Chemical and Electrophysiological Characterisation of Headspace Volatiles from Yeasts Attractive to Drosophila suzukii.

Chemical control of Drosophila suzukii (Diptera: Drosophilidae) based on the use of insecticides is particularly challenging as the insect attacks ripening fruits shortly before harvest. An alternative strategy may rely on the use of yeasts as phagostimulants and baits, applied on canopy as attract-and-kill formulations. The aim of this research was to identify the most attractive among six yeast species for D. suzukii: Saccharomyces cerevisiae, Hanseniaspora uvarum, Clavispora santaluciae, Saccharomycopsis vini, Issatchenkia terricola, and Metschnikowia pulcherrima. The volatile profile of C. santaluciae was described for the first time. Behavioural experiments identified H. uvarum and S. vini as the most attractive yeasts. The characterization of yeast headspace volatiles using direct headspace (DHS) and solid-phase microextraction (SPME) revealed several strain-specific compounds. With DHS injection, 19 volatiles were characterised, while SPME revealed 71 compounds constituting the yeast headspace. Both analyses revealed terpenoids including ß-ocimene, citronellol, (Z)-geraniol (nerol), and geranial as distinct constituents of S. vini. H. uvarum and S. vini were further investigated using closed-loop stripping analysis (CSLA) and electroantennography. Out of 14 compounds quantified by CSLA, ethyl acetate, isoamyl acetate, ß-myrcene, benzaldehyde and linalool were detected by D. suzukii antennae and might generate the strong attractiveness of S. vini and H. uvarum. Our results highlight a strong attraction of D. suzukii to various yeasts associated with both the flies and their habitat and demonstrate how different sampling methods can impact the results of volatile compound characterization. It remains to be demonstrated whether the distinct attraction is based on special adaptations to certain yeasts and to what extent the metabolites causing attraction are interchangeable.

Chemical imitation of yeast fermentation by the drosophilid-pollinated deceptive trap-flower Aristolochia baetica (Aristolochiaceae).

Deceptive flowers, unlike in mutualistic pollination systems, mislead their pollinators by advertising rewards which ultimately are not provided. Although our understanding of deceptive pollination systems increased in recent years, the attractive signals and deceptive strategies in the majority of species remain unknown. This is also true for the genus Aristolochia, famous for its deceptive and fly-pollinated trap flowers. Representatives of this genus were generally assumed to be oviposition-site mimics, imitating vertebrate carrion or mushrooms. However, recent studies found a broader spectrum of strategies, including kleptomyiophily and imitation of invertebrate carrion. A different deceptive strategy is presented here for the western Mediterranean Aristolochia baetica L. We found that this species is mostly pollinated by drosophilid flies (Drosophilidae, mostly Drosophila spp.), which typically feed on fermenting fruit infested by yeasts. The flowers of A. baetica emitted mostly typical yeast volatiles, predominantly the aliphatic compounds acetoin and 2,3-butandiol, and derived acetates, as well as the aromatic compound 2-phenylethanol. Analyses of the absolute configurations of the chiral volatiles revealed weakly (acetoin, 2,3-butanediol) to strongly (mono- and diacetates) biased stereoisomer-ratios. Electrophysiological (GC-EAD) experiments and lab bioassays demonstrated that most of the floral volatiles, although not all stereoisomers of chiral compounds, were physiologically active and attractive in drosophilid pollinators; a synthetic mixture thereof successfully attracted them in field and lab bioassays. We conclude that A. baetica chemically mimics yeast fermentation to deceive its pollinators. This deceptive strategy (scent chemistry, pollinators, trapping function) is also known from more distantly related plants, such as Arum palaestinum Boiss. (Araceae) and Ceropegia spp. (Apocynaceae), suggesting convergent evolution. In contrast to other studies working on floral scents in plants imitating breeding sites, the present study considered the absolute configuration of chiral compounds.

Identification, synthesis, and field evaluation of components of the female-produced sex pheromone of Helopeltis cinchonae (Hemiptera: Miridae), an emerging pest of tea.

BACKGROUND: Helopeltis cinchonae (Hemiptera: Miridae) is a major pest of tea plantations in Asia. Conventional control of pests with pesticides is unsustainable. Therefore, safe and eco-friendly alternatives, such as pheromones, are required to manage the pest. RESULTS: In gas chromatography-electroantennographic detection (GC-EAD) analysis of whole-body extracts of virgin female H. cinchonae, two compounds elicited electroantennogram (EAG) responses from male antennae. These were identified as hexyl (R)-3-acetoxybutyrate and (R)-1-acetoxy-5-butyroxyhexane using gas chromatography-mass spectrometry (GC-MS) analysis compared to synthetic compounds. This is the first study to report 1-acetoxy-5-butyroxyhexane as an insect pheromone component. The synthetic compounds elicited dose-dependent EAG responses from the antennae of male H. cinchonae. In two field trapping experiments, the individual compounds were highly attractive to male H. cinchonae when dispensed from polyethylene vials. However, higher catches were obtained with blends of the two compounds in a 1:10 ratio. The blend of racemic compounds was as attractive as the blend of (R)-enantiomers. CONCLUSIONS: We reported that 1-acetoxy-5-butyroxyhexane and hexyl 3-acetoxybutyrate are components of the female-produced sex pheromone of H. cinchonae, but further work is required on the blend and loading of pheromone and on trap design to provide an optimized system for monitoring and control of this pest. The results may also facilitate the identification of the pheromones of other Helopeltis species, which are major pests in many crops. © 2024 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Functional analysis of three odorant receptors in Plutella xylostella response to repellent activity of 2,3-dimethyl-6-(1-hydroxy)-pyrazine.

Plutella xylostella is an important pest showing resistance to various chemical pesticides, development of botanical pesticides is an effective strategy to resolve above problem and decrease utilization of chemical pesticides. Previous study showed that 2,3-dimethyl-6-(1-hydroxy)-pyrazine has significant repellent activity to P. xylostella adult which mainly effect to the olfactory system, however the molecular targets and mechanism are still unclear. Based on the RNA-Seq and RT-qPCR data, eight ORs (Odorant receptor) in P. xylostella were selected as candidate targets response to repellent activity of 2,3-dimethyl-6-(1-hydroxy)-pyrazine. Here, most of the ORs in P. xylostella were clustered into three branches, which showed similar functions such as recognition, feeding, and oviposition. PxylOR29, PxylOR31, and PxylOR46 were identified as the potential molecular targets based on the results of repellent activity and EAG response tests to the adults which have been injected with dsRNA, respectively. Additionally, the three ORs were higher expressed in antenna of P. xylostella, followed by those in the head segment. Furthermore, it was found that the bindings between these three ORs and 2,3-dimethyl-6-(1-hydroxy)-pyrazine mainly depend on the hydrophobic effect of active cavities, and the binding to PxylOR31 was more stabler and easier with an energy of -16.34 kcal/mol, together with the π-π T-shaped interaction at PHE195 site. These findings pave the way for the complete understanding of pyrazine repellent mechanisms.

Molecular and Functional Characterization of Pheromone Binding Protein 2 from Cyrtotrachelus buqueti (Coleoptera: Curculionidae).

Pheromone-binding proteins (PBPs) play important roles in binding and transporting sex pheromones. However, the PBP genes identified in coleopteran insects and their information sensing mechanism are largely unknown. Cyrtotrachelus buqueti (Coleoptera: Curculionidae) is a major insect pest of bamboo plantations. In this study, a novel PBP gene, CbuqPBP2, from C. buqueti was functionally characterized. CbuqPBP2 was more abundantly expressed in the antennae of both sexes than other body parts, and its expression level was significantly male-biased. Fluorescence competitive binding assays showed that CbuqPBP2 exhibited the strongest binding affinity to dibutyl phthalate (Ki = 6.32 µM), followed by styrene (Ki = 11.37 µM), among twelve C. buqueti volatiles. CbuqPBP2, on the other hand, showed high binding affinity to linalool (Ki = 10.55), the main volatile of host plant Neosinocalamus affinis. Furthermore, molecular docking also demonstrated the strong binding ability of CbuqPBP2 to dibutyl phthalate, styrene, and linalool, with binding energy values of -5.7, -6.6, and -6.0 kcal/mol, respectively, and hydrophobic interactions were the prevailing forces. The knockdown of CbuqPBP2 expression via RNA interference significantly reduced the electroantennography (EAG) responses of male adults to dibutyl phthalate and styrene. In conclusion, these results will be conducive to understanding the olfactory mechanisms of C. buqueti and promoting the development of novel strategies for controlling this insect pest.

Identification of odorant receptors of Tribolium confusum in response to limonene repellent activity.

Tribolium confusum is an important storage pest showing significant resistance to various chemical pesticides, development of botanical pesticides is an effective strategy to resolve above problem and decrease utilization of chemical pesticides. Present study attempts to explore the molecular mechanism about the repellent activity of limonene. When treatment concentration of limonene was 200.00 µg/cm2, the repellent level remained at grade V after 24 hours. Our study showed that limonene could be distinguished by T. confusum antenna with a maximal electroantennography test value of 0.90 mV. Simultaneously, 88 upregulated and 98 downregulated genes were sequenced in limonene-repellent T. confusum, and RT-qPCR analysis showed that four down-regulated and one up-regulated OR genes play an important role in the response to limonene. The repellent rate was decreased by 22.13% mediated with a knockdown of dsTconOR93, while the EAG value of the female and male adults was reduced to 0.26 mV (49.06%) and 0.20 mV (54.05%), respectively. In conclusion, limonene had a strong repellent activity against T. confusum and TconOR93 gene was determined to be a major effector in perception of limonene. This study provides a basis for the development of limonene as a novel botanical pesticide for the control to storage pests, which will reduce the utilization of chemical pesticides and postpone the development of resistance.

Functional Characterization of Pheromone Receptors in the Beet Webworm, Loxostege sticticalis (Lepidoptera: Pyralidae).

Lepidopteran insects mainly rely on sex pheromones to complete sexual communications. Pheromone receptors (PRs) are expressed on the olfactory receptor neurons (ORNs) of the sensilla trichodea and play an essential role in sexual communication. Despite extensive investigations into the mechanisms of peripheral recognition of sex pheromones in Lepidoptera, knowledge about these mechanisms in L. sticticalis remains limited. In this study, five candidate LstiPRs were analyzed in a phylogenetic tree with those of other Lepidopteran insects. Electroantennography (EAG) assays showed that the major sex pheromone component E11-14:OAc elicited a stronger antennal response than other compounds in male moths. Moreover, two types of neurons in sensilla trichodea were classified by single sensillum recordings, of which the "a" neuron specifically responded to E11-14:OAc. Five candidate PRs were functionally assayed by the heterologous expression system of Xenopus oocytes, and LstiPR2 responded to the major sex pheromone E11-14:OAc. Our findings suggest that LstiPR2 is a PR sensitive to L. sticticalis's major sex pheromone compound, E11-14:OAc. Furthermore, this study offers valuable insights into the sexual communication behavior of L. sticticalis, forming a foundation for further analysis of the species' central nervous system.

Chronic larval exposure to thiacloprid impairs honeybee antennal selectivity, learning and memory performances.

The use of agricultural neonicotinoid insecticides has sub-lethal chronic effects on bees that are more prevalent than acute toxicity. Among these insecticides, thiacloprid, a commonly used compound with low toxicity, has attracted significant attention due to its potential impact on the olfactory and learning abilities of honeybees. The effect of sub-lethal larval exposure to thiacloprid on the antennal activity of adult honeybees (Apis mellifera L.) is not yet fully understood. To address this knowledge gap, laboratory-based experiments were conducted in which honeybee larvae were administered thiacloprid (0.5 mg/L and 1.0 mg/L). Using electroantennography (EAG), the impacts of thiacloprid exposure on the antennal selectivity to common floral volatiles were evaluated. Additionally, the effects of sub-lethal exposure on odor-related learning and memory were also assessed. The results of this study reveal, for the first time, that sub-lethal larval exposure to thiacloprid decreased honeybee antenna EAG responses to floral scents, leading to increased olfactory selectivity in the high-dose (1.0 mg/L) group compared to the control group (0 mg/L vs. 1.0 mg/L: p = 0.042). The results also suggest that thiacloprid negatively affected odor-associated paired learning acquisition, as well as medium-term (1 h) (0 mg/L vs. 1.0 mg/L: p = 0.019) and long-term memory (24 h) (0 mg/L vs. 1.0 mg/L: p = 0.037) in adult honeybees. EAG amplitudes were dramatically reduced following R-linalool paired olfactory training (0 mg/L vs. 1.0 mg/L: p = 0.001; 0 mg/L vs. 0.5 mg/L: p = 0.027), while antennal activities only differed significantly in the control between paired and unpaired groups. Our results indicated that exposure to sub-lethal concentrations of thiacloprid may affect olfactory perception and learning and memory behaviors in honeybees. These findings have important implications for the safe use of agrochemicals in the environment.

Influence of RVFV Infection on Olfactory Perception and Behavior in Drosophila melanogaster.

In blood-feeding dipterans, olfaction plays a role in finding hosts and, hence, in spreading pathogens. Several pathogens are known to alter olfactory responses and behavior in vectors. As a mosquito-borne pathogen, Rift Valley Fever Virus (RVFV) can affect humans and cause great losses in livestock. We test the influence of RVFV infection on sensory perception, olfactory choice behavior and activity on a non-biting insect, Drosophila melanogaster, using electroantennograms (EAG), Y-maze, and locomotor activity monitor. Flies were injected with RVFV MP12 strain. Replication of RVFV and its persistence for at least seven days was confirmed by quantitative reverse transcription-PCR (RT-qPCR). One day post injection, infected flies showed weaker EAG responses towards 1-hexanol, vinegar, and ethyl acetate. In the Y-maze, infected flies showed a significantly lower response for 1-hexanol compared to uninfected flies. At days six or seven post infection, no significant difference between infected and control flies could be found in EAG or Y-maze anymore. Activity of infected flies was reduced at both time points. We found an upregulation of the immune-response gene, nitric oxide synthase, in infected flies. An infection with RVFV is able to transiently reduce olfactory perception and attraction towards food-related odors in Drosophila, while effects on activity and immune effector gene expression persist. A similar effect in blood-feeding insects could affect vector competence in RVFV transmitting dipterans.

Discovering Aethina tumida responses to attractant and repellent molecules: A potential basis for future management strategies.

Small hive beetle (Aethina tumida) management has been highly dependent upon chemical and mechanical control over the past two decades; however, many of these methods have not been consistently effective or safe for European honey bee (Apis mellifera) colonies. Here we explore the behavioral and physiological effects of the attractants isopentyl acetate and pollen patty upon A. tumida adults, and also investigate the mixture of attractants with repellent compounds, which were previously untested against A. tumida. Electroantennograms established sensitivity of A. tumida antennae to both attractants and all repellents with the exception of DEET, with antennae displaying greatest sensitivity to the repellent pyrrolidine. A walking-response olfactometer, designed specifically for A. tumida, was used for all behavioral experiments. It was found that both pollen patty and isopentyl acetate were attractive to A. tumida adults; conversely, mixes of attractants and repellent volatiles led to less attraction or avoidance of what was previously a significantly attractive source. Of all repellents tested, pyrrolidine was found to be the most repelling molecule, with significant avoidance of the attractive source at a 10 mg treatment of pyrrolidine. The results of this study indicate that, at the behavioral level, the repellent compounds pyrrolidine and 1,4-dimethylpiperazine resulted in a negative preference index indicating a repellent behavioral response. By strategically implementing a repellent source in an apiary environment, A. tumida adults could be deterred from entering and invading hives.

Laboratory Evidence of 2-Isobutyl-3-methoxypyrazine as a Male-Released Aggregative Cue in Labidostomis lusitanica (Germar) (Coleoptera: Chrysomelidae).

In spite of its incidence on pistachio trees, the chemical ecology of Labidostomis lusitanica (Germar) (Coleoptera: Chrysomelidae) has been neglected so far. In this work, we provide the first evidence of a biologically active male-specific compound that may be promoting field aggregation. Headspace collections through solid-phase microextraction from feral males and females reported the presence of 2-isobutyl-3-methoxypyrazine exclusively in males. Electroantennographic recordings revealed that males and females responded in a dose-dependent manner to increasing stimuli of 2-isobutyl-3-methoxypyrazine, with females overall displaying a higher response than males. In dual-choice tests, both males and females showed a significant preference for the compound in comparison to a pure air stimulus. In light of these results, the possible role of 2-isobutyl-3-methoxypyrazine as an aggregation cue in L. lusitanica is discussed.

Electrophysiological and Behavioral Responses of Apis mellifera and Bombusterrestris to Melon Flower Volatiles.

As important pollinators, honeybees and bumblebees present a pollination behavior that is influenced by flower volatiles through the olfactory system. In this study, volatile compounds from melon flowers were isolated and identified by headspace solid-phase microextraction (HS-SPME) and gas chromatography-mass spectrometry (GC-MS), and their effects on Apis mellifera and Bombus terrestris were investigated by electroantennogram (EAG) and behavior tests (Y-tube olfactometer). The results showed that 77 volatile compounds were detected in melon flowers, among which the relative content of aldehydes was the highest (61.34%; 82.09%). A. mellifera showed a strong EAG response to e-2-hexenal, e-2-octenal, and 1-nonanal. B. terrestris showed a strong EAG response to e-2-hexenal, e-2-octenal, 2,5-dimethyl-benzaldehyde, benzaldehyde and benzenepropanal. In behavior tests, the volatiles with the highest attractive rate to A. mellifera were e-2-hexenal (200 µg/µL, 33.33%) and e-2-octenal (300 µg/µL, 33.33%), and those to B. terrestris were e-2-hexenal (10 µg/µL, 53.33%) and 2,5-dimethyl-benzaldehyde (100 µg/µL, 43.33%). E-2-hexenal and e-2-octenal were more attractive to A. mellifera than B. terrestris, respectively (10 µg/µL, 10 µg/µL, 200 µg/µL). In conclusion, the volatiles of melon flowers in facilities have certain effects on the electrophysiology and behavior of bees, which is expected to provide theoretical and technical support for the pollination of A. mellifera and B. terrestris in facilities.

Response of Parasitic Wasp Cotesia glomerata L. (Hymenoptera: Braconidae) to Cabbage Plants of Two Varieties: Olfactory Spectra of Males and Females.

The parasitoid Cotesia glomerata L. (Hymenoptera: Braconidae) oviposits in larvae of the large cabbage white butterfly (Pieris brassicae L.). Many parasitoids are attracted by volatile organic compounds (VOCs) emitted by the plant that the host insects feed on. The objectives of the study were to identify the VOCs in leaves of two varieties of cabbage (white cabbage, Brassica oleracea var. capitata f. alba and cauliflower, B. oleracea var. botrytis) damaged by P. brassicae caterpillars which elicit antennal responses of C. glomerata, and characterize the olfactory spectra of females and males. Leaf extracts were analyzed by using gas chromatography equipped with an electroantennographic detector (GC-EAD) and GC-mass spectrometry. In total, 32 olfactory-active compounds for C. glomerata in cauliflower and 24 in white cabbage were revealed. The females perceived more compounds than males. Hexan-1-ol, (E)-3-hexen-1-ol, (E)-2-octenal, benzylcianide, tetradecanal, and two unidentified compounds elicited EAG responses in females but not in males. Females were more sensitive to (E)-3-hexenal, (Z)-3-hexenal, and pentadecenal, whereas males showed higher sensitivity to (Z)-3-hexenyl butanoate, heptanal, (Δ)-2-pentenal, (E)-2-hexenol, and octanal. The olfactory spectrum of C. glomerata was expanded from 18 to 41 VOCs emitted by different varieties of cabbage damaged by P. brassicae caterpillars. Eight EAG-active VOCs were common for all cabbage varieties. In tritrophic interactions, benzylcyanide can serve as an important signal for C. glomerata females indicating damage of cabbage caused by P. brassicae caterpillars. The data are useful for development of a push-pull strategy for P. brassicae control, based on parasitoid behavior regulation by VOCs.

Links Between Feeding Preferences and Electroantennogram Response Profiles in Dung Beetles: The Importance of Dung Odor Bouquets.

The detection of dung odors is a crucial step in the food-searching behavior of dung beetles (Coleoptera: Scarabaeoidea). Yet, whether certain compounds characteristic of a given dung type contribute to a 'choosy generalism' behavior proposed for this taxonomic group is unknown. To address this, we analyzed the chemical composition of three types of dung (cow, horse, and rabbit) and conducted behavioral and electroantennogram (EAG) bioassays on 15 species of dung beetles using 19 volatile organic compounds representing the three dung samples. Chemical analyses revealed substantial qualitative and quantitative differences among dung types. When offered these food options in an olfactometer, 14 species exhibited a feeding preference. Surprisingly, all 19 compounds used in the EAG assays elicited antennal responses, with species displaying different olfactory profiles. The relationship between behavioral preferences and electrophysiological profiles highlighted that species with different food preferences had differences in antennal responses. Moreover, a specific set of EAG-active compounds (nonanal, sabinene, acetophenone, ρ-cresol, 2-heptanone, 1H-indole, and 6-methyl-5-hepten-2-one) were the strongest drivers in the distinct sensory profiles of the trophic preference groups. Our results point to the importance of the whole bouquet of dung-emanating compounds in driving food-searching behavior, but specific volatiles could aid in determining highly marked trophic preferences in certain species.

Differences in olfactory sensitivity and odor detection correlate with foraging task specialization in honeybees Apis mellifera.

Division of labor is central to the ecological success of social insects. Among honeybees foragers, specialization for collecting nectar or pollen correlates with their sensitivity to gustatory stimuli (e.g. sugars). We hypothesize that pollen and nectar foragers also differ in their sensitivity to odors, and therefore in their likelihood to show odor-mediated responses. To assess foragers sensitivity to natural odors, we quantified the conditioning of the proboscis extension reflex (PER) to increasing concentrations (0.001; 0.01; 0.1; 1 M) of linalool or nonanal. Furthermore, we compared electroantennogram (EAG) recordings to correlate bees' conditioned responses with the electrophysiological responses of their antennae. To further explore differences of the antennal response of foragers in relation to task-related odors, we registered EAG signals for two behaviorally ''meaningful'' odors that mediate pollen collection: fresh pollen odors and the brood pheromone (E)-ß-ocimene. Pollen foragers performed better than nectar foragers in PER conditioning trials when linalool and nonanal were presented at low concentrations (0.001, 0.01 M). Consistently, their antennae showed stronger EAG signals (higher amplitudes) to these odors, suggesting that differences in sensitivity can be explained at the periphery of the olfactory system. Pollen and nectar foragers detect pollen odors differently, but not (E)-ß-ocimene. Pollen volatiles evoked EAG signals with hyper and depolarization components. In pollen foragers, the contribution of the hyperpolarization component was higher than in nectar foragers. We discuss our findings in terms of adaptive advantages to learn subtle olfactory cues that influence the ability to better identify/discriminate food sources.

Electroantennographic Responses of Wild and Laboratory-Reared Females of Xyleborus affinis Eichhoff and Xyleborus ferrugineus (Fabricius) (Coleoptera: Curculionidae: Scolytinae) to Ethanol and Bark Volatiles of Three Host-Plant Species.

Chemical ecology studies on ambrosia beetles are typically conducted with either wild or laboratory-reared specimens. Unlike laboratory-reared insects, important aspects that potentially influence behavioral responses, such as age, physiological state, and prior experience are unknown in wild specimens. In this study, we compared the electroantennographic (EAG) responses of laboratory-reared and wild X. affinis and X. ferrugineus to 70% ethanol and bark odors (host kairomones) of Bursera simaruba, Mangifera indica, and Persea schiedeana aged for 2, 24, and 48 h. Chemical analyses of each odor treatment (bark species x length of aging) were performed to determine their volatilome composition. EAG responses were different between laboratory-reared and wild X. ferrugineus when exposed to ethanol, whereas wild X. affinis exhibited similar EAG responses to the laboratory-reared insects. Ethanol elicited the strongest olfactory responses in both species. Among the bark-odors, the highest responses were triggered by B. simaruba at 48 h in X. affinis, and P. schiedeana at 24 and 48 h in X. ferrugineus. Volatile profiles varied among aged bark samples; 3-carene and limonene were predominant in B. simaruba, whereas α-copaene and α-cubebene were abundant in P. schiedeana. Further studies are needed to determine the biological function of B. simaruba and P. schiedeana terpenes on X. affinis and X. ferrugineus, and their potential application for the development of effective lures.

Electroantennographic activity of 21 aliphatic compounds that bind well to a locust odorant-binding protein.

Odorants that bind well to odorant-binding proteins (OBPs) often trigger olfactory responses and have important biological significance. The locust Locusta migratoria (Meyen) (Orthoptera: Acrididae) is a serious agricultural pest. Twenty-one saturated aliphatic compounds with carbon-oxygen bonds and straight chains of 10-17 carbon atoms bind well to an L. migratoria OBP. In this study, olfactory activities of these aliphatic compounds on L. migratoria adult males were tested by electroantennography (EAG) and comparatively analyzed. Four alcohols (undecanol, dodecanol, tridecanol, and tetradecanol), two ketones (2-dodecanone and 2-tridecanone), and two esters (ethyl octanoate and ethyl nonanoate) triggered strong EAG responses, and there was no significant difference between them. The results suggest that the eight compounds are more likely to have important biological significance than the other compounds. Moreover, we found that there is not necessarily a positive correlation between the olfactory activity of odorants and their binding ability with OBP. The study contributes to understanding the odorants with biological significance for L. migratoria and the molecular mechanism of the locust's olfaction.

Volatiles Produced by Yeasts Related to Prunus avium and P. cerasus Fruits and Their Potentials to Modulate the Behaviour of the Pest Rhagoletis cerasi Fruit Flies.

Yeast produced semiochemicals are increasingly used in pest management programs, however, little is known on which yeasts populate cherry fruits and no information is available on the volatiles that modify the behaviour of cherry pests including Rhagoletis cerasi flies. Eighty-two compounds were extracted from the headspaces of eleven yeast species associated with sweet and sour cherry fruits by solid phase micro extraction. Esters and alcohols were the most abundant volatiles released by yeasts. The multidimensional scaling analysis revealed that the odour blends emitted by yeasts were species-specific. Pichia kudriavzevii and Hanseniaspora uvarum yeasts released the most similar volatile blends while P. kluyveri and Cryptococcus wieringae yeasts produced the most different blends. Combined gas chromatographic and electroantennographic detection methods showed that 3-methybutyl acetate, 3-methylbutyl propionate, 2-methyl-1-butanol, and 3-methyl-1-butanol elicited antennal responses of both R. cerasi fruit fly sexes. The two-choice olfactometric tests revealed that R. cerasi flies preferred 3-methylbutyl propionate and 3-methyl-1-butanol but avoided 3-methybutyl acetate. Yeast-produced behaviourally active compounds indicated a potential for use in pest monitoring and control of R. cerasi fruit flies, an economically important pest of cherry fruits.

Impact of Chronic Exposure to Two Neonicotinoids on Honey Bee Antennal Responses to Flower Volatiles and Pheromonal Compounds.

Volatile compounds provide important olfactory cues for honey bees (Apis mellifera L.), which are essential for their ecology, behavior, and social communication. In the external environment bees locate food sources by the use of floral scents, while inside the hive, pheromones such as the queen mandibular pheromone (QMP) and alarm pheromones serve important functions in regulating colony life and inducing aggressive responses against intruders and parasites. Widely reported alterations of various behaviors in- and outside the hive following exposure to pesticides could therefore be associated with a disturbance of odor sensitivity. In the present study, we tested the effects of neonicotinoid pesticides at field concentrations on the ability of honey bees to perceive volatiles at the very periphery of the olfactory system. Bee colonies were subjected to treatments during the summer with either Imidacloprid or Thiacloprid at sublethal concentrations. Antennal responses to apple (Malus domestica L.) flower volatiles were studied by GC-coupled electro-antennographic detection (GC-EAD), and a range of volatiles, a substitute of the QMP, and the alarm pheromone 2-heptanone were tested by electroantennography (EAG). Short-term and long-term effects of the neonicotinoid treatments were investigated on bees collected in the autumn and again in the following spring. Treatment with Thiacloprid induced changes in antennal responses to specific flower VOCs, with differing short- and long-term effects. In the short term, increased antennal responses were observed for benzyl-alcohol and 1-hexanol, which are common flower volatiles but also constituents of the honey bee sting gland secretions. The treatment with Thiacloprid also affected antennal responses to the QMP and the mandibular alarm pheromone 2-heptanone. In the short term, a faster signal degeneration of the response signal to the positive control citral was recorded in the antennae of bees exposed to Thiacloprid or Imidacloprid. Finally, we observed season-related differences in the antennal responses to multiple VOCs. Altogether, our results suggest that volatile-specific alterations of antennal responses may contribute to explaining several behavioral changes previously observed in neonicotinoid-exposed bees. Treatment effects were generally more prominent in the short term, suggesting that adverse effects of neonicotinoid exposure may not persist across generations.

Volatiles Induced from Hypolepis punctata (Dennstaedtiaceae) by Herbivores Attract Sclomina erinacea (Hemiptera: Reduviidae): Clear Evidence of Indirect Defense in Fern.

Plants have evolved various self-defense mechanisms against insect feeding. There are many reports regarding both direct and indirect defense mechanisms in seed-plant. However, only direct defenses on ferns were considered and the indirect defense mechanism has never been reported. In this study, it was observed that the fern Hypolepis punctata can attract the assassin bug Sclomina erinacea in the field. We collected and analyzed volatiles from H. punctata healthy individuals and the ones wounded by Bertula hadenalis, using dynamic headspace and GC-MS. We recorded the electroantennogram responses of antennae of S. erinacea to different standards of volatile compounds identified from the GC-MS analysis. We also analyzed the behavior of male and female S. erinacea adults in response to volatiles collected from H. punctata using a Y-tube olfactometer. The results showed that a number of volatile compounds were produced when the fern was damaged by B. hadenalis. Electroantennography and Y-tube olfactometer results showed that some herbivore-induced volatiles and volatiles from undamaged leaves could attract S. erinacea. Our research suggests that H. punctata can attract insect predators by releasing herbivory-induced volatile organic compounds, and for the first time we found ferns may also have indirect defense mechanisms using volatile organic compounds.