Specialized odorant receptors in social insects that detect cuticular hydrocarbon cues and candidate pheromones.

Eusocial insects use cuticular hydrocarbons as components of pheromones that mediate social behaviours, such as caste and nestmate recognition, and regulation of reproduction. In ants such as Harpegnathos saltator, the queen produces a pheromone which suppresses the development of workers' ovaries and if she is removed, workers can transition to a reproductive state known as gamergate. Here we functionally characterize a subfamily of odorant receptors (Ors) with a nine-exon gene structure that have undergone a massive expansion in ants and other eusocial insects. We deorphanize 22 representative members and find they can detect cuticular hydrocarbons from different ant castes, with one (HsOr263) that responds strongly to gamergate extract and a candidate queen pheromone component. After systematic testing with a diverse panel of hydrocarbons, we find that most Harpegnathos saltator Ors are narrowly tuned, suggesting that several receptors must contribute to detection and discrimination of different cuticular hydrocarbons important in mediating eusocial behaviour.Cuticular hydrocarbons (CHC) mediate the interactions between individuals in eusocial insects, but the sensory receptors for CHCs are unclear. Here the authors show that in ants such as H. saltator, the 9-exon subfamily of odorant receptors (HsOrs) responds to CHCs, and ectopic expression of HsOrs in Drosophila neurons imparts responsiveness to CHCs

Male-Produced Aggregation Pheromones of the Cerambycid Beetles Xylotrechus colonus and Sarosesthes fulminans

Adults of both sexes of the cerambycid beetles Xylotrechus colonus (F.) and Sarosesthes fulminans (F.) were attracted to odors produced by male conspecifics in olfactometer bioassays. Analyses of headspace volatiles from adults revealed that male X. colonus produced a blend of (R)- and (S)-3-hydroxyhexan-2-one and (2 S,3 S)- and (2R,3R)-2,3-hexanediol, whereas male S. fulminans produced (R)-3-hydroxyhexan-2-one and (2 S,3R)-2,3-hexanediol. All of these compounds were absent in the headspace of females. Two field bioassays were conducted to confirm the biological activity of the synthesized pheromones: (1) enantiomerically enriched pheromone components were tested singly and in species-specific blends and (2) four-component mixture of racemic 3-hydroxyhexan-2-one plus racemic 2-hydroxyhexan-3-one and the four-component blend of the stereoisomers of 2,3-hexanediols were tested separately and as a combined eight-component blend. In these experiments, adult male and female X. colonus were captured in greatest numbers in traps baited with the reconstructed blend of components produced by males, although significant numbers were also captured in traps baited with (R)-3-hydroxyhexan-2-one alone or in blends with other compounds. Too few adult S. fulminans were captured for a statistical comparison among treatments, but all were caught in traps baited with lures containing (R)-3-hydroxyhexan-2-one. In addition to these two species, adults of two other species of cerambycid beetles, for which pheromones had previously been identified, were caught: Neoclytus a. acuminatus (F.) and its congener Neoclytus m. mucronatus (F.). Cross-attraction of beetles to pheromone blends of other species, and to individual pheromone components that are shared by two or more sympatric species, may facilitate location of larval hosts by species that compete for the same host species

Male-Produced Aggregation Pheromones of the Cerambycid Beetles Xylotrechus colonus and Sarosesthes fulminans

Adults of both sexes of the cerambycid beetles Xylotrechus colonus (F.) and Sarosesthes fulminans (F.) were attracted to odors produced by male conspecifics in olfactometer bioassays. Analyses of headspace volatiles from adults revealed that male X. colonus produced a blend of (R)- and (S)-3-hydroxyhexan-2-one and (2 S,3 S)- and (2R,3R)-2,3-hexanediol, whereas male S. fulminans produced (R)-3-hydroxyhexan-2-one and (2 S,3R)-2,3-hexanediol. All of these compounds were absent in the headspace of females. Two field bioassays were conducted to confirm the biological activity of the synthesized pheromones: (1) enantiomerically enriched pheromone components were tested singly and in species-specific blends and (2) four-component mixture of racemic 3-hydroxyhexan-2-one plus racemic 2-hydroxyhexan-3-one and the four-component blend of the stereoisomers of 2,3-hexanediols were tested separately and as a combined eight-component blend. In these experiments, adult male and female X. colonus were captured in greatest numbers in traps baited with the reconstructed blend of components produced by males, although significant numbers were also captured in traps baited with (R)-3-hydroxyhexan-2-one alone or in blends with other compounds. Too few adult S. fulminans were captured for a statistical comparison among treatments, but all were caught in traps baited with lures containing (R)-3-hydroxyhexan-2-one. In addition to these two species, adults of two other species of cerambycid beetles, for which pheromones had previously been identified, were caught: Neoclytus a. acuminatus (F.) and its congener Neoclytus m. mucronatus (F.). Cross-attraction of beetles to pheromone blends of other species, and to individual pheromone components that are shared by two or more sympatric species, may facilitate location of larval hosts by species that compete for the same host species

Novel Lepidopteran Sex Pheromone Components From Marmara gulosa (Lepidoptera: Gracillariidae)

Marmara gulosa Guillén & Davis (Lepidoptera: Gracillariidae) is a sporadic pest of citrus and a number of other crops in southern and central California. Coupled gas chromatographic-electroantennogram detection analyses of headspace volatiles collected by solid phase microextraction from virgin female moths revealed at least four related compounds in the extracts that elicited significant antennal responses from antennae of male moths. These compounds were identified as (8E,10E)-tetradecadien-1-ol, and the corresponding aldehyde, acetate, and formate ester, representing the first report of a formate as a lepidopteran pheromone component. The four compounds were consistently found in headspace volatiles collected from virgin female moths from different regions of the state and from M. gulosa collected from different host plants (citrus and squash). Repeated field trials determined that the formate ester alone was as attractive or more attractive than any blend of the formate with one or more of the remaining compounds. Although large numbers of moths were caught in some field trials, trap catches were not consistent. Thus, the pheromone may be useful for detection of the moth and setting an initial biofix, but it remains unclear whether the pheromone can be used as a reliable and accurate tool for monitoring densities of M. gulosa populations

North American Species of Cerambycid Beetles in the Genus Neoclytus Share a Common Hydroxyhexanone-Hexanediol Pheromone Structural Motif

Many species of cerambycid beetles in the subfamily Cerambycinae are known to use male-produced pheromones composed of one or a few components such as 3-hydroxyalkan-2-ones and the related 2,3-alkanediols. Here, we show that this pheromone structure is characteristic of the cerambycine genus Neoclytus Thomson, based on laboratory and field studies of 10 species and subspecies. Males of seven taxa produced pheromones composed of (R)-3-hydroxyhexan-2-one as a single component, and the synthetic pheromone attracted adults of both sexes in field bioassays, including the eastern North American taxa Neoclytus caprea (Say), Neoclytus mucronatus mucronatus (F.), and Neoclytus scutellaris (Olivier), and the western taxa Neoclytus conjunctus (LeConte), Neoclytus irroratus (LeConte), and Neoclytus modestus modestus Fall. Males of the eastern Neoclytus acuminatus acuminatus (F.) and the western Neoclytus tenuiscriptus Fall produced (2S,3S)-2,3-hexanediol as their dominant or sole pheromone component. Preliminary data also revealed that males of the western Neoclytus balteatus LeConte produced a blend of (R)-3-hydroxyhexan-2-one and (2S,3S)-2,3-hexanediol but also (2S,3S)-2,3-octanediol as a minor component. The fact that the hydroxyketone-hexanediol structural motif is consistent among these North American species provides further evidence of the high degree of conservation of pheromone structures among species in the subfamily Cerambycinae

Identification of Pheromone Components of Plagionotus detritus (Coleoptera: Cerambycidae), and Attraction of Conspecifics, Competitors, and Natural Enemies to the Pheromone Blend

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