OLFACTORY RESPONSES OF COTTON BOLLWORM,HELICOVERPA ARMIGERA TO SEX PHEROMONE AND PLANT VOLATILE COMPONENTS*

Abstract Electroantennograms (EAGs) were recorded from unmated, laboratory-reared male and female Helicover pa armigera (Lepidoptera:Noctuidae) adults to two sex pheromone components and a range of plant volatile components, some of which are known volatiles from leaves and fruits. The female-produced sex pheromone was not detected by other female moths. The sex pheromone components to which male moths responded previously show to possess biological activity. As for plant volatile components, a large degree of EAG response uniformity between male and female moth was observed. The greatest EAG responses of all plant volatiles tested were elicited by monoenic C-6 alcohol and aldehyde. They are constituents of the “general green-leaf odor” that emanates from most plants. The potential adaptive benefit of selective perception to sex pheromone and green-leaf volatile components is discussed.     

棉铃虫Helicverpa armigera对性外激素和植物气味物质的嗅觉反应(英文)

本文用EAG技术研究棉铃虫对性外激素和植物气味物质的嗅觉反应。供试的气味物质有棉铃虫性外激素的二种主要组分和36种与棉铃虫寄主植物有关的植物气味物质。试验结果表明：1）只有棉铃虫雄蛾对性外激素组分产生反应,二种性外激素的组分中,Z-11-16：AL能引起较强的EAG反应。2）对植物气味物质,雌雄棉铃虫之间的嗅觉反应没有明显的差异。棉铃虫对性外激素和植物气味组分的嗅觉感受过程是一个有选择性的感受过程：不同种类的气味物质能引起不同的EAG反应。只有棉铃虫雄蛾才对性外激素组分起反应,在二种性外激素组分中：主要组分：Z-11-16：AL引起的EAG反应最高,高于大多数植物气味物质。次要组分Z-9-16：AL引起的EAG反应很弱。雌、雄棉铃虫对植物气味组分的嗅觉反应的选择性表现出一致的规律：对苯甲醛和绿叶气味物质的反应最高。单萜类中的萜烯衍生物（萜类醇、醛、酯等）的触角电位反应次之。棉铃虫对大多数的供试化合物的EAG反应都很小。最后,作者讨论了性外激素和植物气味物质对棉铃虫行为的作用。     

Characterization of olfactory receptor neurons for pheromone candidate and plant volatile compounds in the clover root weevil,Sitona lepidus

Antennal olfactory receptor neurons (ORNs) for pheromone and plant volatile compounds were identified and characterized in male and female clover root weevil, Sitona lepidus (Gyllenhal), using the single sensillum recording technique with five pheromone-related compounds, and 40 host and non-host plant volatile compounds. Overall, seven different types of olfactory sensilla containing specialized ORNs were identified in each sex of S. lepidus. Among them, three different types of sensilla in the males and two types in the females housed ORNs specialized for pheromone-related compounds. The ORNs in males were specialized for 4-methyl-3,5-heptanedione or one or more of four stereoisomers of 5-hydroxy-4-methyl-3-heptanone. In contrast, female sensilla did not contain ORNs sensitive to 4-methyl-3,5-heptanedione while they contained ORNs sensitive to and specialized for the stereoisomers of (4S,5S)-5-hydroxy-4-methyl-3-heptanone. In addition to the pheromone-related ORNs, four types of olfactory sensilla contained ORNs responsive to plant volatile compounds in male S. lepidus, and five types in females. Most of the ORNs identified in S. lepidus showed a high degree of specificity to specific volatile compounds although some of the active compounds showed overlapping response spectra in the ORNs across different types of sensilla. The most active plant volatile compounds were the four green leaf volatile compounds, (E)-2-hexenol, (Z)-2-hexenol, (Z)-3-hexenol and (E)-2-hexenal, and isomers of two monoterpenols, (±)-linalool and (±)-α-terpineol, all eliciting strong responses from relatively large numbers of ORNs in male and female S. lepidus. Our study indicates that S. lepidus has a set of highly sensitive and selective ORNs for pheromone and plant volatile compounds. Further work is needed to elucidate the behavioral implications of these findings.     

Odorant receptor phylogeny confirms conserved channels for sex pheromone and host plant signals in tortricid moths

The search for mates and food is mediated by volatile chemicals. Insects sense food odorants and sex pheromones through odorant receptors (ORs) and pheromone receptors (PRs), which are expressed in olfactory sensory neurons. Molecular phylogenetics of ORs, informed by behavioral and functional data, generates sound hypotheses for the identification of semiochemicals driving olfactory behavior. Studying orthologous receptors and their ligands across taxa affords insights into the role of chemical communication in reproductive isolation and phylogenetic divergence. The female sex pheromone of green budworm moth Hedya nubiferana (Lepidoptera, Totricidae) is a blend of two unsaturated acetates, only a blend of both elicits male attraction. Females produce in addition codlemone, which is the sex pheromone of another tortricid, codling moth Cydia pomonella. Codlemone also attracts green budworm moth males. Concomitantly, green budworm and codling moth males are attracted to the host plant volatile pear ester. A congruent behavioral response to the same pheromone and plant volatile in two tortricid species suggests co‐occurrence of dedicated olfactory channels. In codling moth, one PR is tuned to both compounds, the sex pheromone codlemone and the plant volatile pear ester. Our phylogenetic analysis finds that green budworm moth expresses an orthologous PR gene. Shared ancestry, and high levels of amino acid identity and sequence similarity, in codling and green budworm moth PRs offer an explanation for parallel attraction of both species to the same compounds. A conserved olfactory channel for a sex pheromone and a host plant volatile substantiates the alliance of social and habitat signals in insect chemical communication. Field attraction assays confirm that in silico investigations of ORs afford powerful predictions for an efficient identification of behavior‐modifying semiochemicals, for an improved understanding of the mechanisms of host plant attraction in insect herbivores and for the further development of sustainable insect control.     

Antennal responses of Cydia pomonella (L.) exposed to surfaces treated with methoxyfenozide

Electroantennogram (EAG) measurements were recorded from the antennae of male and female codling moth, Cydia pomonella L., to determine whether adult moths exposed to surfaces treated with the ecdysteroid agonist methoxyfenozide experience a decline in their antennal reception and thus olfactory sensitivity. Such a phenomenon would offer a possible mechanism for the previously reported decreased responsiveness from moths treated with methoxyfenozide to pheromone‐ and plant volatile‐based monitoring lures. Mean EAG data revealed that the antennae from methoxyfenozide‐treated male moths appear to be just as sensitive to various doses of synthetic codlemone as the antennae from the control and surfactant‐treated moths, but they appeared to be less sensitive to the pheromone component 12OH (collected from female effluvia) than the control male antennae. Mean male EAG responses to the pheromone components E8,E10‐12Al and codlemone collected from methoxyfenozide‐treated females were significantly less than the responses towards those two pheromone components collected from the control and surfactant‐treated females. Female moth exposure to methoxyfenozide did not negatively impact the sensitivity of female antennae to the plant volatile pear ester, but it did towards the apple volatile butyl hexanoate. Data from this study show that adult C. pomonella exposure to methoxyfenozide‐treated surfaces does appear to negatively impact, in a minor way, the (i) olfactory sensitivity (or detection) of male antennae towards some components of the female sex pheromone, (ii) the female antennal sensitivity towards a key apple volatile and (iii) the attractiveness of female pheromone effluvia.     

植物气味化合物对棉铃虫产卵及田间诱蛾的影响

利用触角电位技术（EAG）研究棉铃虫Helicoverpa armigera对17种植物气味物质的嗅觉反应,实验结果表明：棉铃虫雄蛾、雌蛾对所试各种植物气味物质均无明显的嗅觉反应差异,不同种类的气味物质能引起不同的EAG反应。对其中的几种植物气味物质进行了引诱产卵实验及田间诱蛾实验,结果表明：某些寄主植物的挥发性次生物质显示出较好的引诱产卵活性,且对棉铃虫性诱剂具有增效和协同作用。田间实验数据显示,增加了植物气味物质的棉铃虫性诱剂与单一的棉铃虫性诱剂相比差异显著。     

Changes in Odor Background Affect the Locomotory Response to Pheromone in Moths

Many animals rely on chemical cues to recognize and locate a resource, and they must extract the relevant information from a complex and changing odor environment. For example, in moths, finding a mate is mediated by a sex pheromone, which is detected in a rich environment of volatile plant compounds. Here, we investigated the effects of a volatile plant background on the walking response of male Spodoptera littoralis to the female pheromone. Males were stimulated by combining pheromone with one of three plant compounds, and their walking paths were recorded with a locomotion compensator and analyzed. We found that the addition of certain volatile plant compounds disturbed the orientation toward the sex pheromone. The effect on locomotion was correlated with the capacity of the plant compound to antagonize pheromone detection by olfactory receptor neurons, suggesting a masking effect of the background over the pheromone signal. Moths were more sensitive to changes in background compared to a constant background, suggesting that a background odor also acts as a distracting stimulus. Our experiments show that the effects of odorant background on insect responses to chemical signals are complex and cannot be explained by a single mechanism.     

蛾类昆虫性信息素受体及其作用机理

蛾类昆虫性信息素受体首先从烟芽夜蛾Heliothis virescens和家蚕Bombyx mori中鉴定出来, 到目前为止已经克隆得到了19种蛾类昆虫的几十种性信息素受体基因, 并且这些基因在系统发育树中聚成一个亚群。性信息素受体从蛾类蛹期开始表达, 主要表达在雄性触角的毛形感器中, 少部分受体在雌性触角、 雄性触角其他感器以及身体其他部位中也有表达。大部分蛾类性信息素受体的配体并不是单一的, 而是能够对多种性信息素组分有反应, 部分性信息素受体还能够识别性信息素以外的其他物质, 还有一部分性信息素受体的识别配体目前尚不清楚。另外发现在雌性蛾类触角中也存在一些嗅觉受体能够识别雄性分泌的性信息素。在蛾类性信息素受体与性信息素识别的过程中, 性信息素结合蛋白不仅能够特异性地运送配体到嗅觉神经元树状突上, 还能够提高性信息素与性信息素受体之间的结合效率。另外, OrCo类受体与性信息素受体共表达在嗅觉神经元中, 在蛾类性信息素受体与配体的识别过程中扮演了重要角色。但是蛾类信息素对神经元刺激的终止并非由性信息素受体控制, 而是由细胞中的气味降解酶等其他因子调控。蛾类性信息素受体研究中还有很多疑问需要解答, 其过程可能比我们想象的更为复杂。     

Pheromone production and mating behaviour by allatectomized males of the cockroach, Nauphoeta cinerea

Males of Nauphoeta cinerea produce a volatile pheromone which attracts the female for mating. Allatectomy of males either 5 days prior to or within 12 hr following the imaginal ecdysis does not impair pheromone production, pheromone release, nor any observable aspect of mating behaviour. It is proposed that in N. cinerea, and in other cockroach species where the male releases a volatile pheromone to attract the female, pheromone production is not controlled by the corpora allata, and pheromone release is under direct motor control.     

Antennal olfactory sensory neurones responsive to host and nonhost plant volatiles in gorse pod moth Cydia succedana

We investigate the response profiles of the antennal olfactory sensory neurones (OSNs) in male and female gorse pod moth Cydia succedana to host and nonhost volatiles, using the single sensillum recording technique. Eight different classes of olfactory sensilla are identified in female C. succedana and five different classes of olfactory sensilla in males. Nineteen different classes of OSNs are identified from the sensilla in females, and nine different classes of OSNs in the male sensilla. All classes of sensilla, except class F7 and class M1 sensilla, co‐compartmentalize two or three OSNs in each sensillum, and the OSNs present in the same sensillum are specialized for different volatiles. Most plant‐volatile OSNs exhibit phasic‐tonic type of temporal responses, whereas the pheromone OSNs in male C. succedana show rather phasic responses. The majority of OSNs identified in C. succedana display highly specialized responses to a narrow range of volatiles, whereas only a small proportion of OSNs show broad response spectra. Two most abundant classes of OSNs exhibit highly specialized responses to β‐myrcene and (E)‐β‐ocimene, two major volatiles released by gorse (Ulex europaeus), the main host of C. succedana. By contrast, several other classes of OSNs exhibit highly specialized responses to geraniol, (Z)‐3‐hexen‐1‐ol, (±)‐α‐terpineol, citral and benzyl acetate, which are produced by various nonhost plants. Taking the results of the present study together, we suggest that C. succedana use the combinational input from a set of highly specialized OSNs for host plant volatiles and another set of highly specialized OSNs for nonhost volatiles to discriminate between hosts and nonhosts.     

No Linkage between Genes Controlling Female Pheromone Production and Male Pheromone Response in the European Corn Borer, Ostrinia Nubilalis Hubner (Lepidoptera; Pyralidae)

The E and Z pheromonal strains of the European corn borer, Ostrinia nubilalis, are characterized by female production of and male preference for opposite blends of (E)-11-and (Z)-11-tetradecenyl acetate. It is known that the pheromone production is controlled by an autosomal gene and that the males' behavior is determined by a sex-linked gene. A third gene, autosomally inherited, has been shown to determine the organization of the male pheromone receptors. In the present study the linkage relationship between the autosomal genes controlling sex pheromone production and male olfactory sensilla was investigated. A recombination experiment showed unequivocally that the genes determining the variation in pheromone production and male pheromone receptors are not closely linked and are most likely inherited independently.     

Volatile constituents of male and female boll weevils and their frass

The volatile constituents of male and female boll weevils, Anthonomus grandis, and their frass were analysed by GLC-MS. The 4 previously identified components of the male pheromone were present only in the male volatile oil (3·9%) and in the male frass volatile oil (38·9%). The compounds found in one or both sexes and their frass include 26 carbonyls, 23 hydrocarbons, 12 alcohols, 6 phenols, 4 esters, 3 furans, 1 ether, and 1 lactone. Also found were 2 compounds containing nitrogen, 1 halogen, and 1 sulphur. There were 33 terpenes and 24 aromatic compounds. 3,7-Dimethyl-1-octanol comprised 15·6 per cent of the male frass oil. Carvone was found only in females (5·4%) and in female frass (6·8%). A series of monoterpene aldehydes (M⁺ 152) were found only in the female frass oil. A pheromone rôle for these components was suggested.     

Synergism of pheromone and host‐plant volatile blends in the attraction of Grapholita molesta males

Control of Grapholita molesta (Busck) (Lepidoptera: Tortricidae), a major pest of stone and pome fruits, is successfully achieved by mating disruption. Under these conditions, tools other than conventional pheromone dispensers are needed for flight monitoring. The objective of the present work was to determine whether plant volatiles synergize male G. molesta attraction to a suboptimal dose of synthetic sex pheromone. The plant blend (referred to as 5VB), a mixture of three green leaf volatiles [(Z)‐3‐hexenyl acetate, (Z)‐3‐hexenol, and (E)‐2‐hexenal] and two aromatics [benzaldehyde (BZA) and benzonitrile (BZN)], was added to the suboptimal pheromone dose (2 ng on filter paper) in log steps (up to 10 000× the pheromone dose) to test synergism of pheromone and plant blends. In addition, the effect of individual plant volatiles on male responses was investigated by adding to the suboptimal pheromone dose each of the four‐compound plant‐volatile blends, resulting from eliminating one volatile from the 5VB at a time, or each plant volatile alone. Flight behaviour and the time to reach the source were recorded. The 5VB alone was not attractive to G. molesta males, but at a ratio of 1:1 000 (Ph:5VB) or higher, the attractiveness of the suboptimal pheromone dose increased, to a level similar to that of the optimal pheromone dose (10 ng). All tested plant volatiles, except BZA, synergized the response to the pheromone when added individually, but only (Z)‐3‐hexenol and BZN did so to a level not significantly different from the Ph:5VB blend. Aromatics had a stronger effect than green leaf volatiles (GLVs), because their removal, but not the removal of GLVs, decreased landing responses. The addition of the 5VB decreased significantly the time males needed to reach the odour source. The observed enhanced male attraction to mixtures of pheromone and plant volatiles will facilitate the development of lures for G. molesta adult flight monitoring.     

Male housefly (Musca domestica L.) genital system as a source of mating pheromone

Male volatile mating pheromone is released by male houseflies placed in close proximity to females or in contact with female hydrocarbon extract. The pheromone induces the aggregation of virgin females in the vicinity of the male. Preparations of macerated testicular and ejaculatory duct tissue exert a similar effect, and these organs are, apparently, the source of the pheromone. Such preparations are not attractive to males or mated females. The pheromone also appears to induce female receptivity, since females that are exposed to emanations from male system preparations show a quicker rate of pairing with males. The pheromone effect results from the testes and ejaculatory ducts acting together, as preparations of each of these alone are inactive. Pheromone activity is age dependent and the greatest female reaction is observed when tissues of 6–10-day old males are used.     

Field and laboratory evidence for a volatile pheromone produced by parous females of the Scottish biting midge, Culicoides impunctatus

Abstract. Electrophysiological, behavioural and field evidence is presented for a volatile attractant produced by host-seeking, parous female Culicoides impunctatus Goetghebuer. Attraction of host-seeking females to other live parous females or to extracts of parous females was demonstrated in Y-tube bioassays, as well as in field trials using baited sticky traps. Threshold numbers were 100–200 females, both in the laboratory and field. Antennal receptors to this pheromone were revealed by recording electroantennograms (EAGs) in response to odour from live parous females and to the extracts of female midges. The female-produced attractant did not alter field catches of male C. impunctatus.     

Interference of plant volatiles on pheromone receptor neurons of male Grapholita molesta (Lepidoptera: Tortricidae)

In moths, sex pheromone components are detected by pheromone-specific olfactory receptor neurons (ph-ORNs) housed in sensilla trichodea in the male antennae. In Grapholita molesta, ph-ORNs are highly sensitive and specific to the individual sex pheromone components, and thus help in the detection and discrimination of the unique conspecific pheromone blend. Plant odors interspersed with a sub-optimal pheromone dose are reported to increase male moth attraction. To determine if the behavioral synergism of pheromone and plant odors starts at the ph-ORN level, single sensillum recordings were performed on Z8-12:Ac and E8-12:Ac ph-ORNs (Z-ORNs and E-ORNs, respectively) stimulated with pheromone–plant volatile mixtures. First, biologically meaningful plant-volatile doses were determined by recording the response of plant-specific ORNs housed in sensilla auricillica and trichodea to several plant odorants. This exploration provided a first glance at plant ORNs in this species. Then, using these plant volatile doses, we found that the spontaneous activity of ph-ORNs was not affected by the stimulation with plant volatiles, but that a binary mixture of sex pheromone and plant odorants resulted in a small (about 15%), dose-independent, but statistically significant, reduction in the spike frequency of Z-ORNs with respect to stimulation with Z8-12:Ac alone. The response of E-ORNs to a combination of E8-12:Ac and plant volatiles was not different from E8-12:Ac alone. We argue that the small inhibition of Z-ORNs caused by physiologically realistic plant volatile doses is probably not fully responsible for the observed behavioral synergism of pheromone and plant odors.     

Release of male courtship display in Periplaneta americana: evidence for female contact sex pheromone

Male American cockroaches (Periplaneta americana) are attracted to virgin females by volatile sex pheromones. After antennal contact with the female they turn through 180° and spread their wings in courtship display. A chemical contact stimulus releasing male courtship is demonstrated in the female cuticle. Experiments with standardized olfactory stimulation by volatile sex pheromones revealed that the contact stimulus is sex-specific and species-specific. It can be washed off the cuticle with non-polar solvents and was successfully transferred to glass dummies. However, it is not effective in the absence of volatile sex pheromones. Thus volatile sex pheromones are responsible for male attraction and sexual motivation, while mate recognition is accomplished through the contact pheromone.     

Olfactory and Behavioral Mechanisms Underlying Enhanced Mating Competitiveness Following Exposure to Ginger Root Oil and Orange Oil in Males of the Mediterranean Fruit Fly, Ceratitis capitata (Diptera: Tephritidae)

Males of the Mediterranean fruit fly, Ceratitis capitata, are strongly attracted to various plant odors, and previous work has demonstrated that male exposure to certain odors, including the scent of orange oil (OO) and ginger root oil (GRO), increases their mating success relative to non-exposed males. However, the mechanism(s) underlying this mating increase is not known. Here, we describe several experiments that further investigate the association between GRO- and OO-exposure and male signaling activity, pheromone attractiveness, and mating success in male medflies. Exposure to GRO or OO increased time spent pheromone calling but did not accelerate the rate of male sexual maturation. Using a wind tunnel, we compared female attraction to the pheromone of control, non-exposed males versus males previously exposed to OO or GRO. There was no evidence that GRO exposure enhanced the attractiveness of the male pheromone. The data for OO were inconclusive: females tended to spend more time on spheres emanating pheromone from OO-exposed males than on spheres emanating pheromone from non-exposed males, but the number of female landings did not differ between the two types of pheromone sources. Female choice tests confirmed that GRO- and OO-exposure boost male mating success relative to non-exposed males. Application of GRO directly to the abdomen reduced male mating success, whereas similar application of OO boosted male mating success. The potential role and mode of action of plant chemicals in the mating behavior of male medflies are evaluated in light of these findings.     

Pheromone pre-exposure and mating modulate codling moth (Lepidoptera: Tortricidae) response to host plant volatiles

Abstract 1 Two codling moth Cydia pomonella kairomonal attractants, ethyl (E,Z)‐2,4‐decadienoate (pear ester) and (E)‐β‐farnesene, were tested in an insecticide‐sprayed apple orchard and an orchard treated for mating disruption with synthetic pheromone (E,E)‐8,10‐dodecadienol (codlemone). Male captures with pear ester were higher in the pheromone‐treated than in the insecticide‐treated orchard, whereas captures with (E)‐β‐farnesene were not different. Subsequent wind tunnel experiments confirmed that pre‐exposure to sex pheromone codlemone increased the behavioural response of codling moth males to pear ester. This supports the idea that male attraction to the plant volatile pear ester and sex pheromone codlemone is mediated through the same sensory channels. 2 Pear ester is a bisexual codling moth attractant and even captures of female moths were significantly increased in the pheromone‐treated orchard. In the laboratory wind tunnel, pheromone pre‐exposure had no effect on female response to pear ester, but significantly more mated than unmated codling moth females flew upwind towards a pear ester source. Differences in mating status in insecticide‐treated vs. pheromone‐treated orchards may thus account for the differences in female trap captures with pear ester. 3 These findings are important with respect to monitoring of codling moth with pear ester in mating disruption orchards. They also emphasize the importance of host plant volatiles in pheromone‐mediated mating disruption, which has been neglected to date.