Gas Chromatographic-electroantennographic Detection Research Articles

Electrophysiological and Behavioral Responses of <I>Dendroctonus frontalis</I> (Coleoptera: Curculionidae) to Volatiles Isolated from Conspecifics

Olfactory sensitivity of the southern pine beetle, Dendroctonus frontalis Zimmermann, to compounds isolated from the mid/hindguts of newly emerged conspecific adults was assayed with coupled gas chromatography-electroantennographic detection. All previously reported pheromones for D. frontalis plus eight additional compounds (fenchyl alcohol, myrtenal, cis-verbenol, trans-pinocarveol, acetophenone, trans-myrtanol, cis-myrtanol, and 2-phenylethanol) consistently elicited antennal responses from at least one sex. The eight additional compounds were assayed individually at three release rates (0.4-0.8, 3-9, and 25-100 mg/d) for the ability to alter D. frontalis responses to traps baited with D. frontalis attractant (4 mg/d frontalin and 17 mg/d alpha-pinene). At the high release rate, cis-verbenol enhanced attraction of D. frontalis females, whereas the other seven compounds significantly reduced attraction of one or both sexes. Acetophenone significantly reduced attraction of male D. frontalis at the low release rate, and five compounds (fenchyl alcohol, trans-pinocarveol, acetophenone, cis-myrtanol, and 2-phenylethanol) reduced attraction of one or both sexes at the intermediate rate. Only acetophenone significantly altered the sex ratio of beetles trapped, decreasing the proportion of males. Attraction of predatory checkered beetles (Cleridae) was enhanced by cis-verbenol released at the high rate but was not altered by any compound inhibitory to D. frontalis. Analyses of volatiles from individual D. frontalis indicated that the majority of the eight compounds were produced in greater quantities by newly emerged beetles than ones attacking pine bolts. Five of the compounds were associated predominantly with one sex. Possible ecological roles of these compounds in the biology of D. frontalis are discussed.

1,4-Dimethoxybenzene, a Floral Scent Compound in Willows that Attracts an Oligolectic Bee

Many bees are oligolectic and collect pollen for their larvae only from one particular plant family or genus. Here, we identified flower scent compounds of two Salix species important for the attraction of the oligolectic bee Andrena vaga, which collects pollen only from Salix. Flower scent was collected by using dynamic-headspace methods from Salix caprea and S. atrocinerea, and the samples were subsequently analyzed by coupled gas chromatographic-electroantennographic detection (GC-EAD) to detect possible attractants of A. vaga. EAD active compounds were identified by gas chromatography coupled to mass spectrometry. Both Salix species had relatively similar scent profiles, and the antennae of male and female bees responded to at least 16 compounds, among them different benzenoids as well as oxygenated monoterpenoids and sesquiterpenoids. The strongest antennal responses were triggered by 1,4-dimethoxybenzene, and in field bioassays, this benzenoid attracted females of A. vaga at the beginning of its flight period, but not at the end.

Sex Pheromone of the Dogwood Borer, Synanthedon scitula

The sex pheromone of female dogwood borers (DWB) Synanthedon scitula (Harris) (Lepidoptera: Sesiidae) was determined to be an 88:6:6 ternary blend of (Z,Z)-3,13-octadecadienyl acetate (Z,Z-3,13-ODDA), (E,Z)-2,13-octadecadienyl acetate (E,Z-2,13-ODDA), and (Z,E)-3,13-octadecadienyl acetate (Z,E-3,13-ODDA) by gas chromatography-electroantennographic detection (GC-EAD) and gas chromatography-mass spectrometry (GC-MS). The major sex pheromone component, Z,Z-3,13-ODDA, was attractive as a single component. A blend of Z,Z-3,13-ODDA with 1-3% of E,Z-2,13-ODDA (binary blend) was more attractive than the single component. A third component, Z,E-3,13-ODDA, was sometimes observed in GC-EAD analyses, and enhanced attraction to the binary blend in some field bioassays. Lures containing 1 mg of binary and ternary blends attracted 18 and 28 times more male DWB moths, respectively, than caged virgin females in field trials. Attraction was strongly antagonized by addition of as little as 0.5% of E,Z-3,13-octadecadienyl acetate (E,Z-3,13-ODDA). In a period of 12 wk in 2004, more than 60,000 males were captured in sticky traps baited with synthetic pheromone blends in six apple orchards in Virginia, West Virginia, and North Carolina. Lure longevity trials showed that approximately 76% of the pheromone remained in rubber septum lures after 12 wk in the field.

Cuticular Hydrocarbons as Sex Pheromone of the Bee Colletes cunicularius and the Key to its Mimicry by the Sexually Deceptive Orchid, Ophrys exaltata

Male Colletes cunicularius bees pollinate the orchid, Ophrys exaltata, after being sexually deceived by the orchid's odor-mimicry of the female bee's sex pheromone. We detected biologically active volatiles of C. cunicularius by using gas chromatographic-electroantennographic detection (GC-EAD) with simultaneous flame ionization detection. After identification of the target compounds by coupled gas chromatography mass spectrometry (GC-MS), we performed behavioral tests using synthetic blends of the active components. We detected 22 EAD active compounds in cuticular extracts of C. cunicularius females. Blends of straight chain, odd-numbered alkanes and (Z)-7-alkenes with 21-29 carbon atoms constituted the major biologically active compounds. Alkenes were the key compounds releasing mating behavior, especially those with (Z)-7 unsaturation. Comparison of patterns of bee volatiles with those of O. exaltata subsp. archipelagi revealed that all EAD-active compounds were also found in extracts of orchid labella. Previous studies of the mating behavior in C. cunicularius showed linalool to be an important attractant for patrolling males. We confirmed this with synthetic linalool but found that it rarely elicited copulatory behavior, in accordance with previous studies. A blend of active cuticular compounds with linalool elicited both attraction and copulation behavior in patrolling males. Thus, linalool appears to function as a long-range attractant, whereas cuticular hydrocarbons are necessary for inducing short-range mating behavior.

Methyl Salicylate, a Soybean Aphid-Induced Plant Volatile Attractive to the Predator Coccinella septempunctata

Induced volatiles provide a signal to foraging predatory insects about the location of their prey. In Iowa, early in the growing season of soybean, Glycine max, many predacious seven-spotted lady beetles, Coccinella septempunctata, were observed on plants with heavy infestations of soybean aphid, Aphis glycines. We studied whether the attraction of this beetle is caused by the release of specific volatile compounds of soybean plants infested by aphids. Volatile compounds emitted by soybean plants infested by aphids were compared with those of undamaged, uninfested, and artificially damaged plants. Gas chromatography-mass spectrometry analyses revealed consistent differences in the profiles of volatile compounds between aphid-infested soybean plants and undamaged ones. Significantly more methyl salicylate was released from infested plants at both the V1 and V2 plant growth stages. However, release patterns of two other induced plant volatiles, (D)-limonene and (E,E)-alpha-farnesene, differed between the two plant growth stages. Gas chromatographic-electroantennographic detection of volatile extracts from infested soybean plants showed that methyl salicylate elicited significant electrophysiological responses in C. septempunctata. In field tests, traps baited with methyl salicylate were highly attractive to adult C. septempunctata, whereas 2-phenylethanol was most attractive to the lacewing Chrysoperla carnea and syrphid flies. Another common lady beetle, the multicolored Asian lady beetle, Harmonia axyridis, showed no preference for the compounds. These results indicate that C. septempunctata may use methyl salicylate as the olfactory cue for prey location. We also tested the attractiveness of some selected soybean volatiles to alate soybean aphids in the field, and results showed that traps baited with benzaldehyde caught significantly higher numbers of aphids.

Identification of the Sex Pheromone of the Swede Midge, Contarinia nasturtii

Coupled gas chromatographic-electroantennographic detection analyses of ovipositor extracts of calling Contarinia nasturtii females revealed two compounds that elicited responses from antennae of male midges. Using synthetic reference samples, these components were identified by gas chromatography mass spectrometry and enantioselective GC as (2S,9S)-diacetoxyundecane and (2S,10S)-diacetoxyundecane. In addition, trace amounts of 2-acetoxyundecane were found in ovipositor extracts, and the (S)-enantiomer was synthesized. When tested in the wind tunnel, a blend of 5 ng (2S,9S)-diacetoxyundecane and 10 ng (2S,10S)-diacetoxyundecane (mimicking the ratio found in the extracts) did not attract any of the males tested, but when 0.1 ng (S)-2-acetoxyundecane was added to the blend, 86.8% of the males were attracted to the bait. Three-component blends with lower or higher relative concentrations than 1% of (S)-2-acetoxyundecane [relative to (2S,10S)-diacetoxyundecane] were less attractive. In a field trapping experiment with released laboratory-reared C. nasturtii adults, traps baited with 500:1000:10 ng of (2S,9S)-diacetoxyundecane/(2S,10S)-diacetoxyundecane/(S)-2-acetoxyundecane applied to rubber septa or dental cotton rolls were tested. Traps without dispensers were used as controls. All three treatments were tested at 20 and 50 cm above ground. The estimated recapture rate was 30-50%, and 81.9% of the recaptured males were caught in traps positioned at 20 cm above ground, and 88.4% in traps with dental cotton rolls as dispensers.

(7Z,9E)-2-Methyl-7,9-Octadecadiene: A Sex Pheromone Component of Lymantria Bantaizana

Our objective was to identify the sex pheromone of Lymantria bantaizana (Lepidoptera: Lymantriidae) whose larvae feed exclusively on walnut, Juglans spp., in China, and Japan. Coupled gas chromatographic-electroantennographic detection (GC-EAD) analyses of pheromone gland extracts revealed a single EAD-active component. Retention index calculations of this compound on four GC columns suggested that it was a methyl-branched octadecadiene with conjugated double bonds. In GC-EAD analyses of 2-methyloctadecenes, (Z)-2-methyl-7-octadecene and (E)-2-methyl-7-octadecene elicited the strongest antennal responses, suggesting that the double bond positions were at C7 and C9. In comparative GC-EAD analyses of pheromone gland extract and stereoselectively synthesized isomers (E,E; E,Z; Z,E; Z,Z) of 2-methyl-7,9-octadecadiene, the (E,Z)- and (Z,E)-isomer had retention times identical to that of the candidate pheromone, but only the latter isomer elicited strong EAD activity. Results of field experiments in Japan substantiated that (7Z,9E)-2-methyl-7,9-octadecadiene is the L. bantaizana sex pheromone, a compound previously unknown in the Lepidoptera. Detection surveys in North America for exotic Eurasian forest defoliators could include traps baited with the L. bantaizana pheromone.

Sex Pheromones of Five Olethreutine Species (Lepidoptera: Tortricidae) Associated with the Seedlings and Fruits of Mangrove Plants in the Ryukyu Islands, Japan: Identification and Field Evaluation

The sex pheromones of three Cryptophlebia, one Centroxena, and one Eucosma species (Lepidoptera: Olethreutinae) inhabiting mangroves in the Ryukyu Islands, Japan, were studied with coupled gas chromatography-electroantennographic detection, and GC-mass spectrometry (GC-MS). The larvae of each Cryptophlebia species are specifically associated with viviparous seedlings from one of three mangrove Rhizophoraceae plants. Whereas three EAG-active alcohol components, (Z)-8-dodecen-1-ol, (E)-8-dodecen1-ol, and dodecan-1-ol, in a ratio of 100:12:4, were identified from the pheromone gland extract of female of C. horii (host: Bruguiera gymnorrhiza), two other sibling species produced the corresponding acetates, i.e., (Z)-8-dodecenyl acetate, (E)-8-dodecenyl acetate, and dodecyl acetate, in a 100:2:3 ratio from Cryptophlebia palustris (host: Rhizophora stylosa in Iriomote-jima Island) and in a 100:7:13 ratio from C. amamiana (host: Kandelia candel in Amami-oshima Island). The double bond positions of the monounsaturated components were confirmed by GC-MS analyses of their adducts with dimethyl disulfide. On the other hand, the larvae of Centroxena sp. feed on fruits of Sonneratia alba, another mangrove plant in the Sonneratiaceae, and the extract of the female pheromone glands contained (8E,10E)-dodecadienyl acetate and dodecyl acetate in a ratio of 100:5. The double bond position of the diunsaturated compound was confirmed by GC-MS analysis of its adduct with 4-methyl-1,2,4-triazoline-3,5-dione. (E)-9-Dodecenyl acetate w sively identified in the pheromone gland extract of Eucosma coniogramma females reared from seedlings of B. gymnorrhiza. Although the roles of minor components have not been revealed by field tests, synthetic lures baited with the main pheromone component of each species successfully attracted the target males, confirming that the sex pheromone is one of the most important factors for their reproductive isolation.

Flight Activity and Further Evidence for a Female-Produced Sex Pheromone of the Apple Leaf Midge, Dasineura mali, in Nova Scotia

The flight activity of the apple leaf midge, Dasineura mali (Kieffer), was determined in the field and laboratory. Males and females began emerging at the onset of the photophase. In the field, delta-shaped traps baited with 6 virgin females caught males exclusively, with the most abundant catches on the ground as compared to those at 1 and 2 m. Most males were captured at 1100 hours and males remained in the vegetation under the trees throughout the day. Sweep netting revealed that conspecifics mate in the vegetation under orchard trees in the morning hours, and females move from the ground to the orchard canopy to oviposit at 1000 hours. Gas chromatographic-electroantennographic detection (GC-EAD) analyses indicated the presence of an electrophysiologically active compound in solid phase microextraction (SPME) effluvial collections and pheromone gland extractions from virgin females.

(Z, Z)-6,9-Heneicosadien-11-One: Major Sex Pheromone Component Of Painted Apple Moth, Teia anartoides

(Z,Z)-6,9-Heneicosadien-11-one (Z6Z9-11-one-21Hy) was identified as the major sex pheromone component of the painted apple moth (PAM), Teia anartoides (Lepidoptera: Lymantriidae), on the basis of (1) comparative gas chromatographic-electroantennographic detection (GC-EAD) analyses, GC-mass spectrometry (MS), high-performance liquid chromatography (HPLC)-MS, and HPLC-UV/visible spectroscopy of pheromone gland extracts and authentic standards; (2) GC-EAD analyses of effluvia of calling females; and (3) wind tunnel and field trapping experiments with a synthetic standard. In field experiments in Australia, synthetic Z6Z9-11-one-21Hy as a single component attracted male moths. Wind tunnel experiments suggested that a 4-component blend consisting of Z6Z9-11-one-21Hy, (6Z,9R,10S)-cis-9,10-epoxy-heneicosene (Z6-9R10S-epo-21 Hy), (E,E)-7,9-heneicosadien-6, 11-dione (E7E9-6,11-dione-21Hy), and 6-hydroxy-(E,E)-7,9-heneicosadien-11-one (E7E9-6-ol-11-one-21Hy) (all present in pheromone gland extracts) might induce more males to orient toward, approach, and contact the source than did Z6Z9-11-one-21Hy as a single component. Additional experiments are needed to determine conclusively whether or not Z6-9R10S-epo-21Hy, E7E9-6,11-dione-21Hy, and E7E9-6-ol-11-one-21Hy might be minor sex pheromone components of PAM. Moreover, attractiveness of synthetic pheromone and virgin PAM females needs to be compared to determine whether synthetic pheromone could replace PAM females as trap baits in the program to monitor eradication of exotic PAM in New Zealand.

(7R,8S)-cis-7,8-EPOXY-2-METHYLOCTADEC-17-ENE: A NOVEL TRACE COMPONENT FROM THE SEX PHEROMONE GLAND OF GYPSY MOTH, Lymantria dispar

Considering the vast Eurasian distribution of gypsy moth, Lymantria dispar (Lepidoptera: Lymantriidae), the many subspecies, and their presence in different lymantriid communities, we tested the hypothesis that L. dispar populations in eastern Asia employ one or more pheromone components in addition to the previously known single component pheromone (7R,8S)-cis-7,8-epoxy-2-methyloctadecane [= (+)-disparlure]. Coupled gas chromatographic-electroantennographic detection (GC-EAD) analyses of pheromone gland extracts of female L. dispar sensu lato (including both AGM and NAGM) on four GC columns (DB-5, DB-23, DB-210, and SP-1000) revealed a new trace component that eluted just before (DB-5; DB-210) or after (DB-23, SP-1000) disparlure, and elicited strong antennal responses. Isolation of this compound by high-performance liquid chromatography and hydrogenation produced disparlure, suggesting that the new component had the molecular skeleton of disparlure, with one or more double bonds. Of all possible monounsaturated cis-7,8-epoxy-2-methyloctadecenes, only cis-7,8-epoxy-2-methyloctadec-17-ene co-chromatographed with the insect-produced compound on all GC columns and elicited comparable antennal responses. In field experiments in Honshu (Japan) with enantioselectively synthesized compounds, (7R,8S)-cis-7,8-epoxy-2-methyloctadec-17-ene (7R8S-epo-2me-17-ene-18Hy) was weakly attractive to male L. dispar, but was less effective as a trap bait than (+)-disparlure, and failed to enhance attractiveness of (+)-disparlure when tested in blends. The antipode, (7S,8R)-cis-7,8-epoxy-2-methyloctadec-17-ene, was not attractive, and when added to (+)-disparlure and/or 7R8S-epo-2me-17-ene-18Hy reduced attractiveness. Thus, the biological role of 7R8S-epo-2me-17-ene-18Hy remains unclear. It may enhance pheromone attractiveness or specificity in other L. dispar populations within their vast Eurasian distribution.

Mastrus ridibundus parasitoids eavesdrop on cocoon-spinning codling moth, Cydia pomonella, larvae

Cocoon-spinning larvae of the codling moth, Cydia pomonella L. (Lepidoptera: Olethreutidae) employ a pheromone that attracts or arrests conspecifics seeking pupation sites. Such intraspecific communication signals are important cues for illicit receivers such as parasitoids to exploit. We tested the hypothesis that the prepupal C. pomonella parasitoid Mastrus ridibundus Gravenhorst (Hymenoptera: Ichneumonidae) exploits the larval aggregation pheromone to locate host prepupae. In laboratory olfactometer experiments, female M. ridibundus were attracted to 3-day-old cocoons containing C. pomonella larvae or prepupae. Older cocoons containing C. pomonella pupae, or larvae and prepupae excised from cocoons, were not attractive. In gas chromatographic-electroantennographic detection (GC-EAD) analyses of bioactive Porapak Q extract of cocoon-derived airborne semiochemicals, ten compounds elicited responses from female M. ridibundus antennae. Comparative GC-mass spectrometry of authentic standards and cocoon-volatiles determined that these compounds were 3-carene, myrcene, heptanal, octanal, nonanal, decanal, (E)-2-octenal, (E)-2-nonenal, sulcatone, and geranylacetone. A synthetic 11-component blend consisting of these ten EAD-active compounds plus EAD-inactive (+)-limonene (the most abundant cocoon-derived volatile) was as effective as Porapak Q cocoon extract in attracting both female M. ridibundus and C. pomonella larvae seeking pupation sites. Only three components could be deleted from the 11-component blend without diminishing its attractiveness to M. ridibundus, which underlines the complexity of information received and processed during foraging for hosts. Mastrus ridibundus obviously "eavesdrop" on the pheromonal communication signals of C. pomonella larvae that reliably indicate host presence.

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.

Identification of a Sex Pheromone Component for the Blueberry Leafminer, Caloptilia porphyretica

Coupled gas chromatographic-electroantennographic detection (GC-EAD) of both gland extracts and effluvial collections from female blueberry leafminer, Caloptilia porphyretica Braun (Lepidoptera: Gracillariidae), showed that females produced a single EAD-active compound. The amount of the compound collected from virgin female C. porphyretica was below GC and mass spectrometry (MS) detection thresholds, even with highly concentrated gland extracts (approximately 150 female equivalent). (E)-11-Hexadecenal (E11-16:Ald) was determined to be a sex pheromone component mainly by comparison of retention times with authentic standards on both polar and nonpolar capillary columns, microreaction-GC-EAD analyses, and field trapping tests. GC-EAD experiments showed that synthetic E11-16:Ald exhibited extraordinarily high electrophysiological activity, stimulating significant male antennal responses at as low as 10 fg. Traps baited with E11-16:Ald alone were attractive to males. Addition of 1 or 3% of its geometric isomer, Z11-16:Ald, to E11-16:Ald did not significantly increase trap captures, but an inhibitory effect was observed at the 10% level. The influence of two kinds of rubber septa on attraction was also evaluated. Male moth captures were higher in traps baited with red rubber septa than with gray rubber septa at 30-300-microg doses. Monitoring of adult flight activity with 3-microg doses of E11-16:Ald indicated at least three distinct flight periods throughout the 2003 season.

SEX PHEROMONE OF MEALYBUG NAILED

THE TWO COMPOUNDS THAT make up the sex pheromone of female pink hibiscus mealybugs ( Maconellicoccushirsutus ) have been identified and synthesized by Agricultural Research Service (ARS) scientists [ Proc. Natl. Acad. Sci. USA , 101 , 9601 (2004)]. Native to the tropics, these sap-sucking insects are recent invaders of Florida and California. They cause $750 million per year in damages as they devour their way through avocados, citrus fruits, grapes, and other crops. The pheromone constituents could be the key to disrupting their mating, allowing this new pest to be managed. Chemist Aijun Zhang, working at an ARS lab in Beltsville, Md., and coworkers have shown that the pheromone consists of ( R )-lavandulyl ( S )-2-methylbutanoate (top) and ( R )-maconelliyl ( S )-2-methylbutanoate. Using a coupled gas chromatography-electroantennographic detection method, they fully characterized the esters. Maconelliol, the alcohol component of the second compound, is a cyclobutanoid monoterpene containing a methylethy...

Female sex pheromone of Ostrinia orientalis ? throwing a light on the relationship between O. orientalis and the European corn borer, O. nubilalis

The sex pheromone of Ostrinia orientalis (Lepidoptera: Crambidae) was analyzed by gas chromatography–electroantennographic detection (GC–EAD), GC–mass spectrometry and a series of bioassays. Three EAD-active compounds were detected in the female sex pheromone gland extract, and identified as tetradecyl acetate (14:OAc), (Z)-11-tetradecenyl acetate (Z11-14:OAc) and (E)-11-tetradecenyl acetate (E11-14:OAc). The titers (ratio) of 14:OAc, Z11-14:OAc and E11-14:OAc in 3-day-old virgin females were 0.49 ng (10), 4.86 ng (98) and 0.10 ng (2), respectively. In a wind-tunnel bioassay, the 98:2 blend of Z11- and E11-14:OAc, but not Z11-14:OAc alone, elicited the same male behavioral responses as virgin females and crude gland extracts. 14:OAc was inactive by itself, and did not show any synergistic effect on the binary blend. Field trapping experiments also confirmed the attractiveness of the binary blend to O. orientalis males. Based on these results, we concluded that the sex pheromone of O. orientalis is a 98:2 mixture of Z11-14:OAc and E11-14:OAc. This sex pheromone is very similar to that of the Z-type European corn borer, O. nubilalis. The present finding raises the question of whether O. orientalis , which is indistinguishable from O. nubilalis based on external morphology, is a biologically distinct species independent from O. nubilalis.

Antennal responses of four species of tree-killing bark beetles (Coleoptera: Scolytidae) to volatiles collected from beetles, and their host and nonhost conifers

Host selection in tree-killing bark beetles (Coleoptera: Scolytidae) is mediated by a complex of semiochemical cues. Using gas chromatographic-electroantennographic detection (GC-EAD) and GC-mass spectrometric analyses, we conducted a comparative study of the electrophysiological responses of four species of tree-killing bark beetles, the Douglas-fir beetle, Dendroctonus pseudotsugae, Hopkins, the mountain pine beetle, D. ponderosae Hopkins, the spruce beetle, D. rufipennis Kirby, and the western balsam bark beetle, Dryocoetes confusus Swaine, to volatiles captured by aeration of 1) bole and foliage of four sympatric species of conifers, Douglas-fir, Pseudotsuga menziesii (Mirb.) Franco, lodgepole pine, Pinus contorta var. latifolia Engelm., interior spruce, Picea engelmannii x glauca, and interior fir, Abies lasiocarpa x bifolia, and 2) con- and heterospecific beetles at three stages of attack. We identified 13 monoterpenes in the conifers and nine compounds in the volatiles of beetles that elicited antennal responses. There was no qualitative difference in the terpene constitution of the four species of conifers and very little difference across beetle species in their antennal response to compounds from conifers or beetles. The lack of species-specific major or minor components in conifers suggests that beetles would need to detect differences in the ratios of different compounds in conifers to discriminate among them. Attraction to hosts and avoidance of nonhost conifers may be accentuated by perception of compounds emitted by con- and heterospecific beetles, respectively. The 22 compounds identified are candidate semiochemicals with potential behavioural roles in host location and discrimination.

( Z,Z )-11,13-Hexadecadienyl acetate and ( Z,E )-11,13,15-hexadecatrienyl acetate: synergistic sex pheromone components of oak processionary moth, Thaumetopoea processionea (Lepidoptera: Thaumetopoeidae)

Our objective was to identify sex pheromone components of the oak processionary moth, Thaumetopoea processionea (Lepidoptera: Thaumetopoeidae), whose larvae defoliate oak, Quercus spp., forests in Eurasia and impact human health. Coupled gas chromatographic-electroantennographic detection (GC-EAD) and GC-mass spectrometric (MS) analyses of pheromone gland extract of female T. processionea revealed two consistently EAD-active compounds. They were identified as (Z,Z)-11,13-hexadecadienyl acetate (Z11,Z13-16:OAc) and (Z,E)-11,13,15-hexadecatrienyl acetate (Z11,E13,15-16:OAc) by comparative GC, GC-MS and GC-EAD analyses of insect-produced compounds and authentic standards. In replicated field experiments (2000, 2001) in Nordbaden, Sudbaden and Sachsen-Anhalt (Germany), Z11,Z13-16:OAc and Z11,E13,15-16:OAc in combination, but not singly, attracted significant numbers of male moths. It will now be intriguing to investigate whether Z11,E13,15-16:OAc, or its corresponding alcohol or aldehyde, serves as a pheromone component also in other species of the Thaumetopoeidae.

Antennal response of cotton bollworm (Heliocoverpa armigera) to volatiles in transgenic Bt cotton

Abstract: Volatile compositions in transgenic Bt cotton variety ‘GK‐97’ and its parental variety ‘Simian No. 3’ were analysed, and antennal response of cotton bollworm (Heliocoverpa armigera) to the volatiles were investigated. Volatile components in Bt cotton and regular cotton were almost same, except for absence of a sesquiterpene and a minor unknown compound in regular cotton. The results of gas chromatography‐electroantennographic detection (GC‐EAD) experiments showed that cotton bollworm antennae responded to seven compounds and two minor unknown compounds in Bt volatiles, among which concentrations of α‐pinene and β‐pinene in Bt cotton were relatively higher than those in regular cotton, and one minor unknown compound was absent in regular cotton.

Volatiles associated with preferred and nonpreferred hosts of the Nantucket pine tip moth, Rhyacionia frustrana.

Ovipositing female Nantucket pine tip moth, Rhyacionia frustrana, prefer loblolly pine, Pinus taeda L., to slash pine, Pinus elliottii Engelm, except during the first spring following planting of seedlings. Host discrimination by R. frustrana increases as seedlings develop, suggesting that changes in the chemical composition of seedlings may mediate the moth's host preferences. Volatile compounds from slash and loblolly pine seedlings were collected using solid-phase microextraction (SPME) during the first year following planting. Four collection periods coincided with adult emergence and oviposition during each of four annual generations of R. frustrana in the Georgia Coastal Plain. Infestation of slash pine peaked during the second tip moth generation and was similar to the loblolly pine infestation level. By the fourth tip moth generation, slash pine infestation levels had declined and diverged considerably from those of loblolly pine. Significant differences in relative quantities of beta-pinene, alpha-phellandrene, limonene, beta-phellandrene, bornyl acetate, beta-caryophyllene, and an unidentified sesquiterpene occurred between slash and loblolly pine during the fourth generation. However, no strong correlation was observed between any individual compound and host damage that could readily explain the temporal changes in R. frustrana host preference. Gas chromatographic-electroantennographic detection (GC-EAD) analyses of standards identified 19 different seedling-associated compounds that elicited antennal responses from R. frustrana females, indicating that a blend of terpenoids may mediate host discrimination.