in all H. zea strains was controlled by a single, recessive and autosomal gene. However, there were three distinct genetic loci associated with Vip3Aa resistance in the five resistant H. zea strains. The information generated from this study is valuable for exploring mechanisms of Vip3Aa resistance, monitoring the evolution of Vip3Aa resistance, and devising effective strategies for managing Vip3Aa resistance in H. zea. © 2024 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Evaluation of Cotton Fleahopper (Pseudatomoscelis seriatus (Reuter)) Feeding on Mpp51Aa2-Traited Cotton Utilizing Electrical Penetration Graph (EPG) Waveforms.

Prior to the recent implementation of the Mpp51Aa2 pesticidal protein (ThryvOn), transgenic cotton cultivars have historically offered no control of the cotton fleahopper (Pseudatomocelis seriatus (Reuter)). To evaluate the feeding behavior of cotton fleahoppers on ThryvOn cotton, electropenetrography (EPG) using a Giga-8 DC instrument was used to monitor the probing activity of fourth- and fifth-instar cotton fleahopper nymphs on both ThryvOn and non-ThryvOn cotton squares. Nymphs were individually placed on an excised cotton square for 8 h of EPG recording, after which resulting waveforms were classified as non-probing, cell rupturing, or ingestion. Although there were significantly more cell rupturing events per insect on ThryvOn (mean ± SEM, 14.8 ± 1.7) than on non-ThryvOn squares (mean ± SEM, 10.3 ± 1.6), there was no difference attributable to ThryvOn in the average number of ingestion events per insect. However, the average duration of ingestion events was significantly shorter on squares with ThryvOn (mean ± SEM, 509 ± 148 s) than on squares without (mean ± SEM, 914 ± 135 s). This suggests that cotton fleahoppers continued to probe despite their inability to sustain ingestion. These results provide conclusive evidence that the Mpp51Aa2 pesticidal protein affects the feeding behavior of cotton fleahopper nymphs.

Resistance Allele Frequency of Helicoverpa zea to Vip3Aa Bacillus thuringiensis Protein in the Southeastern U.S.

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.

A Modified F2 Screen for Estimating Cry1Ac and Cry2Ab Resistance Allele Frequencies in Helicoverpa zea (Lepidoptera: Noctuidae).

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.

Cross-resistance and redundant killing of Vip3Aa resistant populations of Helicoverpa zea on purified Bt proteins and pyramided Bt crops.

BACKGROUND: Pyramiding Bt proteins is a key strategy to delay insect resistance development. However, the durability of pyramided Bt crops for controlling insect pests is threatened by cross-resistance among Bt proteins, which can ultimately contribute to resistance development. The corn earworm, Helicoverpa zea, is a major agricultural pest of pyramided Bt crops. Previous studies have examined cross-resistance and redundant killing of Cry resistance in H. zea, but such information is lacking for Vip3Aa resistance in this pest. Here, we evaluated cross-resistance and redundant killing of Vip3Aa-resistant H. zea to purified Bt proteins, as well as Bt corn and Bt cotton. RESULTS: Diet bioassays demonstrated high susceptibility of Vip3Aa-resistant H. zea to Cry1Ac, Cry1A.105, and Cry2Ab2 purified proteins. No Vip3Aa-susceptible, -heterozygous, or -resistant H. zea could survive on pyramided Bt corn containing Cry1 and/or Cry2 proteins. Complete redundant killing was observed in pyramided Bt corn containing Cry1 and/or Cry2 proteins against Vip3Aa resistance in H. zea. Vip3Aa-susceptible, -heterozygous, and -resistant H. zea exhibited survival rates ranging from 0.0% to 22.5% on pyramided Bt cotton with Cry1 and/or Cry2 proteins. Incomplete to complete redundant killing was observed for Vip3Aa-resistant H. zea on pyramided Bt cotton containing Cry1 and/or Cry2 proteins. CONCLUSION: Our findings indicate that Vip3Aa-resistant H. zea does not exhibit positive cross-resistance to Cry1 or Cry2 proteins. In addition, most pyramided Bt crops showed complete or nearly complete redundant killing of Vip3Aa-resistant H. zea. These results indicate that a pyramiding strategy would often be effective for managing Vip3Aa resistance in regions of the United States where H. zea has not evolved resistance to Cry1 and Cry2 toxins. © 2023 Society of Chemical Industry.

Field Evaluation of Cotton Expressing Mpp51Aa2 as a Management Tool for Cotton Fleahoppers, Pseudatomoscelis seriatus (Reuter).

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.

The Spatiotemporal Distribution, Abundance, and Seasonal Dynamics of Cotton-Infesting Aphids in the Southern U.S.

Cotton leafroll dwarf virus (CLRDV) is an emerging aphid-borne pathogen infecting cotton, Gossypium hirsutum L., in the southern United States (U.S.). The cotton aphid, Aphis gossypii Glover, infests cotton annually and is the only known vector to transmit CLRDV to cotton. Seven other species have been reported to feed on, but not often infest, cotton: Protaphis middletonii Thomas, Aphis craccivora Koch, Aphis fabae Scopoli, Macrosiphum euphorbiae Thomas, Myzus persicae Sulzer, Rhopalosiphum rufiabdominale Sasaki, and Smynthurodes betae Westwood. These seven have not been studied in cotton, but due to their potential epidemiological importance, an understanding of the intra- and inter-annual variations of these species is needed. In 2020 and 2021, aphids were monitored from North Carolina to Texas using pan traps around cotton fields. All of the species known to infest cotton, excluding A. fabae, were detected in this study. Protaphis middletonii and A. gossypii were the most abundant species identified. The five other species of aphids captured were consistently low throughout the study and, with the exception of R. rufiabdominale, were not detected at all locations. The abundance, distribution, and seasonal dynamics of cotton-infesting aphids across the southern U.S. are discussed.

Resistance of Spodoptera frugiperda to Cry1, Cry2, and Vip3Aa Proteins in Bt Corn and Cotton in the Americas: Implications for the Rest of the World.

The fall armyworm, Spodoptera frugiperda, is an economically important pest of corn, cotton, and soybean, and a major target of transgenic crops expressing Bacillus thuringiensis (Bt) proteins. In recent years, this insect has invaded most countries in Africa, Southeastern Asia, and Oceania, posing a great threat to food security. Successful use of Bt crops in the U.S. indicates that Bt technology can be an effective tool for management of S. frugiperda in other countries. Evolution of insect resistance is the primary threat to the long-term efficacy of Bt technology. There are many factors that may affect the rate of evolution of insect resistance to Bt crops, which include initial resistance allele frequency, the dose of Bt protein in Bt crops, cross-resistance, complete/incomplete resistance, and fitness costs associated with resistance. Currently, the high dose/refuge and gene-pyramiding approaches are the two main IRM strategies used in the U.S. to combat evolution of insect resistance. In this paper, we review research on resistance of S. frugiperda to Cry1, Cry2, and Vip3Aa proteins. Specifically, we discuss the resistance allele frequencies of S. frugiperda to these three proteins in the field, the genetic basis of resistance, the patterns of cross-resistance, and the fitness costs associated with resistance. Experience and knowledge gained from these studies provide valuable information for the successful use of Bt crop technology for control of S. frugiperda worldwide.

Successful use of endovascular aneurysm repair in the treatment of acute aortoduodenal syndrome.

Aortoduodenal syndrome is a rare phenomenon, described as a duodenal obstruction secondary to direct compression from an abdominal aortic aneurysm. Although traditionally managed with gastrointestinal bypass and subsequent open aortic aneurysm repair, such procedures hold high mortality. Therefore, there is great value in treating this syndrome with an endovascular, minimally invasive technique that is both consistent and successful. This case report recounts how an endovascular aneurysm repair successfully treated a patient with acute aortoduodenal syndrome. No postoperative complications occurred and the duodenal obstruction was relieved without the need for surgical re-intervention. This conveys the advantageous utility of endovascular aneurysm repair in treating patients with aortoduodenal syndrome.

Effects of cross-pollination among non-Bt and pyramided Bt corn expressing cry proteins in seed mixtures on resistance development of dual-gene resistant Helicoverpa zea.

BACKGROUND: Seed mixture strategy can guarantee the compliance of planting non-Bt crops to host the susceptible insects for resistance management. However, pollen movement between Bt and non-Bt corn in the mixed plantings could reduce the efficacy of this strategy for ear-feeding insects. Few studies have evaluated the effects of cross-pollination among non-Bt and pyramided Bt corn in seed mixtures on the resistance development of insects possessing multiple resistances. Here, we provided the first study to investigate whether cross-pollination in mixed plantings of pyramided Bt corn producing Cry1A.105 and Cry2Ab2 would increase the dominance of resistance of dual-gene resistant populations of Helicoverpa zea, a target of pyramided Bt corn and cotton in the USA. RESULTS: We compared the survival and development of susceptible, dual-gene resistant (resistance to both Cry1 and Cry2 proteins) and heterozygous genotypes of H. zea in the laboratory on non-Bt and pyramided Bt corn ears collected from mixed plantings and structured plantings in the field. We found higher fitness for F1 heterozygous insects than for the susceptible insects of H. zea on both pyramided Bt corn and non-Bt corn in the mixed plantings. CONCLUSION: These results suggest that cross-pollination in mixed plantings will significantly increase the dominance of resistance by supporting survival of heterozygous insects for dual-gene resistant populations of H. zea, and therefore accelerate evolution of resistance to pyramided Bt crops. © 2022 Society of Chemical Industry.

Evaluation of Bt resistance in Helicoverpa zea (Lepidoptera: Noctuidae) strains using various Bt cotton plant tissues.

BACKGROUND: Diet-overlay bioassays suggest that Helicoverpa zea (Lepidoptera: Noctuidae) field populations have developed resistance to some of the Bt insecticidal proteins that are constituents of the pyramids expressed in the second and third generation Bt cotton technologies. Unfortunately, these bioassays are not always a reliable indicator for how a seemingly resistant population will perform in an actual cotton field, and thus, leaf tissue bioassays have been suggested as a method to better assess field performance. However, bollworm larvae typically prefer to feed on floral tissue rather than leaf tissue, and an alternative cotton structure type may be more ideal for use in plant tissue-based bioassays. A series of diet-overlay bioassays using Bt proteins and Bt cotton plant tissue were conducted with laboratory susceptible (Bz-SS) and resistant (Cry-RR, resistant to Cry1Ac and Cry2Ab) H. zea strains to determine if plant tissue overlays could detect resistance and which cotton plant structure type would be most ideal for use in bioassays. RESULTS: Results suggest that diet overlays using lyophilized plant tissue were able to detect resistance. Lyophilized tissue from white flowers was most ideal for use in bioassays, whereas tissue from non-Bt bolls and leaves affected larval health and behavior, confounding assay results. CONCLUSION: Overlays using white flower tissue could potentially be used to supplement Bt protein overlays and provide an improved assessment of larval performance on Bt cotton technologies. © 2021 Society of Chemical Industry.

Effective dominance and redundant killing of single- and dual-gene resistant populations of Helicoverpa zea on pyramided Bt corn and cotton.

BACKGROUND: Pyramided Bacillus thuringiensis (Bt) crops producing multiple Bt proteins with different modes of action are widely planted in the United States. Helicoverpa zea is a major target pest of pyramided Bt crops and has evolved practical resistance to both Cry1 and Cry2 proteins in some regions of U.S. However, little information is available regarding redundant killing and the dominance of resistance for insects possessing multiple resistance on pyramided Bt crops. In this study, we evaluated redundant killing and the dominance of resistance for H. zea strains resistant to Cry1 or Cry1 + Cry2 on pyramided Bt corn and cotton. RESULTS: We found that the Cry1-resistant H. zea was incompletely dominant on Cry1Ac + Cry1F cotton. Pyramided crops producing Cry2 and/or Vip3Aa proteins showed a complete redundant killing against the Cry1-resistant H. zea. The Cry1 + Cry2-resistant H. zea displayed incompletely recessive to completely dominant resistance on pyramided Bt crops containing Cry1 and/or Cry2 proteins. The redundant killing was complete for the Cry1 + Cry2-resistant H. zea on pyramided Bt crops producing Vip3Aa protein. CONCLUSION: The dominant resistance of Cry1 and Cry2 in H. zea on pyramided Bt crops deviates from the assumption of functionally recessive resistance underlying the high-dose refuge strategy. However, the assumptions of complete redundant killing are achieved for both Cry1- and Cry1 + Cry2-resistant H. zea on pyramided Bt crops. These results suggest that the pyramided strategy could be valuable for increasing the durability of Bt technology for managing H. zea, a pest with inherently low susceptibility against Cry proteins. © 2022 Society of Chemical Industry.

Status of Cry1Ac and Cry2Ab2 resistance in field populations of Helicoverpa zea in Texas, USA.

Helicoverpa zea is a major target pest of Bt corn and Bt cotton. Field-evolved resistance of H. zea to Cry1 and Cry2 proteins has been widely reported in the United States. Understanding the frequency of resistance alleles in a target insect is critical for Bt resistance management. Despite multiple cases of practical resistance to Cry proteins having been documented in H. zea, there are no data on the current status of alleles conferring resistance to Cry1Ac and Cry2Ab2 in field populations of this pest. During 2018-2019, a total of 106 F2 families for Cry1Ac and 120 F2 families for Cry2Ab2 were established using mass mating and light trap strategy. We screened 13,568 and 15,360 neonates using a discriminatory dose of Cry1Ac and Cry2Ab2, respectively. The results showed that 93.4% and 35.0% of the F2 families could survive on the discriminatory dose of Cry1Ac and Cry2Ab2, respectively. The estimated resistance allele frequency for Cry1Ac in H. zea ranged from 0.4150 to 0.4975 and for Cry2Ab2 ranged from 0.1097 and 0.1228. These data indicate that the frequency of alleles conferring resistance to Cry1 and Cry2 proteins in H. zea in Texas are high. In addition, our data suggest the resistance to Cry1Ac and Cry2Ab2 in the screened families of H. zea varies from recessive to dominant. The information in this study provides precise estimates of Cry resistance allele frequencies in H. zea and increases our understanding of the risks to the sustainability of Bt crops.