Identification and bioassay of sex pheromone components of carob moth,Ectomyelois ceratoniae (Zeller)

Three sex pheromone components of the carob moth were isolated and identified from the extract of female pheromone glands, using a variety of techniques including coupled gas chromatographic-electroantennographic recordings, coupled gas chromatographic-mass spectrometric analysis, microozonolysis, electroantennographic assays of monounsaturated standards, wind-tunnel bioassays, and field trials. The major component was identified as (Z,E)-9,11,13-tetradecatrienal, a novel lepidopterous pheromone component structure. Two minor components, either one of which improves the upwind flight response of males when blended with the major component, were identified as (Z,E)-9,11-tetradecadienal, and (Z)-9-tetra-decenal.     

(Z,Z)-6,9-heneicosadien-11-one,labile sex pheromone of the whitemarked tussock moth,Orgyia leucostigma

The whitemarked tussock moth (WMTM), Orgyia leucostigma (J. E. Smith), is a major pest of coniferous and deciduous trees in eastern Canada. Chemical identification of its sex pheromone depended primarily on GC-EAD and HPLC analysis, with confirmation of behavioral activity by wind tunnel and field tests. We identified (Z,Z)-6,9-heneicosadien-11-one (Z,Z-6,9-ket) at 4–5 ng/female as the only essential sex pheromone component. Also detected in female extracts were (Z)-6-heneicosen-11-one (Z6-ket) at 2.5 ng/female, (Z,E)-6,8-heneicosadien-11-one (Z,E-6,8-ket) at about 0.5 ng/female, and a trace amount of (Z,E)-6,9-heneicosadien-11-one. Traps containing as little as 1 g of Z,Z-6,9-ket attracted males at low population levels, indicating it is a potent sex attractant. Traps baited with Z6-ket attracted few males, and in wind-tunnel bioassays it was at least 100-fold less attractive to males than Z,Z-6,9-ket. No improvement in trap catch occurred with the addition of Z6-ket in various binary mixtures with Z,Z-6,9-ket, including the female ratio, and a ternary mixture of Z,Z-6,9-ket, Z6-ket, and Z,E-6,8-ket in the 9:5:1 ratio detected in females was no better than Z,Z-6,9-ket alone. We attribute the presence of Z,E-6,8-ket and Z,E-6,9-ket in female extracts to the spontaneous and rapid stereospecific isomerization of Z,Z-6,9-ket at room temperature. Male flight began at sunset but peaked during the second half of the night.     

Novel sex pheromone components from the fall cankerworm moth,Alsophila pometaria

A sex pheromone extract from fall cankerworm moths,Alsophila pometaria, attracted conspecific males in field tests. Four EAG-active components were isolated from the extract and identified by GC-MS, highfield PMR spectroscopy, and microchemical techniques asn-nonadecane (I), (Z,Z,Z)-3,6,9-nonadecatriene (II), (Z,Z,Z,E)-3,6,9,11-nonadecatetraene (III), and (Z,Z,Z,Z)-3,6,9,11-nonadecatetrane (IV). Studies of the behavioral responses of male moths in a flight tunnel to the isolated components showed II, III, and IV were the major components of the sex pheromone. No sex pheromone behavioral responses were observed for I.Lepidoptera: Geometridae.issued as NRCC No. 22964.     

Sex pheromone components of fall cankerworm moth,Alsophila pometaria

(Z,Z,Z,E)-3,6,9,11-Nonadecatetraene and (Z,Z,Z,Z)-3,6,9,11-nonadecatetraene, sex pheromone components ofAlsophila pometaria, were synthesized by stereoselective Wittig reactions and found to be spectroscopically and chromatographically identical to isolated natural material. Flight-tunnel bioassays and field-trapping experiments confirmed that the two tetraenes together with (Z,Z,Z)-3,6,9-nonadecatriene are sex pheromone components. While traps baited with either tetraene individually captured conspecific males in field-trapping experiments, addition of the triene, which captured no males by itself, to either tetraene resulted in synergistic responses.Lepidoptera: Geometridae.Issued as NRCC No. 23462.     

Stereospecific synthesis of (Z,Z)-11,13-hexadecadienal,a female sex pheromone of the navel orangeworm,Amyelois transitella (Lepidoptera: Pyralidae)

A synthesis of (Z,Z)-11,13-hexadecadienal, a compound previously identified as the major component of the sex pheromone of the female navel orangeworm,Amyelois transitella (Walker), is described. The key step is the reduction of an unsymmetrical conjugated diyne with dicyclohexylborane. This reduction produced theZ,Z isomer with less than 0.5% total of other isomers.Mention of a commercial or proprietary product in this paper does not constitute endorsement of that product by the USDA.     

Sex pheromone ofStenoma cecropia Meyrick (Lepidoptera: Elachistidae)

(Z,E)-9,11-tetradecadienal (Z9,E11–14: Ald; 11%), (Z,E)-9,11,13-tetradecatrienal (Z9,E11, 13–14: Ald; 67%), (Z,E)-9,11-tetradecadienyl acetate (Z9,E11–14: Ac, 5.5%), and (Z,E)-9,11,13-tetradecatrienyl acetate (Z9,E11,13–14: Ac; 16.5%) were identified in the extracts of female pheromone glands ofStenoma cecropia (Lepidoptera: Elachistidae). From electroantennograms and single sensillum recordings, receptors toZ9,E11,13–14:Ald andZ9,E11–14: Ald were found on male antenna. Behavioral data were obtained from olfactometric tests in the laboratory and field trapping experiments in Colombia. It appeared that a blend ofZ9,E11,13–14:Ald (83%) andZ9,E11–14:Ald (17%) was attractive to males. These aldehydes are assumed to be components of the sex pheromone ofS. cecropia, whereas the acetates found in gland extracts might be precursors of the pheromone.     

Identification of sex pheromone of browntail moth,Euproctis chrysorrhoea (L.) (Lepidoptera: Lymantriidae)

A unique sex attractant pheromone was isolated and identified from extracts of ovipositor tips from the female browntail moth (Lepidoptera: Lymantriidae). The pheromone compound, (Z,Z,Z,Z)-7,13,16,19-docosatetraen-1-ol isobutyrate, CH₃CH₂CH=CHCH₂CH=CHCH₂CH= CH(CH₂)₄CH=CH(CH₂)₆O₂CCH(CH₃)₂, was identified by a combination of gas chromatography, mass spectrometry, and microreactions and was confirmed by synthesis. Traps baited with 5–50 g of the synthetic pheromone, dispensed from rubber septa treated with an antioxidant and a UV stabilizer, gave male moth captures that were comparable to traps baited with three virgin females. Higher (250 g) and lower (0.04–2.5 g quantities of the synthetic pheromone on septa captured somewhat fewer males. Captures were the same for similar quantities of the natural and synthetic pheromone. Minor, inactive components in the tip extract were identified as a mixture of 7- and 8-pentacosanone.     

(Z,E)-3,5-Tetradecadien-1-ol acetate sex attractant for the carpenterworm moth,prionoxystus robiniae (peck) (Lepidoptera: Cossidae)

Electroantennogram analyses of female gland extract and of male antennal responses to synthetic standards suggested that (Z,E)-3,5-tetradecadien-1-ol acetate is a pheromone component for the carpenterworm moth,Prionoxystus robiniae (Peck). The four 3,5-geometrical isomers were synthesized and bioassayed in the laboratory and the field in 1972, 1973, and 1974. TheZ,E isomer was found to be active in the laboratory and a good attractant in the field. The synthesis of theZ,E isomer also produced considerable quantities of theE,E isomer, which is difficult to remove completely. TheE,E isomer does not inhibit the response of males to theZ,E isomer when it is present in amounts up to 20% of theZ,E isomer. The addition of a keeper, a volatility modifier, or an antioxidant prolonged the activity of the attractant for as much as 43 days. (Z,E)-3,5-Tetradecadien-1-ol acetate may be a natural pheromone, but it has not been chemically defined from female extract. There is EAG evidence that a second pheromonal component may be present. The attractant nevertheless provides a tool for population survey, behavioral studies, evaluation of economic impact, and possibly control.     

(Z,Z)-7,9-Dodecadienyl acetate,sex pheromone ofEpinotia tedella clerck (Lepidoptera: Tortricidae)

(Z,Z)-7,9-dodecadienyl acetate, a reported male attractant for several New World spp. ofEpinotia, was identified as the primary pheromone of femaleE. tedella (European spruce budmoth) through chemical analysis of pheromone gland washes, the electrophysiological study of male antennal receptor types, and field-trapping tests. The washes contained this compound at a level of approximately 0.4 ng per FE, along with a similar amount of the corresponding alcohol, (Z,Z)-7,9-dodecadien-1-ol. Each compound activated its own specialized type of male receptor cell. No traces of stereoisomers or monoenes were found in the washes. In field-trapping tests conducted in stands of Norway spruce in southern Germany, (Z,Z)-7,9-do-decadienyl acetate as a single chemical proved highly attractive to maleE. tedella over a range of lure doses. The corresponding alcohol or aldehyde did not show attractivity; rather, in binary combinations with the primary pheromone, these compounds, and also the (E,Z)-7,9 stereoisomeric acetate, reduced captures. A 101 blend of (E)-9- and (Z)-9-dodecenyl acetates, reported as anE. tedella male attractant, did not reveal significant captures. No otherEpinotia spp. besidesE. tedella responded to the various 7,9-do-decadienyl test baits during this study.     

Role of (Z)- and (E)-11-tetradecenyl acetate pheromone components in the sexual behavior of the (Z) strain of the european corn borer,Ostrinia nubilalis (Lepidoptera: Pyralidae)

A glass tube olfactometer bioassay was used to examine pheromone response of males of the (Z)-pheromone strain ofOstrinia nubilalis (Hubner). The presence of (E)-11-tetradecenyl acetate at the natural ratio to (Z)-11-tetradecenyl acetate (973; ZE) did not consistently elevate wing-fanning, upwind walking, or clasper extrusion over (Z)-11-tetradecenyl acetate alone. This bioassay did not reveal the behavioral role of (E)-11-tetradecenyl acetate.Published as Journal Article No. 10264 of the Michigan State University Agricultural Experiment Station.     

(E)-11,13-tetradecadienal: Major sex pheromone component of the eastern blackheaded budworm,Acleris variana (Fern.) (Lepidoptera: Tortricidae)

(E)-11,13-Tetradecadienal (E11,13–14:Ald) is the major sex pheromone component of the eastern blackheaded budworm (EBB),Acleris variana (Fern.). The compound was identified in female pheromone gland extracts by coupled gas chromatographic-electroantennographic detection (GC-EAD), coupled GC-mass spectrometry in selected ion monitoring mode, and retention index calculations of candidate pheromone components.E11,13–14:Ald alone as trap bait was very attractive to male EBB. Addition of the corresponding diene alcohol or acetate or both did not enhance attraction. (Z)-11,13-Tetradecadienal in binary combination with (E)-11,13–14:Ald neither enhanced nor reduced trap catches. Increasing the amounts of pheromone from 0.01 to 10 µg increased trap catches, but increase of pheromone quantity above 100 µg proportionately reduced attraction. Stabilization of slowly polymerizingE11,13–14:Ald and development of a sustained, adequate release rate is required for pheromone-based monitoring of EBB populations.     

Heliothis virescens: Attraction of males to blends of (Z)-9-tetradecen-1-ol formate and (Z)-9-tetradecenal

In field trapping experiments, 161 and 321 blends of (Z)-9-tetradecen-1-ol formate (Z-9-TDF) and (Z)-9-tetradecenal (Z-9-TDAL) caught as manyHeliothis virescens (F.) as 3 virgin females and virelure, the synthetic pheromone of this species [a 161 mixture of (Z)-11-hexadecenal (Z-11-HDAL) and (Z)-9-tetradecenal]. Z-9-TDF and (Z)-7-dodecen-1-ol formate (Z-7-DDF) are structurally similar to Z-11-HDAL and Z-9-TDAL, respectively. The sensory input elicited by Z-9-TDF appears to substitute for the sensory input of Z-1 1-HDAL. In contrast, Z-7-DDF had no significant effect on catches of maleH. virescens when used alone, in combination with either Z-11-HDAL or Z-9-TDF as a bait in traps, or as a disruptant of pheromone communication via permeation of the atmosphere. Furthermore, Z-9-TDF may be a more stable and economical attractant forH, virescens males than is Z-1 1-HDAL.This paper reports the results of research only. Mention of a pesticide in this paper does not constitute a recommendation for use by the U.S. Department of Agriculture nor does it imply registration under FIFRA as amended. Also, mention of a commercial or proprietary product in this paper does not constitute a recommendation or an endorsement of that product by the USDA.     

Female sex pheromone of the Yunnan pine caterpillar moth Dendrolimus houi: first (E,Z)-isomers in pheromone components of Dendrolimus spp

The Yunnan pine caterpillar Dendrolimus houi Lajonquière is a serious defoliator of coniferous forests in southwestern China. Gas chromatography–electroantennography (GC–EAG) analyses of extracts of female sex pheromone glands of D. houi moths revealed the presence of three compounds eliciting antennal responses. These were identified as (5E,7Z)-5,7-dodecadien-1-ol (E5,Z7-12:OH), (5E,7Z)-5,7-dodecadien-1-yl acetate (E5,Z7-12:OAc), and (5E,7Z)-5,7-dodecadienal (E5,Z7-12:Ald) by comparison of their GC retention indices, mass spectra, and EAG activities with those of synthetic standards. Average amounts of E5,Z7-12:OH, E5,Z7-12:OAc, and E5,Z7-12:Ald per calling virgin D. houi female were 14.7 ± 12.9 ng (± SD), 5.8 ± 5.4 ng, and 0.8 ± 1.4 ng, respectively, in a ratio of 100:39.7:5.6. These three components were also collected from the headspace of calling virgin female moths by solid-phase microextraction (SPME). In addition, trace quantities of (Z)-5-dodecen-1-ol (Z5-12:OH), (5Z,7E)-5,7-dodecadien-1-ol (Z5,E7-12:OH), (5E,7E)-5,7-dodecadien-1-ol (E5,E7-12:OH), (5Z,7E)-5,7-dodecadien-1-yl acetate (Z5,E7-12:OAc), (5Z,7Z)-5,7-dodecadien-1-yl acetate (Z5,Z7-12:OAc), and (5E,7E)-5,7-dodecadien-1-yl acetate (E5,E7-12:OAc) were tentatively identified in female pheromone gland extracts by selected ion monitoring GC-MS. Field trapping experiments showed that E5,Z7-12:OH, E5,Z7-12:OAc, and E5,Z7-12:Ald were essential for attraction of male D. houi moths. Traps baited with a 20:1:1 blend (alcohol/acetate/aldehyde) loaded on gray rubber septa were as effective as traps baited with virgin female moths. The optimum ratio of acetate to aldehyde was 1:1, and this ratio was more critical than the ratio of either compound to the alcohol. This represents the first example of (E,Z)-isomers in pheromone blends of Dendrolimus species.     

Sex pheromone components of three species ofSemiothisa (Geometridae), (Z,Z,Z)-3,6,9-heptadecatriene and two monoepoxydiene analogs

Adult males ofSemiothisa signaria dispuncta (Walker) were attracted to field traps baited with (Z,Z,Z)-3,6,9-octadecatriene and (Z,Z)-6,9-cis-3,4-epoxy-octadecadiene. However, analyses of sex pheromone gland extracts of females of this species by GC-MS and by GC in combination with an electroantennograph detector (GC-EAD) showed the pheromone to be comprised of a mixture of the next lower homologs: (Z,Z,Z)-3,6,9-heptadecatriene and (Z,Z)-6,9-cis-3,4-epoxy-heptadecadiene. Blends of these two C₁₇ compounds were subsequently found to be more attractive to males in the field than the corresponding C₁₈ mixtures. Sex pheromones of two otherSemiothisa species were also found to contain C₁₇ components. (Z,Z,Z)-3,6,9-Heptadecatriene, detected by GC-EAD analysis of a female abdominal tip extract ofS. bicolorata (Fabricius), attracted conspecific males, and this attraction was significantly reduced by additions of (Z,Z)-6,9-cis-3,4-epoxyheptadecadiene, the major pheromone component ofS. signaria dispuncta, to the lure. (Z,Z)-3,9-cis-6,7-Epoxy-heptadecadiene was detected by GC-EAD analysis as the primary male antennal stimulatory component present in abdominal tip extracts ofS. ulsterata (Pearsall), and males of this species were attracted to traps baited with this epoxide. Each of these three C₁₇ compounds constitute previously unknown lepidopteran sex pheromone components. Blends of (Z,Z, Z)-3,6,9-heptadecatriene and (Z,Z)-3,9-cis-6,7-epoxyheptadecadiene attracted males of a fourth species,S. delectata Hulst, but no females of this species were obtained to permit analysis of its sex pheromone. The occurrence of (Z,Z,Z)-3,6,9-heptadecatriene inS. neptaria (Guenee) females was indicated by GC-MS analysis of an abdominal tip extract; however, no males were attracted to any of the fielded mixtures containing this hydrocarbon.     

Components of the sex pheromone of the female spotted stalk borer,Chilo partellus (Swinhoe) (Lepidoptera: Pyralidae): Identification and preliminary field trials

FemaleChilo partellus (Swinhoe) abdominal tip extracts were examined by gas-liquid chromatography (GLC) combined with simultaneous electroantennographic (EAG) recording from the male moth. Two olfactory stimulants were detected and identified as (Z)-11-hexadecenal (I) and (Z)-11-hexadecen-1-ol (II) by their GLC behavior, microchemical reactions, and comparison with synthetic materials. Both compounds were detected in volatiles emitted by the calling female moth. Synthetic (Z)-9-tetradecenyl formate, a structural analog of aldehyde (I), also elicited a significant EAG response from the male moth. Field trials carried out in India using synthetic (I) and (II) as bait in water traps showed that compound (I) was highly attractive to maleC. partellus; compound (II) was not attractive, and its addition to (I) significantly reduced trap catches.     

Female sex pheromone of the pickleworm,Diaphania nitidalis (Lepidoptera: Pyralidae)

Heptane extracts of the ovipositors from pickleworm adults (Diaphania nitidalis) were found to contain (E)-11-hexadecenal along with proportionally smaller amounts of (Z)-11-hexadecenal, (E)- and (Z)-11-hexadecen-1-ol, hexadecanol, hexadecanal, and a trace amount of (E,Z)-10,12-hexadecadienal. Assays conducted in a flight tunnel and in the field showed that a synthetic mixture of the five unsaturated compounds elicited behavioral responses from pickleworm males that were indistinguishable from those elicited by extracts of the female or by mate-calling females. When any component was deleted from the set of five unsaturated compounds, the intensity and extent of male responses to the resulting mixtures were significantly attenuated. The female sex pheromone of the pickleworm resembles the pheromone of a congeneric species,D. hyalinata, but bioassays indicated that (E,E)-10,12-hexadecadienal, produced byD. hyalinata but not by the pickleworm, plays a role in pheromonal specificity.     

(Z,Z)-5,27-Tritriacontadiene: Major sex pheromone ofDrosophila pallidosa (Diptera; Drosophilidae)

A crude cuticular extract from both sexes of 3660 fruit flies (Drosophila pallidosa) was subjected to SiO₂ and AgNO₃/SiO₂ column chromatography, accompanied by bioassay for the sex pheromone activity. After three chromatographic steps, the active fraction was obtained. The main component of the active fraction was determined to be (Z,Z)-5,27-tritriacontadiene [(Z,Z)-5,27-C_33:2, on the basis of gas-liquid chromatographic analysis, chemical derivatization and gas chromatography-mass spectrometry. Synthetic (Z,Z)-5,27-C_33:2 at 5 female equivalents (FE) elicited a clear courtship response with a high courtship index amongD. pallidosa males. Therefore it was concluded that (Z,Z)-5,27-C_33:2 was a major sex pheromone component in this species.     

Identification of a four-component sex pheromone of the female oriental fruit moth,Grapholitha molesta (Lepidoptera: Tortricidae)

The female-emitted pheromone ofGrapholitha molesta (Busck), the Oriental fruit moth, was collected by holding females in glass flasks during calling. Flask washes were found to contain four pheromone components: (Z)-8-dodecenyl acetate and (E)-8-dodecenyl acetate in a 1007 ratio, and (Z)-8-dodecen-1-ol and dodecanol in a 10020 ratio. The ratio of (Z)-8-acetate to (Z)-8-dodecen-1-ol was approx. 10030. Approximately 0.1–0.2 ng of pheromone was recovered per female per hour of calling.Published as Journal Article No. 8598 of the Michigan State University Agricultural Experiment Station.     

Sex pheromone components isolated from china corn borer,Ostrinia furnacalis guenée (lepidoptera: Pyralidae), (E)- and (Z)-12-tetradecenyl acetates

The sex pheromone components from the corn borer spreading widely in China,Ostrinia furnacalis Guenée, have been identified as (E)-and (Z)-12-tetradecenyl acetates (E andZ12–14 Ac). The ratio ofE isomer toZ isomer was 53 47. Traps containing 1 × 10^–7–1 × 10^–5 g of these compounds captured more males than did live females or their tip extract (3–6 female equivalents). Tetradecyl acetate (14 Ac) was also identified in the tip extract. Its quantity was about 1.8 times the sum of the other two isomers. However, including this compound in its natural ratio in pheromone traps resulted in a decrease in trap catches (P< 0.05).     

(Z)-11-Hexadecenal and (3Z,6Z,9Z)-tricosatriene: sex pheromone components of the red banded mango caterpillar Deanolis sublimbalis

The sex pheromone of the red banded mango caterpillar, Deanolis sublimbalis (Lepidoptera: Crambidae), a serious pest of the mango Mangifera indica (Anacardiaceae) in India and Southeast Asia and a recent invader into northern Australia, has been identified. Three candidate compounds were identified from pheromone gland extracts of female moths, using gas chromatography (GC), GC-electroantennographic detection and GC-mass spectrometric analyses, in conjunction with dimethyldisulfide derivatization. Field bioassays established that both (Z)-11-hexadecenal (Z11-16:Ald) and (3Z,6Z,9Z)-tricosatriene (3Z,6Z,9Z-23:Hy) were required for attraction of male D. sublimbalis moths, and 1,000 μg of a 1:1 mix of Z11-16:Ald and 3Z,6Z,9Z-23:Hy was more attractive to male moths than caged virgin females. However, the binary blend was only attractive when the isomeric purity of the monounsaturated aldehyde was >99%, suggesting that the (E)-isomer was inhibitory. Although (Z)-11-hexadecen-1-ol (Z11-16:OH) was tentatively identified in gland extracts, the addition of this compound to the binary blend did not increase the numbers of moths captured. The pheromone can now be used in integrated pest management strategies. Electronic Supplementary Material Supplementary material is available for this article at and is accessible for authorized users.