Candidate Pheromone Components Research Articles

Identification of the Major Sex Pheromone Component of the Click Beetle Agriotes ferrugineipennis

Synthetic sex pheromone lures are useful tools to monitor and control populations of adult click beetles (Coleoptera: Elateridae). However, sex pheromones for Agriotes click beetle species native to North America have yet to be identified. Here we report the identification and field testing of the sex pheromone of Agriotes ferrugineipennis. Headspace volatiles from female beetles were collected on Porapak Q, and aliquots of Porapak extract were analyzed by gas chromatographic-electroantennographic detection (GC-EAD) and GC-mass spectrometry. 7-Methyloctyl 7-methyloctanoate (7Me7Me) emitted by females was more abundant and elicited much stronger responses from male antennae than the aldehydes octanal and nonanal and the ketone 6,10,14-trimethyl-2-pentadecanone. In a field experiment, captures of A. ferrugineipennis males in traps baited with candidate pheromone components exceeded those of unbaited control traps, on average by nearly 1,200 times. Neither the ketone nor the aldehydes as lure constituents appeared to alter captures of males in 7Me7Me-baited traps. We conclude that 7Me7Me is the major, and possibly the only, sex attractant pheromone component of female A. ferrugineipennis.

(Z,E)‐α‐Farnesene – sex pheromone component of female click beetle Selatosomus aeripennis destructor with intra‐ and inter‐sexual communication function

AbstractSex pheromones are known for only a few major click beetle (Coleoptera: Elateridae) pests in North America. These pheromones could be used to monitor, as well as control, adult beetles. Our objective was to identify the sex pheromone of female Selatosomus aeripennis destructor (Brown), a predominant elaterid pest species in farmland in the Canadian Prairie provinces. Headspace volatiles from a group of S. a. destructor females were collected and analyzed by coupled gas chromatography‐electroantennographic detection (GC‐EAD) and gas chromatography‐mass spectrometry (GC‐MS). The GC‐EAD recordings revealed two candidate pheromone components that elicited weak (CPC‐1) or strong (CPC‐2) responses from adult male S. a. destructor antennae. The mass spectra of CPC‐1 and CPC‐2 indicated, and authentic standards confirmed, that they were the isomers (E)‐β‐farnesene and (Z,E)‐α‐farnesene, respectively. In three field experiments at various sites in Alberta, Canada, traps baited with synthetic (Z,E)‐α‐farnesene captured 7.1×, 6.8×, and 2.6× more males than unbaited control traps. In follow‐up experiments, traps baited with (Z,E)‐α‐farnesene captured significantly more males, but significantly fewer females, than unbaited control traps, indicating sex pheromone autodetection by females. (E)‐β‐Farnesene on its own was not attractive as a trap lure, and when added to (Z,E)‐α‐farnesene at a 20:1 ratio, it reduced the lure’s attractiveness. Our data support the conclusion that (Z,E)‐α‐farnesene is the major sex pheromone component of S. a. destructor females, with both an intra‐ and inter‐sexual communication function.

Identification of Components of the Aggregation Pheromone of the Guam Strain of Coconut Rhinoceros Beetle, Oryctes rhinoceros, and Determination of Stereochemistry

The coconut rhinoceros beetle, Oryctes rhinoceros (Linnaeus 1758) (Coleoptera: Scarabaeidae: Dynastinae) (CRB), is endemic to tropical Asia where it damages both coconut and oil palm. A new invasion by CRB occurred on Guam in 2007 and eradication attempts failed using commonly applied Oryctes rhinoceros nudivirus (OrNV) isolates. This and subsequent invasive outbreaks were found to have been caused by a previously unrecognized haplotype, CRB-G, which appeared to be tolerant to OrNV. The male-produced aggregation pheromone of the endemic, susceptible strain of O. rhinoceros (CRB-S) was previously identified as ethyl 4-methyloctanoate. Following reports from growers that commercial lures containing this compound were not attractive to CRB-G, the aim of this work was to identify the pheromone of CRB-G. Initial collections of volatiles from virgin male and female CRB-G adults from the Solomon Islands failed to show any male- or female-specific compounds as candidate pheromone components. Only after five months were significant quantities of ethyl 4-methyloctanoate and 4-methyloctanoic acid produced by males but not by females. No other male-specific compounds could be detected, in particular methyl 4-methyloctanoate, 4-methyl-1-octanol, or 4-methyl-1-octyl acetate, compounds identified in volatiles from some other species of Oryctes. Ethyl 4-methyloctanoate elicited a strong electroantennogram response from both male and female CRB-G, but these other compounds, including 4-methyloctanoic acid, did not. The enantiomers of ethyl 4-methyloctanoate and 4-methyloctanoic acid were conveniently prepared by enzymatic resolution of the commercially-available acid, and the enantiomers of the acid, but not the ester, could be separated by gas chromatography on an enantioselective cyclodextrin phase. Using this approach, both ethyl 4-methyloctanoate and 4-methyloctanoic acid produced by male CRB-G were shown to be exclusively the (R)-enantiomers whereas previous reports had suggested male O. rhinoceros produced the (S)-enantiomers. However, re-examination of the ester and acid produced by male CRB-S from Papua New Guinea showed that these were also the (R)-enantiomers. In field trapping experiments carried out in the Solomon Islands, both racemic and ethyl (R)-4-methyloctanoate were highly attractive to both male and female CRB-G beetles. The (S)-enantiomer and the corresponding acids were only weakly attractive. The addition of racemic 4-methyloctanoic acid to ethyl 4-methyloctanoate did significantly increase attractiveness, but the addition of (R)- or (S)-4-methyloctanoic acid to the corresponding ethyl esters did not. Possible reasons for the difference in assignment of configuration of the components of the CRB pheromone are discussed along with the practical implications of these results.

3-Hydroxyhexan-2-one and 3-Methylthiopropan-1-ol as Pheromone Candidates for the South American Cerambycid Beetles Stizocera phtisica and Chydarteres dimidiatus dimidiatus, and Six Related Species.

Here, we study the pheromone chemistry of two South American cerambycid beetle species, and their behavioral responses to candidate pheromone components. Adult males of Stizocera phtisica Gounelle (subfamily Cerambycinae: tribe Elaphidiini) produced a sex-specific blend of (R)-3-hydroxyhexan-2-one with lesser amounts of 3-methylthiopropan-1-ol. In field bioassays, traps baited with racemic 3-hydroxyhexan-2-one and 3-methylthiopropan-1-ol did not catch conspecific beetles, but did catch both sexes of a sympatric species, Chydarteres dimidiatus dimidiatus (F.) (Cerambycinae: Trachyderini). We found that males of this species also produce (R)-3-hydroxyhexan-2-one and 3-methylthiopropan-1-ol, and small amounts of 2-phenylethanol. Subsequent bioassays with these compounds showed that a blend of 3-hydroxyhexan-2-one and 3-methylthiopropan-1-ol constitutes the aggregation-sex pheromone of C. d. dimidiatus, with 2-phenylethanol not influencing the attraction of conspecifics. During the field bioassays, six other species in the Cerambycinae also were caught in significant numbers, including Aglaoschema ventrale (Germar) (tribe Compsocerini), congeners Chrysoprasis aurigena (Germar), Chrysoprasis linearis Bates, and an unidentified Chrysoprasis species (Dichophyiini), and Cotyclytus curvatus (Germar) and Itaclytus olivaceus (Laporte & Gory) (both Clytini), suggesting that one or more of the compounds tested are also pheromone components for these species.

Identification and Field Testing of Volatile Components in the Sex Attractant Pheromone Blend of Female House Mice

Recently, it was reported (i) that the sex pheromone blend of male house mice, Mus musculus, comprises not only volatile components (3,4-dehydro-exo-brevicomin; 2-sec-butyl-4,5-dihydrothiazole) but also a component of low volatility (the sex steroid testosterone), and (ii) that the sex steroids progesterone and estradiol are sex pheromone components of female house mice. Here we tested the hypothesis that the sex attractant pheromone blend of female mice, analogous to that of male mice, also comprises volatile pheromone components. Analyzing by GC-MS the head space volatiles of bedding soiled with urine and feces of laboratory-kept females and males revealed three candidate pheromone components (CPCs) that were adult female-specific: butyric acid, 2-methyl butyric acid and 4-heptanone. In a two-choice laboratory experiment, adult males spent significantly more time in the treatment chamber baited with both the synthetic steroids (progesterone, estradiol) and the synthetic CPCs than in the paired control chamber baited only with the synthetic steroids. In field experiments, trap boxes baited with both the CPCs and the steroids captured 6.7-times more adult males and 4.7-times more juvenile males than trap boxes baited with the steroids alone. Conversely, trap boxes baited with both the CPCs and the steroids captured 4.3-times more adult males and 2.7-fold fewer adult females than trap boxes baited with the CPCs alone. In combination, these data support the conclusion that butyric acid, 2-methyl butyric acid and 4-heptanone are part of the sex attractant pheromone of female house mice. With progesterone and estradiol being pheromone components of both female brown rats, Rattus norvegicus, and female house mice, these three volatile components could impart specificity to the sexual communication system of house mice, brown rats and possibly other rodent species.

Evidence for a Nest Defense Pheromone in Bald-Faced Hornets, Dolichovespula Maculata, and Identification of Components.

In eusocial insects like Bald-faced hornets, Dolichovespula maculata, nest defense is essential because nests contain a large number of protein-rich larvae and pupae, and thus are attractive to nest predators. Our objectives were to investigate whether D. maculata exhibit pheromone-mediated nest defense, and to identify and field test any pheromone components. We tested for pheromone-mediated nest defense behavior of D. maculata by placing a paired box-apparatus near the entrance of D. maculata nests, and treating both boxes with a solvent control, or one of the two boxes with a solvent control and the other with either venom sac extract, the putative source of nest defense pheromone, or synthetic pheromone. The sound impulses caused by nest mates attempting to sting or strike the boxes were recorded for 3min. Compared to the double-control treatment, the number of strikes increased 27-fold when one of the two boxes was treated with venom sac extract, providing evidence for an alarm response. The box treated with venom sac extract also induced a significantly greater proportion of strikes than the corresponding control box, providing evidence for a target-oriented response. Analyzing venom sac extract by gas chromatographic-electroantennographic detection (GC-EAD) and GC-mass spectrometry resulted in the identification of seven candidate pheromone components: (a) dimethylaminoethanol, (b) dimethylamino ethyl acetate, (c) 2,5-dimethylpyrazine, (d) N-3-methylbutylacetamide, (e) 2-heptadecanone, (f) (Z)-8-heptadecen-2-one, and (g) (Z)-10-nonadecen-2-one. Testing in paired-box bioassays blends of the nitrogen-containing volatile components a-d, the less volatile ketones e-g, or both (a-g), indicated that a-d primarily have an alarm function. The ketones e-g, in contrast, induced target-oriented responses, possibly marking the box, or potential nest predators, for guided and concerted attacks, or enhancing the alarm-inducing effect of the volatile pheromone components, as shown in honey bees. Comparing the behavioral effects of venom sac extract, blends a-d, e-g, and a-g, venom sac extract was most effective in triggering the full complement of alarm and target-oriented responses. These comparisons further suggested that a component is missing in the group of components that triggers the alarm rather than the target-oriented response.

Reproductive Isolation of Ips nitidus and I. shangrila in Mountain Forests of Western China: Responses to Chiral and Achiral Candidate Pheromone Components.

Eastern Palearctic conifers are subject to frequent bark beetle outbreaks. However, neither the species responsible nor the semiochemicals guiding these attacks are well understood. Two high-mountain Ips species on Qinghai spruce, Picea crassifolia, I. shangrila and I. nitidus, are typical in this regard. Six synthetic candidate pheromone components that we earlier identified from hindguts of unmated males of these two Ips species were tested for field activity in Qinghai province, P. R. China. For I. nitidus, racemic ipsdienol ((±)-Id) could replace the naturally-produced blend of enantiomers containing 74% (-)-(S)-Id (74:26 S:R), in attractive ternary or binary blends. In contrast, sympatric I. shangrila were attracted mainly to blends including Id of opposite chirality, 97%-(+)-(R)-Id. Of the verbenols, (-)-trans-verbenol was inactive for I. nitidus or inhibitory for I. shangrila, but (-)-cis-verbenol (cV) was a key component of the pheromone in both species. Two fully factorial experiments demonstrated that (±)-Id, cV, and 2-methyl-3-buten-2-ol (MB) are components of the aggregation pheromone of I. nitidus, whereas only (+)-Id and cV are essential components of the aggregation pheromone of I. shangrila. While MB is not necessary for attraction of I. shangrila, it is an active antagonist and likely functions in species isolation. A review of the pheromone production and responses in Palearctic Ips and Pseudoips showed that cV is more common than methylbutenols, and both elicit qualitatively variable responses. Ipsdienol is the most common component with variable chirality, and is a necessary, but often not sufficient, factor for determining pheromone specificity.

Sex pheromone of the cloaked pug moth,Eupithecia abietaria(Lepidoptera:Geometridae), a pest of spruce cones

The sex pheromone of the cloaked pug moth, Eupithecia abietaria Gotze, an important cone-feeding pest in spruce seed orchards in Europe, was investigated. Chemical and electrophysiological analyses of pheromone gland extracts of female moths and analogous analyses of synthetic hydrocarbons and epoxides of chain length C19 and C21 revealed (3Z,6Z,9Z)-3,6,9-nonadecatriene (3Z,6Z,9Z-19:H) and 3Z,6Z-cis-9,10-epoxynonadecadiene (3Z,6Z-cis-9,10-epoxy-19:H) as candidate pheromone components, which were found in a gland extract in a ratio of 95 : 5. In field trapping experiments, conspecific males were only attracted to a combination of 3Z,6Z,9Z-19:H and the (9S,10R)-enantiomer of 3Z,6Z-cis-9,10-epoxy-19:H. The (9R,10S)-enantiomer was not attractive, which is in agreement with studies on other Eupithecia species, for which males have only been attracted by the (9S,10R)-enantiomer of epoxides. Subsequent experiments showed that E. abietaria males were attracted to a wide range of ratios of the two active compounds and that trap catches increased with increasing dose of the binary blend. A two-component bait containing 300 μg 3Z,6Z,9Z-19:H and 33 μg of the (9S,10R)-enantiomer of 3Z,6Z-cis-9,10-epoxy-19:H was efficient for monitoring E. abietaria in spruce seed orchards in southern Sweden, where this species has probably been overlooked as an important pest in the past. With sex pheromones recently identified for two other moths that are major pests on spruce cones, the spruce seed moth, Cydia strobilella L., and the spruce coneworm, Dioryctria abietella Denis & Schiffermuller, pheromone-based monitoring can now be achieved for the whole guild of cone-feeding moths in European spruce seed orchards. (Less)

(E)-5-Hexadecenyl Acetate: A Novel Moth Sex Pheromone Component From Stathmopoda auriferella

The sex pheromone of Stathmopoda auriferella (Walker), an important pest of kiwifruit in Korea, was studied. Two candidate pheromone components detected in the gland extracts of females were identified as (E)-5-hexadecenyl acetate (E5-16:OAc) and (E)-5-hexadecenol (E5-16:OH) in a ratio of 75:25 by mass spectral analysis of natural pheromone components and dimethyldisulfide adducts, and retention index comparisons with synthetic standards. In the kiwifruit orchards, E5-16:OAc alone was attractive to S. auriferella males and caught significantly more males than live virgin females. However, addition of E5-16:OH strongly inhibited attraction to E5-16:OAc. These results suggest that the major component of the female-produced sex pheromone of S. auriferella is E5-16:OAc. This hexadecenyl acetate is a novel moth sex pheromone component.

Populations of the Gall Midge Dasineura oxycoccana on Cranberry and Blueberry Produce and Respond to Different Sex Pheromones

We identified and field-tested the sex pheromones of Dasineura oxycoccana (Johnson) (Diptera: Cecidomyiidae) midges collected from cranberry, Vaccinium macrocarpon Aiton, and from highbush blueberry, Vaccinium corymbosum L., commonly named cranberry tipworm (CTW) and blueberry gall midge (BGM), respectively. Coupled gas chromatographic-electroantennographic detection (GC-EAD) analyses of pheromone gland extract from the ovipositor of calling CTW females revealed one component (<10pg per ovipositor/pheromone gland) that elicited antennal responses from CTW males. Stepwise identification was based on its mass spectrum in a concentrated sample with 300 pheromone gland equivalents, retention indices (RI) on three GC columns (DB-5, DB-23, and DB 210), RI inter-column differentials, and RIs and double bond positions of other midge pheromones. These analyses indicated that (8Z)-2,14-diacetoxy-8-heptadecene (2,14-8Z-17) was the candidate pheromone of the CTW. GC-EAD analysis of pheromone gland extract from calling BGM females revealed two components that elicited antennal responses from BGM males. Retention times on the three GC columns were consistent with 2,14-8Z-17 and 2,14-17, indicating that these were candidate pheromone components of the BGM. The four stereoisomers of 2,14-8Z-17 were stereoselectively synthesized and field-tested in cranberry. Delta-type traps baited with SS-2,14-8Z-17 captured significantly more CTW males than did traps baited with any other single stereoisomer or with all four stereoisomers combined. In blueberry, delta-type traps baited with RR-2,14-8Z-17 captured significantly more BGM males than did traps baited with any other single stereoisomer or with all four stereoisomers combined. Subsequent field experiments demonstrated that RR-2,14-17 is the major pheromone component of BGM, and that RR-2,14-8Z-17 is a pheromone component that does not enhance attractiveness of RR-2,14-17. The BGM pheromone RR-2,14-17 has no antagonistic effect on the CTW pheromone SS-2,14-8Z-17 and vice versa. Our results substantiate the conclusion that populations of D. oxycoccana on cranberry and blueberry represent two cryptic species.

Sex pheromone of the red clover casebearer moth, Coleophora deauratella, an invasive pest of clover in Canada

AbstractThe red clover casebearer, Coleophora deauratella Lienig &amp; Zeller (Lepidoptera: Coleophoridae), is an invasive pest of Trifolium species (Fabaceae) in Canada. We identified candidate sex pheromone components from female pheromone gland extracts using coupled gas chromatographic–electroantennographic analysis detection. Three compounds elicited an electrophysiological response from antennae and were identified as: (Z)‐7‐dodecenyl acetate, (Z)‐5‐dodecenyl acetate, and (Z)‐7‐dodecen‐1‐ol. Field tests of the candidate pheromone components revealed that males were attracted to a binary mixture of (Z)‐7‐dodecenyl acetate and (Z)‐5‐dodecenyl acetate. Male moth trap capture was greatest in traps baited with lures containing 100:10 or 100:20 ratios of these pheromone components, respectively. Trap capture was reduced when (Z)‐5‐dodecenyl acetate was present below 10 or above 20% of (Z)‐7‐dodecenyl acetate. Equal numbers of male moths were captured in traps baited with 10, 100, and 1 000 μg of the attractive binary mixture. These findings allow for the development of a pheromone‐based monitoring system for this invasive pest of clover in Canada.

New Contact Sex Pheromone Components of the German Cockroach, Blattella germanica, Predicted from the Proposed Biosynthetic Pathway

Upon contacting the cuticle of a sexually mature female, a male German cockroach exhibits a characteristic courtship behavior: he turns away from the female and raises his wings, thereby exposing tergal glands. The glandular secretion stimulates the female to mount the male and feed, thus positioning her appropriately for copulation. A multi-component contact sex pheromone produced by females is responsible for eliciting courtship behavior. The most abundant pheromone components are 3,11-dimethylnonacosan-2-one and 3,11-dimethylheptacosan-2-one, oxidation products of the abundant hydrocarbon analogs 3,11-dimethylnonacosane and 3,11-dimethylheptacosane, respectively. The C(29)-dimethyl ketone is thought to be further metabolized to two less abundant pheromone components, 29-hydroxy-3,11-dimethylnonacosan-2-one and 29-oxo-3,11-dimethylnonacosan-2-one. Based on this proposed biosynthetic pathway of pheromone production, we hypothesized that 3,11-dimethylheptacosan-2-one also would be oxidized to give two candidate pheromone components, 27-hydroxy-3,11-dimethylheptacosan-2-one, and 27-oxo-3,11-dimethylheptacosan-2-one. By using bioassay-guided fractionation and chemical analyses of cuticular extracts of virgin females and synthesis of the (3S,11S)-isomer of each of the two predicted pheromone components, we showed that the epicuticle of the German cockroach does indeed contain these two compounds. The contact sex pheromone of the female German cockroach, thus may consist of at least six biosynthetically related components.

(S,S)-2,12-, (S,S)-2,13-, and (S,S)-2,14-Diacetoxyheptadecanes: Sex Pheromone Components of Red Cedar Cone Midge, Mayetiola thujae

We identified, synthesized, and field-tested the sex pheromone of female red cedar cone midge Mayetiola thujae (Hedlin) (Diptera: Cecidomyiidae), a pest insect in red cedar Thuja plicata seed orchards. Coupled gas chromatographic (GC)-electroantennographic detection analyses of pheromone extract revealed three components (A, B, C) that elicited responses from antennae of males, all of which occurred below the detection threshold of the mass spectrometer and thus had to be identified without spectroscopic data. Taking into account (1) their retention indices (RI) on three GC columns (DB-5, DB-23, and DB-210), (2) intercolumn RI differentials, and (3) the molecular structures of known cecidomyiid pheromones, we synthesized seven candidate pheromone components: 2,10-, 2,11-, 2,12-, 2,13-, 2,14-, 2,15- and 2,16-diacetoxyheptadecanes. Of these, 2,12-, 2,13-, and 2,14-diacetoxyheptadecane had RIs on all columns consistent with those of A, B, and C and elicited strong antennal responses when tested at picogram levels. In field experiments with the twelve stereoselectively synthesized stereoisomers, only the SS-stereoisomers of 2,12-, 2,13-, and 2,14-diacetoxyheptadecane attracted male M. thujae. The three-component SS-stereoisomer blend was more attractive than the 12-component blend of all stereoisomers, suggesting that one or several nonnatural stereoisomers are inhibitory. One-, two-, and three-component lures of the SS-stereoisomers were equally effective in attracting male M. thujae, indicating redundancy in the pheromone. Identification of the M. thujae sex pheromone will allow development of pheromone-based monitoring, and possibly control, of M. thujae populations in T. plicata seed orchards.

Pheromone Components from Body Scales of Female Anarsia lineatella Induce Contacts by Conspecific Males

Pheromonal communication of adult peach twig borers, Anarsia lineatella Zeller (Lepidoptera: Gelechiidae), was reinvestigated based on recent findings that virgin female-baited traps were more attractive to mate-seeking males than a two-component synthetic sex pheromone consisting of (E)-5-decen-1-yl acetate (1000 microg) and (E)-5-decen-1-ol (100 microg), suggesting that females use additional pheromone components. Hypothesizing that these additional components may be released from body parts other than abdominal sex pheromone glands, we extracted female body scales and analyzed aliquots by coupled gas chromatographic-electroantennographic detection (GC-EAD) and GC-mass spectrometry. Eight straight-chain and four methylated aliphatic hydrocarbons, as well as two acetates, all elicited responses from excised male antennae. In laboratory experiments with synthetic candidate pheromone components, a combination of octadecyl acetate, (R)-11-methyltricosane, and (S)-11-methyltricosane in the presence of gland-derived sex pheromone components were shown to elicit contact of female decoys by males. However, body pheromone components did not enhance attractiveness of sex pheromone components in field trapping experiments, suggesting that they are effective only at close range and that other stimuli are responsible for superior attractiveness of female-baited traps.

Communication ecology of webbing clothes moth: 4. Identification of male- and female-produced pheromones

We investigated the hypothesis that aggregation signals produced by male webbing clothes moths (WCM), Tineola bisselliella (Hum.) (Lepidoptera: Tineidae), and close-range male attractant signals produced by females have a pheromonal basis, at least in part. Gas chromatographic-electroantennographic detection (GC-EAD) and GC-mass spectrometric analyses of bioactive methanolic extracts of male WCM disclosed three candidate pheromone components: hexadecanoic acid methyl ester (16:Ester), (Z)-9-hexadecenoic acid methyl ester (Z9—16:Ester), and octadecanoic acid methyl ester (18:Ester). In bioassay experiments in a large Plexiglas™ arena, a blend of synthetic 16:Ester plus Z9—16:Ester was attractive to male and virgin (but not mated) female WCM; the 18:Ester was inactive. GC-EAD analyses of pheromone gland extracts from female WCM revealed (E,Z)-2,13-octadecadienal (E2Z13—18:Ald) and (E,Z)-2,13-octadecadienol (E2Z13—18:OH) as candidate sex pheromone components. In arena bioassay experiments, 1—5 female equivalents of synthetic E2Z13—18:Ald (0.2 ng) and E2Z13—18:OH (0.1 ng) were more attractive to male WCM than were two virgin female WCM. We anticipate that the combination of aggregation and sex pheromones, male-produced sonic aggregation signals, and habitat-derived semiochemicals will be highly effective in attracting male and female WCM to commercial traps.

Sex pheromone of tomato fruit borer, Neoleucinodes elegantalis.

Five candidate pheromone components were identified by analyzing pheromone gland extracts by gas chromatography (GC), coupled GC-electroantennographic detection (EAD), and coupled GC-mass spectrometry (MS) : (E)-11-hexadecenol(E11-16 : OH), (Z)-11-hexadecenol (Z11-16 : OH), (E)-11-hexadecenal, (E)-11-hexadecenyl acetate, and (Z)-3,(Z)-6,(Z)-9-tricosatriene (Z3,Z6,Z9-23 : Hy). In electroantennogram (EAG) recordings, synthetic E11-16 : OH elicited stronger antennal responses at low doses than other candidate pheromone components. Field tests demonstrated that synthetic E11-16 : OH as a trap bait was effective in attracting males, whereas addition of Z11-16 : OH inhibited the males' response. Z3,Z6,Z9-23 : Hy strongly enhanced attractiveness of E11-16 : OH, but was not attractive by itself. A pheromone blend with synergistic behavioral activity of an alcohol (E11-16 : OH) and hydrocarbon (Z3,Z6,Z9-23 : Hy) component is most unusual in the Lepidoptera. The synthetic two-component pheromone is approximately 60 times more attractive than the female-produced blend and might facilitate the control of this pest.

Aggregation pheromones of two geographical isolates of the new guinea sugarcane weevil, Rhabdoscelus obscurus

The aggregation pheromones were studied from two geographical isolates (Hakalau, Hawaii, and Silkwood, Queensland, Australia) of the New Guinea sugarcane weevil, Rhabdoscelus obscurus. Coupled gas chromatographic–electroantennographic detection (GC-EAD) and GC–mass spectrometric (MS) analyses of Porapak Q volatile extract from male and from female Hawaiian R. obscurus revealed a single EAD-active, male-specific candidate pheromone, which was identified as 2-methyl-4-octanol (1). Corresponding volatile analyses from male and from female Australian R. obscurus consistently revealed three EAD-active, male-specific candidate pheromone components that were identified as 1, (E2)-6-methyl-2-hepten-4-ol (rhynchophorol) (2), and 2-methyl-4-heptanol (3). In field experiment 1 in Hakalau, Hawaii, traps baited with a stereoisomeric mixture of synthetic 1 (3 mg/day) plus sugarcane captured more weevils than did traps baited with 1 or sugarcane alone or no bait, indicating that 1 is the pheromone of the Hawaiian R. obscurus population. In field experiment 2, conducted in Silkwood, Australia, traps baited with stereoisomeric mixtures of synthetic 1, 2, and 3 (3 mg/day each) plus sugarcane caught more weevils than did unbaited traps or traps baited with 1, 2, and 3 or sugarcane. Testing candidate pheromone components 1, 2, and 3 in experiments 2–5 in all possible binary, ternary, and quaternary combinations with sugarcane, indicated that 1 and 2 in combination, but not singly, are pheromone components of the Australian R. obscurus population. Weevils from several locations in Australia and Hawaii could not be differentiated using traditional morphological characters or ultrastructural comparisons with scanning electron microscopy (SEM). However, comparisons of mtDNA sequences (cytochrome oxidase I; regions I1 to M4; 201 base pairs) revealed 5.5% variation between the Hawaiian (N = 2) and the Australian (N = 4) samples. There was no intrapopulation variation in sequence data from the weevils from Hawaii versus Australia, suggesting that they are sibling species.

Sex Pheromone Components of Nettle Caterpillar, Setora nitens

Gas chromatographic–electroantennographic detection (GC-EAD) analyses of pheromone gland extracts of female nettle caterpillars, Setora nitens, revealed four compounds that consistently elicited responses from male moth antennae. Retention indices on three fused silia columns (DB-5, DB-23, and DB-210) of two EAD-active compounds were almost identical to those of (E)-9-dodecenal (E9–12 : Ald) and (E)-9,11-dodecadienal (E9,11–12 : Ald), two pheromone components previously identified in congeneric Setothosea asigna. However, comparative GC, GC-EAD, and GC-mass spectrometry of extracted S. nitens compounds and authentic standards revealed that the candidate pheromone components were (Z)-9-dodecenal (Z9–12 : Ald) and (Z)-9,11-dodecadienal (Z9,11–12 : Ald). The two other EAD-active compounds in pheromone gland extracts proved to be the corresponding alcohols to these aldehydes. In field-trapping experiments in Tawau, Malaysia, synthetic Z9–12 : Ald and Z9,11–12 : Ald at a 1 : 1 ratio, but not singly, attracted male S. nitens. Attractiveness of these two aldehydes could not be enhanced through the addition of their corresponding alcohols. Whether these differences in pheromone biology and chemistry between S. nitens and S. asigna are sufficient to prevent cross-attraction of heterospecific males or whether nonpheromonal mechanisms are required to maintain reproductive isolation is currently being studied.

Decadienoates : Sex pheromone components of nettle caterpillars Darna trima and D. bradleyi

This study was undertaken to identify sex pheromone components of nettle caterpillars Darna trima and Darna bradleyi (Lepidoptera: Limacodidae) whose larvae defoliate oil palm, Elaeis guineensis, in southeast Asia. Coupled gas chromatographic–electroantennographic detection (GCEAD) analyses of pheromone gland extracts revealed two antennally active compounds produced by female D. trima and two by female D. bradleyi. Molecular structures of these candidate pheromone components were identified by electron-impact and chemical-ionization mass spectrometry; retention-index calculations on DB-5, DB-23, and DB-210 columns; microanalytical treatments, as well as syntheses of "auxilliary" compounds that facilitated identification of the compounds. The compounds from D. trima were 2-methylbutyl (E)-7,9-decadienoate (A) and (E)-2-hexenyl (E)-7,9decadienoate (B); from D. bradleyi we identified methyl (E)-7,9-decadienoate (C), and isobutyl (E)-7,9-decadienoate (D). In field experiments in Malaysia, (S)-2-methylbutyl (E)-7,9-decadienoate (SA) in combination with B proved to be essential and synergistic pheromone components for attraction of male D. trima. (R)-2-Methylbutyl (E)-7,9-decadienoate (RA) had no behavioral activity. Compound D singly attracted male D. bradleyi, but addition of C to D at a 1 : 10 ratio significantly enhanced attractiveness of the bait. Synthetic pheromone blends were more effective trap baits than unmated female moths and could be developed for monitoring populations of D. trima and D. bradleyi in Asian oil palm plantations.

Sex Pheromone Components of Pink Gypsy Moth, Lymantria mathura

Pheromone extract of female pink gypsy moth, Lymantria mathura, was analyzed by coupled gas chromatographic-electroantennographic detection (GC-EAD) and coupled GC-mass spectrometry (MS), employing fused silica columns coated with DB-5, DB-210, or DB-23 and a custom-made GC column that separated enantiomers of unsaturated epoxides. These analyses revealed (9R,10S)-cis–9,10-epoxy-Z3,Z6-nonadecadiene [termed here (+)-mathuralure] and (9S,10R)-cis–9,10-epoxy-Z3,Z6-nonadecadiene [termed here (–)-mathuralure] at a 1 : 4 ratio as major candidate pheromone components. In field experiments in northern Japan (Morioka, Iwate Prefecture and Bibai, Hokkaido Prefecture), (+)- and (–)-mathuralure at a ratio of 1 : 4, but not 1 : 1 or singly, were attractive to male L. mathura. This is the first demonstration that attraction of male moths required the very same ratio of pheromone enantiomers as produced by conspecific females. Whether L. mathura employ different blend ratios in different geographic areas, and the role of five additional candidate pheromone components identified in this study remains to be investigated.