Search Results (60)

The bollworm/corn earworm, Helicoverpa zea (Boddie) (Lepidoptera: Noctuidae), is one of the most economically damaging crop pests in North America. Colonies of H. zea are notoriously difficult to maintain and frequently collapse in laboratory rearing. The persistent difficulty in maintaining healthy H. zea colonies has become a major obstacle to performing many research activities on the insect. To optimize colony maintenance, six populations were evaluated across three trials and six tests examining pupal maturity at diet removal, adult emergence synchrony, and cohort size at mating and reproduction. Females emerging from mature pupae produced more eggs than those from mid-aged (5–7 d) or young pupae (0–2 d). Synchronizing male and female emergence within one day yielded higher mating frequency, spermatophore transfer, and progeny, whereas a two-day difference reduced these metrics by 45–67%. Adult cohort size also influenced the outcomes, with ≥10 males and ≥10 females per cage enhancing reproductive success. Most matings occurred on nights 2–3, peaking within 2.5 h after lights off. Positive correlations were observed among mating frequency, spermatophore transfer, and progeny production. Overall, optimal performance was achieved by removing pupae only at maturity, synchronizing adult emergence within one day, and maintaining larger adult cohorts. These findings should establish key conditions to improve the mating success, reproduction, and laboratory rearing of H. zea. Full article

Field-evolved resistance of Helicoverpa zea to crops expressing Cry insecticidal proteins from Bacillus thuringiensis (Bt) is widespread across the United States. To comparatively evaluate physiological factors associated with Bt susceptibility, we analyzed two laboratory strains (Benzon and SIMRU) and one field colony obtained from a commercial corn field near Pickens, Arkansas. Biochemical assays of larval midgut extracts showed that Pickens exhibited significantly altered activities of chymotrypsin-like proteases, aminopeptidase N (APN), and alkaline phosphatase (ALP) compared with the SIMRU or Benzon colonies, with differences varying by larval instar. In contrast, trypsin-like protease activities did not differ significantly among the three colonies. Gene expression analyses of ten serine protease genes and seven candidate Cry receptor genes (including cadherin, ATP-binding cassette family C2, ALP, and four APN genes) revealed significant transcriptional differences in the Pickens relative to the lab colonies. Collectively, these results suggest that chymotrypsin-like proteases may play an important role in the activation of Cry toxins in H. zea. Altered chymotrypsin and APN activities, together with differential gene expressions in the Pickens population, likely contribute to reduced Bt susceptibility. The biochemical and molecular differences provide insight into potential physiological factors underlying reduced Bt susceptibility and may inform future Bt resistance monitoring and management strategies. Full article

Plant nutrient content is spatially and temporally dynamic, exposing insect herbivores to substantial nutritional variability. Such variability can constrain insects to feeding on sub-optimal diets, but it can also allow them to regulate their intake towards an optimal nutrient balance. Nutrient regulation is important in pest management, as the nutritional state of insects may alter their susceptibility to insecticides. Diet macronutrient balance has been shown to significantly affect the susceptibility of Helicoverpa zea larvae to endotoxins produced by transgenic crops containing Bacillus thurigiensis (Bt) genes. However, this was demonstrated using a highly inbred laboratory strain, limiting extrapolation to field populations. Here, we test the impact of field-relevant macronutrient variability on the efficacy of two Bt toxins across three field populations to increase the relevance to resistance monitoring and management. While differences in susceptibility were limited across populations, dietary effects were highly population specific. The Bt toxin that was most affected by diet and the diet that supported optimal survival and performance varied across populations. These findings indicate that nutrition can strongly influence Bt susceptibility, but these effects are influenced by population-level differences. To accurately assess Bt susceptibility in the field, bioassay diets should be tailored to the nutritional ecology of local populations. Full article

Invertebrate iridescent viruses (IIVs, family Iridoviridae) are icosahedral double-stranded DNA viruses that infect a wide range of invertebrates, particularly in humid and aquatic environments. During field trials in Chiapas, southern Mexico, larvae of the fall armyworm, Spodoptera frugiperda (Lepidoptera: Noctuidae), displayed an unexpected lavender iridescence, leading to the discovery of novel IIV isolates from this major agricultural pest. Restriction endonuclease analysis revealed evident diversity among isolates from individual infected larvae. Although one field experiment yielded inconclusive results, a second experiment revealed a positive association between IIV disease and SfMNPV infection, compared to a negative association with the prevalence of parasitoids, and no association with entomopathogenic nematodes (probably Hexamermis sp.). These findings require further investigation to determine the distinct ecological routes through which the virus may transmit across host species. IIV infection of S. frugiperda was also identified in Veracruz State, Mexico, and northern Argentina, revealing a previously unrecognized geographic and host range for these viruses. The genomic and evolutionary features of the three isolates from S. frugiperda were compared with those of two other lepidopteran isolates from Helicoverpa zea (IIV30C obtained from CSIRO) and Anticarsia gemmatalis (AgIIV). Genome sizes ranged between 196.1 and 205.4 kbp (~28% GC content), with several large inversions, and were rich in tandem repeats. The average amino acid identity of the complete genomes and phylogenetic analyses of 26 core gene sequences placed all five isolates within the genus Chloriridovirus, closely related to IIV22 and IV22a isolated from blackflies (Diptera) in Wales and a previously sequenced isolate of IIV30 from the USA. We conclude that these lepidopterans are all infected by closely related strains of the virus species Chloriridovirus simulium1 across their native geographical range. These findings highlight the unexpected ecological breadth and evolutionary adaptability of IIVs. Full article

Pheromone-baited traps are commonly used to monitor and detect moths. Traps and lures are often compared to each other to identify the optimum set up and lure dosage. However, it is also important to understand the efficiency of a trap in capturing the moths that are attracted to it. In the present study, three pheromone-baited traps were placed at the edge of a cornfield to determine the one with the highest capture rate of Helicoverpa zea (Lepidoptera: Noctuidae): Scentry Heliothis traps, clear Unitraps, and green Unitraps. Once it was determined that green Unitraps captured more H. zea, field observations determined the number of moths captured in the traps compared to the number approaching, i.e., trap efficiency. Green Unitraps had a capture efficiency ranged from 5 to 11%, with an average of 11%. Unitraps, although useful for monitoring existing populations, may not be effective in detecting an invasive incursion. The implications of low capture efficiency in the surveillance of H. zea are considered, including possible explanations and next steps to improve monitoring efforts of heliothine moths. Full article

Fall armyworm (FAW), Spodoptera frugiperda (J. E. Smith, 1797) (Lepidoptera: Noctuidae), is a major pest in the U.S. and has spread globally, causing severe agricultural losses in different countries. Due to its high mobility and potential for long-distance dispersal, understanding FAW migration is a key tool for forecasting outbreaks and implementing timely management measures. Recent studies using stable hydrogen isotopes indicated reverse (southward) migration of Helicoverpa zea Boddie (Lepidoptera: Noctuidae). Here, we tested the reverse migration hypothesis for FAW in North America. Estimation of the hydrogen isotopic ratio on 324 samples collected in Florida, an intermixing zone at the edge of the continental U.S., indicated evidence of reverse migration in samples of FAW moths. They showed a high probability of origin from the U.S. Corn Belt, with a greater probability of origin in Nebraska, South Dakota, Minnesota, Kansas and Wisconsin. This southward movement provides new insights into the risk of spreading pesticide resistance alleles in this species to southern regions and contributes to the improvement of integrated pest management and insect resistance management programs. Full article

The corn earworm, Helicoverpa zea (Boddie), causes persistent ear damage to corn grown in the southeastern United States region. Increased levels of ear damage have been associated with mycotoxin contamination in addition to yield loss. Corn hybrids expressing proteins from the Bacillus thuringiensis (Bt) may provide corn earworm control. A selection of hybrids expressing various Bt traits were evaluated in field experiments across Georgia over two years to assess their efficacy for corn earworm control, grain yield and quality protection, and grain mycotoxin mitigation. Ear damage was significantly reduced only by Bt hybrids expressing the Vip3Aa20 protein. The remaining Bt hybrids expressing Cry proteins provided only marginal control. Ear damage had a variable effect on grain yield and was not correlated with grain aflatoxin contamination. In contrast, grain fumonisin contamination was positively associated with earworm damage. These results indicate Bt hybrids that effectively reduce corn earworm damage may also assist in reducing fumonisin contamination and possibly yield loss. Full article

Blended refuges for corn-expressing toxins from Bacillus thuringiensis (Bt) Berliner controls have been approved in the United States as an alternative resistance management approach to structured refuge. This study examined the impact of blended refuges up to 30% non-Bt seed on the corn earworm, Helicoverpa zea (Boddie) (Lepidoptera: Noctuidae), larval numbers, and kernel injury, and its effect on corn grain yield and test weights. The percentage of infested ears, larval numbers, and kernel injury of the 20% blend were not significantly different from the non-Bt and nonblended Bt for the Cry1A.105 + Cry2Ab + Cry1Fa2 treatment, but infested ears, larval numbers, and kernel injury of the nonblended Cry1A.105 + Cry2Ab treatment was lower than the comparable non-Bt hybrids, with the 20% blend being intermediate. The nonblended Cry1A.105 + Cry2Ab + Vip3Aa20 had virtually no larvae in ears and no kernel injury. Ear infestation, larval numbers, and kernel injury of the non-Bt blends with Vip3Aa20 were proportional to the percentage of non-Bt seed in the blend, and all infested ears were from the non-Bt refuge plants. Grain yield and test weight were not significantly different among nonblended or blended treatments of any Bt product tested. Results indicate losses of grain yield and test weight by corn earworm in seed blends up to 30% non-Bt seed are unlikely with infestation levels observed in this study. Full article

Insect control traits are a key component of improving the efficacy of insect pest management and maximizing crop yields for growers. Insect traits based on proteins expressed by the bacteria Bacillus thuringiensis (Bt) have proven to be very effective tools in achieving this goal. Unfortunately, the adaptability of insects has led to resistance to certain proteins in current commercial products. Therefore, new insecticidal traits representing a different mode of action (MoA) than those currently in use are needed. Cry1Ja has good insecticidal activity against various lepidopteran species, and it provides robust protection against insect feeding with in planta expression. For Bt proteins, different MoAs are determined by their binding sites in the insect midgut. In this study, competitive binding assays are performed using brush border membrane vesicles (BBMVs) from Helicoverpa zea, Spodoptera frugiperda, and Chrysodeixis includens to evaluate the MoA of Cry1Ja relative to representatives of the various Bt proteins that are expressed in current commercial products for lepidopteran insect protection. This study highlights differences in the shared Cry protein binding sites in three insect species, Cry1Ja bioactivity against Cry1Fa resistant FAW, and in planta efficacy against target pests. These data illustrate the potential of Cry1Ja for new insect trait development. Full article

The occurrence of insect pests in crops directly affects the yield of plants and grains. This scenario led to the mass investigation of chemical products that overcome these adversities and provide control potential. Nonetheless, over the years, this strategy resulted in high production costs, generation of waste harmful to the environment, and resistance of target insects. The adoption of alternative practices, such as the formulation and production of products of microbial origin, emerges as an encouraging tool compared to control alternatives, indicating a sustainability bias, and allowing a reduction in the risks of human and animal contamination. The purpose of this study was to perform bioprospecting for microbial agents with potential insecticidal effects. The isolated microorganisms were submitted to submerged fermentation, at 28 °C and 120 rpm, for seven days. The fermented broth was filtered using a vacuum pump and centrifuged at 3200× g and 10 °C for 10 min. Initially, 163 microbial agents were collected. Subsequently, a pre-selection of the 50 most promising bioagents was conducted, based on the mortality rates (%) of the applied isolates to target pests. Furthermore, a global mathematical modeling design was created, indicating the best potential microorganisms. Moreover, to stipulate the difference between treatments, dilutions of the fermented broths of each microorganism were conducted (n × 10⁻⁵–n × 10⁻⁸). Mortality was maximum (100%) for Helicoverpa zea and Euschistus heros. Other encouraging results were indicated in the control of Anticarsia gemmatalis and Chrysodeixis includens (up to 87.5%) and Elasmopalpus lignosellus (up to approximately 83.5%). Fungal isolates were identified as Talaromyces piceae. Among the bacteria, based on sequencing of the 16S ribosomal gene, the isolates were identified as Lysinibacillus fusiformis, Paenibacillus ottowii, and Clostridium sphenoides. The results obtained are relevant to the scientific community and, especially, are interesting for companies that are operating in this field in the agricultural sector. Full article

Experiments were conducted in 2020 and 2021 in College Station, TX; Stoneville, MS; and Blackville, SC, to evaluate the potential of grain sorghum to serve as a trap crop for Helicoverpa zea (Boddie), a nursery crop for natural enemies of H. zea, and a source of Helicoverpa armigera nucleopolyhedrovirus (HearNPV) for H. zea management in cotton. The experiments consisted of three treatments, including cotton-only, non-treated cotton–sorghum, and HearNPV-treated cotton–sorghum. Variables, including percent injury to fruiting forms, parasitized H. zea larvae, egg density, H. zea larval density, beneficial arthropod numbers, and HearNPV prevalence, were compared between the treatments. Growing cotton in an intercropping system with grain sorghum did not result in a consistent increase in H. zea control and beneficial arthropod density relative to the cotton-only treatment. Additionally, our results did not show sufficient evidence that grain sorghum interplanted with cotton can serve as a source of HearNPV that can favor H. zea control in cotton. However, we found that, if maintained in the cotton canopy, HearNPV may favor some level of H. zea suppression in cotton. Based on our HearNPV infection analyses using PCR, chrysopids, coccinellids, pentatomids, reduviids, formicids, anthocorids, and spiders appeared to be carrying HearNPV. The virus was detected consistently in specimens of coccinellids, pentatomids, and reduviids across both years of the study. We suggest that further investigation on virus efficacy against H. zea in cotton using the sorghum–cotton system as well as the ability of grain sorghum to serve as a H. zea trap crop and source of H. zea natural enemies be considered in future studies. Full article

Helicoverpa armigera is among the most problematic agricultural pests worldwide due to its polyphagy and ability to evolve pesticide resistance. Molecular detection methods for H. armigera have been developed to track its spread, as such methods allow for rapid and accurate differentiation from the native sibling species H. zea. Droplet digital PCR (ddPCR) is a preferred method for bulk screening due to its accuracy and tolerance to PCR inhibitors; however, real-time PCR is less expensive and more widely available in molecular labs. Improvements to DNA extraction yield, purity, and throughput are crucial for real-time PCR assay optimization. Bulk DNA extractions have recently been improved to where real-time PCR sensitivity can equal that of ddPCR, but these new methods require significant time and specialized equipment. In this study, we improve upon previously published bulk DNA extraction methods by reducing bench time and materials. Our results indicate that the addition of caffeine and RNase A improves DNA extraction, resulting in lower Cq values during real-time PCR while reducing the processing time and cost per specimen. Such improvements will enable the use of high throughput screening methods across multiple platforms to improve the probability of detection of H. armigera. Full article

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. Full article

Insect viruses have been described from numerous lineages, yet patterns of genetic exchange and viral prevalence, which are essential to understanding host–virus coevolution, are rarely studied. In Helicoverpa zea, the virus HzNV-2 can cause deformity of male and female genitalia, resulting in sterility. Using ddPCR, we found that male H. zea with malformed genitalia (agonadal) contained high levels of HzNV-2 DNA, confirming previous work. HzNV-2 was found to be prevalent throughout the United States, at more than twice the rate of the baculovirus HaSNPV, and that it contained several host-acquired DNA sequences. HzNV-2 possesses four recently endogenized lepidopteran genes and several more distantly related genes, including one gene with a bacteria-like sequence found in both host and virus. Among the recently acquired genes is cytosolic serine hydroxymethyltransferase (cSHMT). In nearly all tested H. zea, cSHMT contained a 200 bp transposable element (TE) that was not found in cSHMT of the sister species H. armigera. No other virus has been found with host cSHMT, and the study of this shared copy, including possible interactions, may yield new insights into the function of this gene with possible applications to insect biological control, and gene editing. Full article

The wide occurrence of resistance to Cry1A and Cry2A insecticidal toxins from Bacillus thuringiensis (Bt) in the corn earworm/bollworm Helicoverpa zea (Boddie) leaves the Vip3A toxin produced during the vegetative stage of Bt as the only fully active toxin expressed in transgenic crops to control H. zea in the U.S.A. During 2021, the first unexpected survival of H. zea and injury (UXI) on a maize hybrid expressing Cry1A.105, Cry2Ab2, and Vip3Aa in Louisiana, U.S.A. were observed in two sentinel plots used for resistance monitoring. A follow-up intensive investigation was conducted with two H. zea populations established from larvae collected from the two UXI plots. The main goal of this study was to reveal if the unexpected damage was due to resistance development in the insect to the Bt toxins expressed in the maize hybrid. Diet-overlay bioassays showed that the two populations were highly resistant to Cry1A.105, moderately resistant to Cry2Ab2, but still highly susceptible to Vip3Aa when compared to a reference susceptible strain. In 10 d assays with detached ears, the larvae of the two UXI populations exhibited survival on ears expressing only Cry toxins but presented near 100% mortality on maize hybrids containing both cry and vip3A transgenes. Multiple field trials over three years demonstrated that natural H. zea populations in Louisiana were highly resistant to maize expressing only Cry toxins but remained susceptible to all tested hybrids containing cry and vip3A genes. Altogether, the results of this study suggest that the observed UXIs in Louisiana were associated with a resistance to Cry toxins but were not due to a resistance to Vip3A. The possible causes of the UXIs are discussed. The results generated and procedures adopted in this study help in determining thresholds for defining UXIs, assessing resistance risks, and documenting field resistance. Full article