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.

Sexual repurposing of juvenile aposematism in locusts.

Adaptive plasticity requires an integrated suite of functional responses to environmental variation, which can include social communication across life stages. Desert locusts (Schistocerca gregaria) exhibit an extreme example of phenotypic plasticity called phase polyphenism, in which a suite of behavioral and morphological traits differ according to local population density. Male and female juveniles developing at low population densities exhibit green- or sand-colored background-matching camouflage, while at high densities they show contrasting yellow and black aposematic patterning that deters predators. The predominant background colors of these phenotypes (green/sand/yellow) all depend on expression of the carotenoid-binding "Yellow Protein" (YP). Gregarious (high-density) adults of both sexes are initially pinkish, before a YP-mediated yellowing reoccurs upon sexual maturation. Yellow color is especially prominent in gregarious males, but the reason for this difference has been unknown since phase polyphenism was first described in 1921. Here, we use RNA interference to show that gregarious male yellowing acts as an intrasexual warning signal, which forms a multimodal signal with the antiaphrodisiac pheromone phenylacetonitrile (PAN) to prevent mistaken sexual harassment from other males during scramble mating in a swarm. Socially mediated reexpression of YP thus adaptively repurposes a juvenile signal that deters predators into an adult signal that deters undesirable mates. These findings reveal a previously underappreciated sexual dimension to locust phase polyphenism, and promote locusts as a model for investigating the relative contributions of natural versus sexual selection in the evolution of phenotypic plasticity.

Anisotropic interaction and motion states of locusts in a hopper band.

Swarming locusts present a quintessential example of animal collective motion. Juvenile locusts march and hop across the ground in coordinated groups called hopper bands. Composed of up to millions of insects, hopper bands exhibit aligned motion and various collective structures. These groups are well-documented in the field, but the individual insects themselves are typically studied in much smaller groups in laboratory experiments. We present, to our knowledge, the first trajectory data that detail the movement of individual locusts within a hopper band in a natural setting. Using automated video tracking, we derive our data from footage of four distinct hopper bands of the Australian plague locust, Chortoicetes terminifera. We reconstruct nearly 200 000 individual trajectories composed of over 3.3 million locust positions. We classify these data into three motion states: stationary, walking and hopping. Distributions of relative neighbour positions reveal anisotropies that depend on motion state. Stationary locusts have high-density areas distributed around them apparently at random. Walking locusts have a low-density area in front of them. Hopping locusts have low-density areas in front and behind them. Our results suggest novel insect interactions, namely that locusts change their motion to avoid colliding with neighbours in front of them.

Probing Transcriptional Crosstalk between Cryptochromes and Iron-sulfur Cluster Assembly 1 (MagR) in the Magnetoresponse of a Migratory Insect.

Many organisms can sense and respond to magnetic fields (MFs), with migratory species in particular utilizing geomagnetic field information for long-distance migration. Cryptochrome proteins (Crys) along with a highly conserved Iron-sulfur cluster assembly protein (i.e., MagR) have garnered significant attention for their involvement in magnetoresponse (including magnetoreception). However, in vivo investigations of potential transcriptional crosstalk between Crys and MagR genes have been limited. The brown planthopper, Nilaparvata lugens, is a major migratory pest insect and an emerging model for studying MF intensity-related magnetoresponse. Here, we explored in vivo transcriptional crosstalk between Crys (Cry1 and Cry2) and MagR in N. lugens. The expression of Crys and MagR were found to be sensitive to MF intensity changes as small as several micro-teslas. Knocking down MagR expression led to a significant downregulation of Cry1, but not Cry2. The knockdown of either Cry1 or Cry2 individually did not significantly affect MagR expression. However, their double knockdown resulted in significant upregulation of MagR. Our findings clearly indicate transcriptional crosstalk between MagR and Crys known to be involved in magnetoresponse. This work advances the understanding of magnetoresponse signaling and represents a key initial step towards elucidating the functional consequences of these novel in vivo interactions.

Plant-associated fungi affect above- and belowground pest responses to soybean plants.

AIM: The objective of this research was to screen fungal isolates originally isolated from cotton plants and measure their effects on the interactions between soybean and two aboveground pests (cabbage looper; Trichoplusia ni and soybean looper; Chrysodeixis includens) as well as a belowground pest (soybean cyst nematode; Heterodera glycines). METHODS AND RESULTS: For aboveground pests, we measured the leaf area consumed and larval weight. For our belowground pest tests, we measured shoot height, shoot fresh weight, root fresh weight and number of cysts. Out of the 50 fungal isolates tested, we tested 30 fungi in the interaction with cabbage looper, 36 for soybean looper, 41 for soybean cyst nematode. We tested 23 isolates against all pests and identified multiple isolates that significantly changed the response of pests on inoculated soybean plants versus controls. CONCLUSIONS: We identified three fungal isolates that significantly reduced both leaf area consumed aboveground by caterpillars and number of cysts produced belowground by nematodes. These isolates were an Epicoccum italicum, a Chaetomium undulatum and a Stemphylium majusculum. SIGNIFICANCE AND IMPACT OF STUDY: Overall, this study provides important insights into plant-fungal interactions and their effect on both above- and belowground pests. This study also highlights an important first step towards harnessing the potential of microbial inoculates as a tool for integrated pest management in soybeans.

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.

Change in geomagnetic field intensity alters migration-associated traits in a migratory insect.

Geomagnetic field (GMF) intensity can be used by some animals to determine their position during migration. However, its role, if any, in mediating other migration-related phenotypes remains largely unknown. Here, we simulated variation in GMF intensity between two locations along the migration route of a nocturnal insect migrant, the brown planthopper Nilaparvata lugens, that varied by approximately 5 µT in field intensity. After one generation of exposure, we tested for changes in key morphological, behavioural and physiological traits related to migratory performance, including wing dimorphism, flight capacity and positive phototaxis. Our results showed that all three morphological and behavioural phenotypes responded to a small difference in magnetic field intensity. Consistent magnetic responses in the expression of the phototaxis-related Drosophila-like cryptochrome 1 (Cry1) gene and levels of two primary energy substrates used during flight, triglyceride and trehalose, were also found. Our findings indicate changes in GMF intensity can alter the expression of phenotypes critical for insect migration and highlight the unique role of magnetoreception as a trait that may help migratory insects express potentially beneficial phenotypes in geographically variable environments.

Multiscale analyses on a massive immigration process of Sogatella furcifera (Horváth) in south-central China: influences of synoptic-scale meteorological conditions and topography.

Mass landings of migrating white-backed planthopper, Sogatella furcifera (Horváth), can lead to severe outbreaks that cause heavy losses for rice production in East Asia. South-central China is the main infestation area on the annual migration loop of S. furcifera between the northern Indo-China Peninsula and mainland China; however, rice planthopper species are not able to survive in this region over winter. In this study, a trajectory analysis of movements from population source areas and a spatiotemporal dynamic analysis of mesoscale and synoptic weather conditions from 7 to 10 May 2012 were conducted using the weather research and forecasting (WRF) model to identify source areas of immigrants and determine how weather and topographic terrain influence insect landing. A sensitivity experiment was conducted with reduced topography using the WRF model to explain the associations among rainfall, topography, and light-trap catches of S. furcifera. The trajectory modeling results suggest that the source areas of S. furcifera immigrants into south-central China from 8 to 10 May were mainly southern Guangxi, northern Vietnam, and north-central Vietnam. The appearance of enormous catches of immigrant S. furcifera coincided with a period of rainstorms. The formation of transporting southerly winds was strongly associated with the topographic terrain. Additionally, the rainfall distribution and intensity over south-central China significantly decreased when topography was reduced in the model and were directly affected by wind circulation, which was associated with mountainous terrain that caused strong convection. This study indicates that migrating populations of S. furcifera were carried by the southwesterly low-level jets and that topographically induced convergent winds, precipitation, low temperatures, and wind shear acted as key factors that led to massive landings.

Spatio-Temporal, Genotypic, and Environmental Effects on Plant Soluble Protein and Digestible Carbohydrate Content: Implications for Insect Herbivores with Cotton as an Exemplar.

Plant soluble protein and digestible carbohydrate content significantly affect insect herbivore fitness, but studies reporting plant protein and carbohydrate content are rare. Instead, the elements nitrogen and carbon often are used as surrogates for plant protein and digestible carbohydrate content, respectively. However, this is problematic for two reasons. First, carbon is found in all organic molecules, which precludes strong correlations with ecologically important dietary macronutrients (e.g., digestible carbohydrates, the primary energy source for most insect herbivores). Second, some elements (e.g., nitrogen) are present in both macronutrients (e.g., protein) and non-nutritive secondary compounds (e.g., alkaloids, protease inhibitors); in these cases N values would greatly overestimate protein available for an insect herbivore. Thus, the objective of this study was to explicitly document plant protein-carbohydrate content and assess its variation in cotton (Gossypium hirsutum and G. barbadense), which is a nutritional resource for a number of insect herbivores. We did this by measuring plant soluble protein (P) and digestible carbohydrate (C) content across seven plant tissues, five varieties, and two growing environments. Significant differences in P and C concentration, total macronutrient content (P + C), and P:C ratio were observed across plant tissues, plant age and environment; smaller differences were seen across plant genotype. Foliar tissues had higher total P + C content compared to reproductive tissues, except for developing seeds and developing flowers, which contained twice the total P + C content; these two tissues also had the highest P content. Our data show that even agricultural monocultures offer a highly heterogeneous protein-carbohydrate landscape for insect herbivores. Characterizing plant resources using nutritional currencies (e.g., protein and carbohydrates) that are ecologically and physiologically-relevant to insect herbivores can be used to enhance our understanding of plant-insect interactions.

Nutritional ecology beyond the individual: a conceptual framework for integrating nutrition and social interactions.

Over recent years, modelling approaches from nutritional ecology (known as Nutritional Geometry) have been increasingly used to describe how animals and some other organisms select foods and eat them in appropriate amounts in order to maintain a balanced nutritional state maximising fitness. These nutritional strategies profoundly affect the physiology, behaviour and performance of individuals, which in turn impact their social interactions within groups and societies. Here, we present a conceptual framework to study the role of nutrition as a major ecological factor influencing the development and maintenance of social life. We first illustrate some of the mechanisms by which nutritional differences among individuals mediate social interactions in a broad range of species and ecological contexts. We then explain how studying individual- and collective-level nutrition in a common conceptual framework derived from Nutritional Geometry can bring new fundamental insights into the mechanisms and evolution of social interactions, using a combination of simulation models and manipulative experiments.

Modeling spatiotemporal dynamics of outbreaking species: influence of environment and migration in a locust.

Many pest species exhibit huge fluctuations in population abundance. Understanding their large-scale and long-term dynamics is necessary to develop effective control and management strategies. Occupancy models represent a promising approach to unravel interactions between environmental factors and spatiotemporal dynamics of outbreaking populations. Here, we investigated population dynamics of the Australian plague locust, Chortoicetes terminifera, using density data collected between 1988 and 2010 by the Australian Plague Locust Commission over more than 3 million km2 in eastern Australia. We applied multistate and autologistic multi-season occupancy models to test competing hypotheses about environmental and demographic processes affecting the large-scale dynamics of the Australian plague locust. We found that rainfall and land cover predictors best explained the spatial variability in outbreak probability across eastern Australia. Outbreaks are more likely to occur in temperate than tropical regions, with a faster and more continuous response to rainfall in desert than in agricultural areas. Our results also support the hypothesis that migration tends to propagate outbreaks only locally (over distances lower than 400 km) rather than across climatic regions. Our study suggests that locust outbreak forecasting and management systems could be improved by implementing key environmental factors and migration in hierarchical spatial models. Finally, our modeling framework can be seen as a step towards bridging the gap between mechanistic and more phenomenological models in the spatial analysis of fluctuating populations.

The role of spatial aggregation in forensic entomology.

A central concept in forensic entomology is that arthropod succession on carrion is predictable and can be used to estimate the postmortem interval (PMI) of human remains. However, most studies have reported significant variation in successional patterns, particularly among replicate carcasses, which has complicated estimates of PMIs. Several forensic entomology researchers have proposed that further integration of ecological and evolutionary theory in forensic entomology could help advance the application of succession data for producing PMI estimates. The purpose of this essay is to draw attention to the role of spatial aggregation of arthropods among carrion resources as a potentially important aspect to consider for understanding and predicting the assembly of arthropods on carrion over time. We review ecological literature related to spatial aggregation of arthropods among patchy and ephemeral resources, such as carrion, and when possible integrate these results with published forensic literature. We show that spatial aggregation of arthropods across resources is commonly reported and has been used to provide fundamental insight for understanding regional and local patterns of arthropod diversity and coexistence. Moreover, two suggestions are made for conducting future research. First, because intraspecific aggregation affects species frequency distributions across carcasses, data from replicate carcasses should not be combined, but rather statistically quantified to generate occurrence probabilities. Second, we identify a need for studies that tease apart the degree to which community assembly on carrion is spatially versus temporally structured, which will aid in developing mechanistic hypotheses on the ecological factors shaping community assembly on carcasses.

Fungal seed treatments of cotton affect boll weevil development.

BACKGROUND: Nonpathogenic fungi associated with plants can enhance plant defenses against stress factors, including herbivory. This study assessed whether cotton plants grown from seeds treated with different fungi affected boll weevil, Anthonomus grandis grandis Boheman, development and reproduction along with plant tolerance. We used whole plants grown from seeds treated with different fungi (Chaetomium globosum TAMU520 and TAMU559, Phialemonium inflatum TAMU490, and Beauveria bassiana) versus non-treated controls to test insect growth, reproduction, and plant tolerance assays in a greenhouse. RESULTS: Regarding boll weevil reproduction, fewer larvae hatched and fewer adults emerged from fungal-treated plants. In addition, the developmental time from oviposition to adult emergence was delayed in the plants treated with all fungi. For plant tolerance, B. bassiana-treated plants attacked by boll weevils shed fewer squares than nonfungal-treated plants. CONCLUSION: Fungal treatments can affect boll weevil performance and reproduction on cotton plants, with potentially negative effects on population growth. Collectively, these results support the potential for cottonseed treatments with fungi as a novel tool for boll weevil management in the field. © 2023 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Using genetic comparisons of populations from Arizona, Mexico, and Texas to investigate fall armyworm migration in the American southwest.

Fall armyworm (FAW) is a global agricultural pest, causing substantial economic losses in corn and many other crops. Complicating efforts to control this pest is its capacity for long distance flights, which has been described in greatest detail for the central and eastern sections of the United States. FAW infestations are also routinely found in agricultural areas in southern Arizona, which lie beyond the western limits of the mapped migratory pathways. Climate suitability analysis found that the affected Arizona locations cannot support permanent FAW populations, indicating that these FAW most likely arise from annual migrations. A better understanding of this migration would provide insights into how large moth populations can move across desert habitats as well as the degree of gene flow occurring between FAW populations across the North American continent. In this study the Arizona populations were genetically characterized and compared to a selection of permanent and migratory FAW from multiple sites in the United States and Mexico. The results are consistent with migratory contributions from permanent populations in the states of Texas (United States) and Sinaloa (Mexico), while also providing evidence of significant barriers to gene flow between populations within Mexico. An unexpected finding was that two genetically distinct FAW subpopulations known as "host strains" have a differential distribution in the southwest that may indicate significant differences in their migration behavior in this region. These findings indicate that the combination of mitochondrial and Z-linked markers have advantages in comparing FAW populations that can complement and extend the findings from other methods.

A New qPCR Assay for the Rapid Diagnosis of Anthonomus grandis Subspecies.

Rapid and accurate identification of Anthonomus grandis subspecies is crucial for effective management and eradication. Current diagnostic methods have limitations in terms of time to diagnosis (up to seven days) and can yield ambiguous results. Here, we present the validation of a custom TaqMan SNP Genotyping Assay for the rapid and accurate identification of A. grandis grandis (boll weevil) and A. g. thurberiae (thurberia weevil) subspecies. To validate the assay, we conducted three main experiments: (1) a sensitivity test to determine the DNA concentration range at which the assay performs, (2) a non-target specificity test to ensure no amplification in non-target weevils (false positives), and (3) an accuracy test comparing the results of the new assay to previously established methods. These experiments were carried out in parallel at three independent facilities to confirm the robustness of the assay to variations in equipment and personnel. We used DNA samples from various sources, including field-collected specimens, museum specimens, and previously isolated DNA. The assay demonstrated high sensitivity (PCR success with ≥0.05 ng/µL DNA template), specificity (0.02 false positive rate), and accuracy (97.7%) in diagnosing boll weevil and thurberia weevil subspecies. The entire workflow, including DNA extraction, assay preparation, PCR run time, and data analysis, can be completed within a single workday (7-9 h) by a single technician. The deployment of this assay as a diagnostic tool could benefit boll weevil management and eradication programs by enabling same-day diagnosis of trap-captured or intercepted weevil specimens. Furthermore, it offers a more reliable method for identifying unknown specimens, contributing to the overall effectiveness of boll weevil research and control efforts.

Cannibalism can drive the evolution of behavioural phase polyphenism in locusts.

During outbreaks, locust swarms can contain millions of insects travelling thousands of kilometers while devastating vegetation and crops. Such large-scale spatial organization is preceded locally by a dramatic density-dependent phenotypic transition in multiple traits. Behaviourally, low-density 'solitarious' individuals avoid contact with one another; above a critical local density, they undergo a rapid behavioural transition to the 'gregarious phase' whereby they exhibit mutual attraction. Although proximate causes of this phase polyphenism have been widely studied, the ultimate driving factors remain unclear. Using an individual-based evolutionary model, we reveal that cannibalism, a striking feature of locust ecology, could lead to the evolution of density-dependent behavioural phase-change in juvenile locusts. We show that this behavioural strategy minimizes risk associated with cannibalistic interactions and may account for the empirically observed persistence of locust groups during outbreaks. Our results provide a parsimonious explanation for the evolution of behavioural plasticity in locusts.

Mitochondrial genomes reveal the global phylogeography and dispersal routes of the migratory locust.

The migratory locust, Locusta migratoria, is the most widely distributed grasshopper species in the world. However, its global genetic structure and phylogeographic relationships have not been investigated. In this study, we explored the worldwide genetic structure and phylogeography of the locust populations based on the sequence information of 65 complete mitochondrial genomes and three mitochondrial genes of 263 individuals from 53 sampling sites. Although this locust can migrate over long distances, our results revealed high genetic differentiation among the geographic populations. The populations can be divided into two different lineages: the Northern lineage, which includes individuals from the temperate regions of the Eurasian continent, and the Southern lineage, which includes individuals from Africa, southern Europe, the Arabian region, India, southern China, South-east Asia and Australia. An analysis of population genetic diversity indicated that the locust species originated from Africa. Ancestral populations likely separated into Northern and Southern lineages 895 000 years ago by vicariance events associated with Pleistocene glaciations. These two lineages evolved in allopatry and occupied their current distributions in the world via distinct southern and northern dispersal routes. Genetic differences, caused by the long-term independent diversification of the two lineages, along with other factors, such as geographic barriers and temperature limitations, may play important roles in maintaining the present phylogeographic patterns. Our phylogeographic evidence challenged the long-held view of multiple subspecies in the locust species and tentatively divided it into two subspecies, L. m. migratoria and L. m. migratorioides.

Physiological and transcriptional immune responses of a non-model arthropod to infection with different entomopathogenic groups.

Insect immune responses to multiple pathogen groups including viruses, bacteria, fungi, and entomopathogenic nematodes have traditionally been documented in model insects such as Drosophila melanogaster, or medically important insects such as Aedes aegypti. Despite their potential importance in understanding the efficacy of pathogens as biological control agents, these responses are infrequently studied in agriculturally important pests. Additionally, studies that investigate responses of a host species to different pathogen groups are uncommon, and typically focus on only a single time point during infection. As such, a robust understanding of immune system responses over the time of infection is often lacking in many pest species. This study was conducted to understand how 3rd instar larvae of the major insect pest Helicoverpa zea responded through the course of an infection by four different pathogenic groups: viruses, bacteria, fungi, and entomopathogenic nematodes; by sampling at three different times post-inoculation. Physiological immune responses were assessed at 4-, 24-, and 48-hours post-infection by measuring hemolymph phenoloxidase concentrations, hemolymph prophenoloxidase concentrations, hemocyte counts, and encapsulation ability. Transcriptional immune responses were measured at 24-, 48-, and 72-hours post-infection by quantifying the expression of PPO2, Argonaute-2, JNK, Dorsal, and Relish. This gene set covers the major known immune pathways: phenoloxidase cascade, siRNA, JNK pathway, Toll pathway, and IMD pathway. Our results indicate H. zea has an extreme immune response to Bacillus thuringiensis bacteria, a mild response to Helicoverpa armigera nucleopolyhedrovirus, and little-to-no detectable response to either the fungus Beauveria bassiana or Steinernema carpocapsae nematodes.

Olfactometer Responses of Convergent Lady Beetles Hippodamia convergens (Coleoptera: Coccinellidae) to Odor Cues from Aphid-Infested Cotton Plants Treated with Plant-Associated Fungi.

Microbes have the potential to affect multitrophic plant-insect-predator interactions. We examined whether cotton plants treated with potentially beneficial fungi affect interactions between cotton aphids Aphis gossypii and predatory lady beetles Hippodamia convergens. We used Y-tube olfactometer assays to test lady beetle behavioral responses to stimuli emitted by aphid-infested and non-infested cotton plants grown from seeds treated with either Phialemonium inflatum (TAMU490) or Chaetomium globosum (TAMU520) versus untreated control plants. We tested a total of 960 lady beetles (480 males and 480 females) that had been deprived of food for approximately 24 h. In the absence of any fungal treatments, males preferred stimuli from aphid-infested plants, and females spent more time associated with stimuli from aphid-infested versus non-infested plants. When fungal treatments were added, we observed that lady beetles preferred non-aphid-infested P. inflatum plants, and males responded slower to plants treated with P. inflatum in the absence of aphids. We found some evidence to suggest that lady beetle behavioral responses to plants might vary according to the fungal treatment but not strongly impact their use as part of an insect pest management strategy.

Patterns of genomic and allochronic strain divergence in the fall armyworm, Spodoptera frugiperda (J.E. Smith).

Speciation is the process through which reproductive isolation develops between distinct populations. Because this process takes time, speciation studies often necessarily examine populations within a species that are at various stages of divergence. The fall armyworm, Spodoptera frugiperda (J.E. Smith), is comprised of two strains (R = Rice & C = Corn) that serve as a novel system to explore population divergence in sympatry. Here, we use ddRADSeq data to show that fall armyworm strains in the field are largely genetically distinct, but some interstrain hybridization occurs. Although we detected F1 hybrids of both R- and C-strain maternal origin, only hybrids with R-strain mtDNA were found to contribute to subsequent generations, possibly indicating a unidirectional barrier to gene flow. Although these strains have been previously defined as "host plant-associated," we recovered an equal proportion of R- and C-strain moths in fields dominated by C-strain host plants. As an alternative to host-associated divergence, we tested the hypothesis that differences in nightly activity patterns could account for reproductive isolation by genotyping temporally collected moths. Our data indicates that strains exhibit a significant shift in the timing of their nightly activities in the field. This divergence in phenology creates a prezygotic reproductive barrier that likely maintains the genetic isolation between strains. Thus, we conclude that it may be ecologically inaccurate to refer to the C- and R- strain as "host-associated" and they should more appropriately be considered "allochronic strains."