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 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     

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.     

Reevaluation of the sex pheromone of the lesser date moth, Batrachedra amydraula, using autosampling SPME-GC/MS and field bioassays

In a previous study of the sex pheromone of the lesser date moth, Batrachedra amydraula, using laboratory females, (Z4,Z7)-4,7-decadien-1-yl acetate, Z4-decen-1-yl acetate, Z5-decen-1-yl acetate and decyl acetate were identified by sequential SPME-GC/MS analysis. Traces of Z5-decen-1-ol were detected only in airborne collections. Concomitant field tests and re-evaluation of the composition of the sex pheromone of B. amydraula with feral female moths, using an improved sequential SPME-GC/MS procedure revealed a complex mixture of candidate pheromonal compounds. The same unsaturated acetates and the corresponding alcohols: (Z4,Z7)-4,7-decadien-1-ol, Z4-decen-1-ol, Z5-decen-1-ol were positively identified. In addition, the corresponding aldehydes, octanol and octyl acetate were also detected. All compounds were found to be released in a circadian rhythm, in a narrow time window of 2 h, approximately 1 h before sunrise. Comprehensive field bioassays indicated that the optimal attractive blend is a three-component mixture of (Z4,Z7)-4,7-decadien-1-yl acetate, Z5-decen-1-yl acetate and Z5-decen-1-ol in a ratio of 1:2:2. This blend gave about fivefold higher trap catch of B. amydraula males as compared to the previously published binary blend of (Z4,Z7)-4,7-decadien-1-yl acetate and Z5-decen-1-yl acetate in a ratio of 1:2. The alcohol Z5-decen-1-ol is an essential synergistic component of the sex pheromone of B. amydraula. All other identified compounds are inert, being neither synergists nor inhibitors of the pheromone. The optimal pheromone blend will be a useful tool in monitoring and control of B. amydraula, which is a serious pest of date plantations throughout the Middle East and northern Africa.     

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.     

Sex pheromone of the large aspen tortrix, Choristoneura conflictana(Lepidoptera: Tortricidae)

Summary. Three components that elicited antennal response from male Choristoneura conflictana were found from female gland extracts analyzed using a coupled gas chromatographic-electroantennographic detector system. The main component in gland extracts was (Z)-11-tetradecenal (Z11-14:Ald). Two minor components also elicited antennal response: (E)11-tetradecenal (E11-14:Ald) and (Z)-11- tetradecen-1-ol (Z11-14:OH). Analysis of effluvia indicated that calling virgin females release mostly Z11-14:Ald and trace amounts of Z11-14:OH. Field and wind tunnel behavioral studies showed that Z11-14:Ald alone attracted male moths in a dose response pattern. Tests comparing male response to blends of components detected in gland extracts showed that addition of 1.8% of E11-14:Ald to Z11-14:Ald did not influence male moths in the wind tunnel, but resulted in significantly lower trap captures in the field. The threecomponent blend [Z11-14:Ald (100), E11-14:Ald (1.8), Z11-14:OH (11)], was less attractive than Z11-14:Ald alone in both field and wind tunnel studies. Traps baited with two virgin female moths were equally attractive to males as traps baited with the three-component synthetic blend but less attractive than traps baited with Z11-14:Ald alone. Field tests of various blends of the two components (Z11-14:Ald, Z11-14:OH) detected in the females’ effluvia showed that the addition of 1–10% Z11-14:OH to Z11-14:Ald did not affect the males’ response to Z11-14:Ald. Our data demonstrate that female C. conflictana release sex pheromone components in a different ratio than they are stored in the pheromone gland. The sex pheromone is comprised of a single component, Z11-14:Ald, that can be used to monitor mated and virgin male C. conflictana throughout their flight period.     

Identification of the sex pheromone of Conogethes pluto: a pest of Alpinia

In recent years, Conogethes pluto (Lepidoptera: Crambidae) has become a major pest of Alpinia and other ornamental gingers in the Northern Territory and Queensland, Australia. This pest damages the flowers and bores into the stems, causing substantial losses to production. Currently, no synthetic sex pheromone is available to monitor or control this pest. This work aims at the identification of the sex pheromone of this pest. Analysis of the sex pheromone gland of female C. pluto by gas chromatography/electroantennogram detector revealed the presence of seven candidate pheromone compounds that elicited electroantennogram responses. Using gas chromatography/mass spectrometry analysis and micro-derivatization reactions, six compounds were identified as (E)-10-hexadecenal, as the main pheromone compound, (Z)-10-hexadecenal, hexadecanal, (E)-10-hexadecen-1-ol, (10E,12E)-hexadeca-10,12-dienal and (3Z,6Z,9Z)-tricosa-3,6,9-triene as minor pheromone compounds. In two-field trapping experiments, C. pluto responded to the six-component blend, and three of six compounds, i.e., (E)-10-hexadecenal, (3Z,6Z,9Z)-tricosa-3,6,9-triene, and (10E,12E)-hexadeca-10,12-dienal were shown to be necessary for attraction. In a subsequent experiment testing various doses (i.e., 0.01, 0.1, and 1 mg) of the six-component blend, the largest number of males was captured in traps baited with a lure loading of 1 mg. The availability of the sex pheromone of C. pluto will enable monitoring and provides the basis for additional control options for this pest.     

Sex pheromone composition and reproductive isolation in two Busseola species (Lepidoptera: Noctuidae) in Kenya

Closely related species provide an interesting and useful model to understand mate communication diversification. Pre-mating isolation is the result of selection processes and is achieved by utilization of different sex pheromones, but also by different ecological and life traits such as geographic distribution, host-plant specialization and allochrony of sexual behavior. Here, we report sex pheromone identifications and pre-mating isolation of two closely related species of stemborers: Busseola segeta and the yet undescribed Busseola nov. sp., developing on different host-plants and in different geographical locations in Kenya. GC–MS analyses showed that the two species produced the same pheromone components, but in slight different ratios. The respective synthetic pheromone mixtures tested in the field were attractive for males of each species. The low inter-specific variation in the pheromone mixture would allow possible interbreeding in case of accidental introduction of both species in the same environment. Under laboratory conditions, allochrony of the mating period is total and might prevent interbreeding if confirmed in natural conditions. This point would ensure the reproductive isolation between the two species in case of modifications of ecological factors. This study and the chemical structures of the identified pheromone components clearly demonstrate that these Busseola species are closely related although they differ in morphology.     

Egg antimicrobials, embryo sex and chick phenotype in the yellow-legged gull

Maternal effects through albumen quality are largely unexplored, despite the fundamental role that albumen exerts as source of proteins and water, as well as for antimicrobial defence of the embryo. We analysed the variation of two major albumen antimicrobials, avidin and lysozyme, by extracting samples from freshly laid eggs of the yellow-legged gull (Larus michahellis) and by correlating their levels to egg features. Lysozyme concentration increased along the laying sequence, while avidin concentration decreased. Both antimicrobials declined during the season. In addition, avidin concentration declined from first- to last-laid male eggs, whereas the opposite was true among the female eggs. We also analysed chick body mass and size and immune response, in relation to albumen antimicrobial levels in their original egg while controlling for potential covariation between egg quality and rearing conditions by cross-fostering eggs between nests. Tarsus length decreased with avidin concentration, particularly early in the season. Avidin concentration negatively predicted tarsus length of chicks and the phytohaemagglutinin response of females, but not males. However, chick phenotype did not covary with lysozyme albumen concentration. This is the first study where maternal effects mediated by albumen antimicrobials are investigated in relation to both sex and egg features in any wild bird species. Whether the observed patterns of variation in antimicrobial concentration are the by-product of maternal physiological constraints, or reflect adaptive allocation strategies, cannot be ascertained. The covariation between chick cell-mediated immunity and albumen avidin concentration might be causal, according to the documented effects of albumen proteins on immunity in other species.     

(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.     

Identification of 1-methyloctyl butanoate as the major sex pheromone component from females of the saddle gall midge,Haplodiplosis marginata (Diptera: Cecidomyiidae)

The saddle gall midge, Haplodiplosis marginata (von Roser) (Diptera: Cecidomyiidae), has undergone a resurgence recently as a pest of cereals in Belgium and other European countries. An effective monitoring tool of saddle gall midge flights is needed to understand the enigmatic population dynamics of this pest, and to design an integrated management strategy. Therefore, volatile compounds emitted by females (alkan-2-ols and alk-2-yl butanoates) were identified, and the chirality of the emitted esters was determined to be the R absolute configuration. In field-trapping experiments, racemic non-2-yl butanoate attracted substantial numbers of H. marginata males. Thus, this compound will be useful in baited traps for monitoring seasonal flight patterns, and improving integrated management of the saddle gall midge in agricultural systems.     

Modified pheromone communication associated with insecticidal resistance in the obliquebanded leafroller, Choristoneura rosaceana (Lepidoptera: Tortricidae)

Summary. This study reports on the impact of insecticidal resistance on the diel periodicity of the calling behaviour and pheromone production of different-aged virgin females of the obliquebanded leafroller (OBL), Choristoneura rosaceana. While both resistant (R) and susceptible (S) females initiated calling on the first night following emergence, the periodicity of the calling behaviour, as determined by the mean onset time of calling (MOTC) and the mean time spent calling (MTSC) over the first six nights of calling, differed between the two strains. R females started calling significantly later in the night. However, as the MOTC of R females advanced with age but did not do so in S individuals, the difference between strains was more pronounced in younger than older females. Furthermore, R females spent less time calling than S individuals. However, the MTSC increased as a function of age in both R and S females, so the difference between strains remained fairly constant for each night of calling. The major component of OBL sex pheromone, the Z11-14: Ac, determined at peak calling activity, significantly declined with female age. Overall, pheromone production was lower in R females than in S females, with the difference being more pronounced in younger than in older individuals. Thus, resistant females may have a lower mating success. The mating success of both R and S strain males did not vary with the number of previous matings acquired. With regard to males, although there was a significant decline in spermatophore size with successive matings, there was no significant difference between strains. However, R males are smaller and may be disadvantaged through female choice and/or may respond differently to pheromone source compared with S individuals. If the reproductive success of both sexes is affected, this may have a profound influence on the dynamics of insecticidal resistance in the presence or absence of selection in OBL populations. Received 4 July 2001; accepted 19 October 2001.     

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.     

The sex pheromone of the grain aphid,Sitobion avenae (Fab.) (Homoptera,Aphididae)

Summary The sex pheromone of the grain aphid,Sitobion avenae (Fab.), is identified as (4aS,7S,7aR)-nepetalactone, for which specific olfactory cells are located in the secondary rhinaria of the male antenna. The related (1R,4aS,7S,7aR)-nepetalactol was not produced by females in significant amounts (ca 10,000-fold less than the nepetalactone), although cells in the male antenna were found that responded specifically to this compound. Behavioural activity in the linear track ol-factometer showed attraction of males to the nepetalactone, but not to the nepetalactol.