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.     

ELECTROANTENNOGRAM RESPONSES OF LYSIPHLEBIA JAPONICA ASHMEAD (HYMENOPTERA: APHIDIIDAE) TO SOME COTTON PLANT VOLATILES AND COTTON APHID PHEROMONES

Abstract Electroantennogram responses (EAGs) of Lysiphlebia japonica (Ashmead) (Hymenoptera: Aphidiidae) to cotton plant volatiles and cotton aphid pheromones were measured with EAG recording techniques. No sexual difference of EAG responses was found to all test samples under concentration of 10 μl/ml. The most effective chemicals were 1-heptanol, hexanal, heptanal, saturated primary-alcohols with six-carbon to nine-carbon chain and benzaldehyde. Both male and female showed the highest EAGs to green leaf volatiles (GLVs) and moderate EAGs to terpenoids among which geraniol was the most active. Cotton aphid pheromones elicited relative high EAGs. Among the four plant extracts, cotton leaf extract was the most effective. Dose curves were constructed from six female wasps' EAGs to four chosen chemicals, in which GLVs showed lower thresholds than terpenoids. According to the results above, we may draw the following conclusions: GLVs could be more important in longer distance orientation, while terpenoids may be more important in shorter ranges. Aphid pheromones may be effective attractants to parasitoids. Preadult experience can influence the antenna1 sensitivity of the parasitoid.     

Perception of volatiles associated with sex and food by different adult forms of the black bean aphid, Aphis fabae

Abstract. Electroantennograms (EAGs) were recorded from adult male and asexual forms (winged and wingless virginoparae and gynoparae) of the black bean aphid, Aphis fabae , during stimulation with two sex pheromone components, (+)-(4a S ,7 S ,7a R )-nepetalactone and (-)-(1 R ,4a S ,7 S ,7a R )-nepetalactol, as well as six plant volatiles, i.e. ( E )-2-hexenal, ( E )-2-hexenol-1, ( Z )-3-hexenol-1, ( Z )-3-hexenyl acetate, hexanal and allyl isothiocyanate. The male antennae are 1000-10,000 times more sensitive to nepetalactol and nepetalactone than to the plant compound ( E )-2-hexenal. Besides this marked difference of EAG peak responses in males, the EAG rise and decay are slower for the sex pheromone components. Males are also much more sensitive to the sex pheromone components than asexual females. This high sensitivity correlates with a predominance of antennal secondary rhinaria, the major sites of pheromone perception in the male. However, it is the primary rhinaria on the antennae of the wingless asexual females that are responsible for pheromone perception. Male antennae are as responsive as the asexual female antennae to the plant volatiles. The specialization of the male for mate location is discussed.     

Electrophysiological basis for the behavioural response of male and female Trichoplusia ni to synthetic female pheromone

Electroantennogram (EAG) studies demonstrated that antennae of both male and female Trichoplusia ni have: (1) receptor-neurones sensitive to female pheromone, (2) a low response threshold, (3) an identical mean-percentage EAG curve over a broad concentration range of pheromone, and (4) a similar absolute recovery interval from adaptation to pheromonal stimulation. These factors suggest that antennae of male and female T. ni have homologous and homogeneous acceptor sites for the female pheromone. Pheromonal stimulation of female antennae elicited EAGs with only 25% of the amplitude of those elicited in males.     

Electroantennogram responses of the stable fly, Stomoxys calcitrans, to components of host odour

Abstract. Electroantennograms (EAGs) were recorded from laboratory-reared male and female Stomoxys calcitrans (L.) in response to a range of synthetic chemicals known to be electrophysiologically-active for other biting flies. Of the eight compounds initially tested, only two - 1-octen-3-ol and 3-methylphenol - consistently elicited larger electroantennograms (EAGs) than did control treatments; 1-octen-3-ol was the most potent. EAG recovery time was inversely correlated with EAG amplitude. EAGs recorded with primary C₂-C₁₂ carbon chain-length primary aliphatic alcohols peaked at octan-1–ol with pentan-1-ol, hexan-1-ol and heptan-1-ol also eliciting EAG responses significantly larger than the controls. When different C₈ carbon chain compounds and nonane were tested: 1-octen-3-ol elicited the largest EAGs followed by, in decreasing activity, octan-1-ol, 1-bromooctane, octan-3-ol, octanal, 2-octanone, octanoic acid and nonane. The EAG response of 1-octen-3-ol increased sigmoidally with dose, with the threshold at between 2 and 20 ng, and the peak response at 200 μg on filter paper. EAGs larger than control were also elicited by entrained ox odour and ox breath. The behavioural implications are discussed.     

Electroantennogram responses of the stable fly, Stomoxys calcitrans, to carbon dioxide and other odours

ABSTRACT. Electroantennograms (EAGs) were recorded from Stomoxys calcitrans (L.) in response to increasing concentrations of carbon dioxide in an airstream. The magnitude of the EAG increases logarithmically from +0.023% carbon dioxide up to approximately +2.0% where a maximum is reached. Flies deprived of food for 48–50 h are more responsive to small increases in the carbon dioxide concentration than those deprived of food for only 20–23 h. It is concluded that the sensitivity of carbon dioxide receptors on the antennae of S. calcitrans increases as hunger develops. EAGs were also recorded in response to cattle odour, odour from fresh cattle faeces, expired human breath, acetone, and l-octen-3-ol. Acetic acid vapour causes a reversal of the usual EAG response indicating inhibition.     

ACOMPARATIVE STUDY ON THE ELECTROANTENOGRAM RESPONSES OF THREE APHID SPECIES TO PLANT VOLATILES

Abstract  Electroantennogram responses (EAGs) to 39 plant volatiles from Aphis glycines, A. gossypii and Myzus persicae were recorded. The olfactory system of these three aphid species differentially responded to green leaf, aromatic, mono-and sequiterpene chemicals. The EAG depolarizatin to green leaf chemicals and their isomers were more pronounced than to terpenes, alcohols and aldehydes, 6 carbon atom compounds being the most effective. It was shown that there was certain difference in the olfactory spectra of these three aphid species to plant volatiles. Higher responses in A. glycines and A. gossypii were elicited by the leaf aldehydes and esters than by the corresponding alcohols, which are in contrast with those in M. persicue . The activity of aphid olfactory system was related to molecular structures of the compounds. The responses to saturated alcohols and aldehydes were higher than those to unsaturated. The responses of spring emigrants of A. glycines to terpene derivatives (alcohols, aldehydes and esters) were higher than those of A. gossypii , but were lower to terpene hydrocarbons. The dissimilarity in olfactory spectra of the aphid species reflected the evolution of their host plant selection specificity. In the evolution, their chemosensory system adapted to perceive host plants. The shapes of the EAGs evoked by the various components were consistently different, with the slowest recovery for geraniol, 1-decanol and (±)-β-cit-ronellol, and ware related to number of carbon atoms in straight chain saturated alcohols.     

Odor discrimination using insect electroantennogram responses from an insect antennal array

Insects have a highly developed olfactory sensory system, mainly based in their antennae, for the detection and discrimination of volatile compounds in the environment. Electroantennogram (EAG) response profiles of five different insect species, Drosophila melanogaster, Heliothis virescens, Helicoverpa zea, Ostrinia nubilalis and Microplitis croceipes, showed different, species-specific EAG response spectra to 20 volatile compounds tested. The EAG response profiles were then reconstructed for each compound across the five insect species. Most of the compounds could be distinguished by comparing the response spectra. We then used a four-antenna array, called a Quadro-probe EAG, to see if we could discriminate among odorants based on the relative EAG amplitudes evoked when the probe was placed in plumes in a wind tunnel and in a field. Stable EAG responses could be simultaneously and independently recorded with four different insect antennae mounted on the Quadro-probe, and different volatile compounds could be distinguished in real time by comparing relative EAG responses with a combination of differently tuned insect antennae. Regardless of insect species or EAG amplitudes, antennae on the Quadro-probe maintained their responsiveness with higher than 1 peak/s of time resolution.     

Electrophysiological and behavioural identification of host kairomones as olfactory cues for Culicoides impunctatus and C. nubeculosus

Electroantennograms (EAGs) were recorded from wild‐caught parous, female Culicoides impunctatus (Goetghebuer) in response to components of host odour. Nine synthetic compounds were found to be electrophysiologically active, eliciting EAGs which were significantly different from solvent control. An EAG hierarchy was established, in which 1‐octen‐3‐ol elicited the highest amplitude EAGs, followed by acetone, lactic acid and butanone. The overall responses to phenolic compounds were reduced compared to the non‐phenolics. Subsequent behavioural analyses of the effects of these compounds when tested singly revealed 1‐octen‐3‐ol, acetone and butanone to be attractive over specific stimulus doses. Exposure to supra‐optimal doses modified the insects’ behaviour; insects either ceased to respond or were repelled. Lactic acid was attractive at the lowest dose tested but was repellent at high doses. Behavioural responses to the phenolic components of host odour and lactic acid were similar, generally causing arrestment at low doses and repelling at the higher doses tested. A comparison of EAG profiles and behavioural assays between laboratory‐reared Culicoides nubeculosus (Meigen) and C. impunctatus suggested that the same kairomones are utilized by both species, with C. nubeculosus being less sensitive than C. impunctatus. The EAG hierarchy of C. nubeculosus to the four non‐phenolics was identical to that of C. impunctatus.     

Defining Interaction between Electroantennogram Responses of Epiphyas postvittana (lepidoptera: Tortricidae) to Pheromone and Other Volatiles

The potential for electroantennograms (EAGs) to assist in the measurement of atmospheric pheromone concentrations was examined in laboratory and field experiments by using multiple stimuli, the main component of the pheromone of Epiphyas postvittana, (E)-11-tetradecenyl acetate, its Z-isomer (a behavioral antagonist), and alpha-terpineol (a representative host-plant odor) were presented to the antenna simultaneously to simulate field conditions. The EAG results were compared with predictions from two models describing responses to combined stimuli. Responses are defined as log-additive if they can be described with the equation [EAG((total)) =a (log ([P(x)]+[B(y)])+c] and as linear-additive if the EAG follows the equation [EAG((total)) =a (log [P(x)])+c+a'(log [B(y)])+c'] where [B(y)] is the concentration of the stimulant background odor and [P(x)] is the concentration of an additional odor stimulus. The EAGs elicited by the added stimuli were inversely related to the concentration of the volatile in the background. EAGs elicited by all combined stimuli followed the log-additive model. Our laboratory results were validated in field tests; alpha-terpineol represents the volatiles present in orchard air. In spite of this interaction between the perception of pheromone and plant volatiles, the field EAG can be used for the measurement of atmospheric pheromone concentrations, where background odor concentrations are relatively constant.     

Circadian rhythm in olfactory response in the antennae controlled by the optic lobe in the cockroach

The olfactory response in antennae of the cockroach, Leucophaea maderae, was investigated by measuring electroantennograms (EAGs) in restrained animals. The amplitude of the EAG response to pulses of ethyl acetate, octanol, or fenchone, exhibited a robust, light entrained, circadian rhythm that persisted at least 14 days in constant darkness. Dilution-response curves measured at the peak and trough of the rhythm indicated there was a 10-fold change in sensitivity. The EAG rhythm was abolished by severing the optic tracts, while entrainment was abolished by ablation of the compound eyes. The results indicate that the circadian system modulates olfactory sensitivity in the antennae and that the rhythm is driven by a circadian pacemaker in the optic lobes that is entrained by photoreceptors in the compound eyes.     

Using Insect Electroantennogram Sensors on Autonomous Robots for Olfactory Searches

Robots designed to track chemical leaks in hazardous industrial facilities¹ or explosive traces in landmine fields² face the same problem as insects foraging for food or searching for mates³: the olfactory search is constrained by the physics of turbulent transport⁴. The concentration landscape of wind borne odors is discontinuous and consists of sporadically located patches. A pre-requisite to olfactory search is that intermittent odor patches are detected. Because of its high speed and sensitivity^5-6, the olfactory organ of insects provides a unique opportunity for detection. Insect antennae have been used in the past to detect not only sex pheromones⁷ but also chemicals that are relevant to humans, e.g., volatile compounds emanating from cancer cells⁸ or toxic and illicit substances^9-11. We describe here a protocol for using insect antennae on autonomous robots and present a proof of concept for tracking odor plumes to their source. The global response of olfactory neurons is recorded in situ in the form of electroantennograms (EAGs). Our experimental design, based on a whole insect preparation, allows stable recordings within a working day. In comparison, EAGs on excised antennae have a lifetime of 2 hr. A custom hardware/software interface was developed between the EAG electrodes and a robot. The measurement system resolves individual odor patches up to 10 Hz, which exceeds the time scale of artificial chemical sensors¹². The efficiency of EAG sensors for olfactory searches is further demonstrated in driving the robot toward a source of pheromone. By using identical olfactory stimuli and sensors as in real animals, our robotic platform provides a direct means for testing biological hypotheses about olfactory coding and search strategies¹³. It may also prove beneficial for detecting other odorants of interests by combining EAGs from different insect species in a bioelectronic nose configuration¹⁴ or using nanostructured gas sensors that mimic insect antennae¹⁵.     

Electroantennogram responses of aphid nymphs to plant volatiles

Abstract. Electroantennogram (EAG) responses of immature aphids were investigated for 30 plant volatile compounds in third‐ and fourth‐stadium nymphs of the black bean aphid, Aphis fabae. The nymphs were destined to develop into adult alate (winged) virginoparae. The EAG response profiles were similar to those previously reported in the adults. Among the compounds tested, hexanonitrile elicited the largest EAG responses in both nymphal stadia, corresponding to previously reported results with adults. Six‐carbon aliphatic compounds showed relatively higher EAG activities in the nymphs but, in contrast, (E)‐2‐hexenal, benzaldehyde, α‐pinene and β‐pinene, and citronellal elicited relatively smaller EAG responses in nymphs than adults. Although overall EAG response profiles were similar between the third and the fourth stadia for the majority of the volatiles, four aldehyde compounds, hexanal (E)‐2‐heptenal, 2‐hydroxybenzaldehyde and citronellal, showed relatively higher EAG activities in the third than in the fourth stadium. The present study indicates that aphid nymphs possess a functional olfactory receptor system before the antennae are fully developed morphologically and physiologically.     

棉铃虫成虫对性信息素的电生理和行为反应研究

通过EAG和风洞实验,研究了棉铃虫雌雄成虫对性信息素组分和诱芯(Z-11-16Ald∶Z-9-16Ald=97∶3)的电生理反应。其中棉铃虫雌、雄蛾对诱芯的平均EAG反应测定值分别为1.06mV和4.32mV,分别高出对照(无性信息素空白诱芯)0.67mV和0.366mV,差异均达到极显著水平(雌蛾：t＝25.020, P≤0.01;雄蛾：t＝44.269,P≤0.01);棉铃虫雌蛾对性信息素组分(Z-11-16-Ald和Z-9.16Ald)的EAG反应值随浓度增加而增加;雄蛾在被剪除触角后与雌蛾不能正常交配,而雌蛾在被剪除触角后仍有40%的交配率,比正常雌雄蛾的交配率(70%)有所下降;在风洞实验中,雄蛾没有顺风远离诱源的飞行行为,趋向诱源的比率为81.8%,与对照有显著差异。研究表明性信息素组分对棉铃虫的交配活动有明显的影响。     

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.     

Olfactory receptor responses of the nymphal American cockroach to sex pheromones and their mimics

1. EAG responses to highly purified sex pheromones (periplanone-A and -B), sex pheromone mimics [germacrene-D, (+)-verbanyl acetate and (+)-trans-verbenyl acetate] and general odor (camphor) were recorded from both sexes of adult and three nymphal stages (7, 10 and 11th instars) of the American cockroach. 2. The M/F ratios were evaluated for each stage by stimulation with the above chemicals. 3. The ratio values indicated undeveloped sex pheromone receptor on the antennae of 7th male instar and females of all the stages. On the other hand, precursory development of the receptor was expected on the antennae of males of the old-aged nymphal instars and full development on the adult male antennae.     

中红侧沟茧蜂对棉花挥发性物质的触角电位反应

中红侧沟茧蜂Microplitis mediator (Haliday) 是棉铃虫 Helicoverpa amigera (Hübner)的主要寄生蜂之一。在寻找害虫的过程中,害虫的寄主植物释放出的气味起着至关重要的作用。本文测定了中红侧沟茧蜂雌蜂对棉花中17种气味物质的触角电位反应,比较了雌雄茧蜂对6种气味反应的差别,并且测定了雌蜂对4种气味物质的剂量 反应。结果表明,在所测的气味物质中,中红侧沟茧蜂对脂肪族化合物的反应最强,对芳香族化合物的反应居中,而对萜类化合物的反应较弱。对所测的气味物质的反应,雌雄茧蜂之间的差异不显著。雌蜂对正己醛、苯乙酮、正壬醇的反应阈值为25 μL, 对苯甲醛的反应阈值为250 μL,在所测的浓度范围内,对4种化合物的反应均未达到饱和。     

Olfactory antennal responses of the vine weevil Otiorhynchus sulcatus to plant volatiles

Electroantennograms (EAGs) were recorded from the vine weevil, Otiorhynchus sulcatus F. (Coleoptera: Curculionidae) to a broad range of volatile plant compounds. The response profile is restricted to a small number of volatiles that evoke substantial EAGs. Large EAG responses were particularly found among green leaf volatiles (GLV) such as (E)-2-hexenol-1, (Z)-3-hexenol-1, hexanol-1, hexanal, and heptanal. Other plant volatiles eliciting responses in the weevils' antenna are 2,5-dimethylpyrazine, hexylamine, benzylalcohol, 1,2-dimethoxybenzene, o-cresol, myrtenol, 3-methylcyclohexanol, -hexalactone, and -heptalactone. EAG responses to terpenes were generally weak. Many of the monoterpenes are characteristic for the odour of conifers, a group of plants which tend to be avoided by adult vine weevils. The EAG response to several GLV and benzene derivatives in three geographically distinct populations of the vine weevil differed, suggesting between-population variation in receptor sensitivities for several compounds under test. The GLV-composition of the odour profile of potential food plants may be an important criterion for a polyphagous insect like the vine weevil to be used in host-plant selection, since compounds in this odour group dominate so strongly the EAG response profile. In multiple food-choice situations the weevils are known to prefer certain plant species and we hypothesize that they combine GLV information with that of more specific plant volatiles, thereby promoting attraction or avoidance.     

OLFACTORY RESPONSE OF THE GRASSHOPPER MELANOPLUS SANGUINIPES TO PLANT ODOURS AND VOLATILE COMPOUNDS

Abstract Electroantennograms (EAGs1 were recorded from male adults of Melanoplus sanguinipes to foliage odours from 5 plant species and 9 volatile compounds as well as their blends. Higher EAG responses were recorded for Louisianna sage and Alfalfa foliages, compared to other three gramineous plants. Among green odour chelricals, 6-carbon alcohols (e. g. E-2-hexen-1–01 and E-3-hexen-1–01] resulted in the highest EAG responszs. No significant differences among the pentenol, 3-pentanone and blank control were observed. In ternary and quaternary mixture tests of volatile compounds, the combinations with pentenol did not elicit greater amplitudes; thus pentenol and 3-pentanone were proved to be unimportant to the response of this generalist grasshopper. The EAG data are discussed with regard to the role of host plant volatiles in host plant orientation and selection by the grasshopper.     

黑蝗对植物气味及其挥发性化合物的嗅觉反应(英文)

本文使用触角电位技术研究了黑蝗雄虫的嗅觉反应,测试的材料包括5种植物叶片、9种植物挥发性气味化合物及它们的定量混合物。在5种植物材料中,路易斯安纳蒿和紫花苜蓿能引起更大的电位反应,而3种禾本科植物的反应相对较小。在绿色气味化合物中,6-碳醇类如E-2-hexen-l-ol和E-3-hexen-l-ol导致最高的电位反应,而pentenol、3-pentanone与空白对照间并无明显差异。在3种或4种挥发性化合物的混合物中,没有含pentenol的组合总是引起较高的电位。因此,该项研究证明pentenol和3-pentanone在这种多食性黑蝗的嗅觉定位中所起的作用不明显。另外,结合触角电位的数据资料,作者讨论了寄主植物挥发性化合物在该种蝗虫定位和选择寄主植物方面的作用。