Electroantennographic Responses Research Articles

Antennal olfactory sensitivity and its age‐dependence in the hemimetabolous insect Metcalfa pruinosa

AbstractMetcalfa pruinosa is a highly polyphagous, invasive insect in Europe with the capability of transferring phytoplasma. In the present study, we aim to identify volatile organic compounds that may act as semiochemicals for M. pruinosa. Using a dynamic volatile collection system, volatile organic compounds were collected from Ailanthus altissima, Aristolochia clematitis and Tagetes patula. Compounds were identified using gas chromatography–mass spectrometry. We also tested the electrophysiological responses of M. pruinosa to the collected volatile organic compounds using gas chromatography‐electroantennographic detection. In total, 77 volatile organic compounds were identified, and 29 compounds elicited an electroantennographic response. We analysed responses to volatile organic compounds extracted from Tagetes patula to compare antennal sensitivity between developmental stages and sexes. No differences were found in responders' number or response amplitude between males and females or adults and nymphs for any volatile. Based on the detected compounds, M. pruinosa adult females and males seem to have similar ecological needs in the host habitat. However, when we compared the response amplitudes among compounds, significant differences were found. These differences suggest that (E)‐4,8‐dimethyl‐nonatriene, methyl salicylate, and piperitone could have an important role in M. pruinosa's behaviour. This is the first contribution to M. pruinosa olfaction profile providing a base for future semiochemical‐based pest control tactics.

Effects of pear fruit‐derived volatiles and their concentrations on the responses of oriental fruit moths

AbstractGrapholita molesta (Busck) is an important pest of pear trees. Numerous esters accumulate consistently in mature pear fruits. However, little is known about the effects of single esters from pear fruits at different concentrations on the responses of male and female G. molesta. In this study, the responses of virgin males, virgin females, and gravid females to five esters (3‐methylbutyl acetate, ethyl hexanoate, ethyl butanoate, butyl acetate, and hexyl acetate) of pear fruits at three dosage levels (1, 5 and 10 μg/μL) were investigated using electroantennography (EAG), wind tunnel experiments, and field tests. Our results showed that ethyl butanoate (5 μg/μL) elicited higher EAG responses in virgin females and males than a mixture of all five volatiles, and elicited an equal EAG response to that for the mixture in gravid females, with all responses to ethyl butanoate and the mixture being higher than the response to hexane alone. For upwind flight, ethyl butanoate (5 μg/μL) excited equal moth flight–location behavior in virgin and gravid females to that for the mixture, which were both higher than the response to hexane. For close flight, ethyl hexanoate (5 μg/μL) and 3‐methylbutyl acetate (10 μg/μL) appeared to be the activating compounds, eliciting equal responses from virgin females to that of the mixture, and higher responses of gravid females to that of the mixture, both of which were higher than the responses to hexane. Upwind and close flight responses of virgin males were lower than those of virgin and gravid females. In field trials, we verified that traps baited with ethyl butanoate (5 μg/μL) captured more moths than the other esters. Therefore, from the esters and concentrations tested, we recommend ethyl butanoate (5 μg/μL) for potential use as an attractant for G. molesta in orchards.

Characterisation of Antennal Sensilla and Electroantennography Responses of the Dung Beetles Bubas bison, Onitis aygulus and Geotrupes spiniger (Coleoptera: Scarabaeoidea) to Dung Volatile Organic Compounds.

Locating sporadically distributed food resources and mate finding are strongly aided by volatile cues for most insects, including dung beetles. However, there is limited information on the olfactory ecology of dung beetles. We conducted a scanning electron microscopy study on the morphology and distribution of the antennal sensilla of three introduced dung beetle species in Australia: Geotrupes spiniger (Coleoptera: Geotrupidae), Bubas bison and Onitis aygulus (Coleoptera: Scarabaeidae). Three main morphological types of antennal sensilla were identified: sensilla trichodea (ST), sensilla basiconica (SB) and sensilla chaetica (SCh). Distinct variations of SB distribution were observed in B. bison and G. spiniger and on different lamellar surfaces in both sexes of all three species. Sexual dimorphism in antennal sensilla distribution or their abundance was not evident. To complement the morphological characterisation of sensilla, electroantennography (EAG) was carried out to construct EAG response profiles of the three species to selected dung volatiles. An initial study revealed that antennae of all species were sensitive to a mix of phenol, skatole, indole, p-cresol, butanone and butyric acid, common components of livestock dung headspace. In addition to these six compounds, dimethyl sulfide, dimethyl disulfide, eucalyptol and toluene were tested for antennal activity. All compounds evoked measurable EAG responses, confirming antennal sensitivity. Geotrupes spiniger exhibited significant responses to all the compounds compared to the control, whereas B. bison and O. aygulus only responded to a subset of compounds. A comparison of relative EAG amplitudes revealed highly significant responses to p-cresol in G. spiniger and to skatole in B. bison. Geotrupes spiniger displayed differential responses to all the compounds. Pooled EAG data suggest highly significant differences in responses among the three species and among compounds. Our findings suggest that a blend of volatiles may offer potential for the trapping of dung beetles, thereby avoiding the use of dung baits that are inconvenient, inconsistent and may pose a threat to farm biosecurity.

Oviposition substrate location by the invasive woodwasp Sirex noctilio: the combined effect of chemical cues emitted by its obligate symbiont Amylostereum areolatum and different host-tree species.

Sirex noctilio is an invasive forest wasp that affects pines and can result in severe economic losses. The use of semiochemicals offers an opportunity to develop sensitive and specific capturing systems to mitigatenegative impacts. Previous research showed that female S. noctilio would use volatiles emitted by its fungal symbiont, Amylostereum areolatum, but little is known about how these modulate behaviour when combined with pine-wood emissions. Our aim was to understand the relevance of fungal volatiles grown on artificial media and wood from two hosts trees, Pinus contorta and Pinus ponderosa, on behavioural and electroantennographic responses of wasp females. Because background odours can modify an insect's response towards resource-indicating semiochemicals, we propose that the behaviour towards the symbiont (resource) will be modulated by host pine emissions (background odours). Olfactometric assays showed that both host species with fungus were attractive when contrasted against air (P. contorta versus Air, χ2 = 12.19, P < 0.001; P. ponderosa versus Air, χ2 = 20.60, P < 0.001) and suggest a clear hierarchy in terms of female preferences towards the tested stimuli, with response highest towards the fungus grown on P. contorta (olfactory preference index: 5.5). Electrophysiological analyses indicate that females detect 62 volatile compounds from the tested sources. Results indicate a strong synergy between symbiont and host semiochemicals, suggesting that the pine species could play a fundamental role in the interaction. Further understanding of the chemical basis of this, could guide the development of specific and attractive lures, in order to maximize attraction of wasps in surveillance programmes. © 2023 Society of Chemical Industry.

Roles of GOBP1 in perception of host plant volatiles revealed by CRISPR/Cas9‐mediated mutagenesis in Spodoptera litura

AbstractThe general odorant‐binding proteins (GOBPs) in Lepidoptera form a distinct OBP subclass, which exclusively has two members (GOBP1 and GOBP2) in a single species. GOBPs are long thought to function in perception of plant odorants and recently in sex pheromones, but the in vivo evidence is rarely provided. Here, by using CRISPR/Cas9 technique, we demonstrated that GOBP1 in a noctuid moth Spodoptera litura is involved in perception of host plant volatiles but not of the female sex pheromone, and GOBP1 in females is less important than GOBP2 in host plant perception. First, the GOBP1 knockout strain (GOBP1−/−) of S. litura was obtained by CRISPR/Cas9 technique. Second, in the electroantennography (EAG) tests, GOBP1−/− females displayed significant reduction in responses to plant volatile farnesene, and leaf discs of the host plant cabbage (males only to high leaf doses), but GOBP1−/− adults did not show significant EAG reduction to the major sex pheromone component and two pheromone analogues. Third, compared with GOBP2−/− females, GOBP1−/− females were significantly lower in reduction (in relation to WT moths) of EAG response to host plant leaves. Our results provide in vivo evidence that GOBP1 is involved in perception of host plant volatiles but is less important than GOBP2 in S. litura.

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 (0.5mg/L and 1.0mg/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 (1.0mg/L) group compared to the control group (0mg/L vs. 1.0mg/L: p = 0.042). The results also suggest that thiacloprid negatively affected odor-associated paired learning acquisition, as well as medium-term (1h) (0mg/L vs. 1.0mg/L: p = 0.019) and long-term memory (24h) (0mg/L vs. 1.0mg/L: p = 0.037) in adult honeybees. EAG amplitudes were dramatically reduced following R-linalool paired olfactory training (0mg/L vs. 1.0mg/L: p = 0.001; 0mg/L vs. 0.5mg/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.

Attraction of Sitophilus oryzae (L.) (Coleoptera: Curculionidae) to the semiochemical volatiles of stored rice materials

AbstractIn order to explore the influence of stored cereal volatiles on the behavior of Sitophilus oryzae, the olfactory responses of adult rice weevils to the volatiles of different rice cultivars [Red brown rice (RBR), Daohuaxiangmi (DHXM), Baishuigongmi (BSGM), Yashuixinmi (YSXM), and white glutinous rice (WGR)] were studied using electroantennography (EAG) and behavioural bioassays in different types of olfactometers. S. oryzae showed significantly different preferences for these rice cultivars, in the order RBR &gt; DHXM = YSXM ≥ BSGM &gt; WGR. Furthermore, 26 components were identified in the volatile profile of RBR. Nonanal (29.37%), hexanal (16.08%), and 1-octen-3-ol (8.83%) were the most abundant compounds. EAG recordings showed that the antennae of S. oryzae were able to perceive these three compounds in a dose-dependent manner. The compounds elicited significant EAG responses at various concentrations, with the strongest responses at 100 μg μL−1. S. oryzae had a significant positive behavioural response to nonanal, hexanal, and 1-octen-3-ol at various concentrations, with the most attractive being 50, 100, and 100 µg µL−1, respectively. The olfactory preferences of S. oryzae, based on a comparison of these compounds at their optimal concentrations, were nonanal &gt; 1-octen-3-ol = hexanal. These results indicated that the volatiles of the preferred rice cultivar (RBR) were perceived by the peripheral olfactory system of S. oryzae adults and individually elicited positive chemotaxis. These findings offer new insights into the mechanism of host preferences of stored-grain pests. Nonanal showed the greatest potential for use as a novel monitoring and control tool against this storage-beetle pest.

Piperitone (p-Menth-1-En-3-One): A New Repellent for Tea Shot Hole Borer (Coleoptera: Curculionidae) in Florida Avocado Groves.

The tea shot hole borer, Euwallacea perbrevis, has been recently established in Florida, USA, where it vectors fungal pathogens that cause Fusarium dieback in avocado. Pest monitoring uses a two-component lure containing quercivorol and α-copaene. Incorporation of a repellent into IPM programs may reduce the incidence of dieback in avocado groves, particularly if combined with lures in a push-pull system. This study evaluated piperitone and α-farnesene as potential repellents for E. perbrevis, comparing their efficacy to that of verbenone. Replicate 12-week field tests were conducted in commercial avocado groves. Each test compared beetle captures in traps baited with two-component lures versus captures in traps containing lures plus repellent. To complement field trials, Super-Q collections followed by GC analyses were performed to quantify emissions from repellent dispensers field-aged for 12 weeks. Electroantennography (EAG) was also used to measure beetle olfactory response to each repellent. Results indicated that α-farnesene was ineffective; however, piperitone and verbenone were comparable in repellency, achieving 50-70% reduction in captures, with longevity of 10-12 weeks. EAG responses to piperitone and verbenone were equivalent, and significantly greater than response to α-farnesene. Since piperitone is less expensive than verbenone, this study identifies a potential new E. perbrevis repellent.

Isolation and Identification of Volatile Substances with Attractive Effects on Wohlfahrtia magnifica from Vagina of Bactrian Camel.

Vaginal myiasis is one of the most serious parasitic diseases in Bactrian camels. At present, there are no reports on biological control measures of the disease. In this paper, the metabolomic analysis of vaginal secretions from susceptible and non-susceptible camels was performed by ACQUITY UPLC H-Class Ultra Performance Liquid Chromatograph. The results matched in 140 vaginal compounds. Methylheptenone, 1-octen-3-ol, and propyl butyrate and their mixtures were selected for gas chromatography-electroantennography (GC-EAD), electroantennography (EAG), behavioral experiments and trapping experiments of Wohlfahrtia magnifica (W. magnifica). Results showed that the W. magnifica had EAG responses to the three compounds, respectively. The EAG responses of female flies to different concentrations of methylheptenone were significantly different, but to the others had no significant difference, and there was no significant difference in the same compounds between the different sexes. Behavioral and trapping experiments showed that methylheptenone and 1-octen-3-ol have significant attraction to W. magnifica, but there was no significant difference to propyl butyrate. When methylheptenone and 1-octen-3-ol were mixed in different proportions, it was found that a mixture at the ratio of 1:1 and 0.5:1 had extremely significant and significant attraction, respectively, to both male and female W. magnifica. The study showed that, except for propyl butyrate, the higher the concentrations of the other two compounds, the stronger the attractivity to the W. magnifica, and a mixture at the ratio of 1:1 could enhance the attractivity to the W. magnifica.

Sexual competitiveness of sterile Ceratitis capitata males exposed to essential oils from non‐host plant species native to Argentina

AbstractThe sterile insect technique (SIT) is used to control fruit fly pests, such as the Mediterranean fruit fly (medfly), Ceratitis capitata (Wiedemann) (Diptera: Tephritidae). Enhancing sexual competitiveness of mass‐reared sterile males can contribute to making this technique more effective. It has been shown that exposure to volatiles from essential oils (EOs), such as ginger root oil (GRO) and those from host fruits, increases male mating success. We evaluated the effect of EOs from non‐host species native to Argentina, Schinus polygama (Cav.) Cabrera (Anacardiaceae) and Baccharis spartioides (Hook. &amp; Arn.) Remy (Asteraceae), on the sexual competitiveness of sterile C. capitata males. In field cage experiments, sterile males exposed to S. polygama EO increased their mating success. In addition, sterile males exposed to this EO achieved more matings on trees than non‐exposed males, thus suggesting the former are more prone to locate and defend the pheromone‐calling territory. Deprivation of water and/or food significantly reduced males' life span, but exposure to S. polygama EO and GRO did not affect their survival. Schinus polygama EO is composed of mono‐ and sesquiterpenes with behavioral and electroantennographic responses in medflies, indicating that semiochemicals that boost medfly sexual competitiveness combine in this EO.

Differences in EAG Response and Behavioral Choices between Honey Bee and Bumble Bee to Tomato Flower Volatiles.

Bumble bees and honey bees are of vital importance for tomato pollination, although honey bees are less attracted to tomato flowers than bumble bees. Little is known about how tomato flower volatile compounds influence the foraging behaviors of honey bees and bumble bees. In this study, compounds of tomato flower volatiles were detected by gas chromatography-mass spectrometry. Electroantennography (EAG) and a dynamic two-choice olfactometer were used, respectively, to compare the differences of antennal and behavioral responses between Apis mellifera and Bombus terrestris towards selected volatile compounds. A total of 46 compounds were detected from the tomato flower volatiles. Of the 16 compounds tested, A. mellifera showed strong antennal responses to 3 compounds (1-nonanal, (+)-dihydrocarvone, and toluene) when compared with a mineral oil control, and B. terrestris showed 7 pronounced EAG responses (1,3-xylene, (+)-dihydrocarvone, toluene, piperitone, eucarvone, 1-nonanal, and β-ocimene). Additionally, 1-nonanal and (+)-dihydrocarvone elicited significant avoidance behavior of A. mellifera, but not of B. terrestris. In conclusion, bumble bees are more sensitive to the compounds of tomato flower volatiles compared to honey bees, and honey bees showed aversion to some compounds of tomato flower volatiles. The findings indicated that compounds of flower volatiles significantly influenced bee foraging preference for tomato.

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.

Simple SummaryThe ambrosia beetles Xyleborus affinis and Xyleborus ferrugineus are wood borers reported as secondary vectors of pathogenic fungi that cause lethal vascular diseases in mango, cacao, and trees within the laurel family. The use of specific attractants or repellants is one potential method for monitoring or controlling these pests. Chemical ecology studies to develop such tools often use wild or laboratory-reared beetles without first determining whether there are differences in their responses. We compared the antennal olfactory responses of wild and laboratory-reared X. affinis and X. ferrugineus to bark odors of gumbo-limbo (Bursera simaruba), mango (Mangifera indica) and chinini (Persea schiedeana) with different aging times and used GC–MS to analyze the chemical composition of these bark odors. The antennal responses of laboratory-reared and wild females differed in X. affinis and X. ferrugineus when interacting with odors. In addition, both beetle species displayed stronger antennal responses to aged bark odors of gumbo-limbo and chinini, apparently due to changes in volatile emissions over time.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.

Electrophysiological and behavioural responses of Stegobium paniceum to volatile compounds from Chinese medicinal plant materials.

BACKGROUND Stegobium paniceum (Coleoptera, Anobiidae) is an important pest of stored products causing severe damage to dried Chinese medicinal plant materials (CMPMs). Plant volatiles play an important role in host‐searching of insects. The olfactory responses of S. paniceum to the most abundant volatile components of some drugstore attractant CMPMs such as Panax notoginseng, Angelica sinensis, Gastrodia elata and Peucedanum praeruptorum, namely falcarinol, 3‐n‐butylphthalide, p‐cresol and β‐pinene, respectively, were studied by electroantennography (EAG) and behavioural bioassays in six‐ and four‐arm olfactometers.RESULTSEAG recordings showed that male and female antennae are able to perceive the test compounds in a wide range of concentrations and in a dose‐dependent manner. Moreover, for each dose of different compounds tested, no significant differences were found between the mean male and female EAG responses. In six‐arm olfactometer bioassays, S. paniceum exhibited positive responses to falcarinol, 3‐n‐butylphthalide, p‐cresol and β‐pinene at doses of 1, 10, 100, 500 and 1000 μg. The most attractive dose was 500 μg for falcarinol, 100 μg for 3‐n‐butylphthalide, 500 μg for p‐cresol and 1000 μg for β‐pinene. Olfactory preferences of S. paniceum, based on comparison of these four compounds at their optimally attractive concentrations in a four‐arm olfactometer, were 3‐n‐butylphthalide > p‐cresol > falcarinol > β‐pinene.CONCLUSIONThe results indicated that the four volatiles of CMPMs are perceived by the peripheral olfactory system of S. paniceum adults and are able to individually elicit a positive chemotaxis in S. paniceum adults confirming the role of chemical cues in host‐plant detection and selection of this pest. Further field studies are needed to evaluate the potential of the attractive compounds identified in this study, particularly 3‐n‐butylphthalide, to be applied as a novel monitoring and control tool against this storage‐beetle pest. © 2022 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Electroantennographic activity of 21aliphatic 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 10-17carbon atoms bind well to anL. 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.

Antennae of psychodid and sphaerocerid flies respond to a high variety of floral scent compounds of deceptive Arum maculatum L.

Insect-pollinated plants often release complex mixtures of floral scents to attract their pollinators. Yet scent compounds eliciting physiological or behavioural responses in pollinators have only been identified in few plant species. The sapromyiophilous aroid Arum maculatum releases a highly diverse dung-like scent with overall more than 300 different compounds recorded so far to attract its psychodid and other fly pollinators. The volatiles’ role in pollinator attraction is mostly unknown. To identify potential behaviourally active compounds, we recorded electroantennographic responses of four Psychodidae and one Sphaeroceridae species to (1) inflorescence scents of A. maculatum and (2) the scents released by cow dung, likely imitated by the plant species. Here we show that these flies are sensitive to 78 floral volatiles of various chemical classes, 18 of which were also found in cow dung. Our study, which for the first time determined physiologically active compounds in the antennae of Psychoda spp. and Sphaeroceridae, identified various volatiles not known to be biologically active in any floral visitors so far. The obtained results help deciphering the chemical basis that enables A. maculatum and other plants, pollinated by psychodids and sphaerocerids, to attract and deceive their pollinators.

Unusual (2R,6R)-bicyclo[3.1.1]heptane ring construction in fungal α-trans-bergamotene biosynthesis

SummaryBergamotenes are bicyclo[3.1.1]heptane sesquiterpenes found abundantly in plants and fungi. Known bergamotene derivatives all possess (2S,6S)-bergamotene backbone. In this study, two (+)-α-trans-bergamotene derivatives (1 and 2) with unusual (2R,6R) configuration were isolated and elucidated from marine fungus Nectria sp. HLS206. The first (+)-α-trans-bergamotene synthase NsBERS was characterized using genome mining and heterologous expression-based strategies. Based on homology search, we characterized another (+)-α-trans-bergamotene synthase LsBERS from Lachnellula suecica and an (+)-α-bisabolol synthase BcBOS from Botrytis cinerea. We proposed that the cyclization mechanism of (+)-α-trans-bergamotene involved endo-anti cyclization of left-handed helix farnesyl pyrophosphate by (6R)-bisabolyl cation, which was supported by molecular docking. The biosynthesis-based volatiles (3–6) produced by heterologous fungal expression systems elicited significant electroantennographic responses of Helicoverpa armigera and Spodoptera frugiperda, respectively, suggesting their potential in biocontrol of these pests. This work enriches diversity of sesquiterpenoids and fungal sesquiterpene synthases, providing insight into the enzymatic mechanism of formation of enantiomeric sesquiterpenes.

The Odorant Binding Protein, SiOBP5, Mediates Alarm Pheromone Olfactory Recognition in the Red Imported Fire Ant, Solenopsis invicta.

Olfaction is crucial in mediating various behaviors of social insects such as red imported fire ants, Solenopsis invicta Buren. Olfactory receptor (OR) complexes consist of odor-specific ORs and OR co-receptors (Orco). Orcos are highly conserved across insect taxa and are widely co-expressed with ORs. Odorant binding proteins (OBPs) can transport semiochemicals to ORs as protein carriers and thus constitute the first molecular recognition step in insect olfaction. In this study, three OBP genes highly expressed in S. invicta antenna, OBP1, OBP5, OBP6, and Orco were partially silenced using RNA interference (RNAi). RNAi SiOBP5- and Orco-injected ants showed significantly lower EAG (electroantennography) responses to fire ant alarm pheromones and the alkaloid, 2,4,6-trimethylpyridine than water- or GFP-injected ants 72 h post injection. Subsequent qRT-PCR analysis demonstrated that the transcript level of the OBP1, OBP5, OBP6, and Orco significantly decreased 72 h after ants were injected with dsRNAs; however, there were no transcript level or EAG changes in ants fed dsRNAs. Our results suggest that S. invicta Orco and SiOBP5 are crucial to fire ants for their responses to alarm pheromones. RNAi knocking down SiOBP5 can significantly disrupt alarm pheromone communication, suggesting that disrupting SiOBP5 and Orcos could be potentially useful in the management of red imported fire ants.

Electroantennographic Responses of Cerambyx welensii Küster to Host-Related Volatiles

Wood-boring insects, such as Cerambyx welensii Küster, are involved in oak decline in Mediterranean areas. To advance our understanding of the olfactory perception of C. welensii, we recorded electroantennographic (EAG) responses from male and female antennae to 32 tree volatile organic compounds typical of emissions from its main Quercus L. hosts, and also analysed the dose-dependent response. Cerambyx welensii antennae responded to 24 chemicals. Eight odorants elicited the highest EAG responses (normalized values of over 98%): 1,8-cineole, limonene-type blend, β-pinene, pinene-type blend, sabinene, α-pinene, turpentine and (E)-2-hexenal. Cerambyx welensii exhibits a broad sensitivity to common tree volatiles. The high EAG responses to both limonene- and pinene-type blends suggest the detection of specific blends of the main foliar monoterpenes emitted by Q. suber L. and Q. ilex L. (limonene, α- and β-pinene, sabinene and myrcene), which could influence the intraspecific host choice by C. welensii, and in particular, females may be able to detect oak trees with a limonene-type chemotype. In addition, C. welensii showed high antennal activity to some odorants that characterize emissions from non-host tree species (1,8-cineole, β-pinene, α-pinene, turpentine, δ3-carene and camphene). The results obtained may be applicable to optimize monitoring and mass-trapping programmes in an integrated pest management context.

Pollination of the strongly scented Sarcoglottis acaulis (Orchidaceae) by male orchid bees: nectar as resource instead ofperfume.

The Neotropical orchid genus Sarcoglottis comprises ~40 species, which emit strong floral scents, presumably involved in pollinator attraction. Information on basic aspects of its natural history is scant, with the few studies available so far pointing to nectar-seeking orchid bees as pollinators. Here, we investigated the reproductive biology of Sarcoglottis acaulis, addressing the ecological meaning of its floral scent. In Atlantic Forest fragments of NE Brazil, we described the floral biology, determined the breeding system and recorded the pollinators of S. acaulis. Additionally, we chemically characterized its floral scent and assessed its role on pollinator attraction. Although self-compatible, S. acaulis depends on pollinators to boost fruit set. Male orchid bees of Eulaema atleticana and E. niveofasciata were the only recorded pollinators. They foraged for nectar only, in spite of the strong scent emitted by S. acaulis flowers. The floral scent is composed of six compounds, of which geraniol and nerol elicited electroantennographic responses in Eulaema bees. A synthetic mixture of these compounds attracted Eulaema bees in field assays but did not trigger the stereotyped scent-gathering behaviour. The floral scent of S. acaulis acts in signalling and nectar is the sole reward for pollinators. Despite the low pollinator frequency, S. acaulis shows a high fruit set (77%), particularly when compared to other orchids. Attributes such as pollinia that release small pollen loads, allowing a single pollinia-carrying bee to pollinate several flowers, low amount of nectar, steady-state flowering and traplining behaviour of pollinators, might act together to assure this outstanding fruit set.

Cadmium exposure disrupts the olfactory sensitivity of fire ants to semiochemicals

Ants are eusocial insects and have evolved sensitive chemosensory systems for social communication. However, the effect of heavy metal contamination on the olfactory sensitivity of ants remains largely unknown. Here, we investigated the survival and olfactory response of Solenopsis invicta under cadmium (Cd) exposure. As a result, exposure to dietary Cd at different concentrations (100, 300 and 500 mg/L) caused higher Cd accumulation and lower survival of the ants compared with the control (0 mg/L). Cd exposure induced diverse expression patterns of odor binding protein genes (SiOBPs) in S. invicta antenna. Specifically, the expression of SiOBP4, SiOBP11, SiOBP12 and SiOBP16 was increased by 1.84-, 1.14-, 0.83- and 1.76-fold, respectively, at 300 mg/L Cd, while SiOBP7 and SiOBP9 were suppressed as Cd concentration increased. Electroantennography (EAG) and behavioral bioassays were performed to further evaluate the effect of Cd contamination on the olfactory sensitivity of S. invicta workers to 2, 4, 6-trimethylpyridine (TMP) and 2-ethyl-3,6(5)-dimethylpyrazine (EDP), the two frequent functional semiochemicals for S. invicta. The results showed that under no Cd exposure, S. invicta workers exhibited strong EAG response and apparent residing repellence to TMP and EDP, but Cd exposure suppressed EAG response and deprived the behavioral repellence to TMP and EDP of the workers, suggesting that Cd exposure decreases the olfactory sensitivity of S. invicta to these two functional semiochemicals. Further fluorescence competitive binding assay revealed that SiOBP7 had strong binding affinity to TMP and EDP, suggesting that the decrease in olfactory sensitivity may be attributed to the inhibitory effect of Cd exposure on SiOBP7. Overall, our results suggest that Cd exposure may not only directly decrease the survival of ants, but also affect their olfactory recognition.