Microarray and Functional Pathway Analyses Revealed Significantly Elevated Gene Expressions Associated with Metabolic Resistance to Oxamyl (Vydate) in Lygus lineolaris.

The tarnished plant bug (TPB, Lygus lineolaris) remains a major pest for a variety of crops. Frequent sprays on row crops, especially cotton, prompted resistance development in field populations. To maintain chemical control as an effective tool against the pest, knowledge of global gene regulations is desirable for better understanding and managing the resistance. Novel microarray expressions of 6688 genes showed 685 significantly upregulated and 1382 significantly downregulated genes in oxamyl-selected TPBs (Vyd1515FF[R]) from a cotton field. Among the 685 upregulated genes (participated in 470 pathways), 176 genes code 30 different enzymes, and 7 of the 30 participate in 24 metabolic pathways. Six important detoxification pathways were controlled by 20 genes, coding 11 esterases, two P450s, two oxidases, and three pathway-associated enzymes (synthases, reductase, and dehydrogenase). Functional analyses showed substantially enhanced biological processes and molecular functions, with hydrolase activity as the most upregulated molecular function (controlled by 166 genes). Eleven esterases belong to the acting on ester bond subclass of the 166 hydrolases. Surprisingly, only one GST showed significant upregulation, but it was not involved in any detoxification pathway. Therefore, this research reports a set of 20 genes coding 6 enzyme classes to detoxify a carbamate insecticide oxamyl in Vyd1515FF. Together with three previous reports, we have obtained the best knowledge of resistance mechanisms to all four conventional insecticide classes in the economically important crop pest. This valuable finding will greatly facilitate the development of molecular tools to monitor and manage the resistance and to minimize risk to environment.

Early Season Monitoring of Tarnished Plant Bug, Lygus lineolaris, in Wild Hosts Using Pheromone Traps.

The tarnished plant bug, Lygus lineolaris (Hemiptera: Miridae), has a wide host range of over 700 plant species, including 130 crops of economic importance. During early spring, managing the field edges with weeds and other wild hosts is important in preventing early-season infestations of L. lineolaris in cotton to prevent damage to the squares and other fruiting structures. Scouting fields for L. lineolaris is time- and labor-intensive, and end-user variability associated with field sampling can lead to inaccuracies. Insect traps that combine visual cues and pheromones are more accurate, sustainable, and economically feasible in contrast to traditional insect detection methods. In this study, we investigated the application of red or white sticky cards baited with the female-produced sex pheromone to monitor overwintering L. lineolaris populations in early spring. Field experiments demonstrated that the red sticky cards baited with a pheromone blend containing hexyl butyrate, (E)-2-hexenyl butyrate, and (E)-4-oxo-2-hexenal in 4:10:7 ratio are highly effective in trapping L. lineolaris adults in early spring before the row crops are planted, and in monitoring their movement into a cotton crop. The monitoring of L. lineolaris should help growers to make judicious decisions on insecticide applications to control early pest infestations, thereby reducing economic damage to cotton.

Resistance risk assessment of six pyrethroids and acephate toward the resistant adult tarnished plant bug, Lygus lineolaris.

Due to rapidly developed resistance, pest management relies less on pyrethroids to control economically damaging infestations of the tarnished plant bug (TPB), Lygus lineolaris (Palisot de Beauvois) in cotton fields of Mississippi. Yet, pyrethroid resistance remains prevalent in TPB populations. This study assessed the resistance levels in adult TPB to six common pyrethroids and acephate. Resistant TBPs were collected from wild host plants in late October after harvest in the Mississippi Delta region of the United States. Based on LC50 values, the field-resistant TPBs displayed higher resistance to permethrin, esfenvalerate, and bifenthrin (approximately 30 fold) and moderate resistance to λ-cyhalothrin, ß-cyfluthrin, ζ-cypermethrin, and acephate (approximately 15 fold). Further investigations showed that the inhibitors of three detoxification enzyme, triphenyl phosphate (TPP), diethyl maleate (DEM), and piperonyl butoxide (PBO) had synergistic effects on permethrin, λ-cyhalothrin, and bifenthrin in resistant TPBs. Furthermore, elevated esterase, GST, and P450 activities were significantly expressed in field-resistant TPBs. Additionally, GST and esterase were reduced after 48 h exposure to certain pyrethroids at LC50 dose. The synergistic and biochemical assays consistently indicated that P450 and esterase were involved in pyrethroid detoxification in TPBs. This study provides valuable information for the continued use of pyrethroids and acephate in controlling TPBs in cotton fields in the Mississippi Delta region of the United States.

A chromosome scale assembly of the tarnished plant bug, Lygus lineolaris (Palisot de Beauvois), genome.

OBJECTIVE: The tarnished plant bug (TPB), Lygus lineolaris (Palisot de Beauvois) (Hemiptera: Miridae), is a pest damaging many cultivated crops in North America. Although partial transcriptome data are available for this pest, a genome assembly was not available for this species. This assembly of a high-quality chromosome-length genome of TPB is aimed to develop the genetic resources that can provide the foundation required for advancing research on this species. RESULTS: The initial genome of TPB assembled with paired-end nucleotide sequences generated with Illumina technology was scaffolded with Illumina HiseqX reads generated from a proximity ligated (HiC) library to obtain a high-quality genome assembly. The final assembly contained 3963 scaffolds longer than 1 kbp to yield a genome of 599.96 Mbp. The N50 of the TPB genome assembly was 35.64 Mbp and 98.68% of the genome was assembled into 17 scaffolds larger than 1 Mbp. This megabase scaffold number is the same as the number of chromosomes observed in karyotyping of this insect. The TPB genome is known to have high repetitive DNA content, and the reduced assembled genome size compared to flowcytometric estimates of approximately 860 Mbp may be due to the collapsed assembly of highly similar regions.

The mechanisms of metabolic resistance to pyrethroids and neonicotinoids fade away without selection pressure in the tarnished plant bug Lygus lineolaris.

BACKGROUND: Heavy selection pressure prompted the development of resistance in a serious cotton pest tarnished plant bug (TPB), Lygus Lineolaris in the mid-southern United States. Conversely, a laboratory resistant TPB strain lost its resistance to five pyrethroids and two neonicotinoids after 36 generations without exposure to any insecticide. It is worthwhile to examine why the resistance diminished in this population and determine whether the resistance fade away has practical value for insecticide resistance management in TPB populations. RESULTS: A field-collected resistant TPB population in July (Field-R1) exhibited 3.90-14.37-fold resistance to five pyrethroids and two neonicotinoids, while another field-collected TPB population in April (Field-R2) showed much lower levels of resistance (0.84-3.78-fold) due to the absence of selection pressure. Interestingly, after 36 generations without exposure to insecticide, the resistance levels in the same population [laboratory resistant strain (Lab-R)] significantly decreased to 0.80-2.09-fold. The use of detoxification enzyme inhibitors had synergistic effects on permethrin, bifenthrin and imidacloprid in resistant populations of Lygus lineolaris. The synergism was more pronounced in Field-R2 than laboratory susceptible (Lab-S) and Lab-R TPB population. Moreover, esterase, glutathione S-transferase (GST), and cytochrome P450-monooxygenases (P450) enzyme activities increased significantly by approximately 1.92-, 1.43-, and 1.44-fold in Field-R1, respectively, and 1.38-fold increased P450 enzyme activities in Field-R2 TPB population, compared to the Lab-S TPB. In contrast, the three enzyme activities in the Lab-R strain were not significantly elevated anymore relative to the Lab-S population. Additionally, Field-R1 TPB showed elevated expression levels of certain esterase, GST and P450 genes, respectively, while Field-R2 TPB overexpressed only P450 genes. The elevation of these gene expression levels in Lab-R expectedly diminished to levels close to those of the Lab-S TPB populations. CONCLUSION: Our results indicated that the major mechanism of resistance in TPB populations was metabolic detoxification, and the resistance development was likely conferred by increased gene expressions of esterase, GST, and P450 genes, the fadeaway of the resistance may be caused by reversing the overexpression of esterase, GST and P450. Without pesticide selection, resistant gene (esterase, GST, P450s) frequencies declined, and detoxification enzyme activities returned to Lab-S level, which resulted in the recovery of the susceptibility in the resistant TPB populations. Therefore, pest's self-purging of insecticide resistance becomes strategically desirable for managing resistance in pest populations. Published 2023. This article is a U.S. Government work and is in the public domain in the USA.

Population Dynamics of Insect Pests and Beneficials on Different Snap Bean Cultivars.

Snap bean is an important crop in the United States. Insecticides are commonly used against pests on snap bean, but many pests have developed resistance to the insecticides and beneficials are threatened by the insecticides. Therefore, host plant resistance is a sustainable alternative. Population dynamics of insect pests and beneficials were assessed on 24 snap bean cultivars every week for six weeks. The lowest number of sweetpotato whitefly (Bemisia tabaci) eggs was observed on cultivar 'Jade', and the fewest nymphs were found on cultivars 'Gold Mine', 'Golden Rod', 'Long Tendergreen', and 'Royal Burgundy'. The numbers of potato leafhopper (Empoasca fabae) and tarnished plant bug (Lygus lineolaris) adults were the lowest on cultivars 'Greencrop' and 'PV-857'. The highest numbers of adults were found in Week 1 (25 days following plant emergence) for B. tabaci and Mexican bean beetle (Epilachna varivestis); Week 3 for cucumber beetle, kudzu bug (Megacopta cribraria), and E. fabae; Weeks 3 and 4 for thrips; Week 4 for L. lineolaris; and Weeks 5 and 6 for bees. Temperature and relative humidity correlated with B. tabaci, E. varivestis, bee, and predator ladybird beetle populations. These results provide valuable information on the integrated pest management of snap beans.

Mining Lygus hesperus (western tarnished plant bug) transcriptomic data for transient receptor potential channels: Expression profiling and functional characterization of a Painless homolog.

The transient receptor potential (TRP) family of cation channels are evolutionarily conserved proteins with critical roles in sensory physiology. Despite extensive studies in model species, knowledge of TRP channel functional diversity and physiological impact remains limited in many non-model insect species. To assess the TRP channel repertoire in a non-model agriculture pest species (Lygus hesperus), publicly available transcriptomic datasets were mined for potential homologs. Among the transcripts identified, 30 are predicted to encompass complete open reading frames that encode proteins representing each of the seven TRP channel subfamilies. Although no homologs were identified for the Pyrexia and Brivido channels, the TRP complement in L. hesperus exceeded the 13-16 channels reported in most insects. This diversity appears to be driven by a combination of alternative splicing, which impacted members of six subfamilies, and gene expansion of the TRPP subfamily. To validate the in silico data and provide more detailed analyses of L. hesperus TRP functionality, the putative Painless homolog was selected for more in depth analysis and its functional role in thermosensation examined in vitro. RT-PCR expression profiling revealed near ubiquitous expression of the Painless transcript throughout nymphal and adult development. Electrophysiological data generated using a Xenopus oocyte recombinant expression system indicated activation parameters for L. hesperus Painless homolog that are consistent with a role in noxious heat (40°-45 °C) thermosensation.

A Dynamic Threshold Approach for Tarnished Plant Bug (Hemiptera: Miridae) Management in the Midsouthern U.S. Cotton.

One of the most economically important pests of cotton, Gossypium hirsutum L., in the midsouth region of the United States is the tarnished plant bug, Lygus lineolaris (Palisot de Beauvois, Hemiptera: Miridae). Tarnished plant bug populations across the region have exhibited widespread resistance to numerous insecticide classes. To minimize late season resistance development, reducing unwarranted applications during the late flowering period can aid in resistance management and potentially reduce input costs. Trials were conducted during 2019 and 2020 to evaluate the impacts of tarnished plant bug populations in the later flowering period of cotton by modifying or terminating threshold regimes during the later weeks of bloom. Results showed that dynamic thresholds altered at the fourth week of bloom or later can reduce the number of late season applications made with no penalty to yield. Additionally, when utilizing a week of bloom termination approach, no significant yield losses were seen when terminating applications after the fourth week of bloom. These data may offer an alternative method to managing tarnished plant bug populations during the later flowering period of midsouth cotton.

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.

Tarnished Plant Bug (Heteroptera: Miridae) Behavioral Responses to Chemical Insecticides.

The tarnished plant bug (Lygus lineolaris Palisot de Beauvois) is the dominant insect pest of cotton (Gossypium hirsutum L.) in the Mid-South Cotton Belt. This is partly due to the fact that this pest has developed resistance to most insecticides used for control. Laboratory experiments were conducted during 2014 and 2015 to study the behavioral response of tarnished plant bug nymphs to several classes of insecticides. Twenty third-instar nymphs were placed in individual dishes divided into four quadrants with five green bean pieces in each quadrant (10 treated and 10 untreated green beans in each dish). Dishes were checked at 1, 4, 8, and 24 h. Tarnished plant bug nymphs appeared to avoid green beans treated with IGR, pyrethroid, organophosphate, or carbamate insecticides, while there appeared to be an attraction to green bean pieces treated with sulfoxamine and pyridine carboxamide insecticides. No relationship was observed with neonicotinoid insecticides within 24 h.

Biology, Ecology, and Pest Management of the Tarnished Plant Bug, Lygus lineolaris (Palisot de Beauvois) in Southern Row Crops.

The tarnished plant bug, Lygus lineolaris (Palisot de Beauvois), (Hemiptera: Miridae) is considered the most damaging pest of cotton (Gossypium hirsutum L.) in the mid-southern United States, although it is established throughout the United States, southern Canada, and northern Mexico. The introduction of transgenic crops for the control of moths in the Heliothine complex and eradication of the boll weevil, Anthonomus grandis, from much of the United States led to greatly reduced pesticide use in cotton fields, which allowed L. lineolaris to emerge as a new primary pest of cotton in the mid-southern United States. Since the publication of a review by Layton (2000) on damage caused by Lygus lineolaris, many new studies have been published on the changes in host range, population dynamics, sampling methods and thresholds, cultural practices, sex pheromones and attractant blends, novel pesticides and insecticide resistance mechanisms, olfactory and feeding behaviors, introduction of biological control agents, host-plant resistance mechanisms, and new molecular and genetic tools for integrated pest management of Lygus species in cotton and other important crops. Here, we review and discuss the latest developments in L. lineolaris research in the last two decades.

Expression Profiles of Digestive Genes in the Gut and Salivary Glands of Tarnished Plant Bug (Hemiptera: Miridae).

Host plant preference of agricultural pests may shift throughout the growing season, allowing the pests to persist on wild hosts when crops are not available. Lygus Hahn (Hemiptera: Miridae) bugs are severe pests of cotton during flowering and fruiting stages, but can persist on alternative crops, or on weed species. Diversity of digestive enzymes produced by salivary glands and gut tissues play a pivotal role in an organism's ability to utilize various food sources. Polyphagous insects produce an array of enzymes that can process carbohydrates, lipids, and proteins. In this study, the digestive enzyme repertoire of the tarnished plant bug, Lygus lineolaris (Palisot de Beauvois), was identified by high-throughput sequencing followed by cDNA cloning and sequencing. This study identified 87 digestive genes, including 30 polygalacturonases (PG), one ß-galactosidase, three α-glucosidases, six ß-glucosidases, 28 trypsin-like proteases, three serine proteases, one apyrase-like protease, one cysteine protease, 12 lipases, and two transcripts with low similarity to a xylanase A-like genes. RNA-Seq expression profiles of these digestive genes in adult tarnished plant bugs revealed that 57 and 12 genes were differentially expressed in the salivary gland and gut (≥5-fold, P ≤ 0.01), respectively. All polygalacturonase genes, most proteases, and two xylanase-like genes were differentially expressed in salivary glands, while most of the carbohydrate and lipid processing enzymes were differentially expressed in the gut. Seven of the proteases (KF208689, KF208697, KF208698, KF208699, KF208700, KF208701, and KF208702) were not detected in either the gut or salivary glands.

Filling in the gaps: A reevaluation of the Lygus hesperus peptidome using an expanded de novo assembled transcriptome and molecular cloning.

Peptides are the largest and most diverse class of molecules modulating physiology and behavior. Previously, we predicted a peptidome for the western tarnished plant bug, Lygus hesperus, using transcriptomic data produced from whole individuals. A potential limitation of that analysis was the masking of underrepresented genes, in particular tissue-specific transcripts. Here, we reassessed the L. hesperus peptidome using a more comprehensive dataset comprised of the previous transcriptomic data as well as tissue-specific reads produced from heads and accessory glands. This augmented assembly significantly improves coverage depth providing confirmatory transcripts for essentially all of the previously identified families and new transcripts encoding a number of new peptide precursors corresponding to 14 peptide families. Several families not targeted in our initial study were identified in the expanded assembly, including agatoxin-like peptide, CNMamide, neuropeptide-like precursor 1, and periviscerokinin. To increase confidence in the in silico data, open reading frames of a subset of the newly identified transcripts were amplified using RT-PCR and sequence validated. Further PCR-based profiling of the putative L. hesperus agatoxin-like peptide precursor revealed evidence of alternative splicing with near ubiquitous expression across L. hesperus development, suggesting the peptide serves functional roles beyond that of a toxin. The peptides predicted here, in combination with those identified in our earlier study, expand the L. hesperus peptidome to 42 family members and provide an improved platform for initiating molecular and physiological investigations into peptidergic functionality in this non-model agricultural pest.

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.

A Molecular Approach for Detecting Stage-Specific Predation on Lygus hesperus (Hemiptera: Miridae).

A molecular gut analysis technique is described to identify predators of Lygus hesperus (Knight), a significant pest of many crops. The technique is unique because it can pinpoint which life stage of the pest was consumed. Sentinel egg masses designed to mimic the endophytic egg-laying behavior of L. hesperus were marked with rabbit serum, while third instar and adult L. hesperus were marked with chicken and rat sera, respectively. Then, the variously labeled L. hesperus life stages were introduced into field cages that enclosed the native arthropod population inhabiting an individual cotton plant. After a 6-h exposure period, the predator assemblage, including the introduced and native L. hesperus population, in each cage were counted and had their gut contents examined for the presence of the variously marked L. hesperus life stages by a suite of serum-specific enzyme-linked immunosorbent assays (ELISA). The whole-plant sampling scheme revealed that Geocoris punticpes (Say) and Geocoris pallens Stal (Hemiptera: Geocoridae) and members of the spider complex were the numerically dominant predator taxa in the cotton field. The gut content analyses also showed that these two taxa appeared to be the most prolific predators of the L. hesperus nymph stage. Other key findings include that Collops vittatus (Say) (Coleoptera: Melyridae) and Solenopsis xyloni McCook (Hymenoptera: Formicidae) appear to be adept at finding and feeding on the cryptic L. hesperus egg stage, and that L. hesperus, albeit at low frequencies, engaged in cannibalism. The methods described here could be adapted for studying life stage-specific feeding preferences for a wide variety of arthropod taxa.

A Laboratory Diet-Overlay Bioassay to Monitor Resistance in Lygus lineolaris (Hemiptera: Miridae) to Insecticides Commonly Used in the Mississippi Delta.

A laboratory, diet-overlay pesticide bioassay was developed using a susceptible population of the tarnished plant bug, Lygus lineolaris (Palisot de Beauvois), to study its susceptibility to neonicotinoid, sulfoxamine, organophosphate, and pyrethroid insecticides (thiamethoxam, sulfoxaflor, acephate, and permethrin, respectively). The diet-overlay bioassay was compared to the traditional glass-vial surface residue bioassay. We measured LC50 values by feeding tarnished plant bug adults known doses of insecticides dispensed on top of diet in a 10% solution of honey water for thiamethoxam and 10% acetone in water solutions for permethrin, acephate, and sulfoxaflor. Both the diet-overlay and glass-vial bioassays used dose-response (mortality) regression lines to calculate LC50 values for each insecticide at 6-, 24-, 48-, and 72-h post-exposure. Data variability from the glass-vial bioassay was higher for permethrin, sulfoxaflor, and thiamethoxam than the diet-overlay bioassay, for all evaluation times. In contrast, there was lower variability among replicates to acephate in the glass-vial assay compared to the diet-overlay assay. Control mortalities observed on diet-overlay bioassay were lower (0-5%) than those observed on the glass-vial bioassay (4-27%). The use of green beans, floral-foam, rolling glass vials, and insect handling made the existing standard method tedious to manipulate and difficult to handle large numbers of individuals. The nonautoclaved solid diet provides an opportunity to significantly reduce cost and variability associated with procedures of other bioassay methods. In general, the baseline data provide a basis for future comparison to determine changes in resistance over time.

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.

A Novel CO2-Based Insect Sampling Device and Associated Field Method Evaluated in a Strawberry Agroecosystem.

There is high demand for accurate insect sampling methods to inform integrated pest management strategies. Despite widespread application, existing sampling methods, such as portable aspirating and sweep netting, can result in overrepresentation of prominent pests, underrepresentation of natural enemies, and damage to plants. In this study, we test a novel device for insect sampling via anesthetization. Specifically, we test the effect of CO2 (application pressure and duration of exposure) on Lygus hesperus Knight (Hemiptera: Miridae) anesthetization in the laboratory and on insect community density in a strawberry agroecosystem. Carbon dioxide application proves an effective means of anesthetization compared to negative controls, and an increase in net CO2 exposure results in a decrease in time until L. hesperus anesthetization. Field results indicate the CO2 method collects more parasitoids and thrips than a portable aspirator, and at the 50 PSI application pressure and 15-s exposure, the CO2 method results in a comparable number of pests collected as the research standard, a portable aspirator with 8-s aspiration time. Benefits of the CO2 method include minimal plant damage, highly explicit spatial and temporal data, and scalability.

A Gut Analysis Technique for Pinpointing Egg-Specific Predation Events.

A universal food immunomarking technique (UFIT) is described for postmortem gut analysis detection of predation on the egg stage of Lygus hesperus Knight (Hemiptera: Miridae). Collops vittatus Say (Coleoptera: Melyridae) and Hippodamia convergens Guérin-Méneville (Coleoptera: Coccinellidae) were fed a single L. hesperus egg that was marked with rabbit and chicken sera proteins. The protein-marked egg remnants were detectable in the guts of the majority of the predators by each sera-specific enzyme-linked immunosorbent assay (ELISA) for 3 to 6 h after a feeding event. A novel technique was then developed to expose protein-marked eggs to predators that simulated the L. hesperus endophytic oviposition behavior. The procedure entailed embedding L. hesperus eggs in an artificial substrate that mimicked the stem of a plant. A predator feeding choice study was then conducted in cages that contained a cotton plant and artificial stems containing endophytic (concealed) and exophytic (exposed) egg patches. The endophytic and exophytic egg treatments were marked with chicken and rabbit protein, respectively. The gut analyses revealed that higher proportions of both predator populations contained remnants of the exophytic egg treatment and L. hesperus eggs were more vulnerable to C. vittatus than H. convergens. This study shows how the UFIT can be used to pinpoint stage-specific feeding activity on two distinct egg exposure treatments (endophytic and exophytic) of the same species.