Using targeted sequencing and TaqMan approaches to detect acaricide (bifenthrin, bifenazate, and etoxazole) resistance associated SNPs in Tetranychus urticae collected from peppermint fields and hop yards.

Tetranychus urticae (Koch) is an economically important pest of many agricultural commodities world-wide. Multiple acaricides, including bifenazate, bifenthrin, and extoxazole, are currently registered to control T. urticae. However, populations of T. urticae in many different growing regions have developed acaricide resistance through multiple mechanisms. Within T. urticae, single nucleotide polymorphisms (SNPs) have been documented in different genes which are associated with acaricide resistance phenotypes. The detection of these mutations through TaqMan qPCR has been suggested as a practical, quick, and reliable tool to inform agricultural producers of acaricide resistance phenotypes present within their fields and have potential utility for making appropriate acaricide application and integrated pest management decisions. Within this investigation we examined the use of a TaqMan qPCR-based approach to determine genotypes which have been previously associated with acaricide resistance in field-collected populations of T. urticae from peppermint fields and hop yards in the Pacific Northwest of the United States and confirmed the results with a multiplex targeted sequencing. The results suggest that a TaqMan qPCR approach accurately genotypes T. urticae populations for SNPs that have been linked to Bifenazate, Bifenthrin, and Etoxazole resistance. The results also demonstrated that different populations of mites in Washington and Idaho displayed varying frequencies of the examined SNPs. While we were able to detect the SNPs associated with the examined acaricides, the mutation G126S was not an appropriate or accurate indicator for bifenazate resistance.

Multiplex Polymerase Chain Reaction Reveals Unique Trends in Pathogen and Parasitoid Infestations of Alfalfa Leafcutting Brood Cells.

The alfalfa leafcutting bee Megachile rotundata Fabricius (HYMENOPTERA: Megachilidae) is an important pollinator for multiple agricultural seed commodities in the United States. M. rotundata is a solitary cavity nesting bee that forms brood nests where its larvae can develop. During the developmental stages of growth, brood can be preyed upon by multiple different fungal pathogens and insect predators and parasitoids, resulting in the loss of the developing larvae. Larval loss is a major concern for alfalfa (Medicago sativa L.) seed producers because they rely on pollination services provided by M. rotundata. Reduced pollination rates result in lower yields and increased production costs. In the present study, we examined the taxonomic composition of organisms found within M. rotundata brood cells using a multiplex PCR assay which was developed for the detection of bacterial, fungal, and invertebrate pests and pathogens of M. rotundata larvae. Known pests of M. rotundata were detected, including members of the fungal genus Ascosphaera, the causative agent of chalkbrood. The presence of multiple Ascosphaera species in a single brood cell was observed, with potential implications for chalkbrood disease management. The multiplex assay also identified DNA from more than 2,400 total species, including multiple predators and pathogenetic species not previously documented in association with M. rotundata brood cells.

Examining Historical Rates of Leafcutting Bee Brood Cell Pathogens, Parasitoids, and Predators to Establish Baseline Infectivity Rates for Alfalfa Seed Growers.

The alfalfa leafcutting bee Megachile rotundata (Fabricius) (Hymenoptera: Megachilidae) is the primary pollinator for the alfalfa seed industry. It is a solitary cavity nesting bee that utilizes leaf lined brood cells provisioned with pollen for larval development and pupation into the adult stage. During development, multiple pathogens, parasitoids, and predators can prey upon or use the larvae as a host, resulting in the loss of the future adult bee. As such, the presence of invertebrate pests and fungal pathogens presents a major concern for commercial alfalfa seed growers. In the present study, we used historic data from the Parma Cocoon Diagnostic Laboratory to determine baseline rates of pathogens, parasitoids, and predators of Megachile rotundata brood cells and used this analysis to determine cutoffs for management practices to inform growers when the purchase of new bee stocks should be considered. Additionally, we compared the presence of chalkbrood, predators, and parasitoids in samples collected from both grower-produced stocks and newly purchased Canadian bees. The results of the investigation provide historic averages of the presence of chalkbrood, predators, and parasitoids, show a significant increase in chalkbrood and predators in 2007-2011, and find a significant difference in rates of chalkbrood and predators between samples from Canadian suppliers and grower stocks. We speculate that these differences may have resulted from economic conditions that increased the cost of Canadian Megachile rotundata cells and likely resulted in increased reliance on 2nd-year U.S. grower stocks and subsequently increased infection rates during this time period.

Multiple insecticide resistance in onion thrips populations from Western USA.

Thrips tabaci is a key pest of onions, especially in the Pacific Northwestern USA. Management of T. tabaci is dominated by the application of various insecticides. However, T. tabaci is known to develop insecticide resistance which possibly leads to control failures, crop loss, and environmental concern. Here, we evaluated resistance status of T. tabaci populations from conventional and organic commercial onion fields to three widely used insecticides: oxamyl, methomyl, and abamectin with on-field concentration-mortality bioassays. The biochemistry and molecular mechanisms underlying resistance to these insecticides were also investigated by using enzymatic assays and detecting resistance-associated mutations. Field-evolved resistance to oxamyl, methomyl and abamectin were detected in most of the T. tabaci populations collected from conventional onion farms. At the labeled field rate, all the tested insecticides, particularly methomyl and oxamyl, had significantly reduced efficacy. Enzymatic assays of insecticide target and detoxification enzymes indicated that T. tabaci populations in Western USA onions harbor multiple mechanisms of resistance including enhanced activities of detoxification enzymes and target site insensitivity. Our results provide new information in understanding the dynamics of T. tabaci adaptation to multiple insecticides, which will help to design sustainable insecticide resistance management strategies for T. tabaci. Furthermore, this study provides the foundation for future research in identifying the biochemical and molecular markers associated with insecticide resistance in T. tabaci.

Physiological resistance alters behavioral response of Tetranychus urticae to acaricides.

Multiple acaricide resistance in Tetranychus urticae continues to threaten crop production globally, justifying the need to adequately study resistance for sustainable pest management. Most studies on acaricide resistance have focused on the acute contact toxicity of acaricides with little or no information on the behavioral responses elicited after acaricide exposure. Furthermore, the impact of physiological resistance on these behavioral responses remains unknown in most pest species, including T. urticae. We tested the effect of acaricide resistance on contact toxicity, irritancy and repellency of mitochondrial electron transport inhibitor of complex I (MET-I) and mite growth inhibitor (MGI) acaricides on multiple T. urticae strains. We also tested whether acaricides with similar physiological target site/mode of action also elicit similar behavioral effects on T. urticae strains. MET-I acaricides (fenazaquin, fenpyroximate, and pyrabiden) and MGIs (clofentezine, hexythiazox and etoxazole) elicited a dose-dependent irritant and repellent effect on T. urticae. Selection of strains for physiological resistance to these acaricides affected the behavioral response of T. urticae, especially in MET-I resistant strains, that showed reduced irritancy and repellency to MET-I acaricides. Behavioral response also affected the oviposition of T. urticae, where strains generally showed preferential oviposition away from the acaricides. The outcome of this study highlights negative consequences of acaricide resistance that can potentially affect T. urticae management.

Spatial Analysis of Seasonal Dynamics and Overwintering of Drosophila suzukii (Diptera: Drosophilidae) in the Okanagan-Columbia Basin, 2010-2014.

Spotted wing drosophila, Drosophila suzukii (Matsumura) (Diptera: Drosophilidae), was monitored from 2010 to 2014 in 314-828 sites located in interior fruit-growing regions of OR and WA, United States, and BC, Canada, using traps baited with apple cider vinegar or sugar-water-yeast. Seasonal population dynamics and sex ratios were summarized for berry, cherry, stone fruit, grape, non-crop host plants, non-host sites, and for conventional IPM, certified organic, backyard, and feral sites, by region and year. Overwintering was detected in all regions and years, despite winter temperatures below -17°C. A spatial analysis was conducted using a Geographic Information System (GIS), daily weather data, geomorphometric measures of terrain, distance to water, and other variables, at each site. Overwintering success at a site, measured as Julian week of first capture of D. suzukii, was significantly related (R2 = 0.49) in cherry habitats to year, agronomic treatment, and number of winter days with temperatures >-5°C. In berry, cherry, stone fruit and grape habitats, 2011-2014, it was significantly related (R2 = 0.42) to year, agronomic treatment, the logarithm of peak population of D. suzukii in the prior autumn, latitude, elevation, and topographic wetness index. The results show that D. suzukii has adapted to exploit a succession of irrigated crops and feral habitats in mixed landscapes of a semi-arid region with cold winters and hot dry summers, and are shaping strategies for pest management and for biological control.

Trap designs for monitoring Drosophila suzukii (Diptera: Drosophilidae).

Drosophila suzukii (Matsumura), an invasive pest of small and stone fruits, has been recently detected in 39 states of the United States, Canada, Mexico, and Europe. This pest attacks ripening fruit, causing economic losses including increased management costs and crop rejection. Ongoing research aims to improve the efficacy of monitoring traps. Studies were conducted to evaluate how physical trap features affect captures of D. suzukii. We evaluated five colors, two bait surface areas, and a top and side position for the fly entry point. Studies were conducted at 16 sites spanning seven states and provinces of North America and nine crop types. Apple cider vinegar was the standard bait in all trap types. In the overall analysis, yellow-colored traps caught significantly more flies than clear, white, and black traps; and red traps caught more than clear traps. Results by color may be influenced by crop type. Overall, the trap with a greater bait surface area caught slightly more D. suzukii than the trap with smaller area (90 vs. 40 cm(2)). Overall, the two traps with a side-mesh entry, with or without a protective rain tent, caught more D. suzukii than the trap with a top-mesh entry and tent.

Susceptibility of cranberries to Drosophila suzukii (Diptera: Drosophilidae).

Drosophila suzukii Matsumura (Diptera: Drosophilidae), commonly referred to as the spotted wing drosophila, is an exotic species that has proven a troublesome pest of fruit production in the United States. The fly targets small fruit and thus represents a concern for the U.S. cranberry industry. Two studies were conducted to assess whether cranberries may serve as hosts for D. suzukii. In the first study, the suitability of ripe, unripe, and over-ripe cranberries were assayed by examining adult oviposition and larval development in no-choice trials. In the second study, wounded and unwounded fruit were examined as potential hosts in choice and no-choice trials. Our first study showed that ripe, unripe, and over-ripe cranberries were unsuitable hosts (few eggs were laid, with no surviving puparia). In the wounded and unwounded berry study, no larvae survived to adulthood among unwounded berries. Within wounded fruit, D. suzukii readily fed and developed into adults. Together, these results suggest that unwounded cranberries--whether ripe, unripe, or over-ripe--are unsuitable as hosts for D. suzukii. Wounded rotting cranberries, however, can serve as hosts. Across the landscape, cranberry marshes with rotting fruit may contribute to D. suzukii source-sink dynamics.

Evaluation of monitoring traps for Drosophila suzukii (Diptera: Drosophilidae) in North America.

Drosophila suzukii Matsumura (Diptera: Drosophilidae), a recent invasive pest of small and stone fruits, has been detected in more than half of the U.S. states, and in Canada, Mexico, and Europe. Upon discovery, several different trap designs were recommended for monitoring. This study compared the trap designs across seven states/provinces in North America and nine crop types. Between May and November 2011, we compared a clear cup with 10 side holes (clear); a commercial trap with two side holes (commercial); a Rubbermaid container with mesh lid and rain tent (Haviland), and with 10 side holes and no tent (modified Haviland); a red cup with 10 side holes (red); and a white container with mesh lid and rain tent (Van Steenwyk). Although fly catches among traps varied per site, overall, the Haviland trap caught the most D. suzukii, followed by the red, Van Steenwyk, and clear trap. The modified Haviland and commercial trap had low captures. Among five crop types in Oregon, a clear cup with mesh sides (Dreves) also was tested and caught the most flies. Traps with greater entry areas, found in mesh traps, caught more flies than traps with smaller entry areas. In terms of sensitivity and selectivity, traps that caught more flies likewise caught flies earlier, and all traps caught 26-31% D. suzukii out of the total Drosophila captured. Future trap improvements should incorporate more entry points and focus on selective baits to improve efficiency and selectivity with regard to the seasonal behavior of D. suzukii.

Interactions between population density of the Colorado potato beetle, Leptinotarsa decemlineata, and herbicide rate for suppression of solanaceous weeds.

The presence of volunteer potato Solanum tuberosum L., cutleaf nightshade, S. triflorum N., and hairy nightshade, S. physalifolium Rusby (Solanales: Solanaceae), throughout potato crop rotations can diminish the effectiveness of crop rotations designed to control disease and pest problems associated with growing potatoes. In greenhouse bioassays, larvae of the Colorado potato beetle, Leptinotarsa decemlineata Say (Coleoptera: Chrysomelidae) were placed in population densities of 0, 5, 10, and 40 per potato (cv. Russet Burbank) plant and 0, 5, 10, and 15 per cutleaf nightshade and hairy nightshade plant. Plants were treated with different rates of herbicides including fluroxypyr, prometryn, and mesotrione rates, and the physiological response on the potato plants was assessed by weighing shoot biomass 14 days after treatment. Consistently, across all bioassays, rate response functions were shifted as L. decemlineata density increased, such that less herbicide was required to achieve control. For instance, the herbicide rate needed to achieve 90% reduction in potato biomass was reduced from 62 to 0 g fluroxypyr per hectare and 711 to 0 g prometryn per hectare as L. decemlineata density was increased to 40 larvae per plant. Herbivory at higher L. decemlineata population densities and herbicides above certain rates resulted in large reductions in cutleaf and hairy nightshade biomass. Differences in rate response functions among L. decemlineata population densities indicated that L. decemlineata contributed to weed suppression in combination with herbicides. These data suggest that integrated weed management systems targeting volunteer potato, cutleaf nightshade, and hairy nightshade can be more effective when herbicide applications are combined with herbivory by naturally occurring Colorado potato beetles.