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A large proportion of the insects which have invaded new regions and countries are emerging species, being found for the first time outside their native range. Being able to detect such species upon arrival at ports of entry before they establish in non-native countries is an urgent challenge. The deployment of traps baited with broad-spectrum semiochemical lures at ports-of-entry and other high-risk sites could be one such early detection tool. Rapid progress in the identification of semiochemicals for cerambycid beetles during the last 15 years has revealed that aggregation-sex pheromones and sex pheromones are often conserved at global levels for genera, tribes or subfamilies of the Cerambycidae. This possibly allows the development of generic attractants which attract multiple species simultaneously, especially when such pheromones are combined into blends. Here, we present the results of a worldwide field trial programme conducted during 2018–2021, using traps baited with a standardised 8-pheromone blend, usually complemented with plant volatiles. A total of 1308 traps were deployed at 302 sites covering simultaneously or sequentially 13 European countries, 10 Chinese provinces and some regions of the USA, Canada, Australia, Russia (Siberia) and the Caribbean (Martinique). We intended to test the following hypotheses: 1) if a species is regularly trapped in significant numbers by the blend on a continent, it increases the probability that it can be detected when it arrives in other countries/continents and 2) if the blend exerts an effective, generic attraction to multiple species, it is likely that previously unknown and unexpected species can be captured due to the high degree of conservation of pheromone structures within related taxa. A total of 78,321 longhorned beetles were trapped, representing 376 species from eight subfamilies, with 84 species captured in numbers greater than 50 individuals. Captures comprised 60 tribes, with 10 tribes including more than nine species trapped on different continents. Some invasive species were captured in both the native and invaded continents. This demonstrates the potential of multipheromone lures as effective tools for the detection of ‘unexpected’ cerambycid invaders, accidentally translocated outside their native ranges. Adding new pheromones with analogous well-conserved motifs is discussed, as well as the limitations of using such blends, especially for some cerambycid taxa which may be more attracted by the trap colour or other characteristics rather than to the chemical blend.

Corn wireworm, Melanotus communis Gyllenhal (Coleoptera: Elateridae), is an economically important larval pest of root and tuber crops in the United States. Previous work to estimate field-level abundance of M. communis has focused on grain-based larval baits placed in soil. However, this sampling method is labor intensive and may not estimate population size accurately. Recent discovery of the M. communis sex pheromone, 13-tetradecenyl acetate, provides a new method to monitor this pest during the adult stage. Early studies with this pheromone showed that different trapping methods might enhance catch and improve trap servicing. We hypothesized that placing lures on elevated traps would increase M. communis capture relative to the in-ground pitfall trapping that is currently used. We had 2 objectives for this study: (a) to compare pheromone captures among in-ground pitfall traps, on-ground pitfalls, elevated pitfalls (1 m), or elevated sticky cards (1 m) and (b) test lure longevity by aging the lures outdoors at 8-, 6-, 4-, 2-, and 0-wk intervals prior to trap deployment in the field. Experiments were conducted in North Carolina, Virginia, South Carolina, and Florida during the 2021 and 2022 field seasons. Results highlight large variation in M. communis abundance across the 4 states. We showed that 1 m elevated pheromone traps caught the most beetles. The age of the lure prior to deployment had a significant effect on trap catch. The lures that were aged for fewer weeks attracted significantly more beetles, with 0- and 2-wk-old lures capturing the greatest numbers.

Firefly flashes are well-known visual signals used by these insects to find, identify, and choose mates. However, many firefly species have lost the ability to produce light as adults. These “unlighted” species generally lack developed adult light organs, are diurnal rather than nocturnal, and are believed to use volatile pheromones acting over a distance to locate mates. While cuticular hydrocarbons, which may function in mate recognition at close range, have been examined for a handful of the over 2000 extant firefly species, no volatile pheromone has ever been identified. In this study, using coupled gas chromatography - electroantennographic detection, we detected a single female-emitted compound that elicited antennal responses from wild-caught male winter fireflies, Photinus corruscus. The compound was identified as (1S)-exo-3-hydroxycamphor (hydroxycamphor). In field trials at two sites across the species’ eastern North American range, large numbers of male P. corruscus were attracted to synthesized hydroxycamphor, verifying its function as a volatile sex attractant pheromone. Males spent more time in contact with lures treated with synthesized hydroxycamphor than those treated with solvent only in laboratory two-choice assays. Further, using single sensillum recordings, we characterized a pheromone-sensitive odorant receptor neuron in a specific olfactory sensillum on male P. corruscus antennae and demonstrated its sensitivity to hydroxycamphor. Thus, this study has identified the first volatile pheromone and its corresponding sensory neuron for any firefly species, and provides a tool for monitoring P. corruscus populations for conservation and further inquiry into the chemical and cellular bases for sexual communication among fireflies.

Hexanoic acid, 1-octanol, 1,8-octanediol, octyl hexanoate, 1,8-octanediol monohexanoate, and 1,8-octanediol dihexanoate were identified in headspace volatiles collected from the crushed abdomen of a female click beetle of the species Parallelostethus attenuatus (Say) (Elaterinae, tribe Elaterini). In field trials carried out in Illinois, South Carolina, North Carolina, and Virginia, adult male beetles were strongly attracted to 1,8-octanediol dihexanoate alone. Blends of the dihexanoate with one or more of the other compounds proved to be less attractive than the dihexanoate alone, suggesting that the pheromone of this species may consist of a single compound. The symmetrical diester structure of the pheromone is a novel natural product and appears to be structurally unique among insect pheromones. USDA-ARS State Potato Partnership Funds [2092-22000-022-00D, 6080-22000-030-00D] Published version LMH, JGM, ASH, TPK, and LW III appreciate funding support from the USDA-ARS State Potato Partnership Funds projects 2092-22000-022-00D and 6080-22000-030-00D. Public domain – authored by a U.S. government employee

Communication in social insect colonies depends on signals accurately reflecting the identity and physiological state of the individuals. Such information is coded by the products of multiple exocrine glands, and the resulting blends reflect the species, sex, caste, age, task, reproductive status, and health of an individual, and may also contain caste-specific pheromones regulating the behavior and physiology of other individuals. Here we examined the composition of labial gland secretions in females of the bumble bee Bombus impatiens, of different castes, social condition, age, mating status, and ovarian activation. We show that active queens, gynes, and workers each produce caste-specific compounds that may serve different communicative functions. The composition and amounts of wax esters, mostly octyl esters produced by active queens, differed significantly between castes, mating, and social conditions, suggesting a social signaling role. Farnesyl esters were predominant in gynes and peaked at optimal mating age (6-10 days), suggesting their possible roles as sex pheromone components. Reproductive status of females and age across castes was reflected by the ratio between short- and long-chain hydrocarbons, suggesting that these compounds may serve as fertility signals. Our findings overall suggest that the labial gland composition in B. impatiens reflects different facets of female bee physiology. While further bioassays are required to determine the function of these compounds, they are likely to have an important role in the communication between individuals and to the cohesion of the social structure.

Firefly flashes are well-known visual signals used by these insects to find, identify, and choose mates. However, many firefly species have lost the ability to produce light as adults. These “unlighted” species generally lack developed adult light organs, are diurnal rather than nocturnal, and are believed to use volatile pheromones acting over a distance to locate mates. While cuticular hydrocarbons, which may function in mate recognition at close range, have been examined for a handful of the over 2000 extant firefly species, no volatile pheromone has ever been identified. In this study, using coupled gas chromatography - electroantennographic detection, we detected a single female-emitted compound that elicited antennal responses from wild-caught male winter fireflies,Photinus corrusca. The compound was identified as (1S)-exo-3-hydroxycamphor (hydroxycamphor). In field trials at two sites across the species’ eastern North American range, large numbers of maleP. corruscawere attracted to synthesized hydroxycamphor, verifying its function as a volatile sex attractant pheromone. Males spent more time in contact with lures treated with synthesized hydroxycamphor than those treated with solvent only in laboratory two-choice assays. Further, using single sensillum recordings, we characterized a pheromone-sensitive odorant receptor neuron in a specific olfactory sensillum on maleP. corruscaantennae and demonstrated its sensitivity to hydroxycamphor. Thus, this study has identified the first volatile pheromone and its corresponding sensory neuron for any firefly species, and provides a tool for monitoringP. corruscapopulations for conservation, and further inquiry into the chemical and cellular bases for sexual communication among fireflies.

Ants use chemical signals to communicate for various purposes related to colony function. Social organization in the red imported fire ant, Solenopsis invicta, is determined by the Sb supergene, with colonies of the monogyne (single-queen) form lacking the element and colonies of the polygyne (multiple-queen) form possessing it. Polygyne workers accept new reproductive queens in their nest, but only those carrying Sb; young winged queens lacking this genetic element are executed as they mature sexually in their natal nest or as they attempt to enter a foreign nest to initiate reproduction after mating and shedding their wings. It has been suggested that queen supergene genotype status is signaled to workers by unsaturated cuticular hydrocarbons, while queen reproductive status is signaled by piperidines (venom alkaloids). We used high-throughput behavioral assays to study worker acceptance of paper dummies dosed with fractions of extracts of polygyne queens, or blends of synthetic counterparts of queen cuticular compounds. We show that the queen supergene pheromone comprises a blend of monoene and diene unsaturated hydrocarbons. Our assays also reveal that unsaturated hydrocarbons elicit discrimination by polygyne workers only when associated with additional compounds that signal queen fertility. This synergistic effect was obtained with a polar fraction of queen extracts, but not by the piperidine alkaloids, suggesting that the chemical(s) indicating queen reproductive status are compounds more polar than cuticular hydrocarbons but are not the piperidine alkaloids. Our results advance understanding of the role of chemical signaling that is central to the regulation of social organization in an important invasive pest and model ant species.

We conducted field bioassays with several known cerambycid pheromones in two zones of central-southern Chile: (1) Las Trancas (Ñuble region) and Coñaripe (Los Rios region) (Study 1) and (2) Rucamanque and Maquehue (La Araucania region) (Study 2). Up to eight compounds were tested individually, including 3-hydroxy-2-hexanone, (2R*,3S*)- and (2R*,3R*)-2,3-hexanediol, fuscumol, fuscumol acetate, monochamol, 2-methylbutanol, and geranylacetone. Compounds were loaded in plastic sachets placed in either multiple funnel or cross-vane panel traps hung in trees in a randomized block design (n = 3 or 4). The number of treatments and bioassay periods varied depending on the study. A total of 578 specimens belonging to 11 native species were collected, with the three captured in the highest numbers being Eryphus laetus (292 specimens), Calydon submetallicum (n = 234), and Chenoderus testaceus (n = 20). The three species are of economic importance: E. laetus is considered a minor pest in apple orchards, and the other two species infest Nothophagus hosts, including some timber species. Traps baited with 3-hydroxy-2-hexanone collected significant numbers of both sexes of the two most abundant species, and this compound was the only treatment that attracted C. submetallicum. (2R*,3R*)- and (2R*,3S*)-2,3-Hexanediols were also significantly attractive to E. laetus. Our results suggested that 3-hydroxy-2-hexanone and 2,3-hexanediols, which are known pheromone components of cerambycid species worldwide, are also likely to be conserved aggregation pheromone components among some species in western South America.

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.

The leaffooted bugs

An arrayed combination of water-soluble deep cavitands and cationic dyes has been shown to optically sense insect pheromones at micromolar concentration in water. Machine learning approaches were used to optimize the most effective array components, which allows differentiation between small structural differences in targets, including between different diastereomers, even though the pheromones have no innate chromophore. When combined with chiral additives, enantiodiscrimination is possible, dependent on the size and shape of the pheromone.

Municipalities in Arizona and Nevada along the Colorado River are subject to seasonal mass emergences of a nuisance net-spinning caddisfly, Smicridea fasciatella McLachlan (Trichoptera: Hydropsychidae). Here, we describe the characterization and field testing of S. fasciatella extracts to evaluate their potential as lures in baited traps. Solvent extracts of external (i.e., full body—cuticular hydrocarbon, abdominal hexane washes) and internal (i.e., crushed abdomen) parts of adult S. fasciatella were prepared from both sexes, and analyzed by coupled gas chromatography-mass spectrometry (GCMS). Several sex-specific compounds were identified, including (6Z,9Z)-6,9-nonadecadiene and (3Z,6Z,9Z)-3,6,9-nonadecatriene from males, and 2-undecanone, 2-tridecanone, and a heptadecene isomer from females. Extracts from both sexes were also analyzed by coupled gas chromatography-electroantennographic detection (EAD) using antennae of males for detection. Antennae of males weakly responded to 2-undecanone and 2-tridecanone, and to their corresponding alcohols, 2-undecanol and 2-tridecanol, which were included in field tests. Extracts of adult males did not elicit a response from male antennae, suggesting that males do not produce aggregation pheromones attractive to other males. The synchronized, dense populations of lekking males and other possible mating signals (e.g., visual recognition) may have contributed to the minimal attraction seen to test lures deployed in PHEROCON 1C traps. Overall, our results suggest that for this species, attractant pheromones have minimal or no role in bringing the sexes together for mating.

In a previous study, we reported the identification and synthesis of a male-specific sex pheromone component of the stink bug, Pellaea stictica, as the alcohol 2,4,8,13-tetramethyltetradecan-1-ol (1). To establish the correlation between the stereochemistry of the pheromone and its bioactivity, it first was necessary to determine its absolute configuration. For this purpose, a series of syntheses were designed to: (a) furnish a mixture of all possible stereoisomers; (b) a narrowed down group of diastereomers, and (c) one specific enantiomer. A crucial step in the syntheses involved a coupling reaction between two key intermediates: a phosphonium salt and an aldehyde, through a Wittig olefination. Nuclear magnetic resonance data of a mixture of the synthetic pheromone diastereomers and further comparison of GC retention times with that of the natural product by gas chromatography suggested that the methyl branches at C2 and C4 were in a syn relationship, reducing the possibilities to only four of the eight possible stereoisomers. Employing GC analysis, chiral derivatization reagents and synthetic (8R)-2,4-syn-1 it was possible to confirm the configuration of the methyl branch at C8 as R, reducing the number of possible stereoisomers to two. After enantioselective synthesis of (2R,4R,8R)-1, the absolute configurations of all methyl branches of the natural compound were confirmed as R, fully identifying the male-produced sex pheromone of P. stictica as (2R,4R,8R)-2,4,8,13-tetramethyltetradecan-1-ol.

The wood-boring larvae of longhorned beetles (Coleoptera: Cerambycidae) can be important pests of woody plants, particularly as invasive species introduced by international commerce. Previous research has revealed that cerambycid species native to different parts of the world often share the same aggregation-sex pheromones and that pheromones of different species can be combined to create multi-species attractants that would be advantageous for surveillance monitoring for a number of species simultaneously. To explore the extent to which these chemicals can be combined into single lures, we developed four different blends of six to eight compounds and tested their effects as attractants for a community of longhorned beetle species in Iowa. The blends included known pheromones of species native to the study site, as well as pheromones identified from cerambycid species native to other parts of the world. The experiment confirmed that several cerambycid species were attracted by specific blends, in accordance with their known pheromone chemistry, and despite the presence of pheromone components of heterospecifics. This finding lends further support to developing multi-component blends that can effectively monitor for new incursions of multiple exotic species concurrently.

Females of two click beetle species, Cardiophorus tenebrosus and C. edwardsi (Coleoptera: Elateridae), produce methyl (3R,6E)-2,3-dihydrofarnesoate as their sex pheromone. We had serendipitously discovered that males of both species were also strongly attracted to (R)-fuscumol acetate ((E)-6,10-dimethylundeca-5,9-dien-2-yl acetate), a known longhorned beetle (Coleoptera: Cerambycidae) pheromone, due to its structural similarities to the click beetle pheromone. To further investigate the specificity of the responses of Cardiophorus males, additional analogs with different chain lengths and structural relationships compared to the natural pheromone were synthesized and tested. In field and electroantennogram bioassays, only fuscumol propionate ((E)-6,10-dimethylundeca-5,9-dien-2-yl propionate) elicited strong responses from Cardiophorus males, indicating that they were able to distinguish chain length and spatial relationships between the structural elements. In field trials, C. tenebrosus males were attracted equally to the analog and their natural pheromone, but the pheromone elicited stronger antennal responses from males. In contrast, traps baited with fuscumol propionate caught approximately 26 times as many C. edwardsi males compared to traps baited with the natural pheromone, although the analog elicited significantly smaller antennal responses from C. edwardsi males. Thus, in terms of behavioral responses, fuscumol propionate appears to be acting as a hyperactive pheromone mimic, a phenomenon which has rarely been observed in insect semiochemistry.

The Asian eulophid wasp Tetrastichus planipennisi is being released in North America as a biocontrol agent for the emerald ash borer (Agrilus planipennis), a very destructive invasive buprestid beetle that is devastating ash trees (Fraxinus spp.). We identified, synthesized, and tested a female-produced sex pheromone for the wasp. The key component eliciting behavioral responses from male wasps in flight tunnel bioassays was identified as (6S,10S)-(2E,4E,8E)-4,6,8,10-tetramethyltrideca-2,4,8-triene. Female specificity was demonstrated by gas chromatographic (GC) comparison of male and female volatile emissions and whole body extracts. The identification was aided by coupled gas chromatography/mass spectrometry analysis, microchemical reactions, NMR, GC analyses with a chiral stationary phase column, and matching GC retention times and mass spectra with those of synthetic standards. The tetramethyl-triene hydrocarbon was synthesized as a mixture of two enantiomeric pairs of diastereomers, and as the pure insect-produced stereoisomer. In flight-tunnel bioassays, males responded to both the natural pheromone and the chiral synthetic material by upwind flight and landing on the source. In contrast, the mixture of four stereoisomers was not attractive, indicating that one or more of the "unnatural" stereoisomers antagonized attraction. Field trials, using yellow pan traps baited with natural pheromone, captured significantly more male wasps than control traps over a four week trial. The identified pheromone could increase the efficiency and specificity of the current detection methods for Tetrastichus planipennisi and aid in the determination of parasitoid establishment at release sites.

SUMMARYOptimal mating decisions depend on the robust coupling of signal production and perception because independent changes in either could carry a fitness cost. However, since the perception and production of mating signals are often mediated by different tissues and cell types, the mechanisms that drive and maintain their coupling remain unknown for most animal species. Here, we show that in Drosophila, sensory perception and production of an inhibitory mating pheromone are co-regulated by Gr8a, a member of the Gustatory receptor gene family. Specifically, we found that the pleiotropic action of Gr8a independently regulates the perception of pheromones by the chemosensory systems of males and females, as well as their production in the fat body and oenocytes of males. These findings provide a relatively simple molecular explanation for how pleiotropic receptors maintain robust mating signaling systems at the population and species levels.

Cuticular hydrocarbons (CHCs) are the main components of the epicuticular wax layer that in many insects functions as a barrier against desiccation. CHCs also play many other roles, including serving as sex pheromones, kairomones, primer pheromones, and colony-, caste-, species- and sex-recognition signals. In insects, CHC profiles can vary depending upon age, species, sex, and strain. Understanding factors associated with variation in hydrocarbon profiles is important for identifying potential vulnerabilities relating to pest ecology and life histories and for developing tools for pest monitoring and management strategies. In this study, we assessed potential sources of variation in CHC profiles in the navel orangeworm Amyelois transitella (Walker) (Lepidoptera: Pyralidae), an economically important pest of nut crops in California. Using coupled gas chromatography-mass spectrometry, we characterized and compared CHC profiles between adults of pyrethroid-resistant (R347) and susceptible (ALMOND) strains. We further compared CHC profiles from adults differing in age (1, 3, 5, and 7 d post-eclosion) and sex. Hydrocarbon profiles comprised 47 different CHCs in detectable quantities that ranged from C17 to C43 in chain length and included straight-chain alkanes and a variety of mono-, di-, and tri-methylalkanes. Adults from resistant populations had greater quantities of CHCs in total than those from susceptible strains, but relative quantities of individual components were similar. The six most abundant compounds were n-pentacosane, n-heptacosane, n-nonacosane, n-hentriacontane, 11,25 + 13,23 + 15,21-dimethylpentatriacontane, and 13,23 + 11,25 + 9,17-dimethylheptatriacontane. Post-eclosion, total CHCs increased with adult age, with males producing greater quantities than females at all ages. Our results show that CHC profiles vary depending on age, sex, and strain and suggest that CHC profiles may be useful as biomarkers to differentiate between insecticide- resistant and susceptible populations.

Hexanoic acid, 1-octanol, 1,8-octanediol, octyl hexanoate, 1,8-octanediol monohexanoate, and 1,8-octanediol dihexanoate were identified in headspace volatiles collected from the crushed abdomen of a female click beetle of the species Parallelostethus attenuatus (Say) (Elaterinae, tribe Elaterini). In field trials carried out in Illinois, South Carolina, North Carolina, and Virginia, adult male beetles were strongly attracted to 1,8-octanediol dihexanoate alone. Blends of the dihexanoate with one or more of the other compounds proved to be less attractive than the dihexanoate alone, suggesting that the pheromone of this species may consist of a single compound. The symmetrical diester structure of the pheromone is a novel natural product and appears to be structurally unique among insect pheromones.

We describe the identification and field testing of 3-methylthiopropan-1-ol (methionol) as a male-produced aggregation-sex pheromone for the cerambycid beetle Knulliana cincta cincta (Drury) (subfamily Cerambycinae, tribe Bothriospilini). The corresponding sulfoxide, 3-methylsulfinylpropan-1-ol, was also produced sex-specifically by males, but its function remains unclear because the measured release rates of this compound from five different types of release devices were very low to undetectable. Unexpectedly, adults of the cerambycine Elaphidion mucronatum (Say) (Elaphidiini), primarily females, also were attracted by methionol, despite males of this species producing an aggregation-sex pheromone of entirely different structure, (2E,6Z,9Z)-2,6,9-pentadecatrienal.

Simple Summary The longhorn beetle, Plagionotus detritus (L.), occurs throughout Europe and into the Middle East. Our principal aim was to identify the male-produced aggregation-sex attractant pheromone of this species. A pheromone-based monitoring system could support plant protection in areas where it might become a pest and natural conservation in areas where it is endangered. Headspace volatiles were collected from live beetles and analyzed by coupled gas chromatography-electroantennogram detection and gas chromatography-mass spectrometry. Two components of the extracts that elicited responses from antennae of P. detritus, (R)-3-hydroxyhexan-2-one and (S)-2-hydroxyoctan-3-one were identified, (±)-3-hydroxyhexan-2-one was purchased, and (S)-2-hydroxyoctan-3-one was synthesized, and field-tested. The blend of the two components attracted both sexes of P. detritus in field bioassays. Unexpectedly, a predatory clerid beetle, Clerus mutillarius F., was also attracted by both of the synthetic compounds. Another longhorn beetle species, Xylotrechus antilope Schönh. was significantly attracted to traps baited with (S)-2-hydroxyoctan-3-one alone or the blend containing this compound. These results show that the pheromone components and combinations play key roles in longhorn beetle life history and ecology, and the community of species in which they live. Abstract (1) Background: The principal aim of our work was to identify pheromone components for Plagionotus detritus (L.) (Coleoptera: Cerambycidae), which could be exploited for developing a pheromone-based monitoring system for the complementary purposes of plant protection in areas where it might become a pest, and natural conservation in areas where it is rare or endangered. (2) Methods: Collection and analysis of headspace volatiles were carried out with field-collected beetles. Bioactive volatile compounds identified [(R)-3-hydroxyhexan-2-one and (S)-2-hydroxyoctan-3-one] from extracts of males were purchased [(±)-3-hydroxyhexan-2-one], and synthesized [(S)-2-hydroxyoctan-3-one] and field-tested. Electroantennogram assays showed that antennae of the predatory beetle Clerus mutillarius F. (Coleoptera: Cleridae) also responded to the synthetic compounds. (3) Results: A two-component aggregation-sex pheromone consisting of (R)-3-hydroxyhexan-2-one and (S)-2-hydroxyoctan-3-one was identified for P. detritus. (±)-3-hydroxyhexan-2-one and (S)-2-hydroxyoctan-3-one attracted adults of P. detritus in field bioassays. Adults of the clerid C. mutillarius also were attracted to both compounds. The cerambycid Xylotrechus antilope Schönh was significantly attracted to traps baited with (S)-2-hydroxyoctan-3-one alone or the blend containing this compound. (4) Conclusions: Our data confirmed that 3-hydroxyhexan-2-one and 2-hydroxyoctan-3-one are male-produced pheromone components for P. detritus. These results show that both intraspecific and interspecific communication may play key roles in longhorn beetle life history and ecology, with closely and more distantly related species eavesdropping on each other’s signals.

Here, we study the pheromone chemistry of two South American cerambycid beetle species, and their behavioral responses to candidate pheromone components. Adult males of Stizocera phtisica Gounelle (subfamily Cerambycinae: tribe Elaphidiini) produced a sex-specific blend of (R)-3-hydroxyhexan-2-one with lesser amounts of 3-methylthiopropan-1-ol. In field bioassays, traps baited with racemic 3-hydroxyhexan-2-one and 3-methylthiopropan-1-ol did not catch conspecific beetles, but did catch both sexes of a sympatric species, Chydarteres dimidiatus dimidiatus (F.) (Cerambycinae: Trachyderini). We found that males of this species also produce (R)-3-hydroxyhexan-2-one and 3-methylthiopropan-1-ol, and small amounts of 2-phenylethanol. Subsequent bioassays with these compounds showed that a blend of 3-hydroxyhexan-2-one and 3-methylthiopropan-1-ol constitutes the aggregation-sex pheromone of C. d. dimidiatus, with 2-phenylethanol not influencing the attraction of conspecifics. During the field bioassays, six other species in the Cerambycinae also were caught in significant numbers, including Aglaoschema ventrale (Germar) (tribe Compsocerini), congeners Chrysoprasis aurigena (Germar), Chrysoprasis linearis Bates, and an unidentified Chrysoprasis species (Dichophyiini), and Cotyclytus curvatus (Germar) and Itaclytus olivaceus (LaporteGory) (both Clytini), suggesting that one or more of the compounds tested are also pheromone components for these species.

We studied the pheromone chemistry of the cerambycids Anaglyptus mysticus and Xylotrechus antilope ssp. antilope with the goal of identifying attractants that could be used as tools for pheromone-based monitoring of these two species, which are rare and red-listed in parts of northern Europe. Beetles were reared from naturally colonized branches of hazel (Corylus avellana) or oak (Quercus robur), respectively, and used for headspace sampling. The extracts of volatiles were analyzed by gas chromatography–mass spectrometry. Males of A. mysticus consistently produced large quantities of (R)-3-hydroxy-2-hexanone and 2-nonanone, minor amounts of 2,3-hexanedione, and trace amounts of six other compounds. The average proportion of 2-nonanone to (R)-3-hydroxy-2-hexanone was 70:100. Males of X. antilope produced large quantities of (S)-2-hydroxy-3-octanone, and minor quantities of 2,3-octanedione. None of these compounds were present in the corresponding extracts of females from either species. The attractiveness of the dominant compounds produced by each species was determined in field bioassays. Lures with racemic 3-hydroxy-2-hexanone and 2-nonanone as single compounds were not attractive to A. mysticus. However, the compounds acted synergistically in blends of 100:100 and 160:100 of 2-nonanone to the hydroxyketone’s (R)-enantiomer, but not in a 40:100 blend. Similarly (S)-2-hydroxy-3-octanone and its racemate attracted significantly more X. antilope than controls. Males and females of both species were captured in approximately equal numbers. The aggregation-sex pheromones of A. mysticus and X.antilope have high potential to serve as sensitive and efficient tools for detection and monitoring of local populations, and in studies of the species’ ecology and conservation requirements.

Insect pheromones have rarely been exploited in surveys or studies of rare and endangered species, despite their potential as effective and highly selective attractants for target species. Here, we report the identification, synthesis, and field bioassays of a male-produced aggregation-sex pheromone blend of a rare false click beetle species endemic to southern California, Dohrn's elegant eucnemid beetle, Palaeoxenus dohrni Horn (Coleoptera: Eucnemidae). This species is the only extant species in its genus and subfamily. Analyses of extracts of headspace volatiles collected from adult beetles revealed several male-specific compounds. Two of these compounds, identified as (E)-2-nonen-4-one and (R)-2-nonanol, elicited electroantennographic responses from antennae of beetles of both sexes. In field bioassays, a blend of the two compounds attracted both sexes, whereas the individual compounds were not attractive. The identification of an attractant pheromone should provide a useful tool for bioconservation and ecological studies of this iconic species.

Aromia bungii is a serious pest of stone fruit trees including cherries, plums, peaches and apricots. It is native to eastern Asia but has recently been introduced into and has established in Japan, Germany and Italy and has been intercepted in cargo entering the USA and Great Britain. We synthesized the naturally produced enantiomer of the major pheromone component, (E)-2-cis-6,7-epoxynonenal, and in field tests, comparing its attractiveness to that of the racemate. We also tested different ratios of a minor pheromone component, (2E,6Z)-nona-2,6-dienal, on attraction to the major component. Lastly, we conducted a dose–response assay to determine the optimal loading rates. Addition of the minor component at a ratio of 0.31 mg minor to 25 mg major component is more than double the trap captures of males but not females. Using this ratio, we found no difference in trap captures with 10 mg, 32 mg or 100 mg of the major component, but the 10 mg load attracted significantly more females than the 3.2 mg load, and only the 32 mg load attracted significantly more males than the 3.2 mg load. The natural enantiomer was significantly more attractive to females than the racemate. However, the multi-step, low-yielding synthesis likely will prohibit the insect-produced enantiomer from being used for operational trapping. The addition of the minor component to the racemate significantly increased trap catches, and the dose–response assay did not indicate significant differences in loads above 10 mg of the racemic major component when formulated with ~ 1.2% of the minor component.

A number of recent studies have demonstrated the potential for using blends of pheromones of cerambycid beetles to attract several species simultaneously. Here, we tested the effects of adding the pheromones of two invasive species, Trichoferus campestris (Faldermann) and Aromia bungii (Faldermann), on the attraction of native species to a generic blend of synthesized pheromones, in season-long field trials at 12 sites in Pennsylvania. Of the four species attracted in significant numbers, Megacyllene caryae (Gahan), Phymatodes amoenus (Say), and P. testaceus (L.) (all subfamily Cerambycinae) were not significantly affected by the addition of the T. campestris pheromone trichoferone and the A. bungii pheromone (E)-2-cis-6,7-epoxynonenal to the generic blend. In contrast, trap catches of Sternidius alpha (Say) (subfamily Lamiinae) were completely shut down by addition of the pheromones of the two exotic species. In addition, there was no indication that any native species were attracted to trichoferone or (E)-2-cis-6,7-epoxynonenal, suggesting that these pheromones are probably not used by species native to eastern North America.

The transcription factor Doublesex (Dsx) has a conserved function in controlling sexual morphological differences in insects, but our knowledge on its role in regulating sexual behavior is widely limited to Drosophila. Here, we show in the parasitoid wasp Nasonia vitripennis that males whose Dsx gene had been silenced by RNA interference (NvDsx-i) underwent a three-level pheromonal feminization: (1) NvDsx-i males were no longer able to attract females from a distance, owing to drastically reduced titers of the abdominal long-range sex pheromone. (2) NvDsx-i males were courted by wild-type males like females which correlated with a lower abundance of alkenes in their cuticular hydrocarbon (CHC) profiles. Supplementation of NvDsx-i male CHC profiles with realistic amounts of synthetic (Z)-9-hentriacontene (Z9C31), the most significantly reduced alkene in NvDsx-i males, interrupted courtship by wild-type conspecific males. Supplementation of female CHC profiles with Z9C31 reduced courtship and mating attempts by wild-type males. These results prove that Z9C31 is crucial for sex discrimination in Nasonia. (3) Nvdsx-i males were hampered in eliciting female receptivity during courtship and thus experienced severely reduced mating success, suggesting that they are unable to produce the hitherto unidentified oral aphrodisiac pheromone reported in N. vitripennis males. We conclude that Dsx is a multi-level key regulator of pheromone-mediated sexual communication in N. vitripennis. Silencing Dsx by RNA interference provides a new avenue for unraveling the molecular mechanisms underlying the pheromone-mediated sexual communication in insects.

Here, we study the pheromone chemistry of two South American cerambycid beetle species, and their behavioral responses to candidate pheromone components. Adult males of Stizocera phtisica Gounelle (subfamily Cerambycinae: tribe Elaphidiini) produced a sex-specific blend of (R)-3-hydroxyhexan-2-one with lesser amounts of 3-methylthiopropan-1-ol. In field bioassays, traps baited with racemic 3-hydroxyhexan-2-one and 3-methylthiopropan-1-ol did not catch conspecific beetles, but did catch both sexes of a sympatric species, Chydarteres dimidiatus dimidiatus (F.) (Cerambycinae: Trachyderini). We found that males of this species also produce (R)-3-hydroxyhexan-2-one and 3-methylthiopropan-1-ol, and small amounts of 2-phenylethanol. Subsequent bioassays with these compounds showed that a blend of 3-hydroxyhexan-2-one and 3-methylthiopropan-1-ol constitutes the aggregation-sex pheromone of C. d. dimidiatus, with 2-phenylethanol not influencing the attraction of conspecifics. During the field bioassays, six other species in the Cerambycinae also were caught in significant numbers, including Aglaoschema ventrale (Germar) (tribe Compsocerini), congeners Chrysoprasis aurigena (Germar), Chrysoprasis linearis Bates, and an unidentified Chrysoprasis species (Dichophyiini), and Cotyclytus curvatus (Germar) and Itaclytus olivaceus (Laporte & Gory) (both Clytini), suggesting that one or more of the compounds tested are also pheromone components for these species.

The division of labour between reproductive queens and mostly sterile workers is among the defining characteristics of social insects. Queen-produced chemical signals advertising her presence and fertility status, i.e. queen pheromones, are normally used to assert the queen's reproductive dominance in the colony. Most queen pheromones identified to date are chemicals that stop the daughter workers from reproducing. Nevertheless, it has long been suggested that queen pheromones could also regulate reproduction in different ways. In some multiple-queen ants with obligately sterile workers, for example—such as fire ants and pharaoh ants—queen pheromones are thought to regulate reproduction by inhibiting the rearing of new sexuals. Here, we identify the first such queen pheromone in the pharaoh ant Monomorium pharaonis and demonstrate its mode of action via bioassays with the pure biosynthesized compound. In particular, we show that the monocyclic diterpene neocembrene, which in different Monomorium species is produced solely by fertile, egg-laying queens, strongly inhibits the rearing of new sexuals (queens and males) and also exerts a weakly attractive ‘queen retinue’ effect on the workers. This is the first time that a queen pheromone with such a dual function has been identified in a social insect species with obligately sterile workers.

Simple Summary The vine mealybug is a key insect pest of vineyards that currently is controlled by one or more insecticide applications per season. Here, we sought to develop a more sustainable control tool by using the mealybug’s sex pheromone to reduce mating and thereby lower pest damage. The mature female mealybug emits a sex pheromone that the winged adult male uses to find and mate with females. Synthetically produced sex pheromone, specific to the vine mealybug, was enclosed in commercial dispensers and deployed in vineyards in 2004–2007 studies to determine if mating disruption could provide a viable control option. Trials were conducted in commercial vineyards with cooperating farmers. Across all trials, mating disruption reduced pheromone trap captures of adult male mealybugs—an indication that the population numbers were lowered—and there was often a reduction mealybug numbers on vines and/or crop damage. There was not a clear reduction in the proportion of female mealybugs with ovisacs (a cottony-like mass containing mealybug eggs), but this may have resulted from the production of non-viable ovisacs that were not differentiated in the field samples. Pheromone trap captures were never lowered to zero (often called trap shut down), possibly because trials were conducted in vineyards with unusually high mealybug densities. Trap capture patterns commonly began low in April-May, increased in mid-July or August, and often decreased in September–October when post-harvest insecticides were applied. Results over all years suggest season-long coverage or late season coverage may be as or more important than dose per hectare. This research was used to help initiate the commercialization of mating disruption products for the vine mealybug, which are now being successfully used throughout the world’s grape-growing regions where this pest is found. Abstract The vine mealybug (VMB), Planococcus ficus (Hemiptera: Pseudococcidae), is a key insect pest of vineyards, and improvements in sustainable control of this pest are needed to meet increasing consumer demand for organically farmed products. One promising option is mating disruption. In a series of experiments conducted from 2004 to 2007, we tested the effects of mating disruption on trap captures of Pl. ficus males in pheromone-baited traps, on Pl. ficus numbers and age structure on vines, and on damage to grape clusters. From 2004 to 2005, the effects of dispenser load (mg active ingredient per dispenser) were also assessed, and dispensers were compared to a flowable formulation. Across all trials, mating disruption consistently reduced pheromone trap captures and often reduced mealybug numbers on vines and/or crop damage, regardless of the pheromone dose that was applied. Reductions in Pl. ficus densities in mating disruption plots were not accompanied by clear effects on mealybug population age structure; however, production of non-viable ovisacs by unmated females may have obscured differences in proportional representation of ovisacs. Pheromone trap captures were never lowered to zero (often called trap shut down), possibly because trials were conducted in vineyards with unusually high Pl. ficus densities. Trap-capture patterns in both treated and control plots commonly began low in April–May, increased in mid-July or August, and often decreased in September–October when post-harvest insecticides were applied. During the four-year trial, the release rate from plastic sachet dispensers was improved by industry cooperators as pheromone was released too quickly (2004) or not completely released during the season (2005–2006). The flowable formulation performed slightly better than dispensers at the same application dose. Results over all years suggest season-long coverage or late-season coverage may be as or more important than dose per hectare. Development of a dispenser with optimized season-long pheromone emission or targeted seasonal periods should be a future goal.

Reproductive division of labor in insect societies is regulated through multiple concurrent mechanisms, primarily chemical and behavioral. Here, we examined if the Dufour’s gland secretion in the primitively eusocial bumble bee Bombus impatiens signals information about caste, social condition, and reproductive status. We chemically analyzed Dufour’s gland contents across castes, age groups, social and reproductive conditions, and examined worker behavioral and antennal responses to gland extracts.We found that workers and queens each possess caste-specific compounds in their Dufour’s glands. Queens and gynes differed from workers based on the presence of diterpene compounds which were absent in workers, whereas four esters were exclusive to workers. These esters, as well as the total amounts of hydrocarbons in the gland, provided a separation between castes and also between fertile and sterile workers. Olfactometer bioassays demonstrated attraction of workers to Dufour’s gland extracts that did not represent a reproductive conflict, while electroantennogram recordings showed higher overall antennal sensitivity in queenless workers. Our results demonstrate that compounds in the Dufour’s gland act as caste- and physiology-specific signals and are used by workers to discriminate between workers of different social and reproductive status.

Brassicadiene, a novel tricyclic diterpene hydrocarbon, was identified by a combination of mass spectrometry, microchemical tests, and analysis of NMR spectra. The compound constitutes90% of the volatile organic compounds produced by cauliflower seedlings

The original version of this article unfortunately contained a mistake. When filing the final publication details, we failed to include the following statement in our Acknowledgments paragraph: We thank the Almond Board of California for research funding.

The objective of our project was to identify the sex attractant pheromone of queens of the slave-making ant Polyergus samurai, which was characterized as a blend of methyl 6-methylsalicylate and methyl 3-ethyl-4-methylpentanoate. Both compounds were identified in volatiles collected from squashed heads of queens, and in field trials, a blend of the two compounds was highly attractive to males. In contrast, males were not attracted to a blend of methyl 6-methylsalicylate with 3-ethyl-4-methylpentanol. Thus, the pheromone blend of this species is analogous to that of species in the Polyergus lucidus complex from eastern North America, rather than that of the Polyergus breviceps complex from western North America and the European species P. rufescens. These results are discussed in the context of speciation within the genus.

Chemosensory signals play a key role in species recognition and mate location in both invertebrate and vertebrate species. Closely related species often produce similar but distinct signals by varying the ratios or components in pheromone blends to avoid interference in their communication channels and minimize cross-attraction among congeners. However, exploitation of reproductive signals by predators and parasites also may provide strong selective pressure on signal phenotypes. For example, bolas spiders mimic the pheromones of several moth species to attract their prey, and parasitic blister beetle larvae, known as triungulins, cooperatively produce an olfactory signal that mimics the sex pheromone of their female host bees to attract male bees, as the first step in being transported by their hosts to their nests. In both cases, there is strong selection pressure on the host to discriminate real mates from aggressive mimics and, conversely, on the predator, parasite, or parasitoid to track and locally adapt to the evolving signals of its hosts. Here we show local adaptation of a beetle, Meloe franciscanus (Coleoptera: Meloidae), to the pheromone chemistry and mate location behavior of its hosts, two species of solitary bees in the genus Habropoda . We report that M . franciscanus ’ deceptive signal is locally host-adapted in its chemical composition and ratio of components, with host bees from each allopatric population preferring the deceptive signals of their sympatric parasite population. Furthermore, in different locales, the triungulin aggregations have adapted their perching height to the height at which local male bees typically patrol for females.

The longhorned beetle Arhopalus rusticus (Coleoptera: Cerambycidae, Spondylinae) is a common species in conifer forests of the Northern Hemisphere, but with global trade, it has invaded and become established in New Zealand, Australia, and South America. Arhopalus rusticus is a suspected vector of the phytopathogenic nematode, Bursaphelenchus xylophilus, the causative agent of pine wilt disease, which is a major threat to pine forests worldwide. Here, we report the identification of a volatile, male-produced aggregation-sex pheromone for this species. Headspace odours from males contained a major male-specific compound, identified as (2 S, 5E)-6,10-dimethyl-5,9-undecadien-2-ol (common name (S)-fuscumol), and a minor component (E)-6,10-dimethyl-5,9-undecadien-2-one (geranylacetone). Both compounds are known pheromone components for species in the same subfamily. In field trials in its native range in Slovenia, (S)-fuscumol was significantly more attractive to beetles of both sexes, than racemic fuscumol and a blend of host plant volatiles commonly used as an attractant for this species. Fuscumol-baited traps also caught significant numbers of another spondylidine species, Spondylis buprestoides (L.), and a rare click beetle, Stenagostus rufus (De Geer). The pheromone can be exploited as a cost-effective and environmentally safe tool for detection and monitoring of this invasive species at ports of entry, and for monitoring the beetle’s distribution and population trends in both endemic and invasive populations.

In field trials testing attraction of cerambycid beetles to a blend of known pheromone components plus host plant volatiles, several species in the subfamily Spondylidinae were attracted to baited traps, suggesting that one or more components of the blend might constitute their pheromones. Here, we describe laboratory and field experiments aimed at identifying the actual pheromone components produced by these species. Analysis of headspace odors collected from male Tetropium abietis (Fall) (Coleoptera: Cerambycidae) contained (S)-fuscumol as a single component, whereas Asemum nitidum (LeConte) (Coleoptera: Cerambycidae) males produced both (S)-fuscumol and geranylacetone, and Asemum caseyi (Linsley) (Coleoptera: Cerambycidae) produced only geranylacetone. In field trials testing fuscumol, fuscumol acetate, and geranylacetone as individual components or in blends, in combination with host plant volatiles, A. nitidum were attracted to blends of fuscumol and geranylacetone, T. abietis were attracted to fuscumol alone, and A. caseyi were attracted to geranylacetone alone. Fuscumol acetate did not appear to be either attractive or inhibitory. These results, along with previous catches of spondylidine species in traps baited with fuscumol, provide evidence that fuscumol and geranylacetone are likely to be relatively common pheromone structures for species in the subfamily Spondylidinae.

In geographic regions with warm winters, invasive yellowjacket wasp colonies (genus Vespula) often exhibit polygyny (multiple queens) and persist for multiple years, despite these phenomena being rare in the native range. Here, we test the hypothesis that polygyny, caused by foreign queens being accepted into an existing colony, is the result of relaxed nestmate recognition in the invasive range, as has been observed in some supercolonial invasive ants. In bioassays with wild colonies in the field, we found that nestmate discrimination was weak in both invasive (Hawaii) and native (California) populations of Vespula pensylvanica, with significant nestmate discrimination in only ~ 30% of trials. We also found that the diversity and variability of cuticular hydrocarbons, chemical compounds that mediate nestmate recognition, were not reduced in introduced populations, unlike several supercolonial invasive ant species. Our findings suggest that ancestral weak nestmate discrimination behavior of V. pensylvanica may make this species pre-adapted to transition to polygyny and extended colony lifespans when introduced into environments with benign winters that facilitate foreign queens joining existing colonies in late season.