Extended Sentinel Monitoring of Helicoverpa zea Resistance to Cry and Vip3Aa Toxins in Bt Sweet Corn: Assessing Changes in Phenotypic and Allele Frequencies of Resistance.

Transgenic corn and cotton that produce Cry and Vip3Aa toxins derived from Bacillus thuringiensis (Bt) are widely planted in the United States to control lepidopteran pests. The sustainability of these Bt crops is threatened because the corn earworm/bollworm, Helicoverpa zea (Boddie), is evolving a resistance to these toxins. Using Bt sweet corn as a sentinel plant to monitor the evolution of resistance, collaborators established 146 trials in twenty-five states and five Canadian provinces during 2020-2022. The study evaluated overall changes in the phenotypic frequency of resistance (the ratio of larval densities in Bt ears relative to densities in non-Bt ears) in H. zea populations and the range of resistance allele frequencies for Cry1Ab and Vip3Aa. The results revealed a widespread resistance to Cry1Ab, Cry2Ab2, and Cry1A.105 Cry toxins, with higher numbers of larvae surviving in Bt ears than in non-Bt ears at many trial locations. Depending on assumptions about the inheritance of resistance, allele frequencies for Cry1Ab ranged from 0.465 (dominant resistance) to 0.995 (recessive resistance). Although Vip3Aa provided high control efficacy against H. zea, the results show a notable increase in ear damage and a number of surviving older larvae, particularly at southern locations. Assuming recessive resistance, the estimated resistance allele frequencies for Vip3Aa ranged from 0.115 in the Gulf states to 0.032 at more northern locations. These findings indicate that better resistance management practices are urgently needed to sustain efficacy the of corn and cotton that produce Vip3Aa.

Removing neonicotinoid seed treatments has negligible effects on refuge function and crop protection in transgenic maize targeting western corn rootworm (Coleoptera: Chrysomelidae).

Nearly all maize seed sold in the United States includes a neonicotinoid seed treatment (NST), meant to protect seedlings against early-season insect pests. For key pests, including western corn rootworm (Diabrotica virgifera virgifera LeConte) (D.v.v), insecticidal proteins derived from Bacillus thuringiensis (Bt) are expressed in plant tissues as alternatives to soil-applied insecticides. Insect resistance management (IRM) plans use non-Bt "refuges" to encourage survival of Bt-susceptible D.v.v., which maintains susceptible alleles in the population. In non-cotton producing regions, IRM guidelines require a minimum 5% blended refuge for maize expressing more than 1 trait targeting D.v.v. Prior work has shown that 5% blends yield insufficient proportions of refuge beetles to contribute reliably to IRM. Whether NSTs interfere with survivorship of refuge beetles is unknown. Our objective was to determine whether NSTs affect proportions of refuge beetles, and secondarily, to determine whether NSTs provide agronomic advantages over Bt seed alone. To reveal host plant type (i.e., Bt or refuge), we used a stable isotope (15N) to mark refuge plants in plots with 5% seed blends. To assess refuge performance between treatments, we compared proportions of beetles from respective natal hosts. In all site-years, NSTs showed inconsistent effects on proportions of refuge beetles. Treatment comparisons showed inconsistent agronomic benefits of NSTs when combined with Bt traits. Our results demonstrate that NSTs have a negligible impact on refuge performance and reinforces the assertion that 5% blends are serving little benefit for IRM. Plant stand and yield were not improved by NSTs.

Magnitude and Extent of Helicoverpa zea Resistance Levels to Cry1Ac and Cry2Ab2 across the Southeastern USA.

After resistance is first detected, continued resistance monitoring can inform decisions on how to effectively manage resistant populations. We monitored for resistance to Cry1Ac (2018 and 2019) and Cry2Ab2 (2019) from southeastern USA populations of Helicoverpa zea. We collected larvae from various plant hosts, sib-mated the adults, and tested neonates using diet-overlay bioassays and compared them to susceptible populations for resistance estimates. We also compared LC50 values with larval survival, weight and larval inhibition at the highest dose tested using regression, and found that LC50 values were negatively correlated with survival for both proteins. Finally, we compared resistance rations between Cry1Ac and Cry2Ab2 during 2019. Some populations were resistant to Cry1Ac, and most were resistant to CryAb2; Cry1Ac resistance ratios were lower than Cry2Ab2 during 2019. Survival was positively correlated with larval weight inhibition for Cry2Ab. This contrasts with other studies in both the mid-southern and southeastern USA, where resistance to Cry1Ac, Cry1A.105, and Cry2Ab2 increased over time and was found in a majority of populations. This indicates that cotton expressing Cry proteins in the southeastern USA was at variable risk for damage in this region.

Evaluating invasion risk and population dynamics of the brown marmorated stink bug across the contiguous United States.

BACKGROUND: Invasive species threaten the productivity and stability of natural and managed ecosystems. Predicting the spread of invaders, which can aid in early mitigation efforts, is a major challenge, especially in the face of climate change. While ecological niche models are effective tools to assess habitat suitability for invaders, such models have rarely been created for invasive pest species with rapidly expanding ranges. Here, we leveraged a national monitoring effort from 543 sites over 3 years to assess factors mediating the occurrence and abundance of brown marmorated stink bug (BMSB, Halyomorpha halys), an invasive insect pest that has readily established throughout much of the United States. RESULTS: We used maximum entropy models to estimate the suitable habitat of BMSB under several climate scenarios, and generalized boosted models to assess environmental factors that regulated BMSB abundance. Our models captured BMSB distribution and abundance with high accuracy, and predicted a 70% increase in suitable habitat under future climate scenarios. However, environmental factors that mediated the geographical distribution of BMSB were different from those driving abundance. While BMSB occurrence was most affected by winter precipitation and proximity to populated areas, BMSB abundance was influenced most strongly by evapotranspiration and solar photoperiod. CONCLUSION: Our results suggest that linking models of establishment (occurrence) and population dynamics (abundance) offers a more effective way to forecast the spread and impact of BMSB and other invasive species than simply occurrence-based models, allowing for targeted mitigation efforts. Implications of distribution shifts under climate change are discussed. © 2022 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Standardized Field Trials in Cotton and Bioassays to Evaluate Resistance of Tobacco Thrips (Thysanoptera: Thripidae) to Insecticides in the Southern United States.

Foliar-applied insecticide treatments may be necessary to manage thrips in cotton (Gossypium hirsutum L.) under severe infestations or when at-planting insecticide seed treatments do not provide satisfactory protection. The most common foliar-applied insecticide is acephate. Field observations in Tennessee suggest that the performance of acephate has declined. Thus, the first objective was to perform leaf-dip bioassays to assess if tobacco thrips, Frankliniella fusca (Hinds) (Thysanoptera: Thripidae), in cotton production regions have evolved resistance to foliar-applied insecticides. A second objective was to assess the performance of commonly applied foliar insecticides for managing thrips in standardized field trials in Arkansas, Tennessee, Mississippi, and Texas. For both objectives, several insecticides were evaluated including acephate, dicrotophos, dimethoate, lambda-cyhalothrin, imidacloprid, and spinetoram. Field trials and bioassays were completed from 2018 to 2021. Dose-response bioassays with acephate were performed on tobacco thrips field populations and a susceptible laboratory population. Bioassay results suggest that tobacco thrips have developed resistance to acephate and other organophosphate insecticides; however, this resistance seems to be most severe in Arkansas, Tennessee, and the Delta region of Mississippi. Resistance to other classes of insecticides were perhaps even more evident in these bioassays. The performance of these insecticides in field trials was variable, with tobacco thrips only showing consistent signs of resistance to lambda-cyhalothrin. However, it is evident that many populations of tobacco thrips are resistant to multiple classes of insecticides. Further research is needed to determine heritability and resistance mechanism(s).

Row crop fields provide mid-summer forage for honey bees.

Honey bees provide invaluable economic and ecological services while simultaneously facing stressors that may compromise their health. For example, agricultural landscapes, such as a row crop system, are necessary for our food production, but they may cause poor nutrition in bees from a lack of available nectar and pollen. Here, we investigated the foraging dynamics of honey bees in a row crop environment. We decoded, mapped, and analyzed 3459 waggle dances, which communicate the location of where bees collected food, for two full foraging seasons (April-October, 2018-2019). We found that bees recruited nestmates mostly locally (<2 km) throughout the season. The shortest communicated median distances (0.474 and 0.310 km), indicating abundant food availability, occurred in July in both years, which was when our row crops were in full bloom. We determined, by plotting and analyzing the communicated locations, that almost half of the mid-summer recruitment was to row crops, with 37% (2018) and 50% (2019) of honey bee dances indicating these fields. Peanut was the most attractive in July, followed by corn and cotton but not soybean. Overall, row crop fields are indicated by a surprisingly large proportion of recruitment dances, suggesting that similar agricultural landscapes may also provide mid-summer foraging opportunities for honey bees.

Sampling Optimization and Crop Interface Effects on Lygus lineolaris Populations in Southeastern USA Cotton.

Tarnished plant bug, Lygus lineolaris (Hemiptera: Miridae), is an economically damaging pest in cotton production systems across the southern United States. We systematically scouted 120 commercial cotton fields across five southeastern states during susceptible growth stages in 2019 and 2020 to investigate sampling optimization and the effect of interface crop and landscape composition on L. lineolaris abundance. Variance component analysis determined field and within-field spatial scales, compared with agricultural district and state, accounted for more variation in L. lineolaris density using sweep net and drop cloth sampling. This result highlights the importance of field-level scouting efforts. Using within-field samples, a fixed-precision sampling plan determined 8 and 23 sampling units were needed to determine L. lineolaris population estimates with 0.25 precision for sweep net (100 sweeps per unit) and drop cloth (1.5 row-m per unit) sampling, respectively. A spatial Bayesian hierarchical model was developed to determine local landscape (<0.5 km from field edges) effects on L. lineolaris in cotton. The proportion of agricultural area and double-crop wheat and soybeans were positively associated with L. lineolaris density, and fields with more contiguous cotton areas negatively predicted L. lineolaris populations. These results will improve L. lineolaris monitoring programs and treatment management decisions in southeastern USA cotton.

Economic Injury Levels for Bt-resistant Helicoverpa zea (Lepidoptera: Noctuidae) in Cotton.

Thresholds for Helicoverpa zea (Boddie) in cotton Gossypium hirsutum L. have been understudied since the widespread adoption of Bt cotton in the United States. Our study was possible due to the widespread presence of H. zea populations with Cry-toxin resistance. We initiated progressive spray timing experiments using three Bt cotton brands (Deltapine, Stoneville, and Phytogen) widely planted across the U.S. Cotton Belt expressing pyramided toxins in the Cry1A, Cry2, and Vip3Aa19 families. We timed foliar insecticide treatments based on week of bloom to manipulate H. zea populations in tandem with crop development during 2017 and 2018. We hypothesized that non-Bt cotton, cotton expressing Cry toxins alone, and cotton expressing Cry and Vip3Aa19 toxins would respond differently to H. zea feeding. We calculated economic injury levels to support the development of economic thresholds from significant responses. Pressure from H. zea was high during both years. Squares and bolls damaged by H. zea had the strongest negative yield associations, followed by larval number on squares. There were fewer yield associations with larval number on bolls and with number of H. zea eggs on the plant. Larval population levels were very low on varieties expressing Vip3Aa19. Yield response varied across experiments and varieties, suggesting that it is difficult to pinpoint precise economic injury levels. Nonetheless, our results generally suggest that current economic thresholds for H. zea in cotton are too high. Economic injury levels from comparisons between non-Bt varieties and those expressing only Cry toxins could inform future thresholds once H. zea evolves resistance to Vip3Aa19.

Fusarium Hardlock Associated With Lygus lineolaris (Hemiptera: Miridae) Injury in Southeastern Cotton.

The tarnished plant bug, Lygus lineolaris (Palisot de Beauvois), is an important insect pest in cotton that feeds on reproductive fruit, contributing to yield loss. Economically damaging infestations of L. lineolaris have doubled in Virginia since 2013. Escalation of L. lineolaris abundance may increase Fusarium hardlock disease observed in this region, compounding economic losses. Research has linked Fusarium hardlock with fungal species Fusarium verticillioides and F. proliferatum. Field and greenhouse experiments were performed to investigate (i) Fusarium hardlock occurrence in field plots managed and unmanaged for L. lineolaris, (ii) severity of F. verticillioides infection of cotton bolls with and without the presence of L. lineolaris feeding in a greenhouse setting, and (iii) Fusarium species composition and prevalence within field-collected L. lineolaris and cotton lint with and without insect feeding injury and hardlock symptoms present. Nearly twice the amount of hardlock (i.e., proportion of hardlocked locules) occurred in field-collected bolls with L. lineolaris feeding symptoms (0.40 ± 0.02) compared with bolls without (0.21 ± 0.01). Based on real-time quantitative PCR, cotton bolls exposed to F. verticillioides inoculum and caged with L. lineolaris adults had greater levels of F. verticillioides DNA compared with untreated bolls. F. proliferatum, F. verticillioides, and F. fujikuroi were isolated from field-collected L. lineolaris and hardlocked cotton lint at harvest. These findings suggest that the presence of L. lineolaris is associated with an increased risk of Fusarium hardlock in Southeastern cotton, and both should be carefully managed using timely insecticide applications and cultural control practices to minimize yield loss.

Reevaluating the Economic Injury Level for Brown Stink Bug (Hemiptera: Pentatomidae) at Various Growth Stages of Maize.

Economic yield loss and reduction in grain quality from brown stink bug, Euschistus servus (Say), feeding injury in early and late stages of maize, Zea mays (Poales: Poaceae, Linnaeus), development was assessed in Virginia and North Carolina in 2018 and 2019. Varying levels of stink bug infestations were introduced to seedling maize (V2-early stage), and a range of late-stages of maize, including 1) the last stage of vegetative development (V12/V14), 2) prior to tasseling, 3) at tasseling (VT), and 4) across all tested late growth stages. Euschistus servus infestation levels included 33, 67, and 100% of maize seedlings, and 25, 50, 100, and 200% of plants during later stages. Infestations were maintained on seedling maize for 7 d, and 8 or 16 d in reproductive stages. Infestation level in seedling maize had an impact on grain yield. Infestation level and growth stage both had an impact on grain yield in reproductive maize. The percentage of discolored kernels was also affected by infestation level, but not growth stage. Regression analysis between grain yield and infestation level indicated that the average economic injury level is 7% in seedling maize (7 bugs/100 plants) and 12% (12 bugs/100 plants) from the last vegetative stages (V12/V14) through pollination (VT). The economic injury level in the late vegetative stages is only applicable when infestations are present for an extended period of time (16 d), emphasizing the need for continued scouting of maize throughout the season to make informed management decisions.

Systems Approach to Evaluate Tarnished Plant Bug (Hemiptera: Miridae) Management Practices in Virginia and North Carolina Cotton.

Economically damaging infestations of Lygus lineolaris (Palisot de Beauvois), tarnished plant bug, were sporadic in Virginia and North Carolina cotton before 2010. Given the sudden rise of L. lineolaris as an economic pest in these states, regionally specific management practices (i.e., chemical and cultural control) are needed to help producers protect yield potential while minimizing input costs. Field experiments were conducted in Virginia and North Carolina in 2018 and 2019 to 1) determine the effects of various management practices on L. lineolaris density, plant injury (i.e., square retention, internal boll injury), and lint yield, 2) calculate the economic trade-offs between integrated pest management (IPM) systems approach and current management practices for L. lineolaris in these states, and 3) evaluate economic benefits associated with various sampling thresholds. Lygus lineolaris populations peaked mid-season (i.e., August) during cotton flowering in both states. Weekly scouting and applying foliar insecticides when the current University Extension recommended economic threshold was reached was the most critical management treatment in maximizing economic returns. Additional costs among various IPM practices did not translate into significant yield protection and economic gains. Moreover, there were additional economic benefits associated with protecting glabrous and longer maturing varieties in Virginia. Lygus lineolaris density varied significantly between states; therefore, management recommendations should be modified based on the growing region. Results from this study will be used to create an IPM strategy to help cotton producers effectively manage this insect pest in the Southeast.

Resistance monitoring to four insecticides and mechanisms of resistance in Lygus lineolaris Palisot de Beauvois (Hemiptera: Miridae) populations of southeastern USA cotton.

BACKGROUND: Economically damaging infestations of the tarnished plant bug, Lygus lineolaris (Palisot de Beauvois), have become more frequent in Virginia and North Carolina cotton since 2013 and 2010, respectively. Foliar insecticide use has increased dramatically in response. Efficacy data (LC50 ) are needed to evaluate L. lineolaris susceptibility and resistance levels (RR50 ) to commonly used and recommended insecticides for managing this pest in the southeastern USA. RESULTS: Elevated resistance levels to acephate and bifenthrin were measured in L. lineolaris populations collected from wild and cultivated hosts in Virginia, North Carolina and northern Alabama when compared to a susceptible laboratory population. High levels of bifenthrin resistance were observed in 2018 and 2019. Mixed-function oxidase and esterase (EST) inhibitors, piperonyl butoxide and S,S,S-Tributyl phosphotrithioate, respectively, had a synergistic effect on bifenthrin with resistant populations of L. lineolaris. Bifenthrin-resistant L. lineolaris populations expressed elevated levels of cytochrome P450 (CYP450 ) monooxygenase and general EST activity. Results suggest that insecticide resistance is present in some locations and that CYP450 and EST activity in L. lineolaris contribute to pyrethroid resistance in the southeastern USA. CONCLUSIONS: Results can serve as a baseline for continued monitoring of L. lineolaris insecticide resistance and inform insecticide resistance management strategies that help southeastern USA cotton producers to minimize inputs and slow resistance development. © 2020 Society of Chemical Industry.

Effects of Aggregation Lure and Tree Species on Halyomorpha halys (Hemiptera: Pentatomidae) Seasonal Oviposition.

The brown marmorated stink bug, Halyomorpha halys (Stål), is a polyphagous pest that feeds on a wide variety of agricultural commodities including tree fruits, berries, vegetables, field crops, and ornamental trees and shrubs. Accurate knowledge of where H. halys lays eggs is critical to optimize the potential release of Trissolcus japonicus (Ashmead), a scelionid egg parasitoid native to the same host region as H. halys. Ideally, parasitoids should be released in and around areas with high host density. In southwestern Virginia in 2017 and 2018, we searched trees for egg masses in an urban environment and nonmanaged wooded border environment. We also evaluated the effects of a commercial aggregation lure on the number of eggs being deposited. This aggregation lure, when combined with methyl (E,E,Z)-2,4,6-decatrienoate (MDT), has been shown to attract both adult and nymph H. halys and its effects on egg laying were not known. Results of this study showed no difference between the number of eggs laid on trees with and without lures. Catalpa trees, Catalpa bignonioides Walter, had the most egg masses throughout the course of the study; however, the redbud, Cercis canadensis L., had similar numbers in the late July and August. There was an overall trend with more eggs masses found on trees with fruiting structures present. This information can provide insight on where and when to make augmentative releases of egg parasitoids for H. halys.

Evaluating Optimal Spray Timing, Planting Date, and Current Thresholds for Lygus lineolaris (Hemiptera: Miridae) in Virginia and North Carolina Cotton.

Economically damaging infestations of Lygus lineolaris (Palisot de Beauvois) (Hemiptera: Miridae), the tarnished plant bug, have been increasing in North Carolina and Virginia cotton since 2009. We conducted experiments to compare prophylactically timed sprays based on cotton phenology and current action thresholds based on sweep net and drop cloth sampling. In the second year of the study, we included planting date as a factor, with early and late-planted cotton. We found L. lineolaris nymph densities were reduced by at least 60% in North Carolina and 74% in Virginia in threshold plots when compared with the untreated control. Protecting the crop from first square through the fourth week of bloom afforded at least 30% control across planting dates in Virginia and at least 40% control in North Carolina. Economic returns were two to three times greater in early-planted cotton than in late-planted cotton. Treating cotton at action threshold or using prophylactic sprays from first square until the sixth week of bloom in early-planted cotton yielded over $500/ha in net returns in both North Carolina and Virginia. This study supports previous research that shows controlling L. lineolaris infestations during squaring and early weeks of flowering is critical for maximizing yield potential. Our findings also suggest that prebloom and bloom thresholds based on adult and nymphal density devised in the Mid-South may need revision in North Carolina and Virginia.

Long-Term Empirical and Observational Evidence of Practical Helicoverpa zea Resistance to Cotton With Pyramided Bt Toxins.

Evidence of practical resistance of Helicoverpa zea (Boddie) (Lepidoptera: Noctuidae) to Bt cotton in the United States is debatable, supported with occasional reports of boll damage in the field. Our objective was to provide both empirical and long-term observational evidence of practical resistance by linking both in-season and end-of-season measurements of H. zea damage to pyramided Bt cotton bolls and to provide Cry1Ac diet-based bioassay data in support of these damage estimates. In-season boll damage from H. zea was highly correlated to end-of-season damaged bolls. Across North Carolina, Bt cotton fields with end-of-season bolls damaged by H. zea increased during 2016 compared to previous years. Elevated damage was coupled with an increase in field sprays targeting H. zea during 2016, but not related to an increase in H. zea abundance. Bioassay data indicated that there was a range of Cry1Ac susceptibility across the southeastern United States. Given the range of susceptibility to Cry1Ac across the southeastern United States, it is probable that resistant populations are common. Since H. zea is resistant to cotton expressing pyramided Cry toxins, the adoption of new cotton varieties expressing Vip3Aa will be rapid. Efforts should be made to delay resistance of H. zea to the Vip3Aa toxin to avoid foliar insecticide use.

Systemic Imidacloprid Affects Intraguild Parasitoids Differently.

Toxoneuron nigriceps (Viereck) (Hymenoptera, Braconidae) and Campoletis sonorensis (Cameron) (Hymenoptera, Ichneumonidae) are solitary endoparasitoids of the tobacco budworm, Heliothis virescens (Fabricius) (Lepidoptera, Noctuidae). They provide biological control of H. virescens populations in Southeastern US agricultural production systems. Field and greenhouse experiments conducted from 2011-2014 compared parasitism rates of parasitoids that developed inside H. virescens larvae fed on tobacco plants treated with and without imidacloprid. The parasitoids in our study did not have a similar response. Toxoneuron nigriceps had reduced parasitism rates, but parasitism rates of C. sonorensis were unaffected. Preliminary data indicate that adult female lifespans of T. nigriceps are also reduced. ELISA was used to measure concentrations of neonicotinoids, imidacloprid and imidacloprid metabolites in H. virescens larvae that fed on imidacloprid-treated plants and in the parasitoids that fed on these larvae. Concentrations were detectable in the whole bodies of parasitized H. virescens larvae, T. nigriceps larvae and T. nigriceps adults, but not in C. sonorensis larvae and adults. These findings suggest that there are effects of imidacloprid on multiple trophic levels, and that insecticide use may differentially affect natural enemies with similar feeding niches.