Pheromone Release Research Articles

Opaque cells signal white cells to form biofilms in Candida albicans

Upon homozygosis from a/alpha to a/a or alpha/alpha, Candida albicans must still switch from the 'white' to 'opaque' phenotype to mate. It was, therefore, surprising to discover that pheromone selectively upregulated mating-associated genes in mating-incompetent white cells without causing G1 arrest or shmoo formation. White cells, like opaque cells, possess pheromone receptors, although their distribution and redistribution upon pheromone treatment differ between the two cell types. In speculating about the possible role of the white cell pheromone response, it is hypothesized that in overlapping white a/a and alpha/alpha populations in nature, rare opaque cells, through the release of pheromone, signal majority white cells of opposite mating type to form a biofilm that facilitates mating. In support of this hypothesis, it is demonstrated that pheromone induces cohesiveness between white cells, minority opaque cells increase two-fold the thickness of majority white cell biofilms, and majority white cell biofilms facilitate minority opaque cell chemotropism. These results reveal a novel form of communication between switch phenotypes, analogous to the inductive events during embryogenesis in higher eukaryotes.

Comparisons of Standard and Extended-Life Boll Weevil (Coleoptera: Curculionidae) Pheromone Lures

The Southeastern Boll Weevil Eradication Program has proposed reducing maintenance program costs in eradicated zones by using an extended-life "superlure" in traps to detect populations of the boll weevil, Anthonomus grandis grandis Boheman. However, superlure effectiveness has not been extensively evaluated. We compared the superlure (30 mg of eugenol plus 25 mg of grandlure) to a standard lure (10 mg of grandlure) based on captures of weevils and changes in lure pheromone content. Lure treatments (standard and superlure, replaced biweekly or not replaced) were compared in 4-mo-long trapping periods. Captures of weevils did not generally reflect differences among lure treatments indicated by assays of lure contents. During the first 2 wk of exposure, amounts of pheromone released by the superlure were generally comparable with those of the standard lure, but pheromone composition was more stable. During the second 2 wk of exposure, the superlure usually released more pheromone than similarly aged standard lures, but less than half as much as the standard lure replaced biweekly. Based on numbers of captured weevils during the last 2 wk of an extended trapping period, the superlure performed similarly to the standard lure replaced biweekly. However, corresponding pheromone releases by the superlure were less than those by the standard lure replaced biweekly. This inconsistency suggests that numbers of captured weevils alone may be inadequate for evaluation of pheromone formulations. Our results suggest that better understanding of the consequences of reduced pheromone release during an extended trapping period is needed before adoption of the superlure can be recommended.

Larval salivary glands are a source of primer and releaser pheromone in honey bee (Apis mellifera L.)

A brood pheromone identified in honeybee larvae has primer and releaser pheromone effects on adult bees. Using gas chromatography-mass spectrometry (GC-MS) to evaluate fatty acid esters--the pheromonal compounds--in different parts of the larvae, we have localized the source of the esters as the larval salivary glands. A histochemical study describes the glands and confirms the presence of lipids in the glands. Epithelial cells of the gland likely secrete the fatty acids into the lumen of the gland. These results demonstrate the salivary glands to be a reservoir of esters, components of brood pheromone, in honeybee larvae.

Direct determination of the stereoisomer constitution by 2D-HPLC and stereochemistry–pheromone activity relationship of the copulation release pheromone of the cowpea weevil, Callosobruchus maculatus

The copulation release pheromone of the cowpea weevil, Callosobruchus maculatus, was re-isolated from about 30,000 virgin female. The natural pheromone was confirmed to be stereochemically impure by Ohrui–Akasaka-2D-HPLC methodology. The structure–activity relationship of the pheromone was also clarified.

The Mandibular Gland Secretions of the Leaf-Cutting Ants Atta sexdens sexdens and Atta opaciceps Exhibit Intercaste and Intercolony Variations

The mandibular gland secretions of worker castes from wild colonies of the leaf-cutting ants Atta sexdens sexdens and Atta opaciceps were analyzed quantitatively by gas chromatography-mass spectrometry. The secretions comprised a complex mixture of volatile, mainly oxygenated compounds, and their profiles exhibited considerable qualitative and quantitative variations among species and castes. The known alarm pheromone 4-methyl-3-heptanone was common to both species. The elevated relative proportions of this ketone found in the secretions of gardeners and generalists suggest that such castes are primarily responsible for the production and release of the alarm pheromone. Quantitative variations (but no qualitative differences) in the profiles of secretions of soldiers from different colonies of A. sexdens sexdens were detected, supporting the view that intraspecific colony recognition is mediated through mandibular gland secretions. Subsequent laboratory assays showed that, among the compounds identified by GC-MS, 4-methyl-3-heptanone elicited a strong alarm response in workers of A. sexdens sexdens and A. opaciceps.

Attractiveness of year‐old polyethylene Isonet sex pheromone dispensers for Lobesia botrana

AbstractIn this study, we have compared the release of sex pheromone from mating disruption dispensers exposed in the field for 12 months and from calling females. The main pheromone component of the grapevine moth, Lobesia botrana (D. and S.) (Lepidoptera: Tortricidae), is (E)‐7,(Z)‐9‐dodecadienyl acetate, and a minor component is (Z)‐9‐dodecenyl acetate. Aged dispensers from two different years emitted a much higher amount of both pheromone components than calling females. However, the summer temperature during field exposure influenced the release from mating disruption dispensers the following year. In the wind tunnel, male L. botrana were equally attracted to 12‐month, field‐exposed dispensers, a standard monitoring pheromone lure, and to synthetic (E)‐7,(Z)‐9‐dodecadienyl acetate sprayed at the rate of 0.6–60 ng h−1. Field trapping tests confirmed that aged dispensers from both years were at least as attractive to L. botrana males as a standard monitoring pheromone lure. The possible contribution of previously applied dispensers to the mating disruption efficacy during following applications is discussed.

Primer and Short-Range Releaser Pheromone Properties of Premolt Female Urine from the Shore Crab Carcinus maenas

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.

Alarm Pheromone that Aggravates Stress-induced Hyperthermia is Soluble in Water

We previously reported that stressed male Wistar rats released alarm pheromone from the perianal region, which aggravated stress-induced hyperthermia and increased Fos expression in the mitral/tufted cell layer of the accessory olfactory bulb in recipient rats. In this study, we attempted to obtain this pheromone in water using these responses as bioassay parameters. Water droplets were collected from the ceiling of a box in which no animal was placed, or from a box in which an anesthetized donor rat was given electrical stimulation to either the neck or perianal regions in order to induce neck odor or alarm pheromone release, respectively. Then we placed one of the three kinds of water-containing filter papers on the wall of a recipient's home cage and observed heart rate, body temperature and behavioral responses, as well as Fos expression in the main and accessory olfactory bulbs of the recipient. The water collected from the box containing the alarm pheromone was found to generate a reproduction of all of the responses seen in the animal that had been directly exposed to alarm pheromone in our previous studies. These results suggest that the alarm pheromone is soluble in water.

What Is a Pheromone? Mammalian Pheromones Reconsidered

Pheromone communication is a two-component system: signaling pheromones and receiving sensory neurons. Currently, pheromones remain enigmatic bioactive compounds, as only a few have been identified, but classical bioassays have suggested that they are nonvolatile, activate vomeronasal sensory neurons, and regulate innate social behaviors and neuroendocrine release. Recent discoveries of potential pheromones reveal that they may be more structurally and functionally diverse than previously defined.

Dose-Dependent Pheromone Responses ofIps pini,Orthotomicus latidens(Coleoptera: Scolytidae), and Associates in Stands of Lodgepole Pine

We conducted four behavioral choice tests in stands of mature lodgepole pine in British Columbia, from 1988 to 1991, to determine the dose-dependent responses of Orthotomicus latidens (LeConte) and Ips pini (Say) to their respective pheromones. Dose-dependent responses were exhibited by I. pini to (±)-ipsdienol and lanierone, with trap catches directly proportional to pheromone release rates. We found differences in sex ratio of I. pini in their responses to ipsdienol but not lanierone. There was no dose-dependent response exhibited by O. latidens to its pheromone, (±)-ipsenol, with or without the presence of the host kairomone, (−)-&bgr;-phellandrene. The bark beetle predator, Enoclerus lecontei (Wolcott), showed a dose-dependent response to lanierone. Thanasimus undatulus (Say) was attracted to ipsenol in a dose-dependent fashion only when traps were also baited with &bgr;-phellandrene. Neither T. undatulus nor E. sphegeus (F.) exhibited dose-dependent responses to ipsdienol.

Release of sex pheromone and its precursors in the pine sawfly Diprion pini (Hym., Diprionidae)

The first identification of a sex pheromone of a pine sawfly (Hymenoptera, Diprionidae) dates back almost thirty years. Since then, female-produced pheromones of over twenty diprionid species have been investigated by solvent extraction followed by separation and identification. However, no study has shown what the females actually release. Collection of airborne compounds using absorbtion on charcoal filter as well as solid phase microextraction (SPME) followed by analysis employing gas chromatography combined with mass spectrometry (GC-MS), revealed an unusual system in Diprion pini, in which the pheromone precursor alcohol, 3,7-dimethyl-2-tridecanol, is released together with acetic, propionic, butyric and isobutyric acids. The corresponding acetate, propionate and butyrate esters of 3,7-dimethyl-2-tridecanol were also found in the samples. All esters were electrophysiologically active, and the propionate and isobutyrate were attractive in trapping experiments. Based on these and earlier reported results, it seems that at least in part of its range, the pheromone response of D. pini is not very specific with regard to the functional group, as long as this is an ester.

Nocturnal Calling Behavior in Mantids

Females of Mantis religiosa and Empusa pennata were video taped for several 24 h periods to determine if they showed behaviors associated with pheromone release. In the photophase the abdomen of both species was motionless and rested in continuous contact with the wings. However, at the beginning of the scotophase the females bent the abdomen ventrally so that the space between the abdomen and the wings increased significantly with respect to the daytime posture. Calling behavior (abdominal bending) was maintained throughout the 8 h scotophase and ended abruptly at lights on. Females of M. religiosa did not start calling until they were 30 days of age. Calling disappeared in mated females, but it reappeared two weeks later. Males stayed motionless in response to the odors emitted by other males or by noncalling females, but walked when a calling female was placed in the air flow. These observations suggest that female mantids bend their abdomens at night to release a sex pheromone. The adaptive function of nocturnal sex pheromone release in sexually cannibalistic species that rely strongly on visual cues for mating is discussed.

Alarm pheromone mediates production of winged dispersal morphs in aphids

AbstractThe aphid alarm pheromone (E)‐β‐farnesene (EBF) is the major example of defence communication in the insect world. Released when aphids are attacked by predators such as ladybirds or lacewing larvae, aphid alarm pheromone causes behavioural reactions such as walking or dropping off the host plant. In this paper, we show that the exposure to alarm pheromone also induces aphids to give birth to winged dispersal morphs that leave their host plants. We first demonstrate that the alarm pheromone is the only volatile compound emitted from aphid colonies under predator attack and that emission is proportional to predator activity. We then show that artificial alarm pheromone induces groups of aphids but not single individuals to produce a higher proportion of winged morphs among their offspring. Furthermore, aphids react more strongly to the frequency of pheromone release than the amount of pheromone delivered. We suggest that EBF leads to a ‘pseudo crowding’ effect whereby alarm pheromone perception causes increased walking behaviour in aphids resulting in an increase in the number of physical contacts between individuals, similar to what happens when aphids are crowded. As many plants also produce EBF, our finding suggests that aphids could be manipulated by plants into leaving their hosts, but they also show that the context‐dependence of EBF‐induced wing formation may hinder such an exploitation of intraspecific signalling by plants.

Kinetics and molecular properties of pheromone binding and release

Transient kinetic studies have shown that the uptake of the pheromone (bombykol) of the silkworm moth (Bombyx mori), by its pheromone-binding protein (PBP) BmorPBP, proceeds with an "on" rate of 0.068 +/- 0.01 microM(-1).s(-1). With the high concentration of PBP in the sensillar lymph (10 mM), the half-life for the uptake of pheromone in vivo is approximately equal to 1 ms. A pH-dependent conformational change (BmorPBP(B) --> BmorPBP(A)), associated with the release of pheromone, is a first-order reaction (k = 74.1 +/- 0.32 s(-1); t(1/2), 9.3 ms). Under physiological conditions, both reactions proceed with half-life times on the order of milliseconds, as is required for odorant-oriented navigation in insects. Molecular interactions of bombykol with both native and mutated PBPs were analyzed by a novel binding assay. A recombinant protein with the native conformation (BmorPBP) showed high binding affinity (K(D) = 105 nM) at pH 7 but low affinity (K(D) = 1,600 nM) at pH 5, when tested at both low and high KCl concentrations. A protein with a C-terminal segment deleted (BmorPBPDeltaP129-V142) was found to bind bombykol at pH 7 and at pH 5 with the same affinity as the native protein at pH 7, indicating that the C-terminal segment is essential for preventing binding at low pH. Binding studies with three mutated proteins (BmorPBPW37F, BmorPBPW127F, and BmorPBPW37A) showed that replacing Trp-37 (with Phe or Ala) or Trp-127 (with Phe) did not affect the binding affinity to bombykol. Fluorescence studies shed light on the contributions of Trp-37 and Trp-127 emissions to the overall fluorescence.

Evaluation of Pheromone Release from Commercial Mating Disruption Dispensers

Pome fruit growers and crop consultants have expressed concerns about the seasonal release performance of commercial codling moth mating disruption dispenser products. Because of these concerns, we developed a laboratory flow-through volatile collection system (VCS) for measuring the volatile release of the codling moth sex pheromone, codlemone, from commercially available hand-applied dispensers. Under controlled air-flow and temperature conditions, the released vapor was trapped onto a polyurethane foam adsorbent followed by solvent extraction, solvent reduction, and GC/MS determination. Method recovery and breakthrough validations were performed to demonstrate system reliability before determining codlemone release from commercial dispensers field-aged over 140 days. The volatile collection was carried out in a consistent manner among five dispenser types most commonly used by growers, so that direct comparison of performance could be made. The comparison showed differences in the amount of pheromone released and in the patterns of release throughout the season between dispenser types. The variation in release performance demonstrates the need for routine evaluation of commercially marketed mating disruption dispensers. We believe that the simple and cost-effective volatile collection system can assist pheromone dispenser manufacturers in determining seasonal dispenser performance before new products are introduced into the commercial market and in rapidly verifying dispenser release when field-aged dispenser efficacy is in question.

Calling behaviour in the female praying mantis, Hierodula patellifera

Abstract. Virgin mantis females, Hierodula patellifera (Serville) (Dictyoptera: Mantidae), exhibit a characteristic calling posture. When holding the body below a branch or leaf, the female curls the abdomen ventrally, flexing it away from the wings and exposing its dorsal surface. The curling is accompanied by pumping movements. The average age at which females start adopting this calling posture is 14 days after adult moult, and it is related to their nutritional stage. Once initiated, females exhibit the posture everyday until they mate. After mating, the behaviour is completely suppressed. Males are attracted by virgin females adopting the calling posture but are not attracted to mated females. The characteristics of the posture and the responsive behaviour of the males indicate that this female calling involves the release of sex pheromones.

3-Ethyl-2,7-dimethyl octane, a testosterone dependent unique urinary sex pheromone in male mouse ( Mus musculus)

A previous investigation revealed that urine from normal male mice contained five unique volatile constituents; namely: 3-cyclohexene-1-methanol (I); 3-amino triazole (II); 4-ethyl phenol (III); 3-ethyl-2,7-dimethyl octane (IV); 1-iodoundecane (V). The present study was designed to find out whether the production of these male specific urinary compounds was androgen-dependent. Urine of castrated and castrated plus testosterone-treated male mice was analyzed using gas chromatography linked mass spectrometry (GC–MS). Even though castrated male urine contained 10 detectable compounds, the five male specific compounds present in intact males were absent in castrated male mice urine. Only 3-ethyl-2,7-dimethyl octane (IV) reappeared following testosterone treatment into castrated males. Our earlier bioassay revealed that this compound was involved in attracting females. The present study concluded that this compound was a male specific volatile cue that acted as a releaser pheromone and its production was under the control of androgen.

Insect semiochemical research in Korea: Overview and prospects

Rapid progress has been made in research on insect pheromones and other semiochemicals in Korea during the last two decades. Sex pheromone compositions have been identified in several major orchard and other horticultural moth pest species and some other species in Korea, such as Helicoverpa assulta, Grapholita molesta, Carposina sasakii, Adoxophyes orana, Lyonetia prunifoliella, Phyllonorycter ringoniella, Dichocrocis punctiferalis, Ostrinia furnacalis and Peridroma saucia. Some of these identified sex pheromones have already been successfully incorporated into IPM systems, and are now being used as direct and indirect pest control means in Korea. Korean populations of many of these species showed significant differences in pheromone compositions from those reported in the neighboring countries of Japan and China, suggesting that such interpopulational pheromone polymorphism is more common across the insect world than previously thought. Sex pheromone compositions were also identified in some aphid species in Korea such as Aphis spiraecola and Tuberocephalus momonis, and it was soon found that the aphid sex pheromone components were highly attractive to their predator lacewings. The presence of kairomones responsible for attracting egg parasitoids was shown in a pheromone component, hot pepper odor and male scale factor for H. assulta, and in male accessory glands for Lymantria dispar. Studies on the regulation of pheromone biosynthesis and release were carried out simultaneously in some species in Korea, resulting in the characterization of PBAN (pheromone biosynthesis activating neuropeptide) in H. assulta and Adoxophyes sp. A semiochemical research system has been firmly established in Korea through these studies. In some areas, successful monitoring and mating disruption technology have been effectively transferred from researchers to the end users in Korea, which will soon be expanded to wider areas. Continuous attempts to identify and use semiochemicals as well as wider applications of semiochemicals in pest control are expected in Korea in the coming years. Since Korean research on semiochemicals does not cover vast areas due to limited resources, focused research and practical application will be needed for selected target insect species.

Examining disruption of pheromone communication in Choristoneura rosaceana and Pandemis limitata using microencapsulated (Z)‐11‐tetradecenyl acetate applied in a laboratory flight tunnel

AbstractPheromone‐based mating disruption of lepidopteran pests (Tortricidae) of pome fruits using hand‐applied dispensing systems has become standard management practice for many producers in western North America. Sprayable microencapsulated (MEC) pheromone formulations that enable the application of pheromone controls with other orchard sprays and assist in the development of multispecies mating‐disruption systems are currently under development. Responses of male Choristoneura rosaceana (Harris) and Pandemis limitata (Robinson) (Lepidoptera: Tortricidae) to calling females in clean air, and air treated with their major pheromone component (Z)‐11‐tetradecenyl acetate (Z11‐14:OAc), released from a 3M sprayable pheromone formulation containing proprietary 3M Phase I microcapsules, applied at doses of 1, 10, and 100 mg of active ingredient (ai) m−2 to the upwind end of a flight tunnel (equivalent to field rates of 10, 100, and 1000 g ai ha−1) were compared in laboratory flight tunnels. In both species, disorientation was found to be dose‐dependent, because relative to male orientation to calling females in clean air, the orientation of male P. limitata was disrupted 23.3, 46.3, and 71.3%, and orientation by male C. rosaceana was disrupted 31.6, 37.7, and 45.8% by treatment doses of 1, 10, and 100 mg m−2, respectively. Latency of male responses to calling females in a background of Z11‐14:OAc relative to responses in clean air was also dose‐dependent. Albeit short, the disruption lasted 26, 74, and 218 h in P. limitata and 30, 54, and 174 h in C. rosaceana at each application rate, respectively. Disruption by pheromone treatment was greater in P. limitata than in C. rosaceana. This difference may be correlated with species’ differences in the pheromone release rates of females. Mechanisms of disruption invoked by this 3M MEC pheromone formulation are discussed in relation to issues of its longevity and observed differences in the effects against the two species. It appears possible to evaluate relative activity of MEC pheromones in a laboratory setting which may aid in development of new formulations for mating disruption.

Synthesis of the Four Stereoisomers of 2,6-Dimethyloctane-1,8-dioic Acid, a Component of the Copulation Release Pheromone of the Cowpea Weevil,Callosobruchus maculatus

A diastereomeric mixture and the four stereoisomers of 2,6-dimethyloctane-1,8-dioic acid (2), a copulation release pheromone of the cowpea weevil, Callosobruchus maculatus, were synthesized. The stereoisomeric purities of the four synthetic isomers of 2 were determined by the HPLC analyses of their bis-2-(2,3-anthracenedicarboximide)-1-cyclohexyl esters.