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

Summary. 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. Received 12 January 2001; accepted 8 June 2001.     

(<Emphasis Type="Italic">Z,Z</Emphasis>)-11,13-Hexadecadienyl acetate and (<Emphasis Type="Italic">Z,E</Emphasis>)-11,13,15-hexadecatrienyl acetate: synergistic sex pheromone components of oak processionary moth,Thaumetopoea processionea (Lepidoptera: Thaumetopoeidae)

Summary. Our objective was to identify sex pheromone components of the oak processionary moth, Thaumetopoea processionea (Lepidoptera: Thaumetopoeidae), whose larvae defoliate oak, Quercus spp., forests in Eurasia and impact human health. Coupled gas chromatographic-electroantennographic detection (GC-EAD) and GC-mass spectrometric (MS) analyses of pheromone gland extract of female T. processionea revealed two consistently EAD-active compounds. They were identified as (Z,Z)-11,13-hexadecadienyl acetate (Z11,Z13-16:OAc) and (Z,E)-11,13,15-hexadecatrienyl acetate (Z11,E13,15-16:OAc) by comparative GC, GC-MS and GC-EAD analyses of insect-produced compounds and authentic standards. In replicated field experiments (2000, 2001) in Nordbaden, Südbaden and Sachsen-Anhalt (Germany), Z11,Z13-16:OAc and Z11,E13,15-16:OAc in combination, but not singly, attracted significant numbers of male moths. It will now be intriguing to investigate whether Z11,E13,15-16:OAc, or its corresponding alcohol or aldehyde, serves as a pheromone component also in other species of the Thaumetopoeidae.     

A sex pheromone component novel to Ostrinia identified from Ostrinia latipennis (Lepidoptera: Crambidae)

Summary. Extracts from the sex pheromone gland of Ostrinia latipennis (Lepidoptera: Crambidae) were analyzed by gas chromatography-electroantennographic detection (GC-EAD) and GC-mass spectrometry. Only an EAD-active compound was detected in the extract, and it was identified as (E)-11-tetradecenol (E11-14:OH). In a wind-tunnel bioassay, E11-14:OH elicited a series of mate finding behaviors from males, although it was far less active than virgin females and crude extract of the pheromone gland. The attractiveness of E11-14:OH to O. latipennis males was confirmed by field trapping experiments. Based on these findings, we concluded that E11-14:OH, which is novel to the genus Ostrinia, is a major component of the sex pheromone in O. latipennis. The significance of the use of alcohol in place of the usual acetates in Ostrinia is discussed in relation to the pheromone biosynthesis system. Received 9 December 1999; accepted 14 March 2000     

Female sex pheromone of Ostrinia orientalis – throwing a light on the relationship between O. orientalis and the European corn borer, O. nubilalis

Summary. The sex pheromone of Ostrinia orientalis (Lepidoptera: Crambidae) was analyzed by gas chromatography–electroantennographic detection (GC–EAD), GC–mass spectrometry and a series of bioassays. Three EAD-active compounds were detected in the female sex pheromone gland extract, and identified as tetradecyl acetate (14:OAc), (Z)-11-tetradecenyl acetate (Z11-14:OAc) and (E)-11-tetradecenyl acetate (E11-14:OAc). The titers (ratio) of 14:OAc, Z11-14:OAc and E11-14:OAc in 3-day-old virgin females were 0.49 ng (10), 4.86 ng (98) and 0.10 ng (2), respectively. In a wind-tunnel bioassay, the 98:2 blend of Z11- and E11-14:OAc, but not Z11-14:OAc alone, elicited the same male behavioral responses as virgin females and crude gland extracts. 14:OAc was inactive by itself, and did not show any synergistic effect on the binary blend. Field trapping experiments also confirmed the attractiveness of the binary blend to O. orientalis males. Based on these results, we concluded that the sex pheromone of O. orientalis is a 98:2 mixture of Z11-14:OAc and E11-14:OAc. This sex pheromone is very similar to that of the Z-type European corn borer, O. nubilalis. The present finding raises the question of whether O. orientalis , which is indistinguishable from O. nubilalis based on external morphology, is a biologically distinct species independent from O. nubilalis.     

Sex pheromone components of pyralid moths Terastia subjectalis and Agathodes ostentalis feeding on the coral tree, Erithrina variegata: Two sympatric species share common components in different ratios

Summary. Two common components, identified as (E)-11-hexadecenal (E11-16:Ald) and (E,E)-10,12-hexadecadienal (E10E12-16:Ald), were found in the extract of virgin females of two sympatric pyralid moths, Terastia subjectalis Lederer, and Agathodes ostentalis (Geyer). The amount of E11-16:Ald and E10E12-16:Ald was 0.12 and 2.8 ng/female for T. subjectalis and 2.0 and 1.8 ng/female for A. ostentalis, respectively. Hexadecanal (16:Ald) was also found at 0.7 ng/female in the extract of A. ostentalis, but EAG activity was unclear. In the field, T. subjectalis and A. ostentalis males were captured with species specific blends in the proportions of 5:95 and 50:50 blends. Blends attractive to one species did not attract the opposite one. Rubber septa loaded with 1 mg each of 5:95 and 50:50 blends of E11-16:Ald and E10E12-16:Ald were as attractive as two virgin females of T. subjectalis and A. ostentalis , respectively. This demonstrated that different ratios of shared pheromone components could provide species specific cues for critical mate location in the two sympatric pyralid moths. It is suggested that directional selection probably occurs in the pheromone blends in opposite directions in both species.     

Heritable pheromone blend variation in a local population of the butterbur borer moth Ostrinia zaguliaevi (Lepidoptera: Crambidae)

Summary. In a local population of Ostrinia zaguliaevi Mutuura & Munroe (Lepidoptera: Crambidae), extensive variation was found in the blend ratio of three sex pheromone components, (Z)-9-tetradecenyl acetate (Z9-14:OAc, 10.2-63.8%), (Z)-11-tetradecenyl acetate (Z11-14:OAc, 32.2-86.8%), and (E)-11-tetradecenyl acetate (E11-14:OAc, 2.1-11.9%). The variation was observed over a three-year period (2002-2004). Mother-daughter regression analyses have shown that although the heritability of the minor component E11-14:OAc was not significant, the heritability of the proportions of Z9-14:OAc and Z11-14:OAc were substantial (0.5–0.6). In artificial selection experiments, the mean % Z9-14:OAc in the sex pheromone changed significantly within three generations (37% in the control line, 48 and 52% for two lines selected for increase, and 23 and 30% for two lines selected for decrease). Despite these changes, the amounts of fatty acyl pheromone precursors, (Z)-9-, (Z)-11- and (E)-11-tetradecenoate, in the pheromone gland were not significantly affected by the selection. Taken together, variation in the pheromone blend of O. zaguliaevi is likely to be attributable to a few genes involved in the reduction or acetylation of fatty acyl pheromone precursors, the last two steps in pheromone biosynthesis.     

Comparative studies on the sex pheromones of Ostrinia spp. in Japan: the burdock borer, Ostrinia zealis

Summary. To gain insight into the evolution of the sex pheromone communication system in Ostrinia (Lepidoptera Pyralidae), the sex pheromone of the burdock borer, O. zealis was analyzed by means of gas chromatography-electroantennographic detection (GC-EAD), GC-mass spectrometry and a series of bioassays. Four EAD-active compounds were detected in the female sex pheromone gland extract, and these were identified as tetradecyl acetate (14:OAc), (Z)-9-tetradecenyl acetate (Z9–14:OAc), (E)-11-tetradecenyl acetate (E11-14:OAc) and (Z)-11-tetradecenyl acetate (Z11-14:OAc). The average amounts (ratio) of the four compounds in single sex pheromone glands were 2.5 ng (13%), 11.6 ng (61%), 4.1 ng (21%) and 0.9 ng (5%), respectively. In a wind-tunnel bioassay, the ternary blend of Z9-, E11- and Z11-14:OAc at a ratio found in the sex pheromone gland elicited the same behavioral responses from the males as did virgin females. 14:OAc did not show any enhancement or inhibition of the males’ behavioral responses when added to the ternary blend. The attractiveness of the 3-component lure to O. zealis males was also confirmed by field trapping experiments. Based on these results, we concluded that the sex pheromone of O. zealis is composed of Z9-14:OAc, E11-14:OAc and Z11-14:OAc at a ratio of 70:24:6. The evolutionary changes of the sex pheromones in Ostrinia are also discussed based on the presently available information on the sex pheromones and phylogenetic relationships of Ostrinia spp. Received 25 September 1998; accepted 2 December 1998.     

Sex pheromone communication of tentiform leaf-miners <Emphasis Type="Italic">Phyllonorycter insignitella</Emphasis> and <Emphasis Type="Italic">Ph. nigrescentella</Emphasis> from two related species groups

Summary.  Females of both species start their pheromone-releasing activity on the second day after emergence at the beginning of the photophase. During the present work, a peak of calling activity with close to 100% of active Ph. nigrescentella females was registered 1.5 hour after the light had been put on. The high pheromone release behaviour with 50% active females lasted for 3 hours. The calling activity of the group of females was about 6 h/day. The beginning of a photophase under laboratory conditions or an early morning in nature is a common period for sex pheromone release in the genus Phyllonorycter. (8Z,10E)-tetradecadien-1-yl acetate (8Z,10E-14:Ac), (8Z,10E)-tetradecadien-1-ol (8Z,10E-14:OH) and (8E,10Z)-tetradecadien-1-yl acetate (8E,10Z-14:Ac) in the ratio 96:4:traces as well as 8Z,10E-14:Ac and 8Z,10E-14:OH in the ratio 88:12 collected by Solid Phase Micro Extraction (SPME) were found to be specific for the calling periods of virgin Phyllonorycter insignitella and Ph. nigrescentella females respectively. Field trapping experiments demonstrated that all three compounds are important for the attraction of Ph. insignitella males while only 8Z,10E-14:Ac is the essential sex pheromone component for Ph. nigrescentella. The pheromone activity of all three compounds is reported for the first time. Addition of either 8Z,10E-14:OH or 8E,10Z-14:Ac to 8Z,10E-14:Ac did not have a significant effect on the attraction of Ph. nigrescentella males, while the efficiency of the three component blend was 5 times lower as compared to that of 8Z,10E-14:Ac. Our data demonstrate that 8Z,10E-14:OH and 8E,10Z-14:Ac play a dual function, they are minor sex pheromone components of Ph. insignitella essential for attraction of conspecific males and show an allelochemical, antagonistic effect on Ph. nigrescentella males and, thus, ensuring specificity of the mate location signal in two related Phyllonorycter species.     

Differential activity of non-fluorinated and fluorinated analogues of the European corn borer pheromone

Summary.  The differing antagonist activity of (Z)-13-hexadecen-2-one (Z11 – 14 :MK, 1) and its 1,1,1-trifluoro derivative (Z11 –14 :TFMK, 2), two closely related analogues of the European corn borer pheromone Ostrinia nubilalis (Z strain), and their rationale is reported. Both chemicals exhibited some electrophysiological activity, and topical application of 10 pg of pheromone analogue on male antennae was sufficient to induce significantly lower depolarization responses to the pheromone versus untreated insects. In a wind tunnel, the number of European corn borer males attracted to sources containing mixtures of 1 + pheromone in ratios ≥ 1 :1 was significantly lower than the number attracted to a source containing pheromone alone. Source contact behaviour was dramatically impaired when the 1 + pheromone blend reached a ratio of 10 :1, in which only 2% of males displayed source contact in the presence of antagonist. When compound 1 was present at the source, males usually flew upwind with occasional downwind reversals; when compound 2 was present at the lure, males performed wider crosswind reversals, with little progress toward the source. In the field, traps baited with mixtures of both compounds with the pheromone in ratios of 5 :1 and 10 :1 elicited a significantly decreased number of male catches. In esterase inhibition assays, compound 2 was a potent inhibitor (IC₅₀ = 70 nM), whereas the non-fluorinated compound 1 was not. The different activity of both compounds is presumed to be due to different mechanisms of action; considerations for using methyl ketone analogues as new behavioural antagonists of the pheromone are outlined.     

Sex pheromones and their potential role in the evolution of reproductive isolation in small ermine moths (Yponomeutidae)

Summary Sex pheromone communication in the nine European species of small ermine moths (Yponomeuta) is reviewed in regard to the potential role of pheromones in the speciation process. Six of the nine species studied (viz.,Y. evonymellus, Y. cagnagellus, Y. padellus, Y. irrorellus, Y. plumbellus, andY. vigintipunctatus) use a mixture of (E)-11-and (Z)-11-tetradecenyl acetate in different ratios as primary pheromone components, with combinations of tetradecyl acetate, (Z)-9-tetradecenyl acetate, (Z)-11-hexadecenyl acetate and the corresponding alcohols of the acetates as additional pheromone components. Analysis of (Z)- to (E)-11-tetradecenyl acetate ratios produced by individual females of these species demonstrated significant variation among females of all species. However, the ranges of ratios produced byY. cagnagellus, Y. irrorellus, andY. plumbellus, sharing the same host-plant species, spindle tree, did not overlap. Niche separation of all six species mentioned required consideration of at least one additional pheromone component or of temporal aspects. The remaining three species,i.e. Y. malinellus, Y. mahalebellus andY. rorellus, have pheromones that differ qualitatively.Biosynthetic routes to the pheromone components identified are proposed on the basis of fatty acid pheromone precursors found in the pheromone glands. A phylogenetic tree for the genus is constructed based on allozyme frequency data and changes in pheromone composition are superimposed on this tree. We suggest that the ancestral ermine moth pheromone is a mixture of (Z)-11- and (E)-11-tetradecenyl acetate and the corresponding alcohols, and a scenario of how present-day patterns evolved is outlined. The pheromone differences among the three species using spindle tree as their host-plant might have evolved throughreproductive character displacement upon secondary contact between populations that had already diverged genetically in allopatry. Pheromone differences within the so-calledpadellus-complex (includingY. cagnagellus, Y. mahalebellus, Y. malinellus, Y. padellus, andY. rorellus) in which species might have originated sympatrically, may have evolved byreinforcing selection as these species still hybridise and produce viable offspring when confined in cages. The role of pheromones in reproductive isolation amongYponomeuta species is emphasised by (1) the function of pheromone components of some of the species as behavioural antagonists to other species, (2) the cross-attraction under experimental conditions between allochronic species with similar pheromones, and (3) the formation of hybrids in the laboratory between species that are isolated in nature by pheromone differences.     

Host-plant green-leaf volatiles synergize the synthetic sex pheromones of the corn earworm and codling moth (Lepidoptera)

Summary The capture of adult male moths in female sex pheromone traps of two key agricultural pests, the corn earworm (Helicoverpa zea) and the codling moth (Cydia pomonella), is enhanced or synergized by a certain group of host-plant volatiles, the green-leaf volatiles (GLVs). Since female adults of both species call and release their sex pheromones while perched upon the leaves of their host-plants, the volatile constituents from the leaves of a number of host-plants were compared. Sex pheromone traps containing one of the prominent leaf volatiles of certainH. zea hosts, (Z)-3-hexenyl acetate, not only significantly increased the capture ofH. zea males but were preferred over traps baited only with sex pheromone. Similarly, traps baited with synthetic sex pheromome ofC. pomonella plus a blend of GLVs captured significantly more males than traps baited only with sex pheromone. Since male moths are not captured in traps baited only with these GLVs, it appears that these GLVs act as pheromone synergists which increase or enhance the attraction or arrestment of male moths in pheromone traps.     

Identification and field evaluation of the female sex pheromone of Stathmopoda masinissa in Korea

The sex pheromone of Stathmopoda masinissa Meyrick, an important pest of persimmon fruit in East Asia such as Korea, China, and Japan, was investigated. A lure using (E4,Z6)-4,6-hexadecadienyl acetate (E4,Z6-16Ac), which was identified as a sex pheromone compound of Japanese population, did not work at all for Korean population. Therefore, components in the abdominal extract of the moth were identified and their attractiveness was evaluated in the field. Two components, E4,Z6-16Ac and (E4,Z6)-4,6-hexadecadien-1-ol (E4,Z6-16OH) were identified from the extract of female abdominal extract in a ratio of 10–15:90–85 by GC–MS analysis with synthetic standards. E4,Z6-16Ac and E4,Z6-16OH were previously identified as EAG-active components of this moth in Japanese population. However, (E4,Z6)-4,6-hexadecadienal (E4,Z6-16Ald), which is one of the abdominal extract components and EAG-active component in Japanese population, was not detected in our samples. In the persimmon orchard, single component of E4,Z6-16Ac or E4,Z6-16OH was not attractive. However, the 1:1 mixture of the two components significantly increased the captures of male S. masinissa. Interestingly, traps baited with E4,Z6-16Ac captured significantly higher number of Oedematopoda ignipicta (Lepidoptera: Stathmopodidae) than the traps baited with E4,Z6-16OH or blend of the two components. The attractiveness of E4,Z6-16Ac to O. ignipicta is a new finding.     

Identification, synthesis and activity of sex pheromone gland components of the autumn gum moth (Lepidoptera: Geometridae), a defoliator of Eucalyptus

Summary. The autumn gum moth, Mnesampela privata (Guenée) (Lepidoptera: Geometridae), is native to Australia and can be a pest of plantation eucalypts. Field-collected and laboratory-reared female autumn gum moths were dissected to remove glands likely to contain components of the sex pheromone. Using gas chromatography (GC) and combined gas chromatography–mass spectrometry (GC-MS), three compounds were identified from female extracts, namely (3Z,6 Z,9 Z)-3,6,9-nonadecatriene, 1-hexadecanol and 1-octadecanol (confirmed by comparison with synthetic samples). Nonadecatriene elicited an antennal response in male autumn gum moth during gas chromatographic analyses combined with electroantennographic detection (GC-EAD). In electroantennogram (EAG) recording male M. privata antennae responded to the nonadecatriene. Nonadecatriene was synthesised via Kolbe electrolysis, starting with (9Z,12Z,15Z)-octadeca-9,12,15-trienoic acid (linolenic acid) and propanoic acid or via an alternative four-step method also starting from linolenic acid. In field trials (3Z,6Z,9Z)-3,6,9-nonadecatriene proved attractive to male moths. Thus, we conclude that (3Z,6Z,9Z)-3,6,9- nonadecatriene is a sex pheromone component of autumn gum moth. This component has been identified in extracts from other geometrids in the same subfamily, Ennominae. However, to our knowledge this is the first example where (3Z,6Z,9Z)-3,6,9-nonadecatriene has been found in females and also proved attractive to male moths when presented on its own. Our results are discussed in relation to other geometrid pheromones.     

Identification and biological activity of sex pheromone components from females of the plum moth Illiberis rotundata Jordan (Lepidoptera: Zygaenidae: Procridinae)

The plum moth, Illiberis rotundata Jordan (Lepidoptera: Zygaenidae: Procridinae), is a pest of orchards in Japan and China. Few chemical ecological studies have been directed towards the Zygaenidae and particularly the Procridinae. To investigate the sex pheromone of this species, extracts of pheromone glands from adult female I. rotundata were analyzed by coupled gas chromatography-electroantennography (GC-EAG) and coupled gas chromatography-mass spectrometry (GC-MS). Whilst GC-EAG on male moths showed an active peak, identified as 2-butyl (7Z)-dodecenoate, GC-MS also revealed the presence of the homologue 2-butyl (9Z)-tetradecenoate. Electroantennographic investigations, as well as field tests, strongly suggested the natural compounds to have the (R)-configuration at the stereogenic centre. Field results demonstrate 0.2 mg of a 1:1-mixture of (2R)-butyl (7Z)-dodecenoate and (2R)-butyl (9Z)-tetradecenoate to be a powerful lure that may be used in pest control measures against I. rotundata. The chemical structures of the new pheromone components show the same features as those of other zygaenid species: unsaturated fatty acids esterified with a short chain chiral alcohol. This is the first example of a two-component blend constituting the pheromone of a procridinid species.     

A synomone imparting distinct sex pheromone communication channels for Choristoneura rosaceana (Harris) and Pandemis limitata (Robinson) (Lepidoptera: Tortricidae)

Summary. Male obliquebanded leafrollers, Choristoneura rosaceana (Harris), were induced to respond to a pheromone source tainted with a behavioural antagonist, Z9-tetradecenyl acetate, when a source releasing the antagonist was placed 10 cm upwind of the tainted source in a wind tunnel. However, placement of the antagonist upwind of an attractive pheromone source did not interrupt pheromone-mediated responses. Placement of a source releasing Z9-tetradecenyl acetate, a minor pheromone component of the sympatric species, the threelined leafroller, Pandemis limitata (Robinson), upwind of a calling P. limitata female, reduced conspecific male pheromone-mediated response but resulted in upwind flight by male C. rosaceana and contact with heterospecific females. Male P. limitata locked on and flew upwind to but did not contact heterospecific females when a source releasing Z9-tetradecenyl acetate was positioned upwind of a calling C. rosaceana female. In the field, adaptation or habituation to Z9-tetradecenyl acetate caused by atmospheric treatment with this compound apparently resulted in reciprocal heterospecific pheromone responses. More C. rosaceana males were captured in traps baited with their pheromone and the behavioural antagonist in small field plots treated atmospherically with Z9-tetradecenyl acetate than in nontreated control plots. Fewer male P. limitata were captured in traps baited with their own pheromone, or with C. rosaceana pheromone tainted with Z9-tetradecenyl acetate in plots treated atmospherically with Z9-tetradecenyl acetate than in nontreated control plots. We argue that Z9-tetradecenyl acetate is an important synomone which assists in partitioning the sexual chemical communication channels of C. rosaceana and P. limitata. Received 9 February 1999; accepted 22 March 1999.     

Specificity of communication channels in four limacodid moths: Darna bradleyi, Darna trima, Setothosea asigna, and Setora nitens (Lepidoptera: Limacodidae)

Summary. Darna bradleyi Holloway, D. trima Moore, Setothosea asigna van Eecke and Setora nitens Walker are sympatric and coseasonal limacodid moths in plantations of oil palm, Elaeis guineensis Jacq. (Arecales: Palmae), in Borneo, southeast Asia. We tested the hypothesis that these four species maintain reproductive isolation through specificity in diel periods of communication, microlocation for communication and/or communication signal (pheromone). Studying diel periodicity of calling behavior by female moths and response by male moths to traps baited with virgin females or synthetic pheromone, we determined that sexual communication of D. bradleyi and D. trima took place from ˜17:30 to 18:45 hr and that of S. asigna and S. nitens from ˜18:45 to 20:00 hr and from ˜18:30 to 19:30 hr, respectively. Over 80% of male S. asigna and S. nitens were captured in pheromone-baited traps suspended >5 m high, whereas male D. bradleyi and D. trima were captured mostly in traps <5 m high. Synthetic pheromone baits attracted male moths in a species-specific manner. Moreover, baits containing both S. asigna and S. nitens pheromones failed to attract any male moths, indicating that female S. asigna and S. nitens, with overlapping communication periods, use bifunctional pheromone components that attract conspecific males while repelling heterospecifics. Similarly, addition of D. bradleyi pheromone to S. asigna or S. nitens pheromone reduced attraction of male S. asigna and S. nitens. The failure of D. bradleyi and D. trima, which overlap in time and microlocation for communication, to evolve bifunctional pheromones may be attributed to the recent occurrence of sympatry between D. bradleyi and D. trima in Borneo, apparently too recent for bifunctional pheromones to have evolved. We conclude that D. bradleyi, D. trima, S. asigna and S. nitens utilize any or all of diel periodicity, intra and interspecific effects of communication signal and/or microlocation for signaling, allowing these limacodids to co-inhabit the same habitat and remain reproductively isolated. Received 5 May 2000; accepted 18 August 2000     

Aggregation pheromone ofCarpophilus freemani (Coleoptera: Nitidulidae): A blend of conjugated triene and tetraene hydrocarbons

Summary Males ofCarpophilus freemani Dobson (Coleoptera: Nitidulidae) produce an aggregation pheromone to which both sexes fly in a wind-tunnel bioassay. The major pheromone component (ca. 30 ng per male per day in volatile collections) was identified as (2E,4E,6E)-5-ethyl-3-methyl-2,4,6-nonatriene. A minor component, (2E,4E,6E,8E)-7-ethyl-3,5-dimethyl-2,4,6,8-undecatetraene, was 3–10% as abundant as the major triene and was 5–20% as active when compared at relative doses ranging from natural proportions to 1:1. These compounds act synergistically: a mixture of major and minor components in natural proportions attracted more than twice as many beetles as the major component alone, and the mixture fully accounted for the activity of male-derived volatile collections. Six other male-derived conjugated hydrocarbons, ranging from 2% down to 0.04% as abundant as the major component, were also identified. These are (in order of decreasing bioassay activity when compared on an equal-weight basis): (3E,5E,7E)-6-ethyl-4-methyl-3,5,7-decatriene, (2E,4E,6E)-5-ethyl-3-me-thyl-2,4,6-octatriene, (3E,5E,7E,9E)-8-ethyl-4,6-dimethyl-3, 5,7,9-dodecatetraene, (2E,4E,6E,8E)-3,5,7-trimethyl-2,4,6, 8-undecatetraene, (3E,5E,7E)-5-ethyl-7-methyl-3,5,7-undecatriene, and (2E,4E,6E)-3,5-dimethyl-2,4,6-nonatriene. All structure identifications were confirmed by synthesis. In the wind tunnel, the pheromone acted synergistically with host-type volatiles such as propyl acetate, valeric acid, and ethanol. This concept was verified by fields tests in California, in which there was dramatic synergism between the pheromone and fermenting host materials. Pheromone biosynthesis is discussed.     

Neocembrene A, a major component of the trail-following pheromone in the genus Prorhinotermes (Insecta, Isoptera, Rhinotermitidae)

Summary. The diterpene neocembrene A or (1E,5E,9E,12R)-1,5,9-trimethyl-12-(1-methylethenyl)-1,5,9-cyclotetradecatriene, known as the trail-following pheromone of the advanced Termitidae Nasutitermitinae Nasutitermes exitiosus and Trinervitermes bettonianus, has been identified after SPME-GC/MS as the major component of the trail-following pheromone of the Rhinotermitidae Prorhinotermitinae, Prorhinotermes canalifrons and P. simplex. In all the other Rhinotermitidae studied until now, the major component of their trail pheromones is dodecatrienol ((3Z,6Z,8E)-dodeca-3,6,8-trien-1-ol). This biochemical data further add to the anatomical and molecular characteristics that give a special status to the taxon Prorhinotermes among Rhinotermitidae. In Prorhinotermes canalifrons and P. simplex, neocembrene A was the only secretory compound specific to the sternal gland surface that could be detected after SPME. It elicited orientation as well as recruitment behavioral effects. However, the comparison of the respective biological activities triggered by neocembrene A and by sternal gland secretion suggests that minor components of the latter are acting in synergy with neocembrene A.     

(Z)-11-Pentacosene is the major sex pheromone component inDrosophila virilis (Diptera)

Summary A crude cuticular extract from 3450 virgin 9–13 day old female fruit flies(Drosophila virilis), was subjected to chromatography accompanied by bioassay for sex pheromone activity. After three chromatographic steps, fractions containing active monoenes and dienes were obtained. Chemical analysis by infrared absorption, gas liquid chromatography and gas chromatography/mass spectrometry of the active fraction indicated that active monoenes were comprised chiefly of (Z)-11-pentacosene (abbreviated (Z)-11-C_25:1), (Z)-13-C_27:1, (Z)-13- and (Z)-14-C_29:1. Synthetic monoenes were made, and only (Z)-11-C_25:1 elicited good courtship behaviour in maleD. virilis. Therefore it was concluded that (Z)-11-C_25:1 was a major sex pheromone. A total of 16.2±1.32 µg of cuticular hydrocarbons was isolated from 10 day old females, including 5.9±0.56 µg of (Z)-11-C_25:1. An additive effect was suggested from the higher observed courtship response when using a mixture of active dienes with the active monoene.