Antennal resolution of pulsed pheromone plumes in three moth species

Male antennae of Cadra cautella,Pectinophora gossypiella, and Spodoptera exigua were presented with 20-ms-duration pulses of their two-component pheromone at rates of 1 to 33 Hz. Fourier analyses of electroantennograms resolved the temporal structure of trains of pheromone filaments delivered at up to 33 Hz for C. cautella and S. exigua and 25 Hz for P. gossypiella. Pheromone components tested separately for each species were generally equivalent in filament resolution to complete blends. Ambient temperatures of 18, 23 and 28 °C affected filament resolution only slightly, with poorer ability to discriminate rapidly pulsed signals at 18 °C. The question of how, or indeed if, such frequencies are conserved beyond the peripheral nervous system, remains.     

Release mechanism of sex pheromone in the female gypsy moth <Emphasis Type="Italic">Lymantria dispar</Emphasis>: a morpho-functional approach

A morpho-functional investigation of the sex pheromone-producing area was correlated with the pheromone release mechanism in the female gypsy moth Lymantria dispar. As assessed by male electroantennograms (EAG) and morphological observations, the pheromone gland consists of a single-layered epithelium both in the dorsal and ventral halves of the intersegmental membrane between the 8th and 9th abdominal segments. By using the male EAG as a biosensor of real-time release of sex pheromone from whole calling females, we found this process time coupled with extension movements of the ovipositor. Nevertheless, in females in which normal calling behavior was prevented, pheromone release was detected neither in absence nor in presence of electrical stimulation of the ventral nerve cord/terminal abdominal ganglion (TAG) complex. Tetramethylrhodamine-conjugated dextran amine stainings also confirm the lack of any innervation of the gland from nerves IV to VI emerging from the TAG. These findings indicate that the release of sex pheromone from the glands in female gypsy moths is independent of any neural control exerted by the TAG on the glands, at least by way of its three most caudally located pairs of nerves, and appears as a consequence of a squeezing mechanism in the pheromone-producing area.     

松褐天牛触角感器电镜扫描和触角电位反应

应用扫描电镜对松褐天牛的触角感器进行扫描观察,并测定了触角对松树主要挥发物的电生理反应.松褐天牛触角上共有7种感器,即毛形感器、锥形感器、指形感器、棒形感器、瓶形感器、芽形感器以及刺形感器,以毛形和锥形感器的数量最丰富.其中毛形感器和锥形感器各有3个型,指形感器有2个型.毛Ⅱ、毛Ⅲ型和锥Ⅰ、锥Ⅱ型感器表皮结构具有深的纵刻纹,具有嗅觉感器的典型特征.同时对各种感器的数量、长度及分布特点等作了描述.电生理研究结果表明,几种供试化合物及其混合物均能引起松褐天牛雌、雄成虫较强的电生理反应;松褐天牛触角不同部位的EAG反应存在显著差异;剂量试验表明,松褐天牛成虫触角对不同浓度的α-Pinene的EAG反应存在一定的规律性.     

桔小实蝇对几种寄主挥发物的触角电位反应

桔小实蝇寄主范围广,可危害芒果、柑橘、咖啡和红辣椒等250多种水果和蔬菜,现已成为中国、东南亚、印度次大陆和夏威夷群岛一带的危险性果蔬害虫。本文利用触角电位(electroantennogram,EAG)技术,测定了桔小实蝇雌雄成虫对8种寄主挥发物标准化合物(1,8桉叶素、α-蒎烯、β-石竹烯、柠檬醛、香草醛、乙醇、芳樟醇和月桂烯)的EAG反应。结果表明:上述标准化合物均能引起EAG反应,EAG反应值随着浓度增大(0.1μg/μL、1μg/μL、10μg/μL和100μg/μL)而显著不同。当柠檬醛和月桂烯浓度为100μg/μL时,桔小实蝇雌虫交配前、后的EAG反应值存在显著性差异。浓度为100μg/μL的1,8桉叶素对桔小实蝇交配后雌虫和雄虫的刺激反应存在显著性差异。桔小实蝇交配前雌虫和雄虫对浓度为100μg/μL乙醇的EAG反应值存在显著性差异。     

Phorid fly, Pseudacteon tricuspis, response to alkylpyrazine analogs of a fire ant, Solenopsis invicta, alarm pheromone

The phorid fly, Pseudacteon tricuspis Borgmeier, is a parasitoid of the red imported fire ant, Solenopsis invicta Buren. This fly has been reported to use fire ant chemicals, specifically venom alkaloids and possibly alarm pheromone to locate its host. A recent study identified 2-ethyl-3,6-dimethyl pyrazine as a component of the alarm pheromone of S. invicta. To determine the possible involvement of this fire ant alarm pheromone component in mediating fire ant-phorid fly interactions, we tested electroantennogram (EAG) and behavioral responses of P. tricuspis females to the commercially available mixture of 2-ethyl-3,6-dimethyl pyrazine and its 3,5-dimethyl isomer, as well as six structurally related alkylpyrazine analogs at varying doses. Pseudacteon tricuspis females showed significant EAG response to 2-ethyl-3,6(or 5)-dimethyl pyrazine (herein referred to as pheromone-isomer) at all doses, 0.001-10 μg. Among the tested alkylpyrazine analogs, 2,3-diethyl-5-methyl pyrazine showed significant EAG activity at 0.1 and 1 μg. 2,3-dimethyl pyrazine also showed significant EAG activity at 0.1 μg. Results of four-choice olfactometer bioassays demonstrated significant attraction of P. tricuspis females to the pheromone-isomer (2-ethyl-3,6(or 5)-dimethyl pyrazine) at all tested doses (0.01, 0.1, 1 and 10 μg). The analogs, 2,3-diethyl-5-methyl pyrazine and 2,3-dimethyl pyrazine were significantly better than the control at the higher doses (0.1, 1 and 10 μg). The pheromone-isomer was significantly better than both analogs at two doses, 0.1 and 1 μg. These results confirm that the reported fire ant alarm pheromone component plays a role in mediating attraction of phorid flies to host workers. Venom alkaloids were previously shown to attract P. tricuspis; therefore, we propose that fire ant alarm pheromones may act in tandem or synergistically with venom alkaloids to attract phorid fly parasitoids to fire ant workers.     

Behavioural and electrophysiological responses of the European corn borer Ostrinia nubilalis to host‐plant volatiles and related chemicals

The European corn borer Ostrinia nubilalis (Hübner) (Lepidoptera: Crambidae) is a worldwide pest of maize (Zea mays L.) and other crops. The semiochemicals released by maize plants and structurally‐related compounds can be used by adult female O. nubilalis for host‐plant location and oviposition. Headspace volatile compounds emitted by watered and water‐deprived maize plants are collected and identified by their retention indices and mass spectra. The most abundant compounds from watered plants are limonene, linalool, benzoic acid, indole, β‐caryophyllene and acetophenone, whereas, in water‐deprived plants, limonene, acetophenone, hexanoic acid, benzoic acid and indole are dominant. In addition, (E)‐4,8‐dimethyl‐1,3,7‐nonatriene, 6‐methyl‐5‐hepten‐2‐one, anisole and 1‐carvone are undetected in the water‐deprived plants. Some of the identified compounds show electrophysiological activity (electroantennogram) in the antennae of both sexes, with the responses elicited by tridecane, tetradecane, dodecane, nonanal, decanal and 2‐ethylhexanol on males being particularly noteworthy. In a dual‐choice olfactometer, adult females show a preference for 2‐hexanol, heptanal, methyl salicylate, hexyl acetate, nonanal, methyl dodecanoate, β‐pinene and (E)‐2‐hexenyl acetate over hexane controls. Tetradecane, linalool, methyl hexanoate, methyl nonanoate, (Z)‐3‐hexenyl benzoate, tridecane, 2‐cyclopentylcyclopentanone, 3‐methylbutyl acetate, β‐myrcene and (Z)‐3‐hexenyl butanoate result in fewer females in the test arm compared with the control arm. No single compound displays an activity similar to watered maize plants, supporting the hypothesis that blends of volatiles in specific ratios are more effective than single volatile chemicals. The results of the present study suggest that methyl salicylate, which elicits also one of the highest electrophysiological responses in female antennae, plays a role in host preference by O. nubilalis females.     

Identification of host‐plant chemical stimuli for the European grape berry moth Eupoecilia ambiguella

Olfaction is of major importance for survival and reproduction in moths. Males possess highly specific and sensitive olfactory receptor neurones to detect female sex pheromones. However, the capacity of male moths to respond to host‐plant volatiles is relatively neglected and the role that such responses could play in the sensory ecology of moths is still not fully understood. The present study aims to identify host‐plant stimuli for the European grape berry moth Eupoecilia ambiguella Hb. (Tortricidae, Lepidoptera), a major pest of vine in Europe. Headspace volatiles from Vitis vinifera L. cv. Pinot Noir, Vitis vinifera subsp. sylvestris and five other host‐plant species comprising five different families are analyzed by gas chromatography linked to electroantennogram (EAG) recording from male E. ambiguella antennae and by gas chromatography‐mass spectrometry. This procedure identifies 32 EAG‐active compounds, among them the aliphatic compounds 1‐hexanol, (Z)‐3‐hexenol, (Z)‐3‐hexenyl acetate and 1‐octen‐3‐ol; the terpenes limonene, β‐caryophyllene and (E)‐4,8‐dimethyl‐1,3,7‐nonatriene; and the aromatic compounds benzaldehyde and methyl salicylate. Male and female E. ambiguella show similar EAG response amplitudes to individual chemical stimuli and also to mixtures of plant volatiles, as represented by essential oils from ten other plant species. This possibly indicates a common role for plant compounds in the sensory ecology of the two sexes of E. ambiguella.     

How the choice of method influence on the results in electrophysiological studies of insect olfaction

In identifying the volatiles that insects use to locate suitable host plants, electrophysiological recordings of olfactory responses to plant volatiles may give important information. However, divergent results may be obtained with different recording techniques. To illustrate these differences, the results of a previous investigation using single cell recordings linked to a gas chromatograph (SCR-GC) are compared with the results obtained with an electroantennogram linked to a gas chromatograph (EAG-GC). Testing insects of the same species (Hylobius abietis) for the same test sample, 30 potent volatile compounds were identified by SCR-GC and 18 by EAG-GC. Of the 34 different compounds, 14 were identified by both techniques. Furthermore, when the same compound elicited detectable responses by both techniques, the response strength was usually not the same relative to the strongest response recorded by each technique. This shows that both EAG-GC and SCR-GC are important techniques in the identification of potent plant volatiles for insects. However, by using SCR-GC more information was obtained, information that can be crucial for understanding the insect-plant relationship.     

Increased EAG responses of tortricid moths after prolonged exposure to plant volatiles: evidence for octopamine-mediated sensitization

As measured by electroantennograms (EAG), both male and female obliquebanded leafrollers, Choristoneura rosaceana (Harris), and redbanded leafrollers, Argyrotaeniavelutinana (Walker), were similarly sensitive to host-related plant volatiles: trans-2-hexenal, benzaldehyde, 1-hexenol, cis-3-hexen-1-ol, geraniol, linalool, (+)-limonene, hexenal and trans-2-hexenol. Females of both species were similarly sensitive to the shared major component of their sex-attractant pheromone ((Z)11-14:Ac). Continuous 60 min pre-exposure of male and female C. rosaceana and A. velutinana to successively higher concentrations of a mixture of the nine plant volatiles in Teflon chambers with continuous air exchange caused a dosage-dependent increase in subsequent responsiveness (sensitization) to green leaf volatiles, as measured by EAGs. In addition, 60 min of pre-exposure of male C. rosaceana to certain individual volatiles ((+)-limonene, geraniol, benzaldehyde) increased EAGs nearly as much as did the mixture of nine volatiles. Pre-exposures to the nine plant-volatile mixture at concentrations achieved by 100 microg and 1 mg loading dosages in 100 microl of mineral oil significantly increased EAG depolarization to pheromone (cross-sensitization) in males but not females of both moth species. Antennae of male C. rosaceana pre-injected with 100 microg of octopamine (OA) without volatile pre-exposure exhibited sensitization nearly identical to that induced by pre-exposing moths to sensitizing concentrations of the plant-volatile mixture. Moreover, injection of the OA antagonist chlorpromazine (CP) blocked sensitization by the plant-volatile pre-exposure. Collectively, these findings suggest that exposures of tortricid moths to certain host-plant related volatiles may modulate subsequent olfactory sensitivity to behaviorally relevant chemical cues and that plant-volatile induced sensitization may be octopamine mediated.