Cross-pollination in seed-blended refuge and selection for Vip3A resistance in a lepidopteran pest as detected by genomic monitoring.

The evolution of pest resistance to management tools reduces productivity and results in economic losses in agricultural systems. To slow its emergence and spread, monitoring and prevention practices are implemented in resistance management programs. Recent work suggests that genomic approaches can identify signs of emerging resistance to aid in resistance management. Here, we empirically examined the sensitivity of genomic monitoring for resistance management in transgenic Bt crops, a globally important agricultural innovation. Whole genome resequencing of wild North American Helicoverpa zea collected from non-expressing refuge and plants expressing Cry1Ab confirmed that resistance-associated signatures of selection were detectable after a single generation of exposure. Upon demonstrating its sensitivity, we applied genomic monitoring to wild H. zea that survived Vip3A exposure resulting from cross-pollination of refuge plants in seed-blended plots. Refuge seed interplanted with transgenic seed exposed H. zea to sublethal doses of Vip3A protein in corn ears and was associated with allele frequency divergence across the genome. Some of the greatest allele frequency divergence occurred in genomic regions adjacent to a previously described candidate gene for Vip3A resistance. Our work highlights the power of genomic monitoring to sensitively detect heritable changes associated with field exposure to Bt toxins and suggests that seed-blended refuge will likely hasten the evolution of resistance to Vip3A in lepidopteran pests.

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 non-native 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 4Mbp region containing CYP337B3. Next, we identify signatures of selection in the panmictic population of non-admixed 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.

Management of Insect Pests with Bt Crops in the United States.

Genetically engineered corn and cotton that produce insecticidal toxins derived from the bacterium Bacillus thuringiensis (Bt) have been used to manage insect pests in the United States and elsewhere. In some cases, this has led to regional suppression of pest populations and pest eradication within the United States, and these outcomes were associated with reductions in conventional insecticides and increased profits for farmers. In other instances, pests evolved resistance to multiple Bt traits, compromising the capacity of Bt crops to manage pests and leading to increased feeding injury to crops in the field. Several aspects of pest biology and pest-crop interactions were associated with cases where pests remained susceptible versus instances where pests evolved resistance. The viability of future transgenic traits can be improved by learning from these past outcomes. In particular, efforts should be made to delay resistance by increasing the prevalence of refuges and using integrated pest management.

Impact of Caterpillar Increased Feeding Rates on Reduction of Bt Susceptibility.

The use of insect-resistant transgenic crops producing Bacillus thuringiensis protein Cry toxins (Bt) to control caterpillars is wide-spread. Development of a mechanism to prevent Bt from reaching its target site in the digestive system could result in Bt resistance and resistance to other insecticides active per os. Increased feeding rates by increasing temperature in tobacco budworms, Chloridea virescens, and bollworms, Helicoverpa zea, decreased Bt Cry1Ac susceptibility and mortality. The same was found in C. virescens for Bollgard II plant extract containing Bt Cry1Ac and Cry2Ab2 toxins. Furthermore, H. zea from the same inbred laboratory colony that fed faster independent of temperature manipulation were less susceptible to Bt intoxication. A laboratory derived C. virescens Bt resistant strain demonstrated a higher feeding rate on non-Bt artificial diet than the parental, Bt susceptible strain. A laboratory-reared Bt resistant fall armyworm, Spodoptera frugiperda, strain also fed faster on non-Bt diet compared to Bt susceptible caterpillars of the same species, both originally collected from corn. The studies in toto and the literature reviewed support the hypothesis that increased feeding rate is a behavioral mechanism for reducing caterpillar susceptibility to Bt. Its possible role in resistance needs further study.

Whole genome comparisons reveal panmixia among fall armyworm (Spodoptera frugiperda) from diverse locations.

BACKGROUND: The fall armyworm (Spodoptera frugiperda (J.E. Smith)) is a highly polyphagous agricultural pest with long-distance migratory behavior threatening food security worldwide. This pest has a host range of > 80 plant species, but two host strains are recognized based on their association with corn (C-strain) or rice and smaller grasses (R-strain). The population genomics of the United States (USA) fall armyworm remains poorly characterized to date despite its agricultural threat. RESULTS: In this study, the population structure and genetic diversity in 55 S. frugiperda samples from Argentina, Brazil, Kenya, Puerto Rico and USA were surveyed to further our understanding of whole genome nuclear diversity. Comparisons at the genomic level suggest a panmictic S. frugiperda population, with only a minor reduction in gene flow between the two overwintering populations in the continental USA, also corresponding to distinct host strains at the mitochondrial level. Two maternal lines were detected from analysis of mitochondrial genomes. We found members from the Eastern Hemisphere interspersed within both continental USA overwintering subpopulations, suggesting multiple individuals were likely introduced to Africa. CONCLUSIONS: Our research is the largest diverse collection of United States S. frugiperda whole genome sequences characterized to date, covering eight continental states and a USA territory (Puerto Rico). The genomic resources presented provide foundational information to understand gene flow at the whole genome level among S. frugiperda populations. Based on the genomic similarities found between host strains and laboratory vs. field samples, our findings validate the experimental use of laboratory strains and the host strain differentiation based on mitochondria and sex-linked genetic markers extends to minor genome wide differences with some exceptions showing mixture between host strains is likely occurring in field populations.

Extended investigation of field-evolved resistance of the corn earworm Helicoverpa zea(Lepidoptera: Noctuidae) to Bacillus thuringiensis Cry1A.105 and Cry2Ab2 proteins in thesoutheastern United States.

Previous studies have reported that the corn earworm/bollworm, Helicoverpa zea (Boddie), has developed field resistance to pyramided Bacillus thuringiensis (Bt) Cry1A/Cry2A maize and cotton in certain areas of the southeastern United States. The objective of the current study was to determine the current status and distribution of the resistance to Cry1A.105 and Cry2Ab2 in H. zea. In the study, 31 H. zea populations were collected from major maize planting areas across seven southeastern states of the United States during 2018 and 2019 and assayed against the two Bt proteins. Diet over-lay bioassays showed that most of the populations collected during the two years were significantly resistant to the Cry1A.105 protein. Most of the populations collected during 2019 were also resistant to Cry2Ab2, while significant variances were observed in the susceptibility of the populations collected during 2018 to Cry2Ab2. The results showed that Cry1A.105 and Cry2Ab2 resistance in H. zea is widely distributed in the regions sampled. The resistance to Cry1A.105 appeared to have plateaued, while selection for Cry2Ab2 resistance is likely still occurring. Thus, effective measures for mitigating the Cry1A/Cry2A resistance need to be developed and implemented to ensure the sustainable use of Bt crop biotechnology.

Vertical and temporal distribution of Helicoverpa zea (Lepidoptera: Noctuidae) larvae in determinate and indeterminate soybean.

Most oviposition by Helicoverpa zea (Boddie) occurs near the top of the canopy in soybean, Glycine max (L.) Merr, and larval abundance is influenced by the growth habit of plants. However, the vertical distribution of larvae within the canopy is not as well known. We evaluated the vertical distribution of H. zea larvae in determinate and indeterminate varieties, hypothesizing that larval distribution in the canopy would vary between these two growth habits and over time. We tested this hypothesis in a naturally infested replicated field experiment and two experimentally manipulated cage experiments. In the field experiment, flowering time was synchronized between the varieties by manipulating planting date, while infestation timing was manipulated in the cage experiments. Larvae were recovered using destructive sampling of individual soybean plants, and their vertical distribution by instar was recorded from three sampling points over time in each experiment. While larval population growth and development varied between the determinate and indeterminate varieties within and among experiments, we found little evidence that larvae have preference for different vertical locations in the canopy. This study lends support to the hypothesis that larval movement and location within soybean canopies do not result entirely from oviposition location and nutritional requirements.

Brown Stink Bug (Hemiptera: Pentatomidae) Damage to Seedling Corn and Impact on Grain Yield.

Brown stink bugs, Euschistus servus, are an important early-season pest of field corn in the southeastern United States. Feeding in the early stages of corn development can lead to a number of growth deformities and deficiencies and, ultimately, a reduction in yield. An observational and two experimentally manipulated trials were conducted in 2017 and 2018 to 1) determine optimal timing for assessing brown stink bug damage, 2) assess the level of damage from which yield compensation can occur, and 3) examine the relationship between brown stink bug density and early-season damage and yield. Fields were identified with infestations of brown stink bugs and a damage rating system for early stages of corn was established. Varying rates of brown stink bug densities were introduced using field cages and damage was assessed throughout the season. The density and duration of stink bug infestations were critical factors for damage potential, with each day of active feeding per plant resulting in a loss of ~14 kg/ha in yield. The level of damage in early stages of corn was categorized into easily identifiable groups, with only the most severe damage leading to a reduction in yield. Moderate and minimal feeding damage did not result in yield loss. This study emphasizes the need for early and frequent scouting of corn to determine the risk of damage and yield loss from brown stink bugs. Results from this study can be used to help develop management programs for brown stink bugs in the early vegetative stages of field corn.

Location of Helicoverpa zea (Lepidoptera: Noctuidae) larvae on different plant parts of determinate and indeterminate soybean.

Helicoverpa zea (Boddie) is a damaging pest of many crops including soybean, Glycine max (L.), especially in the southern United States. Previous studies have concluded that oviposition and development of H. zea larvae mirror the phenology of soybean, with oviposition occurring during full bloom, younger larvae developing on blooms and leaves, intermediate aged larvae developing on varying tissue types, and older larvae developing on flowers and pods. In a field trial, we investigated the presence of natural infestations of H. zea larvae by instar in determinate and indeterminate soybean varieties. In complementary experiments, we artificially infested H. zea and allowed them to oviposit on plants within replicated cages (one with a determinate variety and two with an indeterminate variety). Plants were sampled weekly during the time larvae were present. In the natural infestation experiment, most larvae were found on blooms during R3 and were early to middle instars; by R4, most larvae were found on leaves and were middle to late instars. In contrast, in the cage study, most larvae were found on leaves regardless of soybean growth stage or larval stage. Determinate and indeterminate growth habit did not impact larval preference for different soybean tissue types. Our studies suggest H. zea larvae prefer specific tissue types, but also provide evidence that experimental design can influence the results. Finally, our finding of larval preference for leaves contrasts with findings from previous studies.

Multiple Known Mechanisms and a Possible Role of an Enhanced Immune System in Bt-Resistance in a Field Population of the Bollworm, Helicoverpa zea: Differences in Gene Expression with RNAseq.

Several different agricultural insect pests have developed field resistance to Bt (Bacillus thuringiensis) proteins (ex. Cry1Ac, Cry1F, etc.) expressed in crops, including corn and cotton. In the bollworm, Helicoverpa zea, resistance levels are increasing; recent reports in 2019 show up to 1000-fold levels of resistance to Cry1Ac, a major insecticidal protein in Bt-crops. A common method to analyze global differences in gene expression is RNA-seq. This technique was used to measure differences in global gene expression between a Bt-susceptible and Bt-resistant strain of the bollworm, where the differences in susceptibility to Cry1Ac insecticidal proteins were 100-fold. We found expected gene expression differences based on our current understanding of the Bt mode of action, including increased expression of proteases (trypsins and serine proteases) and reduced expression of Bt-interacting receptors (aminopeptidases and cadherins) in resistant bollworms. We also found additional expression differences for transcripts that were not previously investigated, i.e., transcripts from three immune pathways-Jak/STAT, Toll, and IMD. Immune pathway receptors (ex. PGRPs) and the IMD pathway demonstrated the highest differences in expression. Our analysis suggested that multiple mechanisms are involved in the development of Bt-resistance, including potentially unrecognized pathways.

Effectiveness of Genes for Hessian Fly (Diptera: Cecidomyiidae) Resistance in the Southeastern United States.

The Hessian fly, Mayetiola destructor (Say) (Diptera: Cecidomyiidae), is the most important insect pest of wheat (Triticum aestivum L. subsp. aestivum) in the southeastern United States, and the deployment of genetically resistant wheat is the most effective control. However, the use of resistant wheat results in the selection of pest genotypes that can overcome formerly resistant wheat. We have evaluated the effectiveness of 16 resistance genes for protection of wheat from Hessian fly infestation in the southeastern United States. Results documented that while 10 of the genes evaluated could provide protection of wheat, the most highly effective genes were H12, H18, H24, H25, H26, and H33. However, H12 and H18 have been reported to be only partially effective in field evaluations, and H24, H25, and H26 may be associated with undesirable effects on agronomic traits when introgressed into elite wheat lines. Thus, the most promising new gene for Hessian fly resistance appears to be H33. These results indicate that identified highly effective resistance in wheat to the Hessian fly is a limited resource and emphasize the need to identify novel sources of resistance. Also, we recommend that the deployment of resistance in gene pyramids and the development of novel strategies for engineered resistance be considered.

Sublethal Effects of Insecticide Exposure on Megacopta cribraria (Fabricius) Nymphs: Key Biological Traits and Acetylcholinesterase Activity.

Megacopta cribraria F. (Hemiptera: Plataspidae), the kudzu bug, is an invasive insect pest of U.S. soybean. At present, insecticide application is the primary and most effective control option for M. cribraria In this study, the potential effects of sublethal and low-lethal concentrations (LC10 and LC40) of three common insecticides on key biological traits and acetylcholinesterase (AChE) activity of the treated nymphal stage of insect were assessed. The results show that the sublethal concentration of imidacloprid significantly reduced adult emergence rate of M. cribraria A low-lethal concentration of imidacloprid significantly increased nymphal development time, but significantly decreased adult emergence rate and adult longevity. Both sublethal and low-lethal concentrations of acephate caused an increase in nymphal development time and a reduction in adult emergence rate and adult longevity. Fecundity of females was significantly reduced only by exposure to low-lethal concentrations of acephate. Sublethal and low-lethal concentrations of bifenthrin increased nymphal development time, but significantly decreased adult emergence rate. In addition, we found that the AChE activity of M. cribraria was significantly increased only by LC40 imidacloprid, but strongly inhibited by acephate.

Biology, Pest Status, Microbiome and Control of Kudzu Bug (Hemiptera: Heteroptera: Plataspidae): A New Invasive Pest in the U.S.

Soybean is an important food crop, and insect integrated pest management (IPM) is critical to the sustainability of this production system. In recent years, the introduction into the United States of the kudzu bug currently identified as Megacopta cribraria (F.), poses a threat to soybean production. The kudzu bug was first discovered in the state of Georgia, U.S. in 2009 and since then has spread to most of the southeastern states. Because it was not found in the North American subcontinent before this time, much of our knowledge of this insect comes from research done in its native habitat. However, since the U.S. introduction, studies have been undertaken to improve our understanding of the kudzu bug basic biology, microbiome, migration patterns, host selection and management in its expanding new range. Researchers are not only looking at developing IPM strategies for the kudzu bug in soybean, but also at its unique relationship with symbiotic bacteria. Adult females deposit bacterial packets with their eggs, and the neonates feed on these packets to acquire the bacteria, Candidatus Ishikawaella capsulata. The kudzu bug should be an informative model to study the co-evolution of insect function and behavior with that of a single bacteria species. We review kudzu bug trapping and survey methods, the development of bioassays for insecticide susceptibility, insecticide efficacy, host preferences, impact of the pest on urban environments, population expansion, and the occurrence of natural enemies. The identity of the kudzu bug in the U.S. is not clear. We propose that the kudzu bug currently accepted as M. cribraria in the U.S. is actually Megacopta punctatissima, with more work needed to confirm this hypothesis.

Action Thresholds for Managing Megacopta cribraria (Hemiptera: Plataspidae) in Soybean Based on Sweep-Net Sampling.

The kudzu bug, Megacopta cribraria (F.), first discovered in the United States in 2009, has rapidly become a pest of commercial soybean, Glycine max (L.) Merrill, throughout much of the southeast. Because of its recent arrival, management practices and recommendations are not well established. To develop action thresholds, we evaluated insecticide applications targeted at different densities of adults and nymphs determined using the standard 38-cm diameter sweep net sampling method in 12 soybean field trials conducted in Georgia, North Carolina, and South Carolina from 2011 to 2013. Average peak densities of M. cribraria in the untreated controls reached as high as 63.5 ± 11.0 adults per sweep and 34.7 ± 8.0 nymphs per sweep. Insecticide applications triggered at densities of one adult or nymph of M. cribraria per sweep, two adults or nymphs per sweep, and one adult or nymph per sweep, with nymphs present, resulted in no yield reductions in most cases compared with plots that were aggressively protected with multiple insecticide applications. A single insecticide application timed at the R3 or R4 soybean growth stages also resulted in yields that were equivalent to the aggressively protected plots. Typically, treatments (excluding the untreated control) that resulted in fewer applications were more cost-effective. These results suggest that a single insecticide application targeting nymphs was sufficient to prevent soybean yield reduction at the densities of M. cribraria that we observed.

Aggregation and Association of NDVI, Boll Injury, and Stink Bugs in North Carolina Cotton.

Sampling of herbivorous stink bugs in southeastern U.S. cotton remains problematic. Remote sensing was explored to improve sampling of these pests and associated boll injury. Two adjacent 14.5-ha cotton fields were grid sampled in 2011 and 2012 by collecting stink bug adults and bolls every week during the third, fourth, and fifth weeks of bloom. Satellite remote sensing data were collected during the third week of bloom during both years, and normalized difference vegetation index (NDVI) values were calculated. Stink bugs were spatially aggregated on the third week of bloom in 2011. Boll injury from stink bugs was spatially aggregated during the fourth week of bloom in 2012. The NDVI values were aggregated during both years. There was a positive association and correlation between stink bug numbers and NDVI values, as well as injured bolls and NDVI values, during the third week of bloom in 2011. During the third week of bloom in 2012, NDVI values were negatively correlated with stink bug numbers. During the fourth week of bloom in 2011, stink bug numbers and boll injury were both positively associated and correlated with NDVI values. During the fourth week of bloom in 2012, stink bugs were negatively correlated with NDVI values, and boll injury was negatively associated and correlated with NDVI values. This study suggests the potential of remote sensing as a tool to assist with sampling stink bugs in cotton, although more research is needed using NDVI and other plant measurements to predict stink bug injury.

Impact of corn earworm injury on yield of transgenic corn producing Bt toxins in the Carolinas.

Transgenic corn, Zea mays L., hybrids expressing insecticidal Cry proteins from Bacillus thuringiensis (Bt) and insecticide applications to suppress injury from Helicoverpa zea (Boddie) were evaluated in Florence, SC, and in Plymouth, NC, in 2012 and 2013. Based on kernel area injured, insecticide applications (chlorantraniliprole) every 3-4 d from R1 until H. zea had cycled out of corn reduced injury by 80-93% in Florence and 94-95% in Plymouth. Despite intensive applications of insecticide (13-18 per trial), limited injury still occurred in all treated plots in 2012, except in DKC 68-03 (Genuity VT Double PRO), based on kernels injured (both locations) and proportion of injured ears (Florence only). In 2013, ear injury was low in Plymouth, with no kernel injury in any insecticide-treated plots, except P1498R (non-Bt) and P1498YHR (Optimum Intrasect). Injury in Florence in 2013 did not occur in treated plots of DKC 68-04 (non-Bt), DKC 68-03 (Genuity VT Double PRO), and N785-3111 (Agrisure Viptera). Yields were not significantly affected by insecticide treatment and were not statistically different among near-isolines with and without Bt traits. Yields were not significantly associated with kernel injury based on regression analyses. The value of using Bt corn hybrids to manage H. zea is discussed.

Yield analysis and corn earworm feeding in Bt and non-Bt corn hybrids across diverse locations.

Corn, Zea mays L. (Poales: Poaceae), growers in the US Cotton Belt are required to plant 20% of total corn acres to non-Bt hybrids for resistance management (non-Bt refuge). Most growers do not meet this requirement, in part, because they perceive non-Bt hybrids to yield less than Bt hybrids. We planted multiple non-Bt and Bt hybrids from a single company in small-plot replicated trials at a single location from 2019 to 2023, as well as in small-plot replicated trials at multiple locations during 2022 and 2023. In the single location, we measured kernel injury from corn earworm, Helicoverpa zea Boddie (Lepidoptera: Noctuidae), and we recorded yield at all locations. In the single location trial, yields only separated among hybrids in 3 out of 5 years. In the multiple location trial, yields were variable between both years. We found that Bt hybrids tended to yield higher than non-Bt hybrids overall, but this was influenced by the inclusion of non-Bt hybrids that had a lower overall genetic yield potential in the environments we tested them in. In both tests, when hybrids were analyzed during each year, both Bt and non-Bt hybrids were among the statistically highest yielders. Our study demonstrates the importance of comparing multiple Bt and non-Bt hybrids to draw yield comparisons. This highlights the need for corn seed company breeders to put effort into improving yield for non-Bt hybrids. Hopefully this effort will translate into increased planting of non-Bt refuge corn for growers in the US Cotton Belt.

Vertical distribution and tissue selection of Helicoverpa zea (Lepidoptera: Noctuidae) adult oviposition and neonates on soybean with an indeterminate or determinate growth habit.

Helicoverpa zea (Boddie) is a polyphagous pest that can cause serious damage to crops, including soybeans (Glycine max L.). In soybeans with both determinate and indeterminate growth habits, H. zea larvae are more commonly found on leaves compared to blooms, stems, and pods. Past research demonstrated that H. zea adults tend to oviposit near the upper part of the plant canopy in soybeans with a determinate growth habit. However, ovipositional selection on soybeans with an indeterminate growth habit is unknown. We hypothesized that H. zea would oviposit more uniformly throughout the canopy on indeterminate soybean growth habits due to more diffuse reproductive tissue. We planted field and greenhouse experiments with varieties sharing a similar relative maturity (2 maturity group [MG] 5.2 varieties and 2 MG 5.4/5.5 varieties) but with different growth habits. To test oviposition selection, adult H. zea females were allowed to oviposit under field and caged conditions. We counted the number of H. zea eggs and neonates on each plant tissue type from each of 3 equal parts in the plant canopy: upper, middle, and lower. In both experiments, eggs and neonates were most common on leaves at the top of the plant regardless of soybean growth habit. Consequently, ovipositional selection is likely independent of reproductive tissue availability, and patterns of oviposition through the canopy are similar in growth habits. An improved understanding of H. zea ecology in soybeans relative to indeterminate growth habits may improve recommendations for managing this significant pest of soybean.

Helicoverpa zea Boddie (Lepidoptera: Noctuidae) pupal success and adult eclosion across variable soil type and moisture.

Helicoverpa zea Boddie (Lepidoptera: Noctuidae) is an important pest in many crops in the southern United States. Upon reaching the final larval instar, H. zea quests for a pupation site in the soil. Pupae are vulnerable to mortality since their movement is limited. Soil type and moisture can influence H. zea emergence, but the interaction of these factors has not been demonstrated. We compared sandy and clay soils in greenhouse and laboratory experiments. In the first experiment, we evaluated the preference of larvae to choose either sandy or clay soil for pupation. In a second experiment, we set the sandy soils at different moisture levels and observed prepupae pupation preference in a choice scenario. In a third experiment, we observed prepupae pupation in different moisture levels in a no-choice scenario. In a 4th experiment, we evaluated adult emergence following pupation when we increased moisture or kept it constant. In a final experiment, we evaluated pupation behavior in sandy or clay soils with a webcam and a glass arena. We found that larvae preferred to pupate in sandy soils over clay soils and that pupal success was highest at intermediate moisture levels. In addition, elevated soil moisture levels did not impact the emergence of H. zea between sandy or clay soil. Finally, H. zea did not take longer to burrow in either sandy or clay soil, but the tunnels of the pupal burrow were larger in sandy soil compared to clay soil. Our results clarify H. zea behavior across soil moisture and soil type.