Landscape-scale spatiotemporal dynamics of Halyomorpha halys (Stål) (Hemiptera: Pentatomidae) populations: implications for spatially-based pest management.

BACKGROUND: Brown marmorated stink bug (BMSB), Halyomorpha halys (Stål), is an invasive and severe pest of specialty and row crops. A 2-year field study conducted in four Mid-Atlantic states in the USA characterized the spatial and temporal dynamics of BMSB populations and its association with landscape elements in commercial agriculture settings. In each state, two 1 km2 sites included typical landscape elements (i.e., tree fruit orchards, annual field and vegetable crops, woodlands, and human-made structures). Twenty-seven georeferenced pheromone traps were deployed per site and the number of BMSB adults and nymphs captured was counted throughout the growing season. RESULTS: Findings from spatial analysis by distance indices, along with time-series maps of BMSB distribution, showed that BMSB exhibited significant spatial aggregation, and that its distribution was spatially consistent between years. Analyses with geographic information systems (GIS) revealed that BMSB 'hot spots' occurred in different landscape elements throughout each season. Most patches (i.e., clusters of significantly higher trap captures) were found near woodlands early in the season, near tree fruit orchards in summer, and on the border of annual field crops in autumn. Buffer analysis with GIS indicated that more BMSB adults were captured closer to woodlands compared with other landscape elements. CONCLUSION: Understanding the spatial and temporal movement and distribution of BMSB is critical to predicting their potential impact and ultimately devising strategies to mitigate this risk to vulnerable crops. The results of this study can be used to design streamlined, spatially-based areawide management of BMSB in heterogeneous and complex agricultural landscapes. © 2023 Society of Chemical Industry. This article has been contributed to by U.S. Government employees and their work is in the public domain in the USA.

Augmentative biological control for squash bug (Hemiptera: Coreidae) using the egg parasitoid, Hadronotus pennsylvanicus (Hymenoptera: Scelionidae).

The squash bug, Anasa tristis (De Geer) (Hemiptera: Coreidae), is a serious pest of cucurbit crops across the United States. Conventional growers commonly use broad-spectrum insecticides to manage squash bugs, however organic growers lack these effective chemical tools and must rely on alternative management strategies. Biological control of A. tristis is largely understudied, specifically the potential of natural enemy, Hadronotus pennsylvanicus (Ashmead) (Hymenoptera: Scelionidae), as an augmentative biological control agent. For this reason, we performed early-season field releases of H. pennsylvanicus on organic farms in southeastern Virginia to test if this would improve A. tristis egg parasitism. We chose organic vegetable farms growing summer squash (Cucurbita pepo L.) as release sites and nearby Virginia Tech Agricultural Research Extension Centers (AREC) as no-release sites. Parasitoids were reared in the lab and deployed as parasitized egg masses (~2-3 females wasps/plant) in June 2020 and 2021. Before parasitoid deployment, host eggs collected from release and no-release sites displayed low levels of H. pennsylvanicus parasitism in 2020 (<21%) and 2021 (<8%). In both years, the percentage of A. tristis eggs parasitized within 2 weeks post deployment was significantly greater at release sites (~60%) than at no-release sites (~14%). High rates of H. pennsylvanicus parasitism (>72%) were further observed at release sites 4, 6, 8, and 10 weeks following parasitoid deployment. Our study demonstrates that releases of lab-reared H. pennsylvanicus can increase A. tristis egg parasitism rates and subsequently decrease successful nymph hatch rates in early summer squash plantings.

Effect of dose and trap type on captures of striped and spotted cucumber beetles using synthetic vittatalactone.

Striped cucumber beetle, Acalymma vittatum (F.) and spotted cucumber beetle (Diabrotica undecimpunctata howardi Barber) are key pests of cucurbits. Pesticide applications directed against cucumber beetles can jeopardize bee pollination which is essential to crop yield. Alternative selective tactics, including behavioral control for managing cucumber beetles, are needed in order to improve pest management. The striped cucumber beetle pheromone vittatalactone is attractive to both male and female adults, and also to spotted cucumber beetles. We tested the response of both species to different doses of synthetic vittatalactone, and to different trap types, in field trapping experiments in Maryland. Both species showed strong dose-response in May-June and October 2020. Among 12 trap types tested during a 2-wk experiment in June, using 1 mg mixed vittatalactone, sticky panel traps caught the most of each species, with vittatalactone-baited traps exceeding unbaited traps by >8 fold for striped and >2 fold for spotted, and with significant response to yellow versus clear color absent in striped, but strong in spotted. Among nonsticky traps, those with yellow or yellow-green exceeded those without these colors for both beetle species, and a ground-placed boll weevil trap captured the most striped cucumber beetles. Bucket-style traps caught excessive nontarget bumblebees (Bombus spp.) if yellow, and a few beetles, if all green. Results allow field monitoring with a choice of sticky or nonsticky traps and suggested lure loading of 1 mg mixed vittatalactone, containing ~90 µg active isomer. These findings contribute to the development of multispecies behavioral monitoring and control as a component of cucurbit IPM.

Vittatalactone is the Male-Produced Aggregation Pheromone of the Western Striped Cucumber Beetle, Acalymma trivittatum.

We found that vittatalactone, specifically (3R,4R)-3-methyl-4-[(1S,3S,5S)-1,3,5,7-tetramethyloctyl]oxetan-2-one, is the male-produced aggregation pheromone of the western striped cucumber beetle, Acalymma trivittatum (Mannerheim), as was previously shown for the striped cucumber beetle, Acalymma vittatum (F.) (Coleoptera: Chrysomelidae). A synthetic mixture containing 9% of the authentic natural pheromone, is attractive to both sexes of both species in the field, as demonstrated by trapping using baited and unbaited sticky panels in California and earlier in Maryland. Females of both species do not produce detectable vittatalactone. This finding expands the usefulness of the synthetic vittatalactone mixture for pest management throughout the range of both A. vittatum and A. trivittatum. Development of vittatalactone time-release formulations combined with cucurbitacin feeding stimulants offer the potential for selective and environmentally-friendly cucurbit pest management tactics.

Ancient origin and conserved gene function in terpene pheromone and defense evolution of stink bugs and hemipteran insects.

Insects use diverse arrays of small molecules such as metabolites of the large class of terpenes for intra- and inter-specific communication and defense. These molecules are synthesized by specialized metabolic pathways; however, the origin of enzymes involved in terpene biosynthesis and their evolution in insect genomes is still poorly understood. We addressed this question by investigating the evolution of isoprenyl diphosphate synthase (IDS)-like genes with terpene synthase (TPS) function in the family of stink bugs (Pentatomidae) within the large order of piercing-sucking Hemipteran insects. Stink bugs include species of global pest status, many of which emit structurally related 15-carbon sesquiterpenes as sex or aggregation pheromones. We provide evidence for the emergence of IDS-type TPS enzymes at the onset of pentatomid evolution over 100 million years ago, coinciding with the evolution of flowering plants. Stink bugs of different geographical origin maintain small IDS-type families with genes of conserved TPS function, which stands in contrast to the diversification of TPS genes in plants. Expanded gene mining and phylogenetic analysis in other hemipteran insects further provides evidence for an ancient emergence of IDS-like genes under presumed selection for terpene-mediated chemical interactions, and this process occurred independently from a similar evolution of IDS-type TPS genes in beetles. Our findings further suggest differences in TPS diversification in insects and plants in conjunction with different modes of gene functionalization in chemical interactions.

Annual and Seasonal Changes in Parasitism Rates by Hadronotus Pennsylvanicus (Hymenoptera: Scelionidae) on the Squash Bug Anasa Tristis (Hemiptera: Coreidae) in Squash Fields: Implications for Augmentative Releases.

This study evaluated parasitism rates by Hadronotus pennsylvanicus (Ashmead) (Hymenoptera: Scelionidae) on the squash bug Anasa tristis DeGeer (Hemiptera: Coreidae) over a six-year period in squash fields in Maryland. From 2016-2021, 2226 wild squash bug egg masses were collected, 2180 (98.0%) A. tristis egg masses and 46 (2.0%) A. armigera egg masses. The mean (±SE) parasitism rate was 10.9 ± 0.16%. Yearly parasitism rates were significantly different with rates in 2017 and 2018 that were significantly lower than in 2019, 2020, and 2021. The significant difference in parasitism rates based on planting date was primarily due to the high parasitism rate observed in 2021. These results suggest that the use of augmentative releases early in the season could result in effective control by increasing parasitism earlier in the season and by causing the parasitism rate in the field to peak at a higher number late in the season.

Both the Squash Bug Anasa tristis and Horned Squash Bug Anasa armigera (Hemiptera: Coreidae) are Attracted to Vittatalactone, the Aggregation Pheromone of Striped Cucumber Beetle.

Vittatalactone, the aggregation pheromone of the striped cucumber beetle, Acalymma vittatum (F.) (Coleoptera: Chrysomelidae), is attractive to two species of squash bugs (Hemiptera: Coreidae), the squash bug Anasa tristis (DeGeer) and horned squash bug Anasa armigera (Say). In field trapping experiments in Maryland and Virginia, clear sticky traps baited with 1 mg of a synthetic 8-isomer mix of vittatalactone captured ~9× more of female A. tristis and of both sexes of A. armigera, whereas male A. tristis were not significantly attracted, compared to unbaited traps. A. armigera showed a distinct dose-response to vittatalactone lure loading in the late season, and this species was more attracted than A. tristis, based on comparison to captures from underneath wooden boards emplaced in adjacent fields. Results suggest that vittatalactone could be a 'keystone semiochemical' in colonization of cucurbit hosts by specialist herbivores, and may offer the opportunity for multi-species behavioral control as a component of integrated pest management in cucurbit crops.

Trapping of Crucifer-Feeding Flea Beetles (Phyllotreta spp.) (Coleoptera: Chrysomelidae) With Pheromones and Plant Kairomones.

Flea beetles (Coleoptera: Chrysomelidae) of the genus Phyllotreta are major pests of cole crops, canola, and related crops in the mustard family (Brassicaceae). Adults may damage seedlings or larger crop plants, impairing crop growth, rendering crops unmarketable, or killing seedlings outright. The two major North American crucifer pest species, Phyllotreta striolata (F.) and Phyllotreta cruciferae (Goeze), have male-produced pheromones attractive to both female and male adults. We tested the racemic synthetic pheromones, himachaladiene and hydroxyhimachalanone, as well as the host-plant-produced allyl isothiocyanate, alone and in combination, with experimental trapping in Maryland, Virginia, and North Dakota, using clear and yellow sticky traps and the ground-based 'rocket' trap (modified from boll weevil trap). Phyllotreta striolata was consistently attracted to the hydroxyketone, and captures were often enhanced by allyl isothiocyanate (AITC), but its response to pheromones, AITC, and trap color were variable from state to state. Phyllotreta cruciferae was strongly attracted to AITC, but its response to pheromone components varied by state, and this species was found rarely at the Maryland site. Phyllotreta bipustulata (F.) was attracted to the diene component, a new finding for this species. Several other genera of flea beetles were captured, some showing response to the semiochemicals and/or color. Results will be helpful in monitoring and possibly population suppression; however, further research is necessary to develop more efficient syntheses, optimal lure loadings, combinations, and controlled release methods.

Host Finding Behavior of the Parasitoid Hadronotus pennsylvanicus (Hymenoptera: Scelionidae) for Egg Masses of the Squash Bugs Anasa tristis and Anasa armigera (Hemiptera: Coreidae) in Squash and Cucumber Fields.

Parasitoid foraging behavior is affected by habitat and host plant differences. Egg parasitoids also use a combination of oviposition-induced and host-derived cues to find host eggs. This study compared parasitism by Hadronotus pennsylvanicus (Ashmead) (Hymenoptera: Scelionidae) on two squash bug species, Anasa tristis (DeGeer) and Anasa armigera Say (Hemiptera: Coreidae), by placing sentinel squash and cucumber plants with egg masses of either of the two squash bug species in squash and cucumber fields in a 3-way factorial design. Host density of wild A. tristis egg masses in squash fields may have influenced parasitoid foraging behavior on sentinel plants. In the 3-way factorial design, parasitism was higher on sentinel squash plants and in squash fields overall. However, parasitism on A. armigera egg masses was highest on sentinel cucumber plants in squash fields and parasitism on A. tristis egg masses was higher on sentinel squash plants in either squash or cucumber fields and lowest on sentinel cucumber plants in cucumber fields. Results suggest that parasitoids were able to specifically orient to the combination of host plant and host cues associated with A. tristis egg masses on sentinel squash plants, but that they were more responsive to plant-induced cues associated with cucumber when searching for A. armigera egg masses. Parasitoids appear to utilize different combinations of host plant and host cues when searching for eggs of the two squash bug species.

Effect of Plastic Mulch Colors on Anasa tristis (Hemiptera: Coreidae) Population Dynamics in Summer Squash, Cucurbita pepo (Cucurbitales: Cucurbitaceae).

The squash bug, Anasa tristis (De Geer), is a serious pest of cucurbit crops across the United States, especially within summer squash (Cucurbita pepo L.) systems. Using their piercing sucking mouthparts, squash bugs feed on both leaf tissue and fruits, often leading to leaf necrosis, marketable fruit loss, and even plant death. To date, the relationship between squash bug presence and plasticulture has not been adequately investigated. This 2-yr study evaluated the effects of white, black, and reflective plastic mulch colors on the occurrence of all squash bug life stages and marketable zucchini yield in Virginia. In both years, A. tristis adults and egg masses were more numerous on zucchini plants grown in white and reflective plastic mulch compared to bare ground plants. Greater nymphal densities and marketable fruit yield were observed in certain plastic mulch treatments versus the bare ground treatment, yet these differences were not consistent in both years. Contrary to the repellency effects reflective mulches have on other cucurbit insect pests, our research suggests that reflective and other plastic mulch colors can negatively impact squash bug management, especially in regions with high A. tristis pressure. Our study offers new insights for cucurbit growers to use when considering whether they should implement plasticulture in their growing systems.

Both male- and female-produced pheromones influence Colorado potato beetle movement in the field.

BACKGROUND: Colorado potato beetle [Leptinotarsa decemlineata (Say); Coleoptera: Chrysomelidae] is a widespread key pest of potatoes and other solanaceous crops of the temperate Northern Hemisphere. Its evolution of multiple pesticide resistance is a continuing threat if chemical control is relied upon for pest management. An integrated approach including semiochemicals and other non-pesticidal tactics is essential to sustainable management. In addition to the known volatile aggregation pheromone produced by male adult beetles that is attractive to both females and males, there is a yet-unidentified female-produced sex pheromone. Using caged 'bait plants' in the field in Maryland, USA, we tested the attraction of male and female beetles to groups of male-only, female-only, larvae, and uninfested bait plants. RESULTS: Results of five field experiments demonstrate that the female-produced sex pheromone is attractive to male beetles, whereas the male-produced pheromone is attractive to both females and males. Male attraction increased with higher densities of females on bait plants, but female attraction did not change with female density on bait plants. CONCLUSION: This is the first study to demonstrate male attraction to the female sex pheromone in the field, and to our knowledge, the first to demonstrate a beetle species using bidirectional communication between males and females. Use of semiochemicals for monitoring and management of Colorado potato beetle should consider male-, female-, as well as plant-produced volatiles as important tools for pest manipulation in potato integrated pest management. Published 2021. This article is a U.S. Government work and is in the public domain in the USA.

Trap Cropping Harlequin Bug: Distance of Separation Influences Female Movement and Oviposition.

To evaluate perimeter trap crops for management of harlequin bug, Murgantia histrionica (Hahn), we undertook greenhouse and field experiments with mustard greens as trap crop for a collard cash crop. We confirmed that harlequin bugs prefer to immigrate to and reside on mustard. Females, however, in greenhouse cage experiments, 'commuted' to collards to lay their eggs. In separate spring and fall field plantings, using replicated 12 m by 12 m collard plots in 1-ha fields, we tested mustard planted as an adjacent perimeter trap crop, or a perimeter trap crop separated by 2 unplanted rows (2.3 m), or with no trap crop. Adults accumulated on the spring mustard crop but overall numbers remained low, with all collards sustaining <1% leaves damaged. In the fall, the separation of 2.3 m reduced oviposition on collards fourfold, and feeding damage approximately 2.5-fold, compared to collards with an adjacent trap crop. Fall control plots with no border trap crop showed even lower foliar damage; likely result of preferential immigration of harlequin bugs to mustard at the field scale, resulting in fewer bugs near the control treatment plots. Thus, the spatial arrangement of the mustard trap crop, and its separation from the cash crop, influences pest abundance and damage. A separated mustard border can reduce bug movement including female commuting and egg-laying, thus better protecting the collard cash crop. Future research should address reduction in area of trap crops, deployment of semiochemicals, and possible changes in timing, to promote trap cropping that is practical for grower implementation.

Parasitism and Predation on Sentinel Egg Masses of Three Stink Bug Species (Hemiptera: Pentatomidae) in Native and Exotic Ornamental Landscapes.

This study evaluated parasitism and predation on sentinel egg masses of three stink bug species, the spined soldier bug, Podisus maculiventris (Say), the brown stink bug, Euschistus servus (Say), and the invasive brown marmorated stink bug (BMSB), Halyomorpha halys (Stål), in ornamental landscapes composed of either native or exotic plants. This study also compared the species composition of parasitoids attacking two native stink bug species (P. maculiventris and E. servus) with those attacking the invasive BMSB on the same tree species in the same habitat. Overall, egg parasitism and predation were much higher on the two native stink bug species compared with BMSB, with an average parasitism rate of 20.6% for E. servus, 12.7% for P. maculiventris, and only 4.2% for H. halys and an average predation rate of 8.2% for E. servus,17.7% for P. maculiventris, and 2.3% for H. halys. Egg predation was also significantly higher on P. maculiventris than on E. servus eggs. Eight parasitoid species attacked sentinel stink bug eggs in the ornamental landscaped plots. Trissolcus euschisti (Ashmead) (Hymenoptera: Scelionidae) was the predominant parasitoid for all three stink bug species. There were no significant differences in parasitism and predation rates on any of the stink bug species between native and exotic plots. Therefore, there is no evidence that ornamental landscapes composed of native plants increased parasitism or predation rates of sentinel egg masses of two native stink bug species or the invasive BMSB, compared with those composed entirely of exotic plants.

Biology and Management of Pest Diabrotica Species in South America.

The genus Diabrotica has over 400 described species, the majority of them neotropical. However, only three species of neotropical Diabrotica are considered agricultural pests: D. speciosa, D. balteata, and D. viridula. D. speciosa and D. balteata are polyphagous both as adults and during the larval stage. D. viridula are stenophagous during the larval stage, feeding essentially on maize roots, and polyphagous as adults. The larvae of the three species are pests on maize, but D. speciosa larvae also feed on potatoes and peanuts, while D. balteata larvae feed on beans and peanuts. None of these species express a winter/dry season egg diapause, displaying instead several continuous, latitude-mediated generations per year. This hinders the use of crop rotation as a management tool, although early planting can help in the temperate regions of the distribution of D. speciosa. The parasitoids of adults, Celatoria bosqi and Centistes gasseni, do not exert much control on Diabrotica populations, or show potential for inundative biocontrol plans. Management options are limited to insecticide applications and Bt genetically modified (GM) maize. Other techniques that show promise are products using Beauveria bassiana and Heterorhabditis bacteriophora, semiochemical attractants for monitoring purposes or as toxic baits, and plant resistance.

Transcriptome Sequencing of the Striped Cucumber Beetle, Acalymma vittatum (F.), Reveals Numerous Sex-Specific Transcripts and Xenobiotic Detoxification Genes.

Acalymma vittatum (F.), the striped cucumber beetle, is an important pest of cucurbit crops in the contintental United States, damaging plants through both direct feeding and vectoring of a bacterial wilt pathogen. Besides providing basic biological knowledge, biosequence data for A. vittatum would be useful towards the development of molecular biopesticides to complement existing population control methods. However, no such datasets currently exist. In this study, three biological replicates apiece of male and female adult insects were sequenced and assembled into a set of 630,139 transcripts (of which 232,899 exhibited hits to one or more sequences in NCBI NR). Quantitative analyses identified 2898 genes differentially expressed across the male-female divide, and qualitative analyses characterized the insect's resistome, comprising the glutathione S-transferase, carboxylesterase, and cytochrome P450 monooxygenase families of xenobiotic detoxification genes. In summary, these data provide useful insights into genes associated with sex differentiation and this beetle's innate genetic capacity to develop resistance to synthetic pesticides; furthermore, these genes may serve as useful targets for potential use in molecular-based biocontrol technologies.

Host Kairomones Influence Searching Behavior of Trissolcus japonicus (Hymenoptera: Scelionidae), a Parasitoid of Halyomorpha halys (Heteroptera: Pentatomidae).

The brown marmorated stink bug, Halyomorpha halys (Stål), is a highly polyphagous species native to Asia that has become a serious invasive agricultural and nuisance pest across North America and Europe. Classical biological control host range evaluations have revealed egg parasitoid Trissolcus japonicus (Ashmead) to be the primary candidate biocontrol agent for field release against H. halys. However, these evaluations only provide us with the physiological host range of T. japonicus. Other Trissolcus species have demonstrated that contact kairomones from different host species elicit varied responses in the parasitoids' host foraging behaviors. To assess T. japonicus response to host kairomones, mated naive females were exposed to leaf surfaces contaminated with adult kairomones from its preferred host, H. halys, or from a native nontarget host, Podisus maculiventris (Say) (Heteroptera: Pentatomidae). Red maple, apple, and soybean were used as plant substrate treatments. The wasp's residence time on the leaf surface, linear walking velocity, and angular walking velocity were observed and measured using Noldus EthoVision XT tracking software. Within each leaf treatment, T. japonicus displayed stronger behavioral responses on leaves contaminated with contact kairomones from H. halys. The parasitoid resided on H. halys contaminated leaves for approximately twice as a long as it did on P. maculiventris contaminated leaves. Further, both species' kairomones elicited significant decreases in parasitoid walking velocity on all tested substrate types. Overall, our study suggests that kairomone-based behavioral studies can be used to further evaluate the host specificity of T. japonicus and can be an invaluable supplement to classical biocontrol host range testing regimes.

Attractiveness of Pheromone Components With and Without the Synergist, Methyl (2E,4E,6Z)-2,4,6-Decatrienoate, to Brown Marmorated Stink Bug (Hemiptera: Pentatomidae).

The brown marmorated stink bug, Halyomorpha halys (Stål) (Hemiptera: Pentatomidae) is attracted to its male-produced aggregation pheromone, a ~3.5:1 mixture of (3S,6S,7R,10S)- and (3R,6S,7R,10S)-10,11-epoxy-1-bisabolen-3-ol (SSRS and RSRS respectively), and also to the pheromone of its Asian sympatric species Plautia stali Scott (Hemiptera: Pentatomidae), methyl (2E,4E,6Z)-2,4,6-decatrienoate (MDT). A stereoisomeric mixture of (7R) 10,11-epoxy-1-bisabolen-3-ols (= mixed murgantiols) is used together in commercialized products with MDT because the latter is a synergist for H. halys attraction to mixed murgantiols. However, the optimal ratio for MDT combination with mixed murgantiols, and the sensitivity of bug captures to variation in ratio of the two pheromone components, have not been tested to date. Using black pyramid traps at two sites (in Maryland and West Virginia, United States), different ratios of mixed murgantiols to MDT were tested over two entire seasons. Also, captures using various ratios of the two active pheromone stereoisomers were undertaken in month-long trials with and without MDT. Results showed that H. halys adult and nymphal captures were relatively insensitive to the ratio of synthetic pheromone (mixed murgantiols) to MDT, as long as each was present in the trap. Captures of adults and nymphs were responsive to the lure loading of the SSRS isomer, but relatively insensitive to levels of the minor component, RSRS. The relative insensitivity of H. halys to these attractant ratios gives flexibility to development of more cost-efficient synthesis and trapping as well as other semiochemical-based management tactics.

Author Correction: An IDS-Type Sesquiterpene Synthase Produces the Pheromone Precursor (Z)-α-Bisabolene in Nezara viridula.

The original version of this article unfortunately contained a mistake. Under the heading "Insects" in "Methods and Materials" the sentence "A colony of N. viridula originated with field collections near Tifton, Georgia, USA" is incorrect.

An IDS-Type Sesquiterpene Synthase Produces the Pheromone Precursor (Z)-α-Bisabolene in Nezara viridula.

Insects use a wide range of structurally diverse pheromones for intra-specific communication. Compounds in the class of terpenes are emitted as sex, aggregation, alarm, or trail pheromones. Despite the common occurrence of terpene pheromones in different insect lineages, their origin from dietary host plant precursors or de novo biosynthetic pathways often remains unknown. Several stink bugs (Hemiptera: Pentatomidae) release bisabolene-type sesquiterpenes for aggregation and mating. Here we provide evidence for de novo biosynthesis of the sex pheromone trans-/cis-(Z)-α-bisabolene epoxide of the Southern green stink bug, Nezara viridula. We show that an enzyme (NvTPS) related to isoprenyl diphosphate synthases (IDSs) of the core terpene metabolic pathway functions as a terpene synthase (TPS), which converts the general intermediate (E,E)-farnesyl diphosphate (FPP) to the putative pheromone precursor (+)-(S,Z)-α-bisabolene in vitro and in protein lysates. A second identified IDS-type protein (NvFPPS) makes the TPS substrate (E,E)-FPP and functions as a bona fide FPP synthase. NvTPS is highly expressed in male epidermal tissue associated with the cuticle of ventral sternites, which is in agreement with the male specific release of the pheromone from glandular cells in this tissue. Our study supports findings of the function of similar TPS enzymes in the biosynthesis of aggregation pheromones from the pine engraver beetle Ips pini, the striped flea beetle Phyllotreta striolata, and the harlequin bug Murgantia histrionica, and hence provides growing evidence for the evolution of terpene de novo biosynthesis by IDS-type TPS families in insects.

Field Attraction of Striped Cucumber Beetles to a Synthetic Vittatalactone Mixture.

The striped cucumber beetle, Acalymma vittatum (F.) (Coleoptera: Chrysomelidae), is a key pest of cucurbits in eastern North America, rapidly colonizing young plantings and vectoring bacterial wilt of cucurbits. Its aggregation pheromone has been identified and synthesized stereospecifically, but has not been field tested to date. Here, we present field bioassays of this pheromone, using mixed vittatalactone made with a novel and cost-efficient semispecific synthesis. This mixture of eight stereoisomers of (2R,3R)-vittatalactone proved highly attractive to both sexes of striped cucumber beetle, using two different trap types and a pilot attract-and-kill combination with watermelon containing the diabroticine feeding stimulant cucurbitacin-E-glycoside, under field conditions in cucurbit vegetable plantings. Availability of mixed vittatalactones could enable highly effective and specific management of striped cucumber beetle.