YDV Resistance in Barley: Phenotyping, Remote Imagery, and Virus-Vector Characterization.

Yellow Dwarf Viruses (YDV) spread by aphids are some of the most economically important barley (Hordeum vulgare L.) virus-vector complexes worldwide. Detection and control of these viruses are critical components in the production of barley, wheat, and numerous other grasses of agricultural importance. Genetic control of plant diseases is often preferable to chemical control to reduce the epidemiological, environmental, and economic cost of foliar insecticides. Accordingly, the objectives of this work were to I) screen a barley population for resistance to YDV under natural infection using phenotypic assessment of disease symptoms, II) implement drone imagery to further assess resistance and test its utility as a disease screening tool, III) identify the prevailing virus and vector types in the experimental environment, and IV) perform a genome-wide association study to identify genomic regions associated with measured traits. Significant genetic differences were found in a population of 192 barley inbred lines regarding their YDV symptom severity and symptoms were moderately to highly correlated with grain yield. The severity of YDV measured with aerial imaging was highly correlated with on-the-ground estimates (r=0.65). Three aphid species vectoring three YDV species were identified with no apparent genotypic influence on their distribution. A QTL impacting YDV resistance was detected on chromosome 2H, albeit undetected using aerial imaging. However, QTL for canopy cover and mean NDVI were successfully mapped using the drone. This work provides a framework for utilizing drone imagery in future resistance breeding efforts for YDV in cereals and grasses, and in other virus-vector disease complexes.

Field-evolved pyrethroid resistance in Tychius picirostris (Coleoptera: Curculionidae) populations in Oregon white clover seed crops.

The clover seed weevil, Tychius picirostris Fabricius, a serious pest of white clover, Trifolium repens L., grown for seed in western Oregon, causing feeding damage to flowers and developing seeds. Since 2017, white clover seed producers have anecdotally reported T. picirostris control failures using foliar pyrethroid insecticide applications. This mode of action (MoA) is an important chemical control option for T. picirostris management. To evaluate insecticide resistance selection to pyrethroids (bifenthrin) and other MoAs labeled for T. picirostris management (malathion and chlorantraniliprole), adult populations were collected from 8 commercial white clover grown for seed fields in the Willamette Valley, OR, in 2022 and 2023. Among collected Oregon populations, very high resistance ratios (RR50 = 178.00-725.67) were observed to technical grade bifenthrin and low to high resistance ratios (RR50 = 7.80-32.80) to malathion in surface contact assays compared to a susceptible Canadian field population. Moreover, >2.73 times the labeled rate of formulated product containing bifenthrin as the sole MoA was required to kill >50% of T. picirostris in topical assays. Synergistic assays with a mixed-function oxidase inhibitor, an esterase inhibitor, and a glutathione-S-transferase inhibitor revealed phase I and II detoxification enzymes are present in Oregon T. picirostris populations and confer metabolic resistance to bifenthrin. This is the first report of T. picirostris insecticide resistance selection to pyrethroid and organophosphate insecticides. Results will inform continued monitoring and insecticide resistance management strategies to slow the evolution of T. picirostris insecticide resistance selection in Oregon's white clover seed production.

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.

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.

Evaluation of 13-Tetradecenyl Acetate Pheromone for Melanotus communis (Coleoptera: Elateridae) Detection in North Carolina Row Crop Agroecosystems.

Melanotus communis Gyllenhal (Coleoptera: Elateridae) larvae are a common soil-dwelling pest of many crops, including sweet potato, grains, and tobacco. Although many studies have focused on the larval stage of this pest, the seasonal activity and ecology of the adults (click beetles) are not well understood. The overarching goal of this study was to relate M. communis adult activity to host crops in the North Carolina row-crop agroecosystem. To do this, we conducted a two-year study documenting male M. communis activity, using a recently identified sex attractant pheromone, 13-tetradecenyl acetate. This project was divided into two parts: 1) a pheromone assessment study testing the efficacy and specificity of 13-tetradecenyl acetate, and two analogs, 13-tetradecenyl butyrate and 13-tetradecenyl hexanoate, and 2) a landscape survey using traps baited with 13-tetradecenyl acetate. Results of the efficacy study showed that 13-tetradecenyl acetate was the most effective M. communis lure when compared to non-baited control traps or traps baited with the two homologs. The landscape study documented a strong association between M. communis catch and the adjacent crop type. We found that adult M. communis abundance was greatest near corn, followed by sweet potato, and then cotton. Analysis of activity over time found that the peak activity occurred during July. Overall, this project demonstrates the usefulness of pheromone-baited traps in providing new information about M. communis activity.

Landscape-level variation in Bt crops predict Helicoverpa zea (Lepidoptera: Noctuidae) resistance in cotton agroecosystems.

BACKGROUND: Helicoverpa zea (Boddie) damage to Bt cotton and maize has increased as a result of widespread Bt resistance across the USA Cotton Belt. Our objective was to link Bt crop production patterns to cotton damage through a series of spatial and temporal surveys of commercial fields to understand how Bt crop production relates to greater than expected H. zea damage to Bt cotton. To do this, we assembled longitudinal cotton damage data that spanned the Bt adoption period, collected cotton damage data since Bt resistance has been detected, and estimated local population susceptibility using replicated on-farm studies that included all Bt pyramids marketed in cotton. RESULTS: Significant year effects of H. zea damage frequency in commercial cotton were observed throughout the Bt adoption period, with a recent damage increase after 2012. Landscape-level Bt crop production intensity over time was positively associated with the risk of H. zea damage in two- and three-toxin pyramided Bt cotton. Helicoverpa zea damage also varied across Bt toxin types in spatially replicated on-farm studies. CONCLUSIONS: Landscape-level predictors of H. zea damage in Bt cotton can be used to identify heightened Bt resistance risk areas and serves as a model to understand factors that drive pest resistance evolution to Bt toxins in the southeastern United States. These results provide a framework for more effective insect resistance management strategies to be used in combination with conventional pest management practices that improve Bt trait durability while minimizing the environmental footprint of row crop agriculture. © 2021 Society of Chemical Industry. This article has been contributed to by US Government employees and their work is in the public domain in the USA.

Computer vision for detecting field-evolved lepidopteran resistance to Bt maize.

BACKGROUND: Resistance evolution of lepidopteran pests to Bacillus thuringiensis (Bt) toxins produced in maize and cotton is a significant issue worldwide. Effective toxin stewardship requires reliable detection of field-evolved resistance to enable the implementation of mitigation strategies. Currently, visual estimates of maize injury are used to document changing susceptibility. In this study, we evaluated an existing maize injury monitoring protocol used to estimate Bt resistance levels in Helicoverpa zea (Lepidoptera: Noctuidae). RESULTS: We detected high interobserver variability across multiple injury metrics, suggesting that the precision and accuracy of maize injury detection could be improved. To do this, we developed a computer vision-based algorithm to measure H. zea injury. Algorithm estimates were more accurate and precise than a sample of human observers. Moreover, observer estimates tended to overpredict H. zea injury, which may increase the false-positive rate, leading to prophylactic insecticide application and unnecessary regulatory action. CONCLUSIONS: Automated detection and tracking of lepidopteran resistance evolution to Bt toxins are critical for genetically engineered crop stewardship to prevent the use of additional insecticides to combat resistant pests. Advantages of this computerized screening are: (i) standardized Bt injury metrics in space and time, (ii) preservation of digital data for cross-referencing when thresholds are reached, and (iii) the ability to increase sample sizes significantly. This technological solution represents a significant step toward improving confidence in resistance monitoring efforts among researchers, regulators and the agricultural biotechnology industry.

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.