Floral synomone of a wild orchid,Bulbophyllum cheiri,lures Bactrocera fruit flies for pollination

The major fruit fly attractant component in the floral fragrance of Bulbophyllum cheiri (fruit fly orchid) is methyl eugenol (ME). In the lowland rain forest of Malaysia, the solitary and nonresupinate flowers of the fruit fly orchid attract only males of the ME-sensitive fruit fly species (Bactrocera carambolae, B. papayae, and B. umbrosa. During the morning, the fruit fly orchid flower is visited by many fruit flies, which can sometimes cover the whole flower. The number of visitors dwindles in the afternoon. Headspace analysis of the flower shows a high ME peak in the morning, a small one between 12:00 and 14:00 hr, and no detectable ME peak after 14:00 hr. The process of pollination in the wild is initiated by attraction of fruit flies to floral ME. The flower, with the aid of its specialized hinged see-saw lip (labellum), temporarily traps (<1 min) a fruit fly pollinator between its lip and column. Just prior to this, the fly is rewarded by the opportunity to feed on the floral attractant found on surfaces of petals, sepals, and lip. The pollinaria borne by two wild B. papayae males (caught on and near the fruit fly orchid flower) are identical in morphology and structure with those obtained from the flower. Many of the B. papayae males (17 of 22 analyzed) attracted to the fruit fly orchid already possessed both ME metabolites, trans-coniferyl alcohol and 2-allyl-4,5-dimethoxyphenol, in their rectal glands, indicating that they had previously consumed ME. In this orchid–fruit fly association, both organisms gain direct reproductive benefits: the orchid flower gets pollinated without having to offer nectar, while the fruit fly boosts its pheromone and defense system, as well as its sexual competitiveness by feeding on the ME produced by the flower.     

Synomone Or Kairomone? – <Emphasis Type="Italic">Bulbophyllum Apertum</Emphasis> Flower Releases Raspberry Ketone To Attract <Emphasis Type="Italic">Bactrocera</Emphasis> Fruit Flies

Bulbophyllum apertum flower (Orchidaceae) releases raspberry ketone (RK) in its fragrance, which attracts males of several fruit fly species belonging to the genus Bactrocera. Besides RK as a major component, the flower contains smaller amounts of 4-(4-hydroxylphenyl)-2-butanol, plus two minor volatile components, veratryl alcohol and vanillyl alcohol. Within the flower, the lip (labellum) had the highest concentration of RK with much smaller quantities present in petals; other flower parts had no detectable RK. Male fruit flies attracted to the flower belong to RK-sensitive species – such as Bactrocera albistragata, B. caudatus, B. cucurbitae (melon fly), and B. tau. Removal and attachment of the pollinarium to a flys thoracic dorsum occurred when a male of B. albistragata was toppled into the floral column cavity, due to an imbalance caused by it shifting its body weight while feeding on the see-saw lip, and then freeing itself after being momentarily trapped between the lip and column. During this process, the stiff hamulus (the pollinia stalk protruding prominently towards the lip) acted as a crowbar when it was brushed downwards by the toppled fly and lifted the pollinia out of the anther. If the fly was big or long for the small triangular lip, it would not be toppled into the column cavity and would just walk across the column, during which time the pollinarium could be accidentally removed by the flys leg, resulting in a failed transport of the pollinarium. This suggests an unstable situation, where the orchid relies only on a particular pollinator species in the complex ecosystem where many RK-sensitive species inhabit.Wild males of B. caudatus (most common visitors) captured on Bulbophyllum apertum flowers were found to sequester RK in their bodies as a potential pheromonal and allomonal ingredient. Thus, RK can act either as a floral synomone (pollinarium transported) or kairomone (accidental removal of pollinarium leading to total pollen wastage), depending on the body size of the male fruit flies visiting the flowers.     

Floral Phenylpropanoid Cocktail and Architecture of Bulbophyllum vinaceum Orchid in Attracting Fruit Flies for Pollination

It is widely believed that most orchid flowers attract insects by using deception or chemical rewards in the form of nectar. Flowers of Bulbophyllum vinaceum produce a large array of phenylpropanoids that lure tephritid fruit fly males and also act as floral reward, which the flies subsequently convert to pheromone components. The major floral volatile components identified are methyl eugenol (ME), trans-coniferyl alcohol (CF), 2-allyl-4,5-dimethoxphenol (DMP), and trans-3,4-dimethoxycinnamyl acetate, whereas the minor components are eugenol, euasarone, trans-3,4-dimethoxy cinnamyl alcohol, and cis-coniferyl alcohol. Among the various floral parts, the lip (which is held in a closed position up against the sexual organs) has the highest concentration of the major compounds. An attracted male fly normally lands on one of the petals before climbing up onto and forcing the “spring loaded” floral lip into the open position, hence exposing the floral sexual organs. The architecture and location of chemical attractants of the lip compel the fly to align itself along the lip’s longitudinal axis in a precise manner. As the fly laps up the compounds and moves towards the base of the lip, it passes the point of imbalance causing the lip to spring back to its normal closed position. The fly is catapulted headfirst into the column cavity, and its dorsum strikes the protruding sticky base of the hamulus and adheres to it. The momentum of the fly and the structural morphology of the long stiff hamulus act to pry out the pollinia from its anther cover. Hence, the pollinarium (pollinia + hamulus) is detached from the flower and adhered to the fly’s dorsum. In this unique mutualistic association, both species receive direct reproductive benefits—the flower’s pollinarium is transported for cross pollination, and the fly is offered a bouquet of phenylpropanoids (synomone) that it consumes, converts, and/or sequesters as sex pheromonal components, thus enhancing sexual attraction and mating success.     

Biology of pheromone release by male caribbean fruit flies,Anastrepha suspensa (Diptera: Tephritidae)

Males of the Caribbean fruit fly,Anastrepha suspensa (Loew), typically form leks and attract females by releasing a multicomponent volatile pheromone. Previous reports have identified two nine-carbon alcohols, three lactones, a sesquiterpene, and a monoterpene in the volatiles. The present report is a study of the physiology of male pheromone release and of ecological and social interactions that influence pheromone release by laboratoryreared flies. Volatiles released by males were trapped on Tenax, eluted, separated, and quantitatively measured by gas chromatography. Experiments showed that the volatiles were primarily released from mouth and anus. Sealing the anal opening or the mouth with melted beeswax resulted in up to 40% or greater reduction in most components, and sealing both mouth and anus further reduced release of volatiles, but some volatiles are possibly still released directly from the cuticle. An anal pouch of everted tissue played a major role as a large evaporative surface for release of some of the volatile components. Male flies entrained to a 1410 light-dark cycle showed a peak release of volatiles at 11–12 hr into the photophase, but smaller quantities of the same volatiles were released over a broad period during the daylight hours. Laboratory-reared males peaked in pheromone release at 7–10 days and production and release continued through 35 days of age. Single males released significantly more of all components measured than did groups of males. The reduction by aggregations of males may be related to lekking behavior in this fruit fly. The pheromone probably serves to attract females to a lek site, but additional parameters are likely to enter into the choice of male made by the arriving female.This is University of Florida Agricultural Experiment Station Journal Series No. 9589.     

Accumulation ofDendrobium superbum (orchidaceae) fragrance in the rectal glands by males of the melon fly,Dacus cucurbitae

4-(4-Hydroxyphenyl)-2-butanone was characterized from flowers of the orchidDendrobium superbum as a specific attractant factor for the male melon fly,Dacus cucurbitae. The male flies compulsively licked the flower surface and sequestered the compound in significant quantities in their rectal glands. The compound was detected within 6 hr after ingestion and was retained for more than six days in the rectal gland sacs.     

Disease Status and Population Origin Effects on Floral Scent<Emphasis Type="SmallCaps">:</Emphasis> Potential Consequences for Oviposition and Fruit Predation in A Complex Interaction Between A Plant,Fungus, and Noctuid Moth

In the Silene latifolia–Hadena bicruris nursery pollination system, the Hadena moth is both pollinator and seed predator of its host plant. Floral scent, which differs among S. latifolia individuals and populations, is important for adult Hadena to locate its host. However, the success of moth larvae is strongly reduced if hosts are infected by the anther smut fungus Microbotryum violaceum, a pathogen that is transmitted by flower visitors. There were no qualitative differences between the scent of flowers from healthy and diseased plants. In addition, electroantennographic measurements showed that Hadena responded to the same subset of 19 compounds in samples collected from healthy and diseased plants. However, there were significant quantitative differences in scent profiles. Flowers from diseased plants emitted both a lower absolute amount of floral scent and had a different scent pattern, mainly due to their lower absolute amount of lilac aldehyde, whereas their amount of (E)-β-ocimene was similar to that in healthy flowers. Dual choice behavioral wind tunnel tests using differently scented flowers confirmed that moths respond to both qualitative and quantitative aspects of floral scent, suggesting that they could use differences in floral scent between healthy and infected plants to discriminate against diseased plants. Population mean fruit predation rates significantly increased with population mean levels of the emission rates of lilac aldehyde per flower, indicating that selection on floral scent compounds may not only be driven by effects on pollinator attraction but also by effects on fruit predation. However, variation in mean emission rates of scent compounds per flower generally could not explain the higher fruit predation in populations originating from the introduced North American range compared to populations native to Europe.     

Identification of Sex Pheromone Components of the Hessian Fly, <Emphasis Type="Italic">Mayetiola destructor</Emphasis>

Coupled gas chromatographic (GC)–electroantennographic detection (EAD) analyses of ovipositor extract of calling Hessian fly, Mayetiola destructor, females revealed that seven compounds elicited responses from male antennae. Four of the compounds—(2S)-tridec-2-yl acetate, (2S,10Z)-10-tridecen-2-yl acetate, (2S,10E)-10-tridecen-2-yl acetate, and (2S,10E)-10-tridecen-2-ol—were identified previously in female extracts. Two new EAD-active compounds, (2S,8Z,10E)-8,10-tridecadien-2-yl acetate and (2S,8E,10E)-8,10-tridecadien-2-yl acetate, were identified by GC–mass spectroscopy (MS) and the use of synthetic reference samples. In a Y-tube bioassay, a five-component blend (1 ng (2S)-tridec-2-yl acetate, 10 ng (2S,10E)-10-tridecen-2-yl acetate, 1 ng (2S,10E)-10-tridecen-2-ol, 1 ng (2S,8Z,10E)-8,10-tridecadien-2-yl acetate, and 1 ng (2S,8E,10E)-8,10-tridecadien-2-yl acetate) was as attractive to male Hessian flies as a similar amount of female extract (with respect to the main compound, (2S,10E)-10-tridecen-2-yl acetate). The five-component blend was more attractive to male flies than a three-component blend lacking the two dienes. Furthermore, the five-component blend was more attractive than a blend with the same compounds but that contained one tenth the concentration of (2S,8E,10E)-8,10-tridecadien-2-yl acetate (more accurately mimicking the ratios found in female extract). This suggests that the ratios emitted by females might deviate from those in gland extracts. In a field-trapping experiment, the five-component blend applied to polyethylene cap dispensers in a 100:10 μg ratio between the main component and each of the other blend components attracted a significant number of male Hessian flies. Also, a small-plot field test demonstrated the attractiveness of the five-component blend to male Hessian flies and suggests that this pheromone blend may be useful for monitoring and predicting Hessian fly outbreaks in agricultural systems.     

Acquisition of Female-Attracting Fragrance by Males of Oriental Fruit Fly from a Hawaiian Lei Flower, Fagraea berteriana

Males of the Oriental fruit fly, Bactrocera dorsalis, are strongly attracted to and compulsively feed on a fragrant lei flower, Fagraea berteriana. A series of phenylpropanoid components, trans-3,4-dimethoxycinnamyl alcohol, its acetate, and trans-3,4-dimethoxycinnamaldehyde were characterized as male attractants. The alcohol stimulated the same level of feeding activity as methyl eugenol. Males that fed on flowers selectively converted the attractant components into trans-coniferyl alcohol and stored it in rectal glands. Males scented with the phenylpropanoids were more successful in mating than unfed males, indicating the advantage of acquiring the fragrance in mating success.     

Sex Pheromone Components in Defense of Melon Fly, Bactrocera cucurbitae Against Asian House Gecko, Hemidactylus frenatus

The Asian house gecko, Hemidactylus frenatus, after being acclimatized to feeding on the melon fly, Bactrocera cucurbitae, consumed fewer numbers of 15-day-old (DO) and older male melon flies compared to sexually immature flies 10 DO and younger. The first male melon fly that performed mating did so on the 10th day after eclosion (DAE), but >80% of the males performed their first mating between 10 and 15 DAE. This confirmed that between 10 and 15 DAE most male melon flies are already sexually mature and producing sex pheromone. Synthetic 1,3-nonanediol, a component of male melon fly sex pheromone produced endogenously, when topically applied onto the thorax of housefly Musca domestica at ca. 80 or 320 ng/fly, reduced consumption by geckos compared with untreated flies in subsequent days after the first day in a four-day feeding test. Raspberry ketone (RK) is consumed and sequestered into the pheromonal gland and later released as a pheromonal component of male melon fly. Houseflies topically treated with 2.6 g RK/fly did not deter predation by geckos. However, houseflies treated with 5.1 g RK/fly caused geckos to consume fewer flies, especially on the third and fourth days during a four-day feeding test, compared with the period when they were offered untreated flies.     

Evidence of Natural Hybridization Between Two Sympatric Sibling Species of <Emphasis Type="Italic">Bactrocera dorsalis</Emphasis> Complex Based on Pheromone Analysis

Bactrocera carambolae and B. papayae are major fruit fly pests and sympatric sibling species of the B. dorsalis complex. They possess distinct differences in male pheromonal components. In the 1990s, wild Bactrocera fruit flies with morphological traits intermediate between those of B. carambolae and B. papayae were often captured in traps baited with methyl eugenol (ME). Chemical analyses of rectal glands of ME-fed males revealed that the laboratory F₁, F₂, and backcross hybrids possessed ME-derived sex pheromonal components ranging from that typical of B. papayae to that of B. carambolae without any specific trend, which included a combination of pheromonal components from both parental species within an individual hybrid. ME-fed hybrids without any ME-derived pheromonal components were also detected. Further chemical analysis of rectal glands from wild Bactrocera males, after ME feeding in the laboratory, showed a combination of pheromonal components similar to that found in the ME-fed, laboratory-bred hybrids. These findings present circumstantial evidence for the occurrence of a natural hybrid of the two Bactrocera species.     

Deterrence of repeated oviposition by fruit-marking pheromone inCeratitis capitata (Diptera: Tephritidae)

During ovipositor dragging on the fruit surface following egg laying in hawthorne fruit,Ceratitis capitata (Wiedemann) females deposit an unidentified pheromone that deters repeated oviposition attempts in that fruit. The pheromone proved water soluble and, when collected and sprayed in aqueous solution onto uninfested fruits in laboratory cages, effectively deterred boring attempts byC. capitata females of wild origin for at least 6 days (termination of test). A laboratory population ofC. capitata cultured on artificial media for more than 200 generations deposited pheromone that proved equally as deterrent to wild fly oviposition as pheromone from wild flies. However, lab fly oviposition was not effectively deterred by the presence of pheromone. The ecological significance of the pheromone is discussed.     

Foraging Behavior of Anastrepha Ludens, A. obliqua, and A. serpentina in Response to Feces Extracts Containing Host Marking Pheromone

Following oviposition, females of many Tephritid flies deposit host marking pheromones (HMPs) to indicate that the host fruit has been occupied. We describe the foraging behavior of these three economically important species (Anastrepha ludens and A. obliqua from the fraterculus species group and A. serpentina from the serpentina species group) when they encounter an artificial fruit (green agar spheres wrapped in Parafilm) marked with intra- and interspecific feces extracts that contain, among other substances, host marking pheromone. When flies encountered fruit treated with either 1 or 100 mg/ml feces extract, there were drastic and statistically significant reductions in tree residence time, mean time spent on fruit, and in the number of oviposition attempts or actual ovipositions when compared to the control treatment (clean fruit). These responses were almost identical irrespective of extract origin (i.e., fly species), indicating complete interspecific HMP cross-recognition by all three Anastrepha species tested. We discuss the ecological and practical implications of our findings.     

Premating Behavior of Bombus Confusus Males and Analysis of Their Labial Gland Secretion

Premating behavior in the bumblebee Bombus confusus was studied. Visual searching for females is not the only premating strategy of this species, as was believed earlier. Males of B. confusus have a normally developed labial gland and its secretion is used to mark a perch from which they visually search for females. The labial gland secretion contains geranylcitronellol and (Z)-9-octadecenyl acetate as the main components.     

Identification of sex pheromone composition of click beetle Agriotes brevis candeze

Geranyl butyrate (GB) and (E,E)-farnesyl butyrate (FB) were identified in the pheromone gland extract of females of the click beetle, Agriotes brevis (Candeze) (Coleoptera: Elateridae) as the major sex pheromone components. Polyethylene vial dispensers containing 20–200 mg of a 1:1 mixture caught high numbers of beetles. Captures did not decrease even after 73 days of field exposure of dispensers. At sites where both Agriotes sputator L. and A. brevis were present, the above baits were selectively catching only A. brevis, despite the fact that GB is also the main pheromone component of A. sputator, suggesting that FB has a role in reproductive isolation. In the early part of the season, traps into which the insects could both crawl and fly captured more A. brevis than designs where the insects could only fly in. Trap design was not important later in the season. This indicates the need for future development of a trap suitable for use throughout the whole season.     

Ability of honeybee,Apis mellifera,to detect and discriminate odors of varieties of canola (Brassica rapa and Brassica napus) and snapdragon flowers (Antirrhinum majus)

Honeybees (Apis mellifera) use odors to identify and discriminate among flowers during foraging. This series of experiments examined the ability of bees to detect and discriminate among the floral odors of different varieties of two species of canola (Brassica rapa and Brassica napus) and also among three varieties of snapdragons (Antirhinnum majus). Individual worker honeybees were trained using a proboscis extension assay. The ability of bees to distinguish a floral odor from an air stimulus during training increased as the number of flowers used during training increased. Bees conditioned to the odor of one variety of flower were asked to discriminate it from the odors of other flowers in two different training assays. Bees were unable to discriminate among flowers at the level of variety in a randomized presentation of a reinforced floral odor and an unreinforced floral odor. In the second type of assay, bees were trained with one floral variety for 40 trials without reinforcement and then tested with the same variety or with other varieties and species. If a bee had been trained with a variety of canola, it was unable to differentiate the odor of one canola flower from the odor of other canola flowers, but it could differentiate canola from the odor of a snapdragon flower. Bees trained with the odor of snapdragon flowers readily differentiated the odor of one variety of a snapdragon from the odor of other varieties of snapdragons and also canola flowers. Our study suggests that both intensity and odor quality affect the ability of honeybees to differentiate among floral perfumes.     

Characterization of a Behaviorally Active,Gender-Specific Volatile Compound from the Male Asparagus Fly <Emphasis Type="Italic">Plioreocepta poeciloptera</Emphasis>

Adult male asparagus flies exhibit typical calling behaviors (suggestive of pheromone production) during which they emit a single volatile compound that was identified as isopropyl (S)-5-hydroxyhexanoate. In laboratory bioassays, synthetic samples elicited an arrestant response in females, but did not appear to attract females. On the other hand, the synthetic material attracted conspecific males in olfactometer bioassays.     

Olfactory Responses of Neotropical Short-Tailed Singing Mice, <Emphasis Type="Italic">Scotinomys teguina</Emphasis>, to Odors of the Mid-Ventral Sebaceous Gland: Discrimination of Conspecifics,Gender, and Female Reproductive Condition

We examined the olfactory communication of the Neotropical short-tailed singing mouse, Scotinomys teguina, by investigating whether S. teguina responded to odors produced by the mid-ventral sebaceous gland of conspecifics. Females spent significantly more time investigating male odor than an odorless stimulus or a female odor. Males spent significantly more time investigating female odor than an odorless stimulus, but not that of a male odor. This latter result does not seem to be explained by differences in age or sexual experience of test subjects, but may be influenced by reproductive condition of the female odor donor. Male S. teguina spent significantly more time (1) investigating and (2) in total proximity to odors of estrous than non-estrous females. Males spent more time (1) investigating and (2) in total proximity to odors of males than non-estrous females. Finally, given the choice between odor of males vs proestrous females, males showed no preference. Thus, the mid-ventral gland in S. teguina seems to convey information about conspecifics, sex, and female reproductive condition. Male odor, compared with proestrous female odor, is equally interesting to males, suggesting that the gland also plays an important role in male–male communication. Sexual dimorphism in the size of the gland and in the amount of secretion produced by the gland may be related to either male–male competition or female choice.     

(S)-2,3-Dihydrofarnesoic Acid, a New Component in Cephalic Glands of Male European Beewolves Philanthus triangulum

The chemical composition and functional significance of pheromones of solitary Hymenoptera is much less well known compared to social species. Males of the genus Philanthus (Sphecidae) are territorial and scent mark their territories to attract females. Because of inconsistent results of earlier studies, we reanalyzed the content of the cephalic glands of male European beewolves, Philanthus triangulum F. Besides a variety of alkanes and alkenes, four major compounds were found. Two of these, (Z)-11-eicosen-1-ol and (Z)-10-nonadecen-2-one, had been previously described as constituents of the cephalic glands. We identified 1-octadecanol as a new component of the cephalic gland, and a fourth compound, enantiopure (S)-2,3-dihydrofarnesoic acid, was identified for the first time in nature. Structural elucidation and enantiomeric analysis were performed by HRGC-MS and HRGC-FTIR as well as enantioselective gas chromatography and by means of authentic reference compounds. Occurrence and     

Consumption and Metabolism of 1,2-Dimethoxy-4-(3-Fluoro-2-Propenyl)Benzene, a Fluorine Analog of Methyl Eugenol, in the Oriental Fruit Fly Bactrocera dorsalis (Hendel)

Methyl eugenol (ME) is a natural phenylpropanoid highly attractive to oriental fruit fly Bactrocera dorsalis (Hendel) males. The flies eagerly feed on ME and produce hydroxylated metabolites with both pheromonal and allomonal functions. Side-chain metabolic activation of ME has long been recognized as a primary reason for hepatocarcinogenicity of this compound on rodents. In an attempt to develop a safer alternative to ME for fruit fly management, we developed a fluorine analog 1,2-dimethoxy-4-(3-fluoro-2-propenyl)benzene (I), which, in earlier field tests, was as active to the oriental fruit fly as ME. Now we report that B. dorsalis males are not only attracted to, but also eagerly consume (up to ∼1 mg/insect) compound I, thus recognizing this fluorinated benzene as a close kin of the natural ME. The flies metabolized the fluorine analog I in a similar fashion producing mostly two hydroxylated products, 2-(3-fluoro-2-propenyl)-4,5-dimethoxyphenol (II) and (E)-coniferyl alcohol (III), which they stored in rectal glands. However, the introduction of the fluorine atom at the terminal carbon atom of the double bond favors the ring hydroxylation over a side-chain metabolic oxidation pathway, by which coniferyl alcohol is produced. It also appears that fluorination overall impedes the metabolism: at high feed rate (10 μl per 10 males), the flies consumed in total more fluorine analog I than ME but were unable to metabolize it as efficiently as ME.     

Attraction of Female Oriental Fruit Fly, Bactrocera dorsalis, to Volatile Semiochemicals from Leaves and Extracts of a Nonhost Plant, Panax (Polyscias guilfoylei) in Laboratory and Olfactometer Assays

Fresh whole leaves and solvent–water leaf extracts of the hedgerow plant panax, Polyscias guilfoylei (Bull), were tested for their attractiveness to male and female Oriental fruit flies, Bactrocera dorsalis, in laboratory flight tunnel and cage olfactometer bioassays. Fresh mature whole panax leaves were found to be attractive to mated female oriental fruit flies in the flight tunnel. Response of males and virgin females was low and in most instances not significantly different from controls. Attraction of mated female flies to the layers resulting from a methylene chloride–water partition or a hexane–water partition of freshly ground leaves using small McPhail traps was greatest in the methylene chloride fraction. When methylene chloride and water layers were tested competitively in a multiple-choice rotating olfactometer, the methylene chloride fraction was more attractive. Tests involving the methylene chloride–water interface (an emulsion of the two partitioned layers) with and without a standard attractant NuLure, showed the emulsion layer to be significantly more attractive than the other fractions or NuLure. In outdoor cage olfactometer assays of methylene chloride and water fractions, activity was greatest in the methylene chloride fraction. The results suggests that volatile semiochemicals from this nonhost plant are attractive to mated female Oriental fruit flies. The results are discussed in relation to the chemical ecology of B. dorsalis and the potential use of this nonhost plant for detection and control of female Oriental fruit flies in the field.