Pheromone receptor system in the females of the greater wax moth Galleria mellonella

Electroantennogram studies of female Galleria mellonella showed a two-acceptor system for the sex pheromones, n-nonanal and n-undecanal, produced by the male. The nine carbon pheromone reacts with both acceptors, however, n-undecanal only reacts with the acceptor for the eleven carbon pheromone.     

棉铃虫齿唇姬蜂对植物挥发物和寄主性信息素腺体化合物的EAG反应

测试了棉铃虫齿唇姬蜂Campoletis chlorideae Uchida对58种植物挥发物和5种寄主性信息素腺体化合物的EAG反应。在100 μg的刺激剂量下,棉铃虫齿唇姬蜂对C₅～C₇脂肪醇的EAG反应最强,碳链延长或缩短都将导致反应的降低;对脂肪醛、酮、酸和酯都有一定的反应。棉铃虫齿唇姬蜂对萜烯的EAG反应很低,对氧化萜的反应普遍升高。3种芳香化合物均可激起棉铃虫齿唇姬蜂较强的EAG反应。剂量反应曲线表明,具有EAG活性的化合物激起的反应强度与化合物的剂量呈正相关,而2种萜烯——α-蒎烯和反式石竹烯,在测定的剂量范围内EAG活性都很低。棉铃虫齿唇姬蜂对寄主性信息素腺体化合物也有EAG反应,用100 μg和1 000 μg的寄主性信息素刺激,雄蜂的反应可比雌蜂的反应高出1倍。对植物挥发物在棉铃虫齿唇姬蜂寻找寄主中的作用进行了讨论。     

Electroantennogram and behavioural responses of different forms of the bird cherry-oat aphid,Rhopalosiphum padi,to sex pheromone and a plant volatile

Winged (alate) virginoparae were induced in the laboratory by crowding the bird cherry-oat aphid, Rhopalosiphum padi (L.) under long-day conditions. Males and gynoparae (the winged female form that produces sexual females) were induced by short days. Electroantennogram (EAG) and behavioural responses were investigated in each of the three forms to two aphid sex pheromone components, (-)-(4aS,7S,7aR)-nepetalactol and (+)-(4aS,7S,7aR)-nepetalactone, and benzaldehyde, a volatile which is released by the winter host plant. All three compounds elicited EAG responses with the males showing the highest sensitivity to each compound. Both the nepetalactol and the nepetalactone elicited larger EAG responses in gynoparae than in the winged virginoparae but antennae from virginoparae were more responsive to benzaldehyde. Although the nepetalactone is not a sex pheromone component in R. padi the EAG responses were similar to those evoked by the nepetalactol, the sex pheromone, in all three aphid forms. In a linear-track olfactometer, significantly more male R. padi moved into air containing nepetalactol, nepetalactone or benzaldehyde than into a simultaneous choice of clean air (i.e. attraction) but nepetalactol was more attractive than nepetalactone. Males, however, showed no response to a mixture of nepetalactol and nepetalactone. Gynoparae were attracted only to the nepetalactol but were less sensitive than the males and showed no response to the nepetalactone or benzaldehyde. In contrast, alate virginoparae showed no behavioural responses to any of the compounds. The present study supports the idea that the male R. padi utilise both sex pheromone and benzaldehyde for mate/host-plant location in autumn. It also demonstrates, for the first time, polyphenic differences in the olfactory responses at the peripheral level between the two female forms. Such differences impact on the life-cycle strategy where winged virginoparae move between graminaceous summer host plants while gynoparae move from the summer hosts to the bird cherry, winter host. The latter move appears to be assisted by the sex pheromone released by sexual females, already present on that host, acting as an aggregation pheromone.     

Electroantennogram (EAG) responses of Microplitis croceipes and Cotesia marginiventris and their lepidopteran hosts to a wide array of odor stimuli: Correlation between EAG response and degree of host specificity?

In order to test whether the electroantennogram (EAG) response spectrum of an insect correlates to its degree of host specificity, we recorded EAG responses of two parasitoid species with different degrees of host specificity, Microplitis croceipes (specialist) and Cotesia marginiventris (generalist), to a wide array of odor stimuli including compounds representing green leaf volatiles (GLVs), herbivore-induced plant volatiles (HIPV), ecologically irrelevant (not used by the parasitoid species and their hosts for host location) plant volatiles, and host-specific odor stimuli (host sex pheromones, and extracts of host caterpillar body and frass). We also tested the EAG responses of female moths of the caterpillar hosts of the parasitoids, Heliothis virescens and Spodoptera exigua, to some of the odor stimuli. We hypothesized that the specialist parasitoid will have a narrower EAG response spectrum than the generalist, and that the two lepidopteran species, which are similar in their host plant use, will show similar EAG response spectra to plant volatiles. As predicted, the specialist parasitoid showed greater EAG responses than the generalist to host-specific odor and one HIPV (cis-3-hexenyl butyrate), whereas the generalist showed relatively greater EAG responses to the GLVs and unrelated plant volatiles. We detected no differences in the EAG responses of H. virescens and S. exigua to any of the tested odor.     

Functional characterization of SlitPBP3 in Spodoptera litura by CRISPR/Cas9 mediated genome editing

Functional gene analysis by using genome editing techniques is limited only in few model insects. Here, we reported an efficient and heritable gene mutagenesis analysis in an important lepidopteran pest, Spodoptera litura, using the CRISPR/Cas9 system. By using this system, we successfully obtained the homozygous S. litura strain by targeting the pheromone binding protein 3 gene (SlitPBP3), which allowed us to elucidate the role of this gene in the olfaction of the female sex pheromones. By co-injection of Cas9 mRNA and sgRNA into S. litura eggs, highly efficient chimera mutation in SlitPBP3 loci was detected both in injected eggs (39.1%) and in the resulting individual moths (87.5%). We used the mutant moths as parents to obtain the G1 offspring and the homozygous mutant strain in G2. The function of SlitPBP3 was explored by Electroantennogram (EAG) recordings with a homozygous mutant strain. The result showed that the EAG responses were significantly decreased in mutant males than in control males when treated with the major sex pheromone component (Z9,E11-14:Ac) and a minor component (Z9-14:Ac) at higher dosages. The results demonstrate that s SlitPBP3 gene plays a minor role in the perception of the female sex pheromones. Furthermore, our study provides a useful methodology with the CRISPR/Cas9 system for gene in vivo functional study, particular for lepidopteran species in which the RNAi approach is not efficient.     

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.     

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.     

Ultrastructure and potential role of integumentary glandular cells in adult male and female Callosobruchus subinnotatus (Pic) and C. maculatus (Fabricius) (Coleoptera : Bruchidae)

Transmission and scanning electron microscopy methods were used to study the ultrastructure of integumentary glandular cells that may be involved in the production of sex pheromones and other semiochemicals in Callosobruchus subinnotatus and C. maculatus (Coleoptera : Bruchidae). Additionally, we measured electroantennogram (EAG) responses of male and female antennae to solvent extracts and glassadsorbed volatiles from both sexes of C. maculatus in order to localize the source of the putative sex pheromone. Both species have numerous cuticular pores on the abdomen and thorax. These pores open via an epicuticular duct into a single type 3 secretory cell. Solvent extracts of the pygidium from the female elicit the highest EAG responses from male antennae, suggesting this area of the abdomen to be the source of the putative sex pheromone in C. maculatus.     

Influence of sex,maturity and host substances on pheromones in the guts of the bark beetles, Ips paraconfusus and Dendroctonus brevicomis

Pheromones and metabolites of host (ponderosa pine) compounds were found in association with the hindgut of both naturally fed and of non-fed, host vapour-exposed bark beetles, Ips paraconfusus and Dendroctonus brevicomis. Much smaller amounts were found in the corresponding heads and mid guts. Sex-specific differences in content of pheromones were observed as in earlier studies. Exposure of I. paraconfusus to vapours of a pheromone component, ipsenol and other monoterpene alcohols resulted in their accumulation in the hindgut but relatively very low amounts in the head. The possible sites of pheromone biosynthesis are discussed. Exposure of male I. paraconfusus to vapours of host compounds, myrcene and α-pinene, revealed that immature adults do not produce the pheromone components, ipsenol and ipsdienol, as mature adults do while both immature and mature sexes produced another pheromone component, cis-verbenol, as well as trans-verbenol and myrtenol. Immature D. brevicomis adults did not contain pheromones until their exposure to vapours of (?)-α-pinene which caused production of trans-verbenol but only about 10% that of mature adults treated similarly. Verbenone, a male-produced inhibitory pheromone of D. brevicomis, apparently was not synthesized from (?)-α-pinene in females nor was its synthesis in males enhanced by exposure to this host compound.     

温度对二化螟性信息素通讯的影响

环境因素影响昆虫两性间的化学通讯,也影响性信息素技术的田间防治效果.本文探讨了温度对二化螟雌蛾性信息素产生以及雄蛾对性信息素触角电位反应的影响,以期为田间二化螟的性信息素防治提供指导.在二化螟蛹期和成虫期进行不同温度处理(15、20、25、30和35 ℃),然后利用气相色谱仪(GC)分析雌蛾性腺内各性信息素组分的含量及比例,同时利用触角电位仪(EAG)测定雄蛾对性信息素组分的电生理反应.结果表明: 25 ℃处理中雌蛾性腺内3个性信息素组分(Z9-16:Ald、Z11-16:Ald和Z13-18:Ald)的含量均显著高于其他温度处理(15、20和30 ℃),且25 ℃处理中Z13-18:Ald的相对比例也显著低于其他温度处理.就雄蛾对性信息素的敏感性而言,对3种性信息素单一组分及特定比例混合物的EAG反应在15～25 ℃间没有显著差异,但在25～35 ℃间(Z13-18:Ald在30～35 ℃间)随温度升高呈下降趋势,且30 ℃较25 ℃显著降低,35 ℃较30 ℃又显著降低.综合分析认为,二化螟性信息素通讯的适宜温度为20～25 ℃,温度过高或过低均不利于二化螟两性间的正常化学通讯.研究结果为二化螟性信息素防治技术的合理应用及极端温度条件下害虫种群发生的预测预报,提供了重要参考.     

Enhancement of attraction to sex pheromone of Grapholita molesta (Busck) (Lepidoptera: Tortricidae) by structurally unrelated sex pheromone compounds of Conogethes punctiferalis (Guenée) (Lepidoptera: Crambidae)

Grapholita molesta (Busck) (the oriental fruit moth; OFM) and Conogethes punctiferalis (Guenée) are both fruit and stem borers with overlapping habitats, occurrences, and outbreak cycles. These two species are in different families and they have completely different sex pheromone components. Here, the effects of the sex pheromone components of C. punctiferalis, (E)-10-hexadecenal (E10-16:Ald) and (Z)-10-hexadecenal (Z10-16:Ald) and their mixture, were evaluated on the sexual communication of OFM by examining electrophysiological (EAG) and behavioral responses. We found that a considerably large amount of E10-16:Ald or Z10-16:Ald and a tiny amount of OFM pheromone elicited comparable EAG responses in OFM males, indicating the low antennal olfactory sensitivity of OFM males to the heterospecific pheromone compounds. In two different peach orchard parcels, captures of OFM by their pheromone lures baited in delta traps were increased by at least 1.5 times when OFM pheromone lures were combined with lures that contained 1000 μg of either E10-16:Ald, Z10-16:Ald or their mixture. In two other pear orchard parcels, both E10-16:Ald and Z10-16:Ald increased captures of OFM in Unitraps in a dose-dependent manner with more than a four-fold increase. Therefore, the enhanced attractiveness of OFM lures by the two interspecific pheromones suggests their potential application to improve mass trapping, population monitoring and mating disruption of OFM.     

植物气味化合物与斜纹夜蛾性信息素的协同作用

为提高现有性信息素对雄蛾的引诱活性, 本研究通过大量的田间试验探索植物气味化合物与斜纹夜蛾Spodoptera litura性信息素（顺9, 反11-十四碳二烯乙酸酯∶顺9, 反12-十四碳二烯乙酸酯=10∶1）的协同作用机制。从斜纹夜蛾寄主植物和花的气味化合物中, 选择9种有代表性的化合物, 并以一定剂量分别加入到斜纹夜蛾性信息素诱芯中, 在田间测试对雄蛾的引诱活性。结果表明: 在测试的9种植源性化合物中, 发现一定剂量(每个诱芯加入0.4 mg)的苯乙醛(PAA), 具有显著提高斜纹夜蛾性信息素的引诱作用, 而高剂量的苯乙醛则强烈抑制性信息素的引诱活性; 此外, 其他各种浓度的测试化合物或混合物对性信息素则没有统计上显著的增效作用。不同剂量的苯乙醛单个化合物及各种植物气味化合物组成的混合物对斜纹夜蛾也有微弱的引诱作用。苯乙醛必须要与性信息素的完整组分(以10∶1比例混合的顺9, 反11-十四碳二烯乙酸酯和顺9反, 12-十四碳二烯乙酸酯)混合才能起作用, 缺少顺9, 反12-十四碳二烯乙酸酯则没有引诱活性。本研究证明, 苯乙醛作为理想的性信息素诱芯增效剂, 可应用于建立更理想的斜纹夜蛾性信息素诱杀技术, 对性诱害虫防治和测报具有应用价值。     

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.     

Modulation of sex pheromone perception in female moths of the eastern spruce budworm, Choristoneura fumiferana by altosid

Topical application of 10 μg of Altosid, a juvenile hormone analogue to adult female moths of the spruce budworm significantly reduced their electroantennogram responses to their own synthetic female sex pheromone provided the EAG recordings were performed at least 10–15 hr after JHA treatment. Newly emerged moths were more sensitive to JHA treatment than older moths. Acetone-treated moths (control) showed an increase in EAG response to sex pheromone with increasing age. These age related responses of moths to their sex pheromone and their reduction by Altosid treatment are discussed in terms of endogenous JH titres.     

笋秀夜蛾雄蛾对性信息素化合物的电生理和行为反应

利用气质联用仪对笋秀夜蛾雌蛾性腺体粗提物成分进行了分析比对,通过触角电位(EAG)试验研究了笋秀夜蛾雄成虫对性信息素组分的电生理反应,以Y形嗅觉仪进行了室内和林间的行为生测.结果表明： 雌蛾性腺体粗提物中顺11-十六碳乙酸酯(Z11-16:Ac)和顺11-十六碳烯醇(Z11-16:OH)为笋秀夜蛾性信息素活性组分,性腺粗提物、两种化合物及其混合物均能激发雄蛾明显的EAG反应,且EAG反应值随性信息素组分浓度增加而增加.Z11-16:Ac和Z11-16:OH按57∶43配置成的二元混合物所激发的EAG反应强于同等剂量下的单一组分.室内的行为生测和林间诱捕试验结果与EAG反应基本一致,Z11-16:Ac+Z11-16:OH(57∶43)混合物对雄蛾引诱力最强,在林间单诱捕器(诱芯浓度为10⁴ng·μL^-1)的平均诱捕量达到(48.5±6.7)头.     

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.     

Moth sex pheromone receptors and deceitful parapheromones

The insect''s olfactory system is so selective that male moths, for example, can discriminate female-produced sex pheromones from compounds with minimal structural modifications. Yet, there is an exception for this “lock-and-key” tight selectivity. Formate analogs can be used as replacement for less chemically stable, long-chain aldehyde pheromones, because male moths respond physiologically and behaviorally to these parapheromones. However, it remained hitherto unknown how formate analogs interact with aldehyde-sensitive odorant receptors (ORs). Neuronal responses to semiochemicals were investigated with single sensillum recordings. Odorant receptors (ORs) were cloned using degenerate primers, and tested with the Xenopus oocyte expression system. Quality, relative quantity, and purity of samples were evaluated by gas chromatography and gas chromatography-mass spectrometry. We identified olfactory receptor neurons (ORNs) housed in trichoid sensilla on the antennae of male navel orangeworm that responded equally to the main constituent of the sex pheromone, (11Z,13Z)-hexadecadienal (Z11Z13-16Ald), and its formate analog, (9Z,11Z)-tetradecen-1-yl formate (Z9Z11-14OFor). We cloned an odorant receptor co-receptor (Orco) and aldehyde-sensitive ORs from the navel orangeworm, one of which (AtraOR1) was expressed specifically in male antennae. AtraOR1•AtraOrco-expressing oocytes responded mainly to Z11Z13-16Ald, with moderate sensitivity to another component of the sex pheromone, (11Z,13Z)-hexadecadien-1-ol. Surprisingly, this receptor was more sensitive to the related formate than to the natural sex pheromone. A pheromone receptor from Heliothis virescens, HR13 ( = HvirOR13) showed a similar profile, with stronger responses elicited by a formate analog than to the natural sex pheromone, (11Z)-hexadecenal thus suggesting this might be a common feature of moth pheromone receptors.     

Evidence for a female‐produced,long range pheromone of Anoplophora glabripennis (Coleoptera: Cerambycidae)

Abstract Volatiles from female Asian longhorned beetle (ALB), Anoplophora glabripennis, were evaluated as candidate sex pheromone components. Previous studies on ALB have revealed several antennally active compounds from virgin females; however the origins and activity of these compounds were not apparent and require further investigation. We tested the hypothesis that one or more of the ALB contact sex pheromones is a precursor that undergoes abiotic oxidation to yield volatile pheromone components, and evaluated the activity of these compounds using laboratory and field bioassays. Gas chromatography coupled electroantennography detection (GC‐EAD) analysis indicated the presence of three antennally active aldehydes (heptanal, nonanal, and hexadecanal) in female cuticular extracts exposed to ozone or UV and visible light. In laboratory bioassays using a Y‐tube olfactometer, males were preferentially attracted to ozonized female body washes over crude body washes. Similarly, synthetic formulations of these compounds were preferred over controls in the olfactometer. Field trapping experiments conducted from 2006 to 2008 in Ningxia, China showed that synthetic lures of the three aldehydes formulated in a ratio simulating that of virgin females attracted more beetles compared to controls, and that combinations of these aldehydes, linalool oxide, and host kairomones captured more beetles than controls, and captured significantly more males.     

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.