Aggregation Pheromone Components Research Articles

A conserved pheromone receptor in the American and the Asian palm weevils is also activated by host plant volatiles

The evolution of chemosensory receptors is key for the adaptation of animals to their environment. Recent knowledge acquired on the tri-dimensional structure of insect odorant receptors makes it possible to study the link between modifications in the receptor structure and evolution of response spectra in more depth. We investigated this question in palm weevils, several species of which are well-known invasive pests of ornamental or cultivated palm trees worldwide. These insects use aggregation pheromones to gather on their host plants for feeding and reproduction. An odorant receptor detecting the aggregation pheromone components was characterised in the Asian palm weevil Rhynchophorus ferrugineus. This study compared the response spectra of this receptor, RferOR1, and its ortholog in the American palm weevil R. palmarum, RpalOR1. Sequences of these two receptors exhibit more than 70 amino acid differences, but modelling of their 3D structures revealed that their putative binding pockets differ by only three amino acids, suggesting possible tuning conservation. Further functional characterization of RpalOR1 confirmed this hypothesis, as RpalOR1 and RferOR1 exhibited highly similar responses to coleopteran aggregation pheromones and chemically related molecules. Notably, we showed that R. ferrugineus pheromone compounds strongly activated RpalOR1, but we did not evidence any response to the R. palmarum pheromone compound rhynchophorol. Moreover, we discovered that several host plant volatiles also activated both pheromone receptors, although with lower sensitivity. This study not only reveals evolutionary conservation of odorant receptor tuning across the two palm weevil species, but also questions the specificity of pheromone detection usually observed in insects.

Comparative Transcriptomics Analysis Reveals Rusty Grain Beetle's Aggregation Pheromone Biosynthesis Mechanism in Response to Starvation.

Pheromones are the basis of insect aggregation, mating, and other behaviors. Cucujoid grain beetles produce macrocyclic lactones as aggregation pheromones, yet research on their biosynthesis at the molecular level remains limited. The rusty grain beetle, C. ferrugineus, is an important economic species in China. Although two aggregation pheromone components have been identified, their suspected biosynthesis via the MVA pathway and the FAS pathway lacks molecular elucidation. Previous evidence supports that starvation affects the production of aggregation pheromones. Therefore, we constructed comparative transcriptome libraries of pheromone production sites in C. ferrugineus under starvation stress and identified genes related to pheromone biosynthesis and hormone regulation. A total of 2665 genes were significantly differentially expressed, of which 2029 genes were down-regulated in starved beetles. Putative C. ferrugineus genes directly involved in pheromone biosynthesis were identified, as well as some genes related to the juvenile hormone (JH) pathway and the insulin pathway, both of which were depressed in the starved beetles, suggesting possible functions in pheromone biosynthesis and regulation. The identification of genes involved in macrolide lactone biosynthesis in vivo holds great significance, aiding in the elucidation of the synthesis and regulatory mechanisms of cucujoid grain beetle pheromones.

(−)‐myrtenol and (−)‐α‐pinene: Aggregation pheromone components of the cypress bark beetle Phloeosinus aubei

AbstractThe cypress bark beetle, Phloeosinus aubei, is of increasing concern in the European market of scale‐leafed conifers. Neither the reproductive behaviour nor the chemical ecology have been studied in detail so far, and no specific attractant traps are available to monitor the flight of adults. We established that stridulation by males is crucial for acceptance by calling females. To unravel the chemical communication, hindgut extracts from unmated females were analysed by GC‐EAD, using male antennae. Gut extracts and Thuja occidentalis leaf extracts were analysed by GC/MS. Two antennally active compounds from gut extracts were identified as (−)‐myrtenol and (−)‐α‐pinene by enantioselective GC/MS. (−)‐Myrtenol was shown to be female‐specific, while (−)‐α‐pinene was found as a 2:1 mixture with the (+)‐enantiomer also among the volatiles of the host tree, T. occidentalis. To prove that the production of the above compounds in the beetles is related to reproduction, we treated females with JHIII. The gut extracts from these sexually mature females showed elevated levels of (−)‐α‐pinene compared to untreated control groups. Synthetic enantiomers of myrtenol and α‐pinene were tested separately by electroantennography, and all were found to be electrophysiologically active. The attractiveness of different mixtures of these compounds was compared using 4‐arm olfactometer tests: 4:1 mixtures of racemic α‐pinene with either (−)‐ or (+)‐myrtenol were significantly attractive to males and females, respectively. In field tests, multifunnel traps baited with a 5:1 mixture of (−)‐α‐pinene and (−)‐myrtenol caught significantly more males and females than unbaited control traps. We concluded that (−)‐α‐pinene and (−)‐myrtenol are components of a female‐produced aggregation pheromone. EAD activity and behaviour assays indicated that α‐pinene from Thuja is used by the beetles as part of a host plant kairomone. The complex communication system of P. aubei is discussed in a chemoecological context.

Even- and odd-numbered fatty acids used as worker arrestant aggregation pheromone components in the Formosan subterranean termite Coptotermes formosanus

Even- and odd-numbered fatty acids used as worker arrestant aggregation pheromone components in the Formosan subterranean termite Coptotermes formosanus

Scalable synthesis of methyl (E,E,Z)-2,4,6-decatrienoate, the aggregation pheromone component of the brown-winged green bug, Plautia stali

Methyl (E,E,Z)-2,4,6-decatrienoate (1) is the aggregation pheromone of the brown-winged green bug, Plautia stali and is also a pheromone synergist of the brown marmorated stink bug, Halyomorpha halys pheromone. A scalable and controllable synthesis of methyl (E,E,Z)-2,4,6-decatrienoate (1) was achieved in 5 steps. The stereoselective formation of the 4E, 6Z conjugated double bond was accomplished by a tandem addition reaction of acrolein with acetylene and the 2E double bond was formed by a Heck reaction. The pheromone was obtained in 99 % stereochemical purity and 40 % overall yield starting from (Z,E)-7-bromo-4,6-heptadienal.

A novel physiological function of pheromone biosynthesis-activating neuropeptide in production of aggregation pheromone

The western flower thrips, Frankliniella occidentalis, is an insect pest, and its aggregation pheromone (AP) plays a crucial role in the recruitment of both sexes. A novel pheromone biosynthesis-activating neuropeptide (PBAN)-like gene is encoded in F. occidentalis genome, but its physiological function has yet to be elucidated. This study hypothesized the physiological role played by PBAN in mediating AP production. AP has been known to be produced only by male adults in F. occidentalis. Surprisingly, our extraction of headspace volatiles contained two AP components in females as well as in males with similar composition. PBAN injection elevated the AP production whereas RNA interference (RNAi) of the gene expression suppressed the AP production in both sexes. A biosynthetic pathway to produce AP components were predicted and the enzymes catalyzing the main steps were confirmed in their expressions. Individual RNAi treatments of these genes significantly suppressed AP production. RNAi of PBAN gene downregulated the expressions of these biosynthesis-associated genes in both sexes. These results suggest that the novel neuropeptide acts as PBAN mediating AP production through stimulating its biosynthetic machinery in F. occidentalis.

Spatial distribution of histamine in bed bug-infested homes

Histamine is a component of the bed bug aggregation pheromone. It was recently identified as an environmental contaminant in homes with active bed bug infestations, posing a potential health risk to humans via skin contact or inhalation. It remains unclear how histamine is distributed in homes and if histamine can become airborne. In the present study, histamine levels in household dust were quantified from multiple locations within bed bug infested and uninfested apartments. Bed bug population levels were quantified using both traps and visual counts. The amount of histamine detected varied significantly with respect to sampling location, with the highest concentration of histamine quantified from bedding material. Infestation severity did not have a significant effect on histamine quantified at any location. Our results indicate that the bedroom should be the primary focus of histamine mitigation efforts, although histamine can be found throughout the home. Histamine quantified from homes without active bed bug infestations suggests that histamine from previous infestations can persist following pest eradication. These findings highlight the importance of histamine as a potential insect allergen and will be important for the development of targeted mitigation strategies of bed bug histamine.

Linoleic Acid Promotes Emission of Bark Beetle Semiochemicals by Fungal Symbionts

Tree-killing bark beetles in conifer forests vector symbiotic fungi that are thought to help the beetles kill trees. Fungal symbionts emit diverse volatile blends that include bark beetle semiochemicals involved in mating and host localization. In this study, all 12 tested fungal isolates emitted beetle semiochemicals when growing in medium amended with linoleic acid. These semiochemicals included the spiroacetals chalcogran, trans-conophthorin and exo-brevicomin, as well as 2-methyl-3-buten-1-ol, the main aggregation pheromone component of the spruce bark beetle Ips typographus. The emission of these compounds was affected by the type of fatty acid present (linoleic vs. oleic acid). Accumulating evidence shows that the fatty acid composition in conifer bark can facilitate colonization by bark beetles and symbiotic fungi, whereas the fatty acid composition of non-host trees can be detrimental for beetle larvae or fungi. We hypothesize that beetles probe the fatty acid composition of potential host trees to test their suitability for beetle development and release of semiochemicals by symbiotic fungi.

Generic Pheromones Identified from Northern Hemisphere Cerambycidae (Coleoptera) Are Attractive to Native Longhorn Beetles from Central-Southern Chile.

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.

Role of 3-Carene in Host Location and Colonization by Dendroctonus pseudotsugae (Coleoptera: Curculionidae).

The Douglas-fir beetle (Dendroctonus pseudotsugae Hopkins) occasionally colonizes western larch [Larix occidentalis Nutt. (Pinales: Pinaceae)] growing in close proximity to its primary host, Douglas-fir [Pseudotsuga menziesii (Mirb.) Franco (Pinales: Pinaceae)], but brood have never been found to survive in live western larch. Western larch produces the monoterpene 3-carene in higher concentrations than Douglas-fir. In this study, the toxicity and repellency of 3-carene to Douglas-fir beetle was evaluated in a series of laboratory and field tests. In a laboratory bioassay, 3-carene was one of the most toxic monoterpenes to Douglas-fir beetles among those tested. In a field trial, addition of 3-carene to multiple-funnel traps baited with frontalin (the primary component of Douglas-fir beetle aggregation pheromone) or frontalin and α-pinene significantly reduced the number of Douglas-fir beetles collected. In another field study, live western larch, felled western larch, live Douglas-fir, felled Douglas-fir, and live Douglas-fir surrounded by 3-carene releasers were baited with Douglas-fir beetle aggregation pheromones. There were significantly fewer Douglas-fir beetle entrance holes and egg galleries excavated on both live western larch and live Douglas-fir surrounded by 3-carene compared with live Douglas-fir. Most egg galleries excavated in live western larch were heavily impregnated with resin and no eggs hatched. There were no significant differences in egg galleries excavated or eggs hatched between felled western larch and felled Douglas-fir. Collectively, these data support the hypothesis that 3-carene slows the colonization process in live western larch allowing more time for host trees to respond to a colonization attempt and a higher likelihood of successfully resisting infestation.

Some Effects of endo-Brevicomin Background on Southern Pine Beetle (Coleoptera: Curculionidae: Scolytinae) Aggregation Behavior

Semiochemical background in the environment can influence insect orientation to release points of the same or different semiochemicals. endo-Brevicomin is a pheromone component of the tree-killing bark beetle Dendroctonus frontalis Zimmermann (Coleoptera: Curculionidae: Scolytinae) that has a biphasic dose-response curve, enhancing attraction at low release rates but reducing attraction at high rates. We investigated the effect of artificial manipulation of background levels of endo-brevicomin on D. frontalis responses to sources of aggregation attractant in the field. Traps baited with the aggregation pheromone component frontalin and the host odor alpha-pinene were deployed either with or without a background of endo-brevicomin produced by three surrounding dispensers of this semiochemical each located 20 m away. Two tested levels of endo-brevicomin background caused catches to increase by an order of magnitude above those in the absence of background. Presence of background also altered the beetles' biphasic dose-response when endo-brevicomin dispensers were added to traps. Background reduced or concealed attraction-enhancement otherwise observed for low-release dispensers added to traps, and it decreased the release rate necessary to produce reductions in catches. We propose that spatial variability in abundance of natural, background sources of endo-brevicomin in the environment (i.e., infested trees) is a cause of the observed variability in effects of endo-brevicomin dispensers on southern pine beetle behavior in the field. Furthermore, our results illustrate the potential complexity of the density-dependent effects of biphasic pheromone components on bark beetle mass attack and colonization behavior.

Identification of male produced compounds in the bark beetle Polygraphus subopacus and establishment of (Z)-2-(3,3-dimethylcyclohexylidene)-ethanol as an aggregation pheromone component

Bark beetles of the genus Polygraphus have recently been involved in large bark beetle outbreaks in central Sweden, together with the European spruce bark beetle Ips typographus. Three species of Polygraphus can be found in this region; Polygraphus poligraphus, Polygraphus punctifrons and Polygraphus subopacus. Efficient pheromone traps would facilitate further investigations of these species and their role in bark beetle outbreaks. Pheromone compounds have previously been identified in P. poligraphus and P. punctifrons, but not in P. subopacus. Thus, we allowed males and females of P. subopacus to bore in the bark of stem sections of Norway spruce (Picea abies) in the laboratory. Volatile organic compounds from boring insects were sampled with SPME and analysed with GC–MS and several male-specific compounds were observed. The male specific compounds were 3-methyl-3-buten-1-ol, 3-methyl-2-buten-1-ol, 3-methyl-2-butenal, grandisol, fragranol, (Z)-2-(3,3-dimethylcyclohexylidene)-ethanol, (E)-2-(3,3-dimethylcyclohexylidene)-ethanol, (Z)-2-(3,3-dimethylcyclohexylidene)-acetaldehyde, (E)-2-(3,3-dimethylcyclohexylidene)-acetaldehyde, geranial and γ-isogeraniol. (Z)-2-(3,3-dimethylcyclohexylidene)-ethanol, [(Z)-DMCHE], was identified from GC–MS analysis to be the major male-specific compound while the (E)-isomer, [(E)-DMCHE], was found as a minor compound. These two compounds gave positive responses in EAG analyses with antennae from males and females of P. subopacus. Thus, (Z)- and (E)-DMCHE were used in a field experiment in central Sweden but only (Z)-DMCHE was found to be attractive to males and females of P. subopacus. Consequently, (Z)-DMCHE was established to be a component of P. subopacus aggregation pheromone.

Rectal gland exudates and emissions of Bactrocera bryoniae: chemical identification, electrophysiological and pheromonal functions

Bactrocera bryoniae is a polyphagous and economically significant fruit fly found in Indonesia, Papua New Guinea and Australia. To understand chemical-mediated sexual communication, and the potential for novel pheromone-based attractants for monitoring and mass-trapping of B. bryoniae, rectal gland exudates and emissions from sexually mature males and females were investigated. Gas chromatography–mass spectrometry showed that male rectal glands contained six compounds, of which 1,7-dioxaspiro[5,5]undecane elicited electroantennographic (EAD) and electropalpographic (EPD) responses in both sexes, ethyl 3-acetoxybutanoate elicited EPD responses in both sexes, N-(3-methylbutyl)acetamide elicited EAD response from males and 4-hydroxy-1,7-dioxaspiro[5.5]undecane elicited EAD responses in males and females and EPD responses in females. Female rectal glands contained 23 compounds with the esters ethyl laurate and ethyl myristate as major components. Amongst the female rectal gland constituents, ethyl laurate, ethyl myristate and ethyl palmitate elicited EAD responses in males and females, N-(3-methylbutyl)acetamide elicited EAD responses in males only, (E,E)-2,8-dimethyl-1,7-dioxaspiro[5.5]undecane elicited EAD responses in males and EPD responses in females, and 2,7-dimethyl-1,6-dioxaspiro[4.5]decane, (E,E)-2-ethyl-7-methyl-1,6-dioxaspiro[4.5]decane, (E,E)-2-ethyl-8-methyl-1,7-dioxaspiro[5.5]undecane, (Z,Z)-2,8-dimethyl-1,7-dioxaspiro[5.5]undecane, (E,E)-2-propyl-8-methyl-1,7-dioxaspiro[5.5]undecane and ethyl caprate elicited EPD responses in females only. Y-tube bioassays indicated that male rectal gland extracts and headspace volatiles attracted females and males, while female rectal gland extracts and headspace volatiles only attracted males. The results suggest that ethyl 3-acetoxybutanoate, 1,7-dioxaspiro[5,5]undecane and 4-hydroxy-1,7-dioxaspiro[5.5]undecane may be components of male-produced sex pheromone in B. bryoniae while (E,E)-2,8-dimethyl-1,7-dioxaspiro[5.5]undecane, N-(3-methylbutyl)acetamide, ethyl laurate, ethyl myristate and ethyl palmitate may be components of female-produced sex pheromone. Ethyl 3-acetoxybutanoate, N-(3-methylbutyl) acetamide, 1,7-dioxaspiro[5,5]undecane and 4-hydroxy-1,7-dioxaspiro[5.5]undecane may be components of male aggregation pheromone. These findings contribute to the understanding of pheromone communication in B. bryoniae and provide a foundation for developing pheromone-based monitoring and control methods.

Trapping Failure Leads to Discovery of Potent Semiochemical Repellent for the Walnut Twig Beetle.

The walnut twig beetle, Pityophthorus juglandis Blackman, and its associated fungal pathogen that causes thousand cankers disease, currently threaten the viability of walnut trees across much of North America. During a 2011 assessment of seasonal flight patterns of P. juglandis with yellow sticky traps baited with the male-produced aggregation pheromone component, 3-methyl-2-buten-1-ol, dramatically reduced catches were recorded when Tree Tanglefoot adhesive was used to coat the traps. In summer 2011, two trap adhesives were tested for potential repellency against P. juglandis in a field trapping bioassay. SuperQ extracts of volatiles from the most repellent adhesive were analyzed by gas chromatography-mass spectrometry, and limonene and α-pinene were identified as predominant components. In field-based, trapping experiments both enantiomers of limonene at a release rate of ~700 mg/d conferred 91-99% reduction in trap catches of P. juglandis to pheromone-baited traps. (+)- and (‒)-α-Pinene reduced trap catch by 40 and 53%, respectively, at the highest release rate tested. While a combination of R-(+)-limonene and (+)-α-pinene resulted in a 97% reduction in the number of P. juglandis caught, the combination did not consistently result in greater flight trap catch reduction than individual limonene enantiomers. The repellent effect of limonene may be valuable in the development of a semiochemical-based tool for management of P. juglandis and thousand cankers disease.

Chemical signal interactions of the bark beetle with fungal symbionts, and host/non-host trees

The symbiosis between the bark beetle (Ips subelongatus) and its fungal symbiont (Endoconidiophora fujiensis) poses a serious threat to larch forests. However, the signaling pathways between these symbiotic partners and their host/non-host trees are not fully understood. Inoculation of the host larch (Larix principis-rupprechtii) with two strains of E. fujiensis induced a rapid and long-term release of monoterpenes. Although the fungi had a level of tolerance to these compounds, many monoterpenes inhibited fungal growth in culture. Moreover, monoterpenes with stronger inhibitory effects on fungal growth exhibited weaker synergistic effects on the attraction of I. subelongatus to aggregation pheromone. Surprisingly, individual isomers of aggregation pheromone components promoted fungal symbiont growth in a culture medium. Non-host volatiles (NHVs) were tested and shown to completely inhibit the growth of fungal symbionts in culture but had no effects on beetle responses to aggregation pheromone, with the exception of (Z)-3-hexen-1-ol. These results reveal convergence and mutualism patterns in the evolution of I. subelongatus and E. fujiensis with respect to host tree volatiles but not in response to NHVs. Ultimately, we put forward a hypothesis that host plants are ecological and evolutionary determinants of bark beetle-fungus symbioses in terms of their complex signaling interactions.

Local, Geographical, and Contextual Variation in the Aggregation Pheromone Blend of the Spruce Beetle, Dendroctonus rufipennis (Coleoptera: Curculionidae)

Prior research from trapping experiments using synthetic pheromone components suggested the existence of local and geographical variation in the pheromone system of the spruce beetle, Dendroctonus rufipennis (Kirby). To test hypotheses concerning variation in the spruce beetle's aggregation pheromone blend, we extracted volatiles from the hindguts of individual spruce beetles sampled from sites in eastern Canada (Nova Scotia and Newfoundland) and western Canada (British Columbia and Alberta) and quantified the amounts of four known aggregation pheromone components within each sample. Chiral analyses were performed on a subset of samples. Frontalin (1,5-dimethyl-6,8-dioxabicyclo[3.2.1]octane) was more dominant in western Canada, whereas MCOL (1-methyl-2-cyclohexen-1-ol) and seudenol (3-methyl-2-cyclohexen-1-ol) were more dominant in the east. Verbenene (4-methylene-6,6-dimethylbicyclo[3.1.1]hept-2-ene) was prevalent only in one of our six sites (Rocky Mountain House, AB). Female beetles in Rocky Mountain House (AB) also produced a higher proportion of (+)-frontalin than females from the other sites, and a comparison to previously published data suggests additional geographic variation that was not captured among our sites. When paired with a male in a gallery, female spruce beetles produced pheromone blends with more frontalin and less MCOL compared to solitary female beetles. Our results show that variation in the spruce beetle's pheromone blend exists at both large and small spatial scales, possibly a consequence of local selective pressures and assortative mating.

Chemical similarity between introduced and native populations of Scots pine can facilitate transcontinental expansion of mountain pine beetle in North America

Introduced forest tree species are frequently attacked by insects in their new range; however, it has been seldom investigated whether the presence of such tree species increases the risk of range expansion of native insect herbivores in the introduced range. European Scots pine has been introduced to North America including within a portion of the range of the mountain pine beetle (MPB). We investigated Scots pine suitability to MPB as a host in the introduced range of the pine populations. We compared chemotypic similarity of foliage between introduced and native Scots pine populations, and then determined the suitability of introduced populations to MPB. Suitability was assessed based on whether beetles produce aggregation pheromone components and complete development in Scots pine bolts. We also assessed whether or not suitability was affected by the host chemotypes. Introduced and native pine populations had the same sesquiterpene chemotypes and shared one of the two monoterpene chemotypes. All introduced populations were suitable for MPB but the suitability varied slightly with host chemotype. This is the first report of chemotypic variations of Scots pine populations outside its native range. Chemotypic similarity between the introduced and native pine populations, ability of beetles to produce pheromones and to complete its life cycle on bolts from all chemotypes, and preferential colonization of Scots pine over native pine species by MPB in field suggest that introduced Scots pine populations could facilitate transcontinental expansion of MPB in North America.

Male-Produced Aggregation Pheromone of the Sloe Bug, Dolycoris baccarum L. (Hemiptera: Heteroptera: Pentatomidae).

The sloe bug, Dolycoris baccarum L. (Hemiptera: Heteroptera: Pentatomidae), is a Palearctic species and an important polyphagous pest. Previous studies have reported that male D. baccarum produce an aggregation pheromone, to which conspecific adults of both sexes respond. We used solid-phase microextraction (SPME) to collect volatiles released by sexually mature virgin males and females, and identified potential components of the male-produced aggregation pheromone. Analysis of the SPME samples by gas chromatography-mass spectrometry (GC-MS) revealed three male-specific components, α-bisabolol, trans-α-bergamotene, and β-bisabolene, at a ratio of 100:10:3. A field trapping experiment showed that (±)-α-bisabolol alone attracted D. baccarum adults, and it was synergized by trans-α-bergamotene, but not (S)-β-bisabolene. However, neither trans-α-bergamotene nor (S)-β-bisabolene was attractive individually. A binary blend of (±)-α-bisabolol and trans-α-bergamotene in a 10:1 ratio was less attractive than caged live males, suggesting that further study is necessary to determine the absolute configuration or steroisomeric composition of the natural pheromone components. This is the first report of α-bisabolol and trans-α-bergamotene as semiochemicals among Pentatomidae, and the first to identify α-bisabolol from insects.

Decanal as a major component of larval aggregation pheromone of the greater wax moth, Galleria mellonella

AbstractLarvae of the greater wax moth (GWM), Galleria mellonella, a destructive pest of the honeybee (Apis mellifera), have been observed to display aggregation behaviours. However, the underlying mechanism by which these larvae come together remains unknown. We hypothesized that the GWM larvae detect, orient towards and utilize conspecific larval chemical cues to aggregate in groups. We used dual‐choice olfactometer assays to investigate the involvement of conspecific larval odours in their aggregation amongst 3–5th instar and 8th instar larvae. The assays revealed that only 8th instar larvae were significantly attracted to their odours and those emanating from newly spun cocoons. Coupled gas chromatography–mass spectrometry (GC‐MS) of larval head space odours analysis revealed the presence of four compounds: nonanal, decanal, tridecane and tetradecane in pupal and mature larval odour extracts. However, using synthetic compounds, behavioural assays showed that only decanal induced significant attraction, therefore, suggesting its role as a major component of the larval aggregation pheromone of GWM. Our findings reveal the involvement of volatile organic compounds in the aggregation behaviour of mature wax moth larvae and thereby offer prospects for the development of an odour‐baited in‐hive trapping management tool for wax moth larva.

Bacterial Associates of a Gregarious Riparian Beetle With Explosive Defensive Chemistry.

Bombardier beetles (Carabidae: Brachininae) are well known for their unique explosive defensive chemistry. These beetles are found in riparian corridors throughout the American Southwest, where they commonly form large diurnal multispecies aggregations in moist areas under rocks, in crevices, and in leaf litter. Using high throughput 16S amplicon sequencing, we provide the first microbiome survey of a bombardier beetle, Brachinus elongatulus, collected from two sites in Arizona. Two bacterial taxa were present in all individuals sampled: Enterococcus and Dysgonomonas. Enterococcus has been implicated in the production of fecal aggregation pheromone components, which have been shown to regulate aggregation in the German cockroach; it is possible that Enterococcus plays a similar role in Brachinus. Dysgonomonas was found in all the secretory cells of the defensive system and gut samples. Additional studies are needed to determine if these microbes play a role in these beetles’ unique chemical defense. Results also show that the majority of B. elongatulus individuals collected from both sites were infected with Spiroplasma. Many Spiroplasma are intracellular, vertically transmitted insect symbionts that may manipulate host reproduction (e.g., cause male-killing) or provide resistance to nematodes and/or parasitoid wasps. Defensive protection could be especially beneficial to B. elongatulus, which are frequently parasitized by horsehair worms (Nematomorpha). In sum, findings suggest several testable hypotheses on the effects bacteria may have on bombardier beetle behavior and physiology.