Male-Released Sex Pheromone of the Stink Bug Piezodorus hybneri

Male-released semiochemicals of the stink bug Piezodorus hybneri (Heteroptera: Pentatomidae) elicit attraction of male and female bugs and homosexual behavior in males. Three active components were isolated from the airborne volatiles of males by flash chromatography, with the activity monitored by GC-EAD and behavioral bioassay. The pheromone system was characterized as a mixture of -sesquiphellandrene, (R)-15-hexadecanolide, and methyl 8-(Z)-hexadecenoate (ratio: 10:4:1), and the activity of the semiochemicals was assessed with authentic samples. Enantiomerically pure samples of the R and S macrolactones were obtained by Yamaguchi's and Mitsunobu's macrolactonization of a key intermediate, (R)-15-hydroxyhexadecanoic acid. The nonnatural S stereoisomer was neither a beneficial nor a behavioral antagonist. Individual constituents or binary mixtures were active, but the optimal male response was elicited only by the full mixture. Behavioral observation and the fact that the onset of pheromone production is coincident with ovarian development strongly suggest that these semiochemicals are, in fact, sex pheromones.     

Identification of male cabbage looper sex pheromone attractive to females

A multicomponent pheromone produced by male cabbage looper moths that is attractive to female moths in a flight tunnel bioassay was isolated and identified. Based on analyses of hairpencil extracts of male cabbage loopers and volatiles emitted by males, the pheromone has been identified as a blend consisting of (S)-(+)-linalool,p-cresol, andm-cresol. The chirality of the major component, (S)-(+)-linalool, is important for behavioral response of females. These pheromonal compounds were also identified as volatiles released by males when males were exposed to the principal pheromone component of female cabbage loopers, (Z)-7-dodecen-1-ol acetate. The amount of male pheromone released was increased significantly when males were exposed to a combination of (Z)-7-dodecen-1-ol acetate and the odor from cabbage. Neither linalool nor the cresols were detected in volatiles from cabbage or from males exposed to cabbage odor.     

Enhancement of Attraction to Sex Pheromones of Spodoptera exigua by Volatile Compounds Produced by Host Plants

We measured the effects of exposure to volatile compounds produced by host plants on the rate of capture of male Spodoptera exigua using synthetic sex pheromones. Exposure to volatile compounds stimulated strong electroantennographic responses of male S. exigua. The behavioral responses of male moths to combinations of sex pheromone and volatile compounds were tested in wind tunnel experiments. When lures were baited with synthetic sex pheromone plus benzaldehyde, phenylacetaldehyde, (Z)-3-hexenyl acetate, or linalool, respectively, the landing rate of S. exigua males was increased by 101.4%, 79.6%, 60.6%, and 34.3%, respectively, compared to sex pheromone alone. In field tests, traps baited with either pheromone + (E)-2-hexenal, pheromone + phenylacetaldehyde, pheromone + (Z)-3-hexenyl acetate, or pheromone + (Z)-3-hexenol enhanced moth catches by 38.8%, 34.6%, 24.6%, and 20.8%, respectively compared to traps baited with pheromone alone. In a second field experiment, more S. exigua males were trapped with a combination of a synthetic sex pheromone blend and several individual host plant volatiles compared to synthetic sex pheromone alone. These results suggest that some host plant volatiles enhance the orientation response of S. exigua male moths to sex pheromone sources.     

Interactions Between Host-plant Volatiles and the Sex Pheromones of the Bird Cherry-oat Aphid, <Emphasis Type="Italic">Rhopalosiphum padi</Emphasis> and the Damson-hop Aphid, <Emphasis Type="Italic">Phorodon humuli</Emphasis>

The bird cherry-oat aphid, Rhopalosiphum padi (L.), and the damson-hop aphid, Phorodon humuli (Schrank), migrate at the same time of year and colonize closely related Prunus spp. as primary hosts, but utilize (1R,4aS,7S,7aR)-nepetalactol and (1RS,4aR,7S,7aS)-nepetalactol, respectively, as sex pheromones. Interactions between these sex pheromones and benzaldehyde and methyl salicylate, plant volatiles common to primary hosts of both species, were investigated to assess whether they confer reproductive isolation between these species. Female autumn migrants (gynoparae) and males of these two species were caught in the field with water traps baited with their respective sex pheromones. Rhopalosiphum padi gynoparae and males also responded positively to benzaldehyde. Release of either benzaldehyde or methyl salicylate with the conspecific sex pheromone increased catches of both species of aphid. However, releasing both plant volatiles with the sex pheromone of R. padi increased catches of gynoparae and males, but reduced those with the sex pheromone of P. humuli. These results support the hypothesis that specific plant volatiles synergize responses of autumn migrating aphids to their sex pheromone. Because these interactions are species-specific, they may be important in allowing males to discriminate between conspecific sexual females (oviparae) and those of other aphid species.     

Ancestral semiochemical attraction persists for adjoining populations of siblingIps bark beetles (Coleoptera: Scolytidae)

Three species ofIps pine bark beetles in Hopping's group IX (S.L. Wood'sgrandicollis group),Ips confusus, I. lecontei, andI. paraconfusus, are parapatrically distributed in the American Southwest. They share post-Pleistocene altitudinal ecotones with their host pines. Adjacent to these areas of host overlap, we tested the hypothesis that aggregation pheromones produced by male beetles and/or host volatiles are sufficient to elicit the speciesspecific colonization behaviors typical of these threeIps species in nature. A more distantly related species,I. pini (Hopping's group IV, S.L. Wood'spini group) was used for outgroup comparison. Under the influence of pheromone, males ofI. confusus andI. paraconfusus do not discern among unin-fested log bolts of host and nonhost pine prior to bark contact. Males responding to pheromones emanating from infested bolts are similarly undiscriminating. Females ofI. confusus andI. lecontei olfactorily discern the combination of conspecific males in host pine from other possible beetlepine combinations; females ofI. paraconfusus do not. FemaleI. pini discerned conspecific pheromone from that ofI. lecontei. The bark beetle predator,Enoclerus lecontei, is attracted by, but does not discriminate among, the male-produced volatiles of these Group IXIps species. These results support a hypothesis that divergence in pheromonal responses by these group IXIps species has evolved following their speciation, having been manifested first in the female sex. Evolutionarily, the derived pheromonal messages have preceded their behavioral discrimination by these beetles. Additional speciesspecific cues may operate between the sexes in the field that may preclude heterospecific pairings.     

Attraction of male beetles to grubs: Evidence for evolution of a sex pheromone from larval odor

Females of the scarabaeid beetleCyclocephala lurida produce a volatile sex pheromone which attracts conspecific males. Field experiments demonstrated that larvae of both sexes also emit volatile chemicals that stimulate similar responses in adult males, including attempts by the attracted males to mate with the nonreproductive immature stage. Significantly more adult males were caught in traps baited with conspecific male or female larvae or adult females than in blank control traps. Hexane extracts of both male and female grubs were at least as effective as live larvae in trapping male adults, demonstrating that the behavioral responses are mediated by volatile chemicals. Sensory and behavioral responses of males to sex pheromones emitted by adult females are part of the functional communication system. However, their response to grubs is not functional, because grubs are normally temporally and spatially inaccessible to mate-seeking males. In theory, the evolution of a communication system is problematic because it requires the development of a signal in one sex and the sensory and behavioral attributes to respond to that signal in the other sex. The ontogeny of sex pheromone communication inC. lurida suggests a partial solution to this evolutionary problem. We propose that this sex pheromone communication system is probably derived from noncommunicative volatile chemicals that are lost in adult males and retained by adult females.     

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.     

Primer and Short-Range Releaser Pheromone Properties of Premolt Female Urine from the Shore Crab <Emphasis Type="Italic">Carcinus maenas</Emphasis>

The European shore crab Carcinus maenas is considered to rely on a female pheromone when mating. Evidence, however, is scarce on how the urine pheromone in itself affects males. We investigated male primer and releaser responses to female pheromones with methods that minimized effects from females, delivering female urine either as a pump-generated plume or deposited on a polyurethane sponge. We delivered the pheromone at different concentrations in far, near, and close/contact range to get a picture of how distance affects behavioral response. Our results show that substances in premolt female urine (PMU) function as primer and potent short-range releaser pheromones. Based on the olfactometer and sponge tests, we conclude that PMU stimulus in itself is sufficient to elicit increased search and mating-specific behaviors such as posing, posing search, cradle carrying, and stroking. Pheromone concentrations do not seem to be important for attenuating search and posing as long as the level is above a certain threshold concentration. Instead, pheromone levels seem to play a role in male acceptance of females, recruiting more males to respond, and generating better responses with increasing concentration.     

Isolation of a Pyrazine Alarm Pheromone Component from the Fire Ant, <Emphasis Type="Italic">Solenopsis invicta</Emphasis>

Alarm pheromones in social insects are an essential part of a complex of pheromone interactions that contribute to the maintenance of colony integrity and sociality. The alarm pheromones of ants were among the first examples of animal pheromones identified, primarily because of the large amount of chemical produced and the distinctive responses of ants to the pheromone. However, the alarm pheromone of the fire ant, Solenopsis invicta, eluded identification for over four decades. We identified 2-ethyl-3,6-dimethylpyrazine as an alarm pheromone component of S. invicta. Worker fire ants detect the pyrazine alarm pheromone at 30 pg/ml, which is comparable to alarm pheromone sensitivities reported for other ant species. The source of this alarm pheromone are the mandibular glands, which, in fire ants, are not well developed and contain only about 300 pg of the compound, much less than the microgram quantities of alarm pheromones reported for several other ant species. Female and male sexuals and workers produce the pyrazine, which suggests that it may be involved in fire ant mating flight initiation, as well as the typical worker alarm response. This is the first report of 2-ethyl-3,6-dimethylpyrazine from a Solenopsis species and the first example of this alkaloid functioning as an alarm pheromone.     

Behavioral responses to food volatiles by two species of stored-product coleoptera,Sitophilus oryzae (curculionidae) andTribolium castaneum (tenebrionidae)

Laboratory experiments were conducted to study the behavioral activity of grain-derived volatiles as attractants and pheromone synergists forSitophilus oryzae, an internal-feeding pest of sound grain, andTribolium castaneum, an external-feeding pest of damaged grains and flour. Behavioral studies with two-choice pitfall bioassays determined that the fresh grain volatiles valeraldehyde, maltol, and vanillin were attractive toS. oryzae at various doses, butT. castaneum were not attracted to any dose of any of these three compounds. When oils from pressed grains were bioassayed, sesame oil was significantly repellent and oat and wheat germ oils were attractive toS. oryzae. However, rice, soybean, oat, wheat germ, and corn oils were all attractive toT. castaneum. A commercial food product composed primarily of soybean oil and wheat germ was highly attractive toT. castaneum, but elicited no response fromS. oryzae. A combination of the three grain volatiles valeraldehyde, maltol, and vanillin with the synthetic pheromone sitophinone was more attractive toS. oryzae than either the pheromone alone or the tripartite grain volatile mix. Similarly, a combination of the commercial food product with the pheromone 4,8-dimethyldecanal was more attractive toT. castaneum than either food alone or pheromone alone. Behavioral responses to grain volatiles may reflect the ecological niche of the granivore:S. oryzae colonizes sound grain and is attracted to volatiles characteristic of fresh grain, whileT. castaneum utilizes damaged or deteriorated grains and responds best to oils characteristic of damaged or fungus-infested grain. Synergism of food odors and pheromones suggests that more effective traps can be devised for management of these pest insects.     

Behaviorally Active Green Leaf Volatiles for Monitoring the Leaf Beetle, Diorhabda elongata, a Biocontrol Agent of Saltcedar, Tamarix spp.

Biological activity and chemistry of host plant volatiles were investigated for Diorhabda elongata, Brullé (Coleoptera: Chrysomelidae), a biological control agent for the invasive tree, saltcedar (Tamarix spp., Tamaricaceae). Gas chromatographic–electroantennographic detection (GC-EAD) analysis of volatiles collected from adult D. elongata feeding on saltcedar foliage or from saltcedar foliage alone showed 15 antennally active compounds. These compounds were more abundant in collections from beetle-infested foliage. Antennally active compounds were identified by GC–mass spectrometry (MS) and confirmed with authentic standards. The emissions of the most abundant GC-EAD-active compounds, green leaf volatiles (GLV), were quantitated by GC-MS. A blend of four GLV compounds, mimicking the natural blend ratio, was highly attractive to male and female D. elongata in the field, and a combination of GLV and male-produced aggregation pheromone attracted significantly greater numbers of D. elongata than did either bait alone. A preliminary experiment with a blend of seven additional GC-EAD-active saltcedar volatiles did not show any behavioral activity. The combination of the pheromone and the green leaf odor blend could be a useful attractant in detecting the presence of the biocontrol agent, D. elongata, in stands of saltcedar newly colonized by the beetle.Mention of trade names or commercial products in this article is solely for the purpose of providing specific information and does not imply recommendation or endorsement by the U.S. Department of Agriculture.     

ROLE OF HOST PLANT VOLATILES IN MATE LOCATION FOR THREE SPECIES OF LONGHORNED BEETLES

Stressed woody plants represent an ephemeral and unpredictable resource for larvae of some species of longhorned beetles (Coleoptera: Cerambycidae) because prime subcortical tissues are rapidly degraded by a guild of xylophagous competitors. Selection favors efficient mechanisms of host and mate location to expedite colonization of hosts by larvae. Based on previous research, we hypothesize that mate location in some species of the subfamily Cerambycinae involves three sequential behavioral stages: (1) both sexes are attracted to larval hosts by plant volatiles; (2) males attract females over shorter distances with pheromones; and (3) males recognize females by contact pheromones in their epicuticular wax layer. We already have evidence of second-stage and third-stage behaviors in three species in this subfamily whose xylophagous larvae feed in hardwood trees: Xylotrechus colonus, Megacyllene caryae, and Neoclytus mucronatus mucronatus. In this report, we evaluate the first behavioral stage of mate location behavior (i.e., independent response of both sexes to host plant volatiles) for the same three species. Supporting our hypothesis, both males and females responded to volatiles emanating from hickory logs in Y-tube olfactometer bioassays.     

2-Methylthiazolidine and 4-ethylguaiacol,male sex pheromone components of the cockroachNauphoeta cinerea (dictyoptera,blaberidae): A reinvestigation

InNauphoeta cinerea, male calling behavior is associated with sex pheromone release by the sternal glands. The male pheromone that attracts females from a distance is a mixture of 2-methylthiazolidine and 4-ethylguaiacol. It is active at very low concentrations, 0.05 and 0.01 ng, respectively. Two other compounds, 3-hydroxy-2-butanone and 2-methyl-2thiazoline, act at close range, keeping the female in the vicinity of the male. The function of the volatile pheromone and those of previously described contact pheromones are discussed in regard to their possible involvement in the establishment of male dominant-subordinate relationships.     

Conspecific scent trailing by garter snakes (Thamnophis sirtalis) during autumn Further evidence for use of pheromones in den location

Adult garter snakes (Thamnophis sirtalis), collected in October near a traditional, communal hibernaculum in central Wisconsin, were acclimated to autumnal conditions and subjected to laboratory tests to determine whether they could follow scent trails of a conspecific. Graded responses were obtained, but 75% of the sample showed at least some inclination to follow scent trails. The results suggest that pheromone cues may be used by male and female garter snakes to locate traditional dens during autumnal migrations. Evidence from this and other studies suggests that pheromones are probably used in conjunction with other homing mechanisms and that the role of pheromones in den location may be more important in younger snakes and in populations inhabiting northern latitudes.     

Evidence for mediation of two releaser pheromones in the aggregation behavior of the gregarious desert locust,Schistocerca gregaria (forskal) (Orthoptera: Acrididae)

The response of nymphal and adult gregarious phase desert locust,Schistocerca gregaria, to a choice of two columns of air, one permeated with airborne volatiles emanating from nymphs or adults and the other untreated, was investigated in a single-chamber bioassay arena. The nymphs, whether released individually or in groups, preferred to be within the precinct of the air column treated with airborne volatiles of the nymphs but were indifferent to volatiles of the adults. Conversely, older adults responded only to their own volatiles but not to those of the nymphs or young adults. The young adults were responsive only to volatiles of the older adults. Charcoal-trapped volatiles from the nymphs and the adults reproduced the effect of living locusts. These results indicate that there are two different aggregation pheromones inS. gregaria: a juvenile pheromone produced by nymphs and an adult pheromone specific to adults.     

<Emphasis Type="Italic">cis</Emphasis>-Vaccenyl Acetate,A Female-Produced Sex Pheromone Component of <Emphasis Type="Italic">Ortholeptura valida</Emphasis>, A Longhorned Beetle in the Subfamily Lepturinae

We report the identification, synthesis, and field bioassays of a female-produced sex attractant pheromone component of the cerambycid beetle Ortholeptura valida (LeConte). Headspace volatiles from females contained a female-specific compound, (Z)-11-octadecen-1-yl acetate, which elicited a strong response from antennae of adult males in coupled gas chromatography-electroantennogram analyses. In field bioassays, significant numbers of males were collected by traps baited with this compound. The pheromone represents a new structural class of cerambycid pheromones, and is the first pheromone identified for a cerambycid species in the subfamily Lepturinae.     

Rust-red grain beetle,Cryptolestes ferrugineus,and flat grain beetle,Cryptolestes pusillus: Antennal and behavioral responses to synthetic components of their aggregation pheromones

Antennal and behavioral responses of the rust-red grain beetle,Cryptolestes ferrugineus, and the flat grain beetle,C. pusillus, to synthetic samples of the macrocyclic lactones reported to comprise their aggregation pheromones were investigated. Electroantennogram (EAG) recordings were obtained successfully from both species for the first time. Females of both species showed larger EAGs than males. The EAGs ofC. ferrugineus showed a high degree of specificity for conspecific aggregation pheromone components;C. pusillus showed much less specificity. Behavioral tests were conducted using two-choice pitfall bioassays. Separation of the results into the two effects of activity stimulation and direction finding showed that both effects contributed to the overall response, although sometimes to different extents. The strain ofC. pusillus studied responded equally well to both components of its pheromone, whereas it had been reported previously that only one was active, the other acting as a Synergist and eliciting no response when tested alone. With both species, behavioral response was elicited with a single lactone, suggesting that it might not be necessary to use both components for field use. Particularly surprising was thatC. pusillus showed a greater response to the pheromone components ofC. ferrugineus than to its own. Aeration of the two species and thermal desorption of the collected volatiles confirmed production of the expected lactones, and aeration of authentic lactones showed that the response was not due to the C.ferrugineus pheromone components being markedly more volatile. This response, which seems to be an actual preference, is the first to be discovered among the cucujid beetles and encourages optimism that a practical lure for various species may not need to be as complex as originally feared.     

A Two-Component Female-Produced Pheromone of the Spider Pholcus beijingensis

Chemical signaling plays an important role in spider sexual communication, yet the chemistry of spider sex pheromones remains poorly understood. Unlike insects and mammals, the identification of spider pheromones has seldom been attempted, and no multicomponent pheromones have been found. Empty webs of sexually receptive females of Pholcus beijingensis were more attractive to male conspecifics as compared to webs of sexually unreceptive females or to mature males. Coincidently, chemical analysis revealed that (E,E)-farnesyl acetate, diisobutyl phthalate, and hexadecyl acetate of the spider webs exhibited higher relative abundances in sexually receptive females than in sexually unreceptive females or males, indicative of possible pheromone components. Two-choice behavioral assays verified that the blend of (E,E)-farnesyl acetate and hexadecyl acetate (w/w: 2:1) attracted males at a dosage equivalent to the amounts of these compounds in one spider web, whereas neither compound alone aroused males. In addition, diisobutyl phthalate (a likely contaminant from contact with plastic) alone or in combination with either of the acetates did not evoke the males’ attraction. The behavioral data suggest that (E,E)-farnesyl acetate and hexadecyl acetate comprise a two-component female-produced sex pheromone in P. beijingensis, the first multicomponent pheromone found in spiders.     

Alarm Pheromone System of the Western Conifer Seed Bug, Leptoglossus occidentalis

The alarm pheromones for adult and nymphal western conifer seed bugs, Leptoglossus occidentalis, were collected from the headspace volatiles of agitated bugs and from extracted adult thoraxes and nymphal abdomens. Adult bugs secreted a blend from the metathoracic glands that consisted of hexyl acetate, hexanal, hexanol, heptyl acetate, and octyl acetate (ratio of 152:103:8:1.5:1). Nymphal alarm pheromone produced by the dorsal abdominal glands consisted of (E)-2-hexenal. Agitated adults emitted 24% of the pheromone contained within the glands, while nymphs released 33% of their constitutive supply. The complete blend from both adults and nymphs, tested in a laboratory headspace bioassay, elicited a dispersal (or alarm) response in >70% of individuals tested. Nymphs in the field exposed to synthetic adult or nymphal pheromones, or a mixture of both, responded with >50% dispersing. When single components were tested on adults reared under summer conditions in a forced-air one-way bioassay, hexanal and hexyl acetate, the major components of the secretion, were responsible for eliciting the alarm response. Adults collected in the fall from the field were unresponsive to the tested blend, suggesting that adults seeking aggregation sites in the fall become refractory to alarm pheromone stimuli that would cause aggregations to disperse. The weak dispersal responses elicited in both adults and nymphs by either nymphal or adult pheromones are consistent with a tradeoff in the advantage gained by avoiding predation and the disadvantage of leaving a food source. Because of these weak responses, use of alarm pheromones as pest management tools against L. occidentalis is unlikely.     

Effects of Sublethal Doses of Malathion on Responses to Sex Pheromones by Male Asian Corn Borer Moths, <Emphasis Type="Italic">Ostrinia furnacalis</Emphasis> (Guenée)

We evaluated the sublethal effects of malathion treatment on the behavioral responses of male Asian corn borers, Ostrinia furnacalis (Guenée), to their sex pheromones. Doses of malathion causing 1–50% mortality were topically applied to Asian corn borer male moths. The survivors were tested for behavioral and electroantennogram (EAG) responses to their sex pheromones. Sublethal doses of malathion affected males’ ability to locate a pheromone source. In wind tunnel tests, male moths treated with sublethal doses of malathion took 2.5 to 12 times as long to take flight as control moths. Malathion-treated male moths were 60–96% less likely to locate a sex pheromone source than control moths. Multiple linear regression analysis revealed that the proportions of males performing each key stage were significantly correlated with the doses of malathion, but not with the time since treatment. Sensitivity and specificity of perception to pheromones in male moths were also disrupted by sublethal doses of malathion. The EAG responses of males treated with malathion decreased 0.13 mV on average compared with control moths. In addition, the ratios of pheromone components that elicited the largest responses shifted from approximately 5:5 to 1:9 (E/Z) and 9:1. We conclude that treatment with sublethal doses of malathion significantly reduced the likelihood that male Asian corn borers could successfully locate a normal female releasing sex pheromones.