Semiochemistry of the Goldeneyed Lacewing Chrysopa oculata: Attraction of Males to a Male-Produced Pheromone

Gas chromatographic-electroantennographic detection (G3C-EAD) experiments showed that antennae of males and females of the goldeneyed lacewing, Chrysopa oculata Say (Co. = Chrysopa), consistently responded to four compounds extracted from the abdominal cuticle of males:nonanal, nonanol, nonanoic acid, and (1R*,2S*,5R*,8R*)-iridodial. These compounds were not detected from abdominal cuticle of females. Thoracic extracts of both sexes contained antennal-stimulatory 1-tridecene and EAD-inactive skatole. Chrysopa oculata adults were most sensitive to (1R,2S,5R,8R)-iridodial standard at an EAD-response threshold between 0.1 and 1 pg, which was 10-100 times lower than thresholds for nonanal and nonanoic acid, and up to 10,000 times lower than thresholds for other compounds tested. A similar EAD response pattern was also found in another Chrysopa sp. (Co. quadripunctata Burmeister). In field-trapping experiments, (1R,2S,5R,8R)-iridodial was the only male-specific compound that attracted Co. oculata males. Males also were weakly attracted to (1R,4aS,7S,7aR)-nepetalactol (an aphid sex pheromone component), probably due to the 5% (1R,2S,5R,8R)-iridodial present in the synthetic sample as an impurity. A herbivore-induced plant volatile, methyl salicylate, increased attraction of males to (1R,2S,5R,8R)-iridodial, whereas 1-tridecene was antagonistic. No females were caught in the entire study. Scanning electron micrographs revealed numerous male-specific, elliptical epidermal glands on the 3rd-8th abdominal sternites of Co. oculata, which are likely the pheromone glands. Another lacewing species, Chrysoperla rufilabris (Burmeister) (Cl. = Chrysoperla), did not produce male-specific volatiles or possess the type of gland presumed to produce pheromone in Co. oculata males, but (Z)-4-tridecene was identified as a major antennal-stimulatory compound from thoracic extracts of both sexes of Cl. rufilabris. Thus, (1R,2S,5R,8R)-iridodial (or its enantiomer) is now identified as a male-produced male aggregation pheromone for Co. oculata, the first pheromone identified for lacewings.     

Defensive Compounds and Male-Produced Sex Pheromone of the Stink Bug, Agroecus griseus

Agroecus griseus is a serious corn pest in Brazil. Contents of the dorsal abdominal glands (DAGs) in nymphs, and the metathoracic gland (MTG) in adults of this species were characterized and quantified. Compounds found were similar to those of other Pentatomidae species and included aliphatic hydrocarbons, aldehydes, oxo-alkenals, and esters. However, two compounds were found in the MTG that have not been described previously for this family. Mass spectrometry, infrared spectroscopy, and gas chromatographic analysis using coinjection with authentic standards confirmed the identities of the compounds as enantiopure (S)-2-methylbutyl acetate and 3-methyl-2-butenyl acetate. The five nymphal instars showed significantly different ratios of components, mainly between those of the first and later instars. No significant differences were detected in MTG contents between sexes. Gas chromatography (GC) analysis of aeration extracts of males and females showed the presence of a compound released exclusively by males. Gas chromatography – electroantennographic detection (GC-EAD) assays indicated that the male-specific compound is bioactive in females, suggesting the presence of an attractant pheromone. The mass spectrum and infrared data for this compound matched with methyl 2,6,10-trimethyltridecanoate, a sex pheromone component previously detected in the stink bugs, Euschistus heros and E. obscurus. The synthetic standard coeluted with the natural pheromone on two different GC stationary phases, confirming the proposed structure. Y-tube olfactometer assays showed that the synthetic standard was strongly attractive to females, and GC-EAD tests produced responses with antennae from females similar to those of the natural pheromone.     

Male-Produced Sex Pheromone of the Stink Bug Edessa meditabunda

Edessa meditabunda is a secondary pest within the piercing-sucking stink bug complex that attacks soybean crops in Brazil. The behavioral responses of males and females to aeration extracts from conspecifics suggested the presence of a male-produced sex pheromone. Gas chromatographic (GC) analysis of male and female aeration extracts revealed the presence of two male-specific compounds in a ratio of 92:8. Gas chromatographic -electroantennographic detection (GC-EAD) assays indicated that the major component is bioactive for females, supporting the behavioral data. Analysis of the mass and infrared spectra of the male-specific compounds suggested that they were both methyl-branched long-chain methyl esters. On the basis of the mass spectra of the respective hydrocarbons obtained by micro derivatizations, the structures of these methyl esters were proposed to be methyl 4,8,12-trimethylpentadecanoate (major) and methyl 4,8,12-trimethyltetradecanoate (minor). An 11 step synthetic route that was based on a sequence of Grignard reactions, starting from cyclopropyl methyl ketone, was developed to obtain synthetic standards with a 7.9 % overall yield for the major compound and a 9.9 % yield for the minor. The synthetic standards co-eluted with the natural pheromones on three different GC stationary phases. Y-tube olfactometer assays showed that the synthetic standards, including the major compound alone and a mixture of the major and minor compounds in the proportion found in natural extracts, were strongly attractive to females.     

Effects of RNAi-Mediated Knockdown of Histone Methyltransferases on the Sex-Specific mRNA Expression of Imp in the Silkworm Bombyx mori

Sexual differentiation in Bombyx mori is controlled by sex-specific splicing of Bmdsx, which results in the omission of exons 3 and 4 in a male-specific manner. In B. mori, insulin-like growth factor II mRNA-binding protein (Imp) is a male-specific factor involved in male-specific splicing of Bmdsx. Male-specific Imp mRNA results from the male-specific inclusion of exon 8. To verify the link between histone methylation and alternative RNA processing in Imp, we examined the effects of RNAi-mediated knockdown of several histone methyltransferases on the sex-specific mRNA expression of Imp. As a result, male-specific expression of Imp mRNA was completely abolished when expression of the H3K79 methyltransferase DOT1L was repressed to <10% of that in control males. Chromatin immunoprecipitation-quantitative PCR analysis revealed a higher distribution of H3K79me2 in normal males than in normal females across Imp. RNA polymerase II (RNAP II) processivity assays indicated that RNAi knockdown of DOT1L in males caused a twofold decrease in RNAP II processivity compared to that in control males, with almost equivalent levels to those observed in normal females. Inhibition of RNAP II-mediated elongation in male cells repressed the male-specific splicing of Imp. Our data suggest the possibility that H3K79me2 accumulation along Imp is associated with the male-specific alternative processing of Imp mRNA that results from increased RNAP II processivity.     

Identification of male-specific chiral compound from the sugarcane weevil Sphenophorus levis

Comparative gas chromatographic analyses of airborne volatiles produced by males and females of the sugarcane weevil Sphenophorus levis, showed one male-specific compound. Gas chromatography–mass spectrometry data indicated an aliphatic alcohol that was identified as 2-methyl-4-octanol. Both optical isomers were synthesized in five steps by employing commercially available (R)- and (S) -2,2-dimethyl-1,3-dioxolane-4-methanol as starting material. Enantiomeric resolution by gas chromatography with a chiral column demonstrated that the natural alcohol possessed the S configuration. Preliminary indoor observations suggested that the alcohol elicited aggregation behavior among adults. The same compound has been previously described as an aggregation pheromone in several other curculionid species.     

Multiple Roles of a Male-Specific Compound in the Sexual Behavior of the Dried Bean Beetle, <Emphasis Type="Italic">Acanthoscelides Obtectus</Emphasis>

Males of Acanthoscelides obtectus (Coleoptera: Chrysomelidae, Bruchinae) emit methyl (E,R)-2,4,5-tetradecatrienoate that attracts females for mating. This study identified further roles for this compound in the sexual behavior of A. obtectus. Earlier observations revealed that males touched females with their antennae while tandem-running with them and initiated mounting and copulation, whereas they showed no such behavior toward other males. A series of subsequent laboratory choice tests were set up to establish if certain cuticular compounds aid contact sex recognition in A. obtectus. Males chose virgin females over other males. The activity toward females could be eliminated by rinsing with hexane, but was regained by application of female extract onto previously rinsed females. Gas chromatographic (GC) comparison of hexane extracts revealed the presence of two male-specific compounds, methyl (E,R)-2,4,5-tetradecatrienoate and octadecanal, which were absent from the behaviorally active female samples. Of the two compounds, methyl (E,R)-2,4,5-tetradecatrienoate was found to be responsible for the inhibition of male sexual behavior, similar to that observed with crude male extracts applied to virgin females. Furthermore, males preferred virgin over mated females. GC analyses revealed the presence of methyl (E,R)-2,4,5-tetradecatrienoate in mated females in amounts sufficient to curtail mating attempts. It appears that methyl (E,R)-2,4,5-tetradecatrienoate, besides being a male-produced sex pheromone, acts as a male-recognition signal in A. obtectus. Males also transfer it onto females during mating, resulting in mated females being avoided by courting males.     

Identification of a Sex Pheromone From Male Yellow MealWorm Beetles, <Emphasis Type="Italic">Tenebrio molitor</Emphasis>

The sex pheromone released by the adult female Tenebrio molitor, 4-methyl-1-nonanol, is well known. In addition, there is evidence that adult males release a pheromone that attracts females. The purpose of the present study was to isolate and identify male-released pheromone(s). Emissions from virgin adult males and females were collected on filter paper and extracted with pentane. Extracts were analyzed by gas chromatography-mass spectrometry. One male-specific compound was detected and identified as (Z)-3-dodecenyl acetate (Z3-12:Ac). In arena bioassays, E3-12:Ac was attractive to females only, at 1 and 10 μg doses. E3-12:Ac was also attractive to females at a 10-μg dose. The presence of both male and female pheromones, each attracting the opposite sex, may contribute to maintaining a high-density population of both sexes.     

Multifunctional communication inRiptortus clavatus (Heteroptera: Alydidae): Conspecific nymphs and egg parasitoidOoencyrtus nezarae use the same adult attractant pheromone as chemical cue

The bean bug,Riptortus clavatus lays scattered eggs (as opposed to the egg masses of pentatomids) on host as well as nonhost plants. Therefore, the first feeding stage (second-instar) nymphs emerging from eggs laid on nonhost plants need a signal that enables them to locate a food source at the lowest energy cost. Male-released (E)-2-hexenyl (E)-2-hexenoate, (E)-2-hexenyl (Z)-3-hexenoate, and myristyl isobutyrate play the double role of attractant pheromone for adults as well as aggregation pheromone, which enables the second-instar nymphs to find the host food plant. These male-specific semiochemicals are released only when foodstuff is available. On the other hand, females ofOoencyrtus nezarae, the most effective parasitoid of the host in Kumamoto, Japan (where the field experiments were conducted), utilize these semiochemicals as kairomones in order to locate the potential host community. Field experiments revealed that the synthetic pheromone rivaled 10 live males in the attraction of adults and second-instar nymphs. Captures of the egg parasitoidO. nezarae females in cylindrical sticky traps were significantly higher in traps baited with the synthetic semiochemicals than in control traps. The number of females captured was significantly higher than the number of males, although the captures in the sticky suction trap system revealed that the populations of male and female were not significantly different.     

Male Phyllotreta striolata (F.) Produce an Aggregation Pheromone: Identification of Male-specific compounds and Interaction with Host Plant Volatiles

The chrysomelid beetle Phyllotreta striolata is an important pest of Brassicaceae in Southeast Asia and North America. Here, we identified the aggregation pheromone of a population of P. striolata from Taiwan, and host plant volatiles that interact with the pheromone. Volatiles emitted by feeding male P. striolata attracted males and females in the field. Headspace volatile analyses revealed that six sesquiterpenes were emitted specifically by feeding males. Only one of these, however, elicited an electrophysiological response from antennae of both sexes. A number of host plant volatiles, e.g., 1-hexanol, (Z)-3-hexen-1-ol, and the glucosinolate hydrolysis products allyl isothiocyanate (AITC), 3-butenyl isothiocyanate, and 4-pentenyl isothiocyanate also elicited clear responses from the antenna. The active male-specific compound was identified as (+)-(6R,7S)-himachala-9,11-diene by chiral stationary phase gas-chromatography with coupled mass spectrometry, and by comparison with reference samples from Abies nordmanniana, which is known to produce the corresponding enantiomer. The pheromone compound was synthesized starting from (-)-α-himachalene isolated from Cedrus atlantica. Under field conditions, the activity of the synthetic pheromone required concomitant presence of the host plant volatile allyl isothiocyanate. However, both synthetic (+)-(6R,7S)-himachala-9,11-diene alone and in combination with AITC were attractive in a two-choice laboratory assay devoid of other natural olfactory stimuli. We hypothesize that P. striolata adults respond to the pheromone only if specific host volatiles are present. In the same laboratory set up, more beetles were attracted by feeding males than by the synthetic stimuli. Thus, further research will be necessary to reveal the components of a more complex blend of host or male-produced semiochemicals that might enhance trap attractiveness in the field.     

Circadian Release of Male-Specific Components of the Greater Date Moth,Aphomia (Arenipses) Sabella,Using Sequential SPME/GC/MS Analysis

The greater date moth (GDM), Aphomia sabella Hampson (Lepidoptera: Pyralidae: Galleriinae), is a serious pest of date palms, Phoenix dactylifera. The release of volatiles from both males and females was investigated using sequential SPME/GC/MS analysis. Males release a complex mixture of compounds in a circadian rhythm during the night between 03:00 and 05:00 hr. Six compounds were identified: benzaldehyde, sulcatol (6-methyl-5-hepten-2-ol), geranyl acetone [(E)-6,10-dimethyl-5,9-undecadien-2-one], phenylacetaldehyde, 2-phenylpropenal, and (R)-fuscumol [(R)-(E)-6, 10-dimethyl-5, 9-undecadien-2-ol]. Benzaldehyde, sulcatol, and geranyl acetone were found only in trace amounts. These compounds were in glands located in the forewing of males only. Small amounts of acetoin and 2,3-butanediol were found sporadically in the SPME/GC/MS analyses of volatiles from females, and these compounds probably originate from microorganisms. This is the first finding of a circadian release of male-specific compounds in moths. GC/EAD analyses with synthetic standards of compounds released by males showed that the female antenna is stimulated by all six compounds, while the male antenna responded only to phenylacetaldehyde. A possible pheromonal role for the male-specific compounds is suggested by the circadian rhythm of their release and the EAD response of females to them. However, trapping tests with the main male-specific compounds in screen cages in the laboratory or in the field did not reveal any significant behavioral responses from females or males. Copulation in the laboratory was observed only in the presence of date palm tissue, thus suggesting that sexual communication and mating of GDM moths probably occurs in the crown of date palms.     

A Male-Predominant Cuticular Hydrocarbon, 7-Methyltricosane, is used as a Contact Pheromone in the Western Flower Thrips Frankliniella occidentalis

In a laboratory bioassay, adult female Frankliniella occidentalis (Pergande) (Thysanoptera: Thripidae) spent more time near filter paper disks that had been exposed to adult males than near unexposed disks; this effect was not observed on disks exposed to adult females. The response could only partly be explained by the known male-produced aggregation pheromone, neryl (S)-2-methylbutanoate, suggesting the presence of an unknown male-produced compound. In gas chromatography/mass spectrometry analyses, 7-methyltricosane was detected on disks exposed to males, but not on disks exposed to females. Extracts of cuticular lipids also showed relatively large amounts of 7-methyltricosane on males, whereas only trace amounts were found on females and none on larvae. Bioassays of synthetic 7-methyltricosane showed that adults responded only after contact. The response to this compound was clearly different from that to n-tricosane or hexane-only controls. Females that contacted 7-methyltricosane on glass beads stayed in the vicinity and frequently raised the abdomen, a behavior that rejects mating attempts by males. Males stayed in the vicinity and wagged the abdomen sideways, a behavior used in fighting between males. This is the first identification of a contact pheromone in the order Thysanoptera.     

Effect of mating on terminating aggregation during host colonization in the bark beetle,Ips paraconfusus

Attraction of maleIps paraconfusus to male-infested ponderosa pine logs was inhibited by volatiles from logs infested with mated males and females over an 8-day period in the field. The response of females during this time was not significantly inhibited by these volatiles. Synthesis of the male-specific pheromones, ipsenol and ipsdienol, appeared negligible after 8 days in males allowed to mate with 3 females in these logs while males alone contained levels of these pheromones at about half their maximum rate. The decline in pheromone production in mated males appears to result, at least in part, from a reduction in the activity of the biosynthetic system that converts the host monoterpene, myrcene, to ipsenol and ipsdienol. Mating and feeding have apparently no effect on the biosynthetic system that converts the host monoterpene, (–)--pinene, to the pheromone,cis-verbenol in either males or females. The reduced production and release of pheromones by males after mating appears to play a major function in the process of terminating the aggregation phase of host colonization.     

Sex Attractant Pheromone from the Neotropical Red-Shouldered Stink Bug, <Emphasis Type="Italic">Thyanta perditor</Emphasis> (F.)

Olfactometer bioassays showed that odors from mature Thyanta perditor males attracted females but not males. Furthermore, odors from females did not attract either sex, indicating that like other phytophagous pentatomid bugs, the males produce a sex pheromone. Attraction appeared to peak in late afternoon to evening. The headspace volatiles collected from male and female T. perditor were analyzed by GC-MS and HPLC. A male-specific compound, methyl (2E,4Z,6Z)-decatrienoate (2E,4Z,6Z-10:COOMe), was identified along with a number of other compounds found in extracts from both sexes. Bioassays carried out with 2E,4Z,6Z-10:COOMe showed it was as attractive to females as the crude extract of male volatiles, suggesting that the male-produced sex pheromone consists of 2E,4Z,6Z-10:COOMe as a single component. Consecutive volatiles collections from males showed that 2E,4Z,6Z-10:COOMe began appearing in extracts from males about 9 d after the final molt, as the males became sexually mature.     

Identification,Synthesis, and Behavioral Activity of 5,11-Dimethylpentacosane,A Novel Sex Pheromone Component of the Greater Wax Moth,Galleria Mellonella (L.)

The greater wax moth, Galleria mellonella (L.), is a serious and widespread pest of the honeybee, Apis mellifera L. In contrast to most moths, for which long-range mate finding is mediated by female-produced sex pheromones, G. mellonella males attract conspecific females over long distances by emitting large amounts of a characteristic scent in combination with bursts of ultrasonic calls. The male scent for this species was previously identified as a blend of nonanal and undecanal. When these compounds were bioassayed, characteristic short-range sexual behavior, including wing fanning, was triggered in conspecific females, but the aldehyde blend failed to elicit attraction over longer distances. We identified, via analysis and synthesis, a third male-specific compound, 5,11-dimethylpentacosane. We show that it acts as a behavioral synergist to the aldehydes. In wind tunnel experiments, very few female moths responded to the aldehyde blend or to 5,11-dimethylpentacosane tested separately, but consistently showed orientation and source contact when a combination of all three compounds was applied. The level of attraction to the three-component mixture was still lower than that to male extract, indicating that the composition of compounds in the synthetic blend is suboptimal, or that additional pheromone components of G. mellonella are yet to be identified. The identification of 5,11-dimethylpentacosane is an important step for the development of an efficient long-range attractant that will be integrated with other environmentally safe strategies to reduce damage to beehives caused by wax moths.     

Using Generic Pheromone Lures to Expedite Identification of Aggregation Pheromones for the Cerambycid Beetles Xylotrechus nauticus, Phymatodes lecontei, and Neoclytus modestus modestus

Males of several species of longhorned beetles in the subfamily Cerambycinae produce sex or aggregation pheromones consisting of 2,3-hexanediols and/or hydroxyhexanones. We tested the hypothesis that this diol/hydroxyketone pheromone motif is highly conserved within the subfamily, and the resulting prediction that multiple cerambycine species will be attracted to compounds of this type. We also tested the concept that live traps baited with generic blends of these compounds could be used as a source of live insects from which pheromones could be collected and identified. Traps placed in a mature oak woodland and baited with generic blends of racemic 2-hydroxyhexan-3-one and 3-hydroxyhexan-2-one captured adults of both sexes of three cerambycine species: Xylotrechus nauticus (Mannerheim), Phymatodes lecontei Linsley, and Phymatodes decussatus decussatus (LeConte). Odors collected from male X. nauticus contained a 9:1 ratio of two male-specific compounds, (R)- and (S)-3-hydroxyhexan-2-one. Field trials with synthetic compounds determined that traps baited with (R)-3-hydroxyhexan-2-one (94% ee), alone or in blends with other isomers, attracted similar numbers of X. nauticus of both sexes, whereas (S)-3-hydroxyhexan-2-one (94% ee) attracted significantly fewer beetles. Phymatodes lecontei and P. d. decussatus also were caught in traps baited with hydroxyhexanones, as well as a few specimens of two other cerambycine species, Neoclytus modestus modestus Fall (both sexes) and Brothylus gemmulatus LeConte (only females). Male N. m. modestus produced (R)-3-hydroxyhexan-2-one, which was not present in extracts from females. Neoclytus m. modestus of both sexes also responded to lures that included (R)-3-hydroxyhexan-2-one as one of the components. The only male-specific compound found in extracts from P. lecontei was (R)-2-methylbutan-1-ol, and adults of both sexes were attracted to racemic 2-methylbutan-1-ol in field bioassays. Surprisingly, P. lecontei of both sexes also were attracted to (R)- and (S)-3-hydroxyhexan-2-ones, although neither compound was detected in extracts from this species. Males of all five beetle species had gland pores on their prothoraces that were similar in structure to those that have been associated with volatile pheromone production in other cerambycine species. The attraction of multiple cerambycine species of two tribes to (R)-3-hydroxyhexan-2-one in this study, and in earlier studies with other cerambycine species, suggests that this compound is a widespread aggregation pheromone component in this large and diverse subfamily. Overall, the attraction of multiple species from different cerambycine tribes to this compound at a single field site supports the hypothesis that the hydroxyketone pheromone structural motif is highly conserved within this subfamily.     

Male-Produced Aggregation Pheromone of the Cerambycid Beetle Neoclytus acuminatus acuminatus

This is the first fully verified report of an aggregation pheromone produced by a cerambycid beetle species. Field bioassays with adult Neoclytus acuminatus acuminatus (F.) (Coleoptera: Cerambycidae) revealed that males produce a pheromone that attracts both sexes. Extracts of odors from males contained a single major male-specific compound, (2S,3S)-hexanediol. Field trials determined that both sexes were attracted by the racemic blend of (2S,3S)- and (2R,3R)-hexanediols and that activity was similar to enantiomerically enriched (2S,3S)-hexanediol (e.e. 80.2%). However, the blend of all four 2,3-hexanediol stereoisomers attracted few beetles, indicating inhibition by one or both of the (2R*,3S*)-stereoisomers. Females of the cerambycid Curius dentatus Newman were attracted to traps baited with the four component blend, suggesting that a male-produced sex pheromone for this species may contain (2R,3S)-hexanediol and/or (2S,3R)-hexanediol. The pheromone of N. a. acuminatus, and presumed pheromone of C. dentatus, bear structural similarities to those produced by males of six other species in the Cerambycinae (straight chains of 6, 8, or 10 carbons with hydroxyl or carbonyl groups at C2 and C3). It is likely that males of other species in this large subfamily produce pheromones that are variations on this structural motif.     

The Aggregation Pheromone of <Emphasis Type="Italic">Phyllotreta striolata</Emphasis> (Coleoptera: Chrysomelidae) Revisited

Aggregations of the striped flea beetle Phyllotreta striolata on their crucifer host plants are mediated by volatiles emitted from feeding males. The male-specific sesquiterpene, (6R,7S)-himachala-9,11-diene (compound A), was shown previously to be physiologically and behaviorally active, but compound A was attractive only when combined with unnaturally high doses of the host plant volatile allyl isothiocyanate (AITC) in field trapping experiments. This indicated that our understanding of the chemical communication in this species is incomplete. Another male-specific sesquiterpenoid, (3S,9R,9aS)-3-hydroxy-3,5,5,9-tetramethyl-5,6,7,8,9,9a-hexahydro-1H-benzo[7]annulen-2(3H)-one (compound G), has been reported from an American P. striolata population. We confirmed the presence of compound G, and investigated its interaction with compound A and AITC in a P. striolata population in Taiwan. Compound G was attractive to Taiwanese P. striolata in laboratory bioassays, but significantly more beetles were attracted to a blend of compounds A and G. Under the same conditions, P. striolata showed no preference for the blend of A and G combined with a range of doses of AITC over the sesquiterpenoid blend alone. The sesquiterpenoid blend was tested further in field trapping experiments and attracted significantly more beetles than traps baited with compound A and ecologically relevant amounts of AITC. We conclude that A and G are components of the male-specific aggregation pheromone of P. striolata in Taiwan, and that the attractiveness of the pheromone is not reliant on the presence of AITC. Our results further indicate that the male-specific sesquiterpenoid blends differ qualitatively between the Taiwanese and American populations of P. striolata.     

Identification of female sex pheromone in alfalfa blotch leafminer,Agromyza frontella (Rondani)(Diptera: Agromyzidae)

Gas chroinatography of hexane extracts of 3-day-old virgin A.frontella adults revealed a branched saturated hydrocarbon present only in females. This compound was isolated by argentation chromotography and preparative GC, identified by GC and GC-MS as 3,7-dimethylnonadecane, and subsequently synthesized. Three-day-old virgin females contained 54.1 ± 3.5 ng (± SEM;N = 48) of 3,7-dimethylnonadecane. Male cadavers do not elicit male courtship behavior, but when treated with 18 ng of 3,7-dimethylnonadecane they were as attractive as cadavers of 3-day-old virgin females. Black cotton knots were less attractive than male cadavers when treated with 3,7-dimethylnonadecane, but all bioassays indicated that this compound is an important semiochemical modulating male mating behavior.     

Behavioral and electrophysiological evidence for volatile sex pheromones in Parcoblatta wood cockroaches

Species within the cockroach genus Parcoblatta are sexually dimorphic for wing length; females have reduced wings and are flightless, while males have long wings that are used in flight. We predicted that Parcoblatta females would release a volatile sex pheromone to attract the more mobile males. Nymphs of the broad wood cockroach, P. lata, and the Caudell's wood cockroach, P. caudelli, were collected in forested areas in North Carolina, USA, and reared in the laboratory for observations of sexual behavior and for pheromone analysis. After several days of sexual maturation, virgin females of both species exhibited distinct calling behaviors. In females of P. lata, calling commenced 6 days after adult emergence. Under a light–dark photoperiod regime, calling behavior in both species was restricted to the scotophase. Calling consisted of a repeated pattern of raising and lowering the abdomen with occasional exposure of the genital vestibulum. To test whether calling behavior is associated with the release of pheromone, volatiles from calling and noncalling females were collected on Super-Q and tested by electroantennogram (EAG) and behavioral assays. Volatile collections from calling females elicited higher male-specific EAG responses than collections from noncalling females of the same physiological stage. In an olfactometer choice test (Y-tube), males preferred volatiles from calling females over those from noncalling females. To determine the anatomical source of the pheromone, solvent extracts of various body parts were analyzed by EAG. The first through seventh tergites were the only body parts that elicited male-specific EAG responses in both species. In P. lata, the activity of the extract increased from 1- to 7-day-old females, but was lower in mated than in virgin females of the same age. The putative pheromone gland appears to consist of numerous class-3 secretory units, each composed of a secretory cell connected to a cuticular pore via a tubular duct. We conclude that female P. lata and P. caudelli produce sex-specific volatile pheromones that are emitted during calling behavior.     

Sex Attractant Pheromone from the Rice Stalk Stink Bug, Tibraca limbativentris Stal

The male-produced sex pheromone from the Brazilian rice stalk stink bug Tibraca limbativentris is reported. Olfactometer bioassays with sexually mature males and females showed that males attracted females, which suggests that males release a sex pheromone. Males were not attracted to either sex, nor were females attractive to conspecific females. Attraction of the females to males was highest at night. The headspace volatiles collected from male and female bugs were analyzed by gas chromatography (GC) and GC–mass spectrometry. Two male-specific compounds were identified as isomers of 1′S-zingiberenol, whereas a series of defensive compounds were identified in extracts from both sexes. Zingiberenol has three chiral centers, and the nonselective syntheses used produced two groups of isomers, zingiberenol I containing four isomers, namely (1RS,4RS,1′R)-4-(1′,5′-dimethylhex-4′-enyl)-1-methylcyclohex-2-en-1-ol, and zingiberenol II containing the other four isomers, namely (1RS,4RS,1′S)-4-(1′,5′-dimethylhex-4′-enyl)-1-methylcyclohex-2-en-1-ol. Both groups of stereoisomers were more attractive than hexane controls. The absolute configuration of the insect-produced pheromonal components remains to be elucidated, but the 1′S stereochemistry was established for at least one of the isomers.