Electrophysiological and Behavioral Responses of Female Beet Armyworm Spodoptera exigua (Hübner) to the Conspecific Female Sex Pheromone

Electroantennogram (EAG) recordings showed that female Spodoptera exigua can detect their own sex pheromones (two single components and their mixture), displaying a similar dose–response pattern to that of males, although intensities of female responses were much less at all doses compared with males. Furthermore, the female calling behavior was inhibited and late-shifted by the presence of the female sex pheromone. When the pheromone components were presented, the calling female proportion in the peak calling period was significantly reduced and the calling peak time and calling termination time postponed, compared with controls. Although the calling behavior was inhibited, the pheromone titer of treated females was not different to the control, implying a reduced pheromone biosynthesis in the pheromone glands of treated moths. However, observations during the olfactometer experiments revealed that there were no obvious behavioral responses of females exposed to sex pheromone stimuli including whole gland extracts, 0.1, 1 or 10 μg binary pheromone mixtures.     

Reduction in responsiveness of male apple moths, Epiphyas postvittana, to sex pheromone following pulsed pheromonal exposure

ABSTRACT. The degree of reduction in sex pheromone-mediated responsiveness of male light-brown apple moth, which results from previous exposure to the pheromone, is increased by presenting the pre-exposure dose as a sequence of short pulses, rather than as a single pre-exposure. This effect is interpreted in terms of adaptation and disadaptation of the pheromone receptors, and of central habituation and dishabituation. When the interval between pulses is of sufficient length for receptor disadaptation, the total amount of excitatory input generated by the pulsed pheromone would be greater than if the same dose were to be applied continuously. It is concluded that the greater resultant reduction in responsiveness following pulsing is directly related to the greater degree of habituation resulting from this presumed increased amount of excitatory input.     

Reduction in responsiveness of male light-brown apple moths, Epiphyas postvittana, to sex pheromone following pulsed pre-exposure to pheromone components

ABSTRACT. A combination of two sex pheromone compounds is required to elicit sexual response in males of Epiphyas postvittana. Sequential exposure to the two compounds failed to elicit sexual response in males, but a pre-exposure sequence of alternated pulses of the two compounds did habituate males to the same extent as did pulsed pre-exposures to the compounds combined.     

Functional specialisation and polyphenism in aphid olfactory sensilla

Electroantennogram (EAG) responses to the aphid sex pheromone components, (1R,4aS,7S,7aR)-nepetalactol and (4aS,7S,7aR)-nepetalactone, and a plant volatile, (E)-2-hexenal, were investigated at three different positions (5/6th, 4/5th and 3/4th inter-segmental regions) along the antennae of four different morphs in two host-alternating aphid species, Aphis fabae Scopoli and Rhopalosiphum padi (L.). Position-dependent and morph-specific EAG responses were elicited in both species. The nepetalactol and nepetalactone isomers elicited large EAG responses at all three recording positions in males of both species, such that primary rhinaria as well as secondary rhinaria appeared to respond. Asexual female morphs showed relatively smaller EAG responses to these compounds. The secondary rhinaria, which have been reported as sex pheromone receptors in males, were not very different in their number and distribution between gynoparae and alate virginoparae, but the gynoparae showed significantly larger EAG responses to nepetalactol and nepetalactone. The alate virginoparae showed EAG responses that were similar to those of apterous virginoparae, which lack secondary rhinaria. Taking the EAG response profiles together with the distribution of the secondary rhinaria, it is suggested that the function of secondary rhinaria differs between the morphs. Secondary rhinaria appear to detect sex pheromone components in males and gynoparae but not in the alate virginoparae. If they are functional in the latter morph, they are likely to play a role in detecting specific, but as yet unknown, volatile compounds. Some 30 plant volatiles were tested but none evoked an EAG response that could be allocated to the secondary rhinaria. In contrast to the very different EAG response profiles to the pheromone compounds between morphs, EAG responses to (E)-2-hexenal were similar in all forms and both species. These findings suggest that this plant volatile was detected only by the two primary rhinaria, which are common to all morphs. The present study showed that EAG responses were not a simple summation of receptor potentials between recording and reference electrodes in aphids. The localised distribution pattern of olfactory receptor neurones around the recording electrode was also likely to contribute to the EAG responses.     

Brief exposure to sensory cues elicits stimulus-nonspecific general sensitization in an insect

The effect of repeated exposure to sensory stimuli, with or without reward is well known to induce stimulus-specific modifications of behaviour, described as different forms of learning. In recent studies we showed that a brief single pre-exposure to the female-produced sex pheromone or even a predator sound can increase the behavioural and central nervous responses to this pheromone in males of the noctuid moth Spodoptera littoralis. To investigate if this increase in sensitivity might be restricted to the pheromone system or is a form of general sensitization, we studied here if a brief pre-exposure to stimuli of different modalities can reciprocally change behavioural and physiological responses to olfactory and gustatory stimuli. Olfactory and gustatory pre-exposure and subsequent behavioural tests were carried out to reveal possible intra- and cross-modal effects. Attraction to pheromone, monitored with a locomotion compensator, increased after exposure to olfactory and gustatory stimuli. Behavioural responses to sucrose, investigated using the proboscis extension reflex, increased equally after pre-exposure to olfactory and gustatory cues. Pheromone-specific neurons in the brain and antennal gustatory neurons did, however, not change their sensitivity after sucrose exposure. The observed intra- and reciprocal cross-modal effects of pre-exposure may represent a new form of stimulus-nonspecific general sensitization originating from modifications at higher sensory processing levels.     

Localization and fine structure of the female sex pheromone-producing glands in Bruchidius atrolineatus (PIC) (Coleoptera : Bruchidae)

The sex pheromone glands of female Bruchidius atrolineatus (Coleoptera: Bruchidae) have been localized by recording the electric response of the antenna of males subjected to a stream of air, containing the volatile sex pheromone (electroanntennography-EAG). Some 50 unicellular glands are distributed irregularly in the ventral and dorsal intersegmental membranes situated at the extremity of the pygidium, each gland containing a short ductule with an evacuation pore, 0.5–1 μm in diameter. The receiving canal is composed of a network of fine epicuticular filaments. The glands are type 3. The ultrastructure of these sex pheromone-producing glands is described in females whose production-emission activity had been previously verified with EAG. Deep basal invaginations and inflated intercellular spaces indicate the transport of substances from the hemolymph to the gland cells. The presence of numerous elongated mitochondria, diverse inclusions, vesicles containing crystalline bodies, and abundant apical microvilli, all reveal elevated cellular activity, which is never observed in young or diapausing females that do not produce sex pheromone. The ultrastructural differences in different types of females (sexually active or diapausing), combined with comparative EAG recordings obtained with intact females or those in which the suspected glandular zone was masked, made it possible to localize the glands.     

Field and laboratory electroantennographic measurements of pheromone plume structure correlated with oriental fruit moth behaviour

Abstract. Peak-to-trough electroantennogram amplitudes (bursts), caused by the individual filaments of a plume of female pheromone, diminish as high-emission-rate sources are approached by male Grapholita molesta , and this reduction is correlated with in-flight arrestment (ceasing to advance upwind). These findings are consistent with the hypothesis that one cause of in-flight arrestment in response to high-concentration point sources is the attenuation of the peak-to- trough amplitudes close to the source. High burst frequency, high pheromone flux, or low levels of continuous neuronal activity all are less well correlated with arrestment. Rather, arrestment appears due to a reduction of chemosensory input to the CNS during flight up the plume, even though the actual molecular concentration continues to increase. In a laboratory wind tunnel, upwind flight initiation by more than 20% of males was elicited only by pheromone source concentrations evoking significant fluctuations in EAG amplitudes at downwind release points. The burst frequencies that evoked high levels of upwind flight initiation ranged from a mean of 0.4-2.2 bursts/s. Because a previous study revealed that flying male G. molesta change their course angle within 0.15 s of losing or contacting pheromone, these EAG burst frequencies indicate that during flight in a pheromone plume, many manoeuvres are probably made in response to contact with individual plume filaments. Thus, upwind flight tracks may be shaped by hundreds of steering reactions in response to encounters with individual pheromone filaments and pockets of clean air. Field-recorded EAGs reveal that burst amplitudes diminish from 3 to 30 m downwind of the source, whereas burst frequencies do not, averaging c. 1/s at 3, 10 and 30 m downwind.     

Effects of sex pheromone in electrostatic powder on mating behaviour by Lobesia botrana males

Abstract:  In laboratory trials, mating behaviour of males of Lobesia botrana Den. and Schiff. (Lep., Tortricidae) was investigated after treatment with an electrostatically chargeable powder, Entostat^TM. Male moths were powdered with either blank Entostat or Entostat containing sex pheromone. Significantly more Entostat adhered to L. botrana males when sex pheromone was added to the powder compared with blank Entostat. Powdering male moths with Entostat, with or without sex pheromone, caused a significant reduction in antennal response when antennae were placed 2 cm away in downwind direction, while there was little effect when antennae were placed 25–100 cm from the EAG in downwind direction. In a flight study, powdering males of L. botrana caused significant increase in time before flight initiation and reduction in proportion of males making contact with calling females compared with untreated males. In a mating experiment in Petri dishes (to minimize required search time to locate female), significantly fewer females mated successfully (based on dissection of bursa copulatrix) when males had been treated with pheromone-loaded powder. Overall, powdering males of L. botrana caused considerable suppression of mating behaviour on various levels, and these suppressing effects were increased after adding sex pheromone to Entostat.     

Ontogeny of electroantennogram responses in the moth, Manduca sexta

The antennae of the moth, Manduca sexta, and the sensilla and sensory neurons they contain, develop during metamorphosis from pupa to adult. To determine when, during their development, antennae become capable of generating electrical responses to various stimuli, we recorded the electroantennogram (EAG), believed to be the summed extracellular record of receptor potentials, from developing and mature antennae. Antennae from male and female moths are similarly responsive to trans-2-hexenal, while only males respond to Manduca sex pheromone; these two odorants presumably stimulate separate receptors. Mechanical stimulation also elicits and EAG response. EAG responses to olfactory and mechanical stimuli are detectable several days before eclosion but not until the neurons are morphologically and biochemically quite mature. Responses increase in magnitude until the end of metamorphosis and then change little during the first 3 days after emergence of the adult. Responses to different stimuli do not develop synchronously.     

Electrophysiological investigations of sex pheromone reception and release in Bruchidius atrolineatus

ABSTRACT. Male antennal sensitivity to female sex pheromone in Bruchidius atrolineatus was investigated electrophysiologically (EAG). The existence of receptors for this pheromone was shown in the antennae of males. (No such receptors exist in the antennae of females.) Young (age < 24h) males gave an EAG of amplitude 25–30% of the response of older males. Diapausing males several days old gave a much smaller response than active males of the same age.
EAGs were used to test the production/release of pheromone by groups of twenty females. Before they are 24 h old, active females do not produce/release the pheromone; older females release it at an almost constant rate. Sexually diapausing females do not produce and/or release the sex pheromone.
In sexually active females, mating inhibits pheromone release almost immediately. There is a correlation between reproductive status of the females (development of ovaries, oogenesis) and the production or release of the pheromone. A corresponding correlation also exists in the males whose antennae show a very low sensitivity when they are young or when they are in reproductive diapause.     

The circadian rhythm of flight activity of Spodoptera exigua males in response to sex pheromone

In many moths, male attraction to the blend of synthetic sex pheromone releasing continuously in the field shows an apparent circadian rhythm similar to that of locomotion activity. In this study, the daily rhythms of electroantennography (EAG) and behavioral responses to sex pheromone, and the daily rhythms of locomotion activity were measured in male beet armyworm, Spodoptera exigua (Hübner) (Lepidoptera: Noctuidae). The peaks of males trapped by light and sex pheromone were all during the latter part of the night in the field. However, there was no significant variation among time intervals in the EAG responses of male antennae to sex pheromone stimuli. The principal period of locomotion activity under L15:D9 (LD) continued to occur during the scotophase and the subjective scotophase in the day of constant darkness (DD1) and the second of two consecutive days of constant darkness (DD2). The majority of males contacted the sex pheromone source in a wind tunnel during the latter part of the scotophase under LD and the subjective scotophase for DD1 and DD2. There were significant associations between the rhythm of the behavioral response to sex pheromone and locomotion activity. These results suggested that the male's behavioral response to sex pheromone in the beet armyworm could be observed only until locomotion activity of the male occurred at the end of the dark period, despite sex pheromone being released continuously from synthetic pheromone‐baited traps in the field.     

Host plant volatiles induce oriented flight behaviour in male European grapevine moths, Lobesia botrana

The European grapevine moth Lobesia botrana relies on a female produced sex pheromone for long-distance mate finding. Grapevine moth males compete heavily during limited time windows for females. The aim of this study was to investigate the perception of host plant volatiles by grapevine moth males and whether such compounds elicit upwind oriented flights. We compared five host plant headspace extracts by means of gas chromatography linked electroantennogram (EAG) recording. We identified 12 common host plant volatiles (aliphatic esters, aldehydes, and alcohols, aromatic compounds and terpenes) that elicit EAG responses from grapevine moth males and that occur in at least three of the host plant volatile headspace extracts tested. Subsequently the behavioural response of grapevine moth males to four these compounds presented singly and in mixtures (1-hexanol, 1-octen-3-ol, (Z)-3-hexenyl acetate and (E)-β-caryophyllene) was recorded in a wind tunnel. Grapevine moth males engaged in upwind flights to all of four compounds when released singly at 10,000 pg/min and to all, except 1-octen-3-ol, when released at 100 pg/min. A blend of the four host plant volatiles released at 10,000 pg/min and mixed at a ratio based on the analysis of Vitis vinifera cv. Solaris volatile emissions attracted significantly more males than any single compound. Grapevine moth males perceive and respond to host plant volatiles at biologically relevant levels indicating that host plant volatiles figure as olfactory cues and that L. botrana males can discern places where the likelihood of encountering females is higher.     

Modification of Male Courtship Motivation by Olfactory Habituation via the GABAA Receptor in Drosophila melanogaster

A male-specific component, 11-cis-vaccenyl acetate (cVA) works as an anti-aphrodisiac pheromone in Drosophila melanogaster. The presence of cVA on a male suppresses the courtship motivation of other males and contributes to suppression of male-male homosexual courtship, while the absence of cVA on a female stimulates the sexual motivation of nearby males and enhances the male-female interaction. However, little is known how a male distinguishes the presence or absence of cVA on a target fly from either self-produced cVA or secondhand cVA from other males in the vicinity. In this study, we demonstrate that male flies have keen sensitivity to cVA; therefore, the presence of another male in the area reduces courtship toward a female. This reduced level of sexual motivation, however, could be overcome by pretest odor exposure via olfactory habituation to cVA. Real-time imaging of cVA-responsive sensory neurons using the neural activity sensor revealed that prolonged exposure to cVA decreased the levels of cVA responses in the primary olfactory center. Pharmacological and genetic screening revealed that signal transduction via GABA_A receptors contributed to this olfactory habituation. We also found that the habituation experience increased the copulation success of wild-type males in a group. In contrast, transgenic males, in which GABA input in a small subset of local neurons was blocked by RNAi, failed to acquire the sexual advantage conferred by habituation. Thus, we illustrate a novel phenomenon in which olfactory habituation positively affects sexual capability in a competitive environment.     

Electroantennogram and behavioural responses of the gypsy moth to enantiomers of disparlure and its trans analogues

The attractive power of both enantiomers of disparlure and its trans analog was tested electrophysiologically (electroantennogram = EAG) and behaviourally (whole organism response). EAG responses correlated very well with the behaviourally determined effectiveness sequence: cis(+)-disparlure was the most effective substance; the authentic racemic disparlure came second; cis(?)-disparlure inhibited the activity of cis(+)-disparlure; and trans enantiomers were not significantly different from the control. These results allow us to conclude that cis(+)-disparlure is the natural sex pheromone of the gypsy moth, and that the male antennae have a chiral receptor system in the antenna for the reception of the sex pheromone.     

Electroantennogram responses of the three migratory forms of the damson-hop aphid, Phorodon humuli, to aphid pheromones and plant volatiles

Electroantennogram (EAG) responses were recorded from alate fundatrigeniae (spring migrants), gynoparae (the winged female form that produces sexual females) and males, the three migratory forms of the damson-hop aphid, Phorodon humuli (Schrank). EAG responses of gynoparae and males showed typical dose response characteristics to (E)-2-hexenal, (-)-R-carvone, hexanenitrile and (1RS,4aR,7S,7aS)-nepetalactol, the sex pheromone of this species. The 34 plant volatiles elicited broadly similar EAG response profiles in the three migratory forms. Green leaf volatiles produced large responses in all forms; however, the relative order of responsiveness varied. EAG responses to isomers of the monoterpene carvone differed between forms, with males being most, and spring migrants least, responsive. The hop-plant volatile and aphid alarm pheromone, (E)-beta-farnesene, evoked similar EAG responses in all forms. By contrast, males were most responsive to the three sex pheromone components, (-)-(4aS,7S,7aR)-nepetalactol, (+)-(4aS,7S,7aR)-nepetalactone and (1RS,4aR,7S,7aS)-nepetalactol. Males were no more responsive to their own sex pheromone, (1RS,4aR,7S,7aS)-nepetalactol, than to the other aphid sex pheromone components tested. Spring migrants and gynoparae also responded to the three sex pheromone components. This study indicates that migratory forms of P. humuli detect a wide range of volatile compounds, and that they are equally well-adapted for the detection of volatiles associated with host and non-host plants and with other species of aphid.     

Electroantennogram responses of Plutella xylostella (L.), to sex pheromone components and host plant volatile semiochemicals

The diamondback moth (DBM), Plutella xylostella (L.) (Lepidoptera: Plutellidae), is a notorious insect pest of cruciferous crops worldwide. Attract-and-kill strategies to manage the DBM based on insect pheromone and plant volatile semiochemicals have been explored and partially applied in the field. However, little is known about whether the attractant effects of insect pheromone and host plant volatile semiochemicals on insects are affected by insect age or volatile concentrations. Therefore, we examined the electroantennogram (EAG) responses of both DBM males and females varying in age and body size to a range of concentrations of seven host plant volatiles and two main female sex pheromone components. Our results showed that DBM age had no influence on EAG responses to trans-2-Hexenal, trans-2-Hexenol, Heptanal, cis-3-Hexen-1-ol and Z11-16:Ald and significantly influenced the EAG responses to Z11-16:Ac and three isothiocyanates. Age and sex had interaction effects on EAG responses of DBMs to some tested semiochemicals. DBM females had significantly stronger responses than males to all tested plant volatiles, and values were enhanced with increasing concentrations. Moreover, the degree of enhancement of EAG responses in females was larger than that in males for trans-2-Hexenal, trans-2-Hexenol, cis-3-Hexen-1-ol, 2-Phenylethyl isothiocyanate and Methyl isothiocyanate treatments. Body size seemed to have no influence on EAG responses. Our results might provide a theoretical basis for optimizing attract-and-kill strategies for insect pests.     

Effects of Trifluoromethyl Ketones and Related Compounds on the EAG and Behavioural Responses to Pheromones in Male Moths

Trifluoromethyl ketones (TFMKs) and analogues affect pheromonedetection and behaviour in male moths. 3-Octylthio-1, 1, 1-trifluoropropan-2-one(OTFP), one of the most effective antiesterase agents, decreasedthe EAG amplitude and increased the repolarization time in spodopteralittoralis. It also modified EAG responses of Marnestra brassicaeand Heliothis zea to their pheromones and analogues, containingan acetate, an alcohol or an aldehyde function. In addition,OTFP also reduced the amplitude of the EAG response to linalool,a monoterpenic alcohol, but not its kinetics. The responsesof male S. littoralis to the pheromone in the wind tunnel weresignificantly reduced after pre-exposure to vapours of OTFP.Comparison of the activity of other TFMKs and analogues withthat of OTFP revealed a good agreement on EAG and behaviour.The effects of TFMKs on the EAG kinetics are discussed consideringthe hypothesis of an inhibition of the pheromone deactivationin the antenna. Chem. Senses 22: 407–416, 1997.     

豆野螟成虫触角对植物挥发性和性信息素化合物的触角电位反应

以顺3-己烯乙酸酯为标准化合物,研究了豆野螟雌、雄成虫触角对不同浓度的植物挥发性和性信息素化合物组分的触角电位（EAG）反应.结果表明：豆野螟雌、雄成虫触角对植物挥发性气味具有强烈的EAG反应,但在大部分浓度下,雌、雄虫的反应没有显著差异.在测试的9种植物挥发性化合物中,雌、雄虫对高浓度反-2-己烯醛刺激的相对EAG反应的最大值分别达到250%和260%,且雌、雄虫触角对这种化合物的EAG反应的形状也有很大差异;但对豇豆花气味中的柏木烯和柏木脑没有明显的EAG反应.雌、雄蛾对性信息素都有明显反应,且雌、雄虫的反应存在显著差异,雄虫触角对性信息素化合物主要组分（反10,反12）-十六碳二烯醛的EAG反应的最大值达到250%,远超过雌虫反应的最大值(53%).雌、雄成虫这种神经感觉反应的差异反映了它们在感器结构、功能和行为上的差异.     

金龟甲对蓖麻叶挥发物的触角电位和行为反应

为了探明金龟甲偏爱选择有害非寄主植物蓖麻的原因,应用触角电位（EAG）仪和“Y”型嗅觉仪,分别测定了华北大黑鳃金龟、暗黑鳃金龟和铜绿丽金龟雌、雄虫对5种蓖麻叶挥发物的触角电位和选择行为反应。EAG测定结果表明,3种金龟甲雄虫对各处理挥发物的EAG反应值均比参照挥发物顺-3-己烯-1-醇强。华北大黑鳃金龟对邻苯二甲酸二丁酯和肉桂醛的EAG反应值相对较高,暗黑鳃金龟对苯甲醇的EAG反应值相对较高,铜绿丽金龟对邻苯二甲酸二丁酯和苯甲醇的EAG反应值相对较高。选择行为反应测定结果表明,当顺-3-己烯-1-醇与其他挥发物配对测试时,华北大黑鳃金龟对邻苯二甲酸二丁酯和肉桂醛2种蓖麻叶挥发物表现出明显的选择偏好,暗黑鳃金龟对蓖麻叶挥发物苯甲醇表现出明显的选择偏好,铜绿丽金龟则更偏好选择蓖麻的绿叶气味物质顺-3-己烯-1-醇。总之,蓖麻叶的挥发性物质与引诱金龟甲偏爱选择密切相关,且偏爱的挥发物因金龟甲的种类而异。     

Production and perception of pheromones by the beetle Tribolium confusum

Adults of Tribolium confusum secrete two pheromones. The first, produced by the male, is attractive to both sexes and the second, produced by the female, is attractive to the male only. Pheromone production and perception was studied in relation to habituation, beetle age, time of day and previous mating. A living source of each pheromone habituates the responding beetles, the male pheromone habituating more strongly; female pheromone habituates only in the absence of the male pheromone. Habituation to one pheromone was always accompanied by an enhanced response to the other.Five days after emergence, production of male pheromone reaches a peak that is maintained. Production of female pheromone peaks after 3 days. Both sexes are responsive to male pheromone immediately upon eclosion, males reaching maximum response at 14 days, females at 8 days. Males are also responsive to female pheromone upon eclosion reaching maximum response at 8 days; female response to female pheromone is imperceptible. Males but not females display a 24 hr rhythm in pheromone production. Mated beetles did not differ significantly from unmated beetles in their ability to perceive pheromones. Alteration in male pheromone production after mating was detected by females but not males; this pheromone may, therefore, act as both a sex and aggregation pheromone.