Gas Chromatographic-electroantennographic Detection Research Articles

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.

(2R,7S)-diacetoxytridecane: sex pheromone of the aphidophagous gall midge, Aphidoletes aphidimyza.

In a recent study, evidence was presented that females of the aphidophagous midge Aphidoletes aphidimyza (Rondi) (Diptera: Cecidomyiidae) release a sex pheromone to attract mates. Our objectives were to identify and bioassay the pheromone. Coupled gas chromatographic-electroantennographic detection(GC-EAD) analyses of untreated and hydrogenated pheromone extract on three fused-silica columns (DB-5, DB-23, DB-210) revealed a single compound that elicited responses from male antennae. Retention index calculations of this candidate pheromone (CP) suggested that it was a di-acetate. Considering that most of the presently identified cecidomyiid pheromones consist of a 13-carbon chain with (at least) one acetate group in C2, we synthesized 2,6-, 2,7-, 2,8-, 2,9-, 2,10-, 2,11-, and 2,12-diacetoxytridecane. In GC analyses of these compounds, only 2,7-diacetoxytridecane cochomatographed with CP on all columns. In laboratory two-choice experiments with stereospecifically synthesized stereoisomers, only (2R,7S)-diacetoxytridecane elicited significant anemotatic responses by male A. aphidimyza. In trapping experiments in greenhouse compartments, only traps baited with (2R,7S)-diacetoxytridecane captured significant numbers of male A. aphidimyza, clearly revealing the absolute configuration of the pheromone. Failure of the stereoisomeric mixture (containing all four stereoisomers including the pheromone) to attract males is due to inhibitory characteristics of the (2R,7R)- and (2S,7R)-stereoisomers. The pheromone of zoophagous A. aphidimyza resembles those from phytophagous cecidomyiid midges, suggesting a common, diet-independent pathway for pheromone biosyntheses.

Equations for nickel–chromium wire heaters of column transfer lines in gas chromatographic–electroantennographic detection (GC–EAD)

Heating of chromatographic columns, transfer lines, and other devices is often required in neuroscience research. For example, volatile compounds passing through a capillary column of a gas chromatograph (GC) can be split, with half exiting the instrument through a heated transfer line to an insect antenna or olfactory sensillum for electroantennographic detector (GC–EAD) recordings. The heated transfer line is used to prevent condensation of various chemicals in the capillary that would otherwise occur at room temperature. Construction of such a transfer line heater is described using (80/20%) nickel–chromium heating wire wrapped in a helical coil and powered by a 120/220 V ac rheostat. Algorithms were developed in a computer program to estimate the voltage at which a rheostat should be set to obtain the desired heater temperature for a specific coil. The coil attributes (radius, width, number of loops, or length of each loop) are input by the user, as well as AWG size of heating wire and desired heater temperature. The program calculates total length of wire in the helix, resistance of the wire, amperage used, and the voltage to set the rheostat. A discussion of semiochemical isolation methods using the GC–EAD and bioassays is presented.

Stimulatory male volatiles for the Neotropical brown stink bug, Euschistus heros (F.) (Heteroptera: Pentatomidae)

Volatiles compounds collected from the male Neotropical brown stink bug, Euschistus heros (F.), a serious Central and South American soybean pest, have been reevaluated. The proportion of three methyl esters is found to be quite different from previous study. A new blend is proposed as the male-produced stimulatory volatiles based on gas chromatographic-electroantennographic detection (GC-EAD) techniques. The three GC-EAD-active components reproducibly found in volatiles collected from males are methyl (2E,4Z)-decadienoate (53%), methyl 2,6,10-trimethyldodecanoate (3%), and methyl 2,6,10-trimethyltridecanoate (44%) respectively. Males of this hemipteran species needed to reach an age of ~15 days to produce enough male-specific compounds to be detected by capillary column GC equipped with flame ionization detector (FID). The amounts of these three stimulatory volatiles increased with age and appeared to reach a maximum at ~35 days old, with the release rate of ~2.5 mug/day/male and the ratio of these three components seemed not to be affected by aging.

Electrophysiological responses of Thaumetopoea pityocampa females to host volatiles: implications for host selection of active and inactive terpenes

AbstractVolatiles from newly cut branches with needles of Pinus sylvestris L. were collected with headspace sampling technique, and then identified and quantified by combined gas chromatographic‐mass spectrometry (GC‐MS). The response of antennae of the female pine processionary moth, Thaumetopoea pityocampa, to these volatiles was recorded by coupled gas chromatographic‐electroantennographic detection (GC‐EAD). Surprisingly, the most common and major monoterpene hydrocarbons (MT), α‐pinene, 3‐carene, and β‐pinene were antennally inactive. Female antennae responded strongly only to four minor MT components, myrcene, β‐phellandrene, trans‐β‐ocimene, and terpinolene. Weaker, but repeatable responses were also found to limonene, cis‐β‐ocimene, and γ‐terpinene. Further EAD recordings with two synthetic MT mixtures supported our findings from the natural material. When separating the two enantiomers of limonene by running different synthetic MT mixtures, the EAD response was found only to the(−)‐enantiomer, but not to the opposite (+)‐enantiomer. EAD‐responses were also found to some less volatile compounds, such as sesquiterpenes (SqT), active at ng‐levels. The sensitivity and specificity of the antenna to a select number of active host MTs and SqTs suggest that these play a role in the host selection process of T. pityocampa females.

Sex pheromone components of male Tirathaba mundella (Lepidoptera: Pyralidae)

During peak calling activity by male oil palm bunch moths, Tirathaba mundella Walker (Lepidoptera: Pyralidae), their hairpencils, wings or entire body were extracted in hexane. Gas chromatographic-electroantennographic detection (GC-EAD) analyses of hair pencil extracts revealed four compounds that consistently elicited responses from female antennae. The NMR spectrum of isolated compound 1, and mass spectra and retention indices of compounds 1–4 suggested that they were (3S,6S)-2,2,6-trimethyl-6-vinyl-tetrahydro-pyran-3-ol (1), 4-hydroxy-3-methoxy-benzaldehyde (2, vanillin), 6,10,14-trimethyl-2-pentadecanone (3), and 6,10,14-trimethyl-2-pentadecanol (4). Comparative GC and GC-MS analyses of hair pencils extract and synthetic standards confirmed these structural assignments. Moreover, comparative chromatography of synthetic and hairpencil-isolated 1 on a Cyclodex-B column (which separated the four stereoisomers with baseline resolution) revealed that male T. mundella produce the SS-stereoisomer (SS-1). In field cage bioassay experiments in Palembang, Indonesia, synthetic SS-1 and vanillin in combination, but not singly, attracted female T. mundella. SS-1 plus vanillin were as effective as male T. mundella in attracting females. Compounds 3 and 4 did not enhance the blend's attractiveness.

Sex pheromone of the cranberry blossom worm, Epiglaea apiata.

The cranberry blossom worm, Epiglaea apiata (Grote) (Lepidoptera: Noctuidae), is a major pest of cranberries in New Jersey. The female sex pheromone of this moth was identified as a blend of (Z)-9-hexadecenyl acetate (Z9-16:Ac), (Z)-9-tetradecenyl acetate (Z9-14:Ac), and tetradecyl acetate (14:Ac) by gas chromatographic-electroantennographic detection and gas chromatography-mass spectrometry. The ratio of the components in extracts of the female pheromone gland was determined to be 65:2:33 of the Z9-16:Ac, Z9-14:Ac, and 14:Ac, respectively. The double bond positions of the pheromone components were confirmed by dimethyl disulfide derivatization. In addition to the above three components, a mixture of C4-C10 aliphatic acids was present in both gland extracts and effluvia collections, and the acids elicited significant EAD responses from male moth antennae. However, addition of the C4-C10 aliphatic acids to the pheromone blend did not significantly increase trap captures. Three-hundred- and 1000-microg doses of a synthetic blend containing Z9-16:Ac, Z9-14:Ac, and 14:Ac (65:2:33), on a rubber septum were more attractive to males than lower doses.

Antennal responses to floral scents in the butterfly Heliconius melpomene.

Floral scent, together with visual floral cues, are important signals to adult butterflies searching for food-rewarding plants. To identify which compounds in a floral scent are more attractive and, thus, of biological importance to foraging butterflies, we applied electrophysiological methods. Antennal responses of male and female adults of the tropical butterfly Heliconius melpomene L. (Lepidoptera: Nymphalidae: Heliconiinae) to individual compounds of natural floral scents and synthetic floral scent mixtures were investigated using gas chromatography-electroantennographic detection (GC-EAD). The natural floral scents included those of two tropical plant species, Lantana camara L. (Verbenaceae) and Warszewiczia coccinea (Vahl) Kl. (Rubiaceae), and two temperate species, Buddleja davidii Franchet (Loganiaceae) and Cirsium arvense (L.) Scop. (Asteraceae). The two synthetic floral scent mixtures contained many of the compounds found in the natural scents, but all in equal quantities. Compounds both present in relatively high abundance in the floral scents and detected exclusively in the floral parts of the plant, such as linalool, linalool oxide (furanoid) I and II, oxoisophoroneoxide, and phenylacetaldehyde, elicited the strongest antennal responses, suggesting that they may reflect adaptations by the plant to attract butterfly pollinators. However, other compounds also present in high abundance in the floral scent, but detected in the vegetative as well as floral plant parts, either elicited strong antennal responses, such as trans-beta-ocimene and benzaldehyde, or failed to elicit antennal responses, such as the sesquiterpenes beta-caryophyllene and alpha-humulene from L. camara. The most volatile monoterpene alkenes in the synthetic scent mixtures elicited only low or no responses. Furthermore, the overall antennal responses were stronger in females than in males. The findings suggest that several floral scent volatiles, especially those of exclusively floral origin, are of high biological significance to H. melpomene butterflies. These include compounds of different biosynthetic origins belonging to the benzenoids, monoterpenoids, and irregular terpenoids.

Enantiomers of (Z,Z)-6,9-heneicosadien-11-ol: sex pheromone components of Orgyia detrita.

(6Z,9Z,11S)-6,9-Heneicosadien-11-ol (Z6Z9-11S-ol-C21) and (6Z,9Z,11R)-6,9-heneicosadien-11-ol (Z6Z9-11R-ol-C21) were identified as major sex pheromone components of female tussock moths, Orgyia detrita Guérin-Méneville (Lepidoptera: Lymantriidae), on the basis of (1) analyses of pheromone gland extracts of female O. detrita by coupled gas chromatographic-electroantennographic detection (GC-EAD) and GC mass spectrometry, and (2) field trapping experiments with synthetic standards. Z6Z9-11S-ol-C21 and Z6Z9-11R-ol-C21 in combination, but not singly, attracted significant numbers of male moths. Racemic Z6Z9-11-ol-C21 was more attractive than the 1:3.5 (R:S) blend ratio found in pheromone gland extracts from female moths. Lower and higher homologues of Z6Z9-11-ol-C21 were also detected in GC-EAD recordings of pheromone extracts, and the racemic compounds enhanced attractiveness of Z6Z9-11-ol-C21 in field experiments. Because of trace amounts of these homologues in extracts, their enantiomeric composition could not be determined. This is the first report of secondary alcohols as pheromone components in the ditrysian (advanced) Lepidoptera.

Identification of odors from overripe mango that attract vinegar flies, Drosophila melanogaster.

Bioassays with a variety of overripe fruits, including mango, plum, pear, and grape, and their extracts showed that odors from overripe mango were most attractive to adult vinegar flies, Drosophila melanogaster. Combined gas chromatography-electroantennographic detection (GC-EAD) analyses of solid-phase microextraction (SPME) and Tenax extracts of overripe mango odors showed that several volatile compounds, including ethanol, acetic acid, amyl acetate, 2-phenylethanol, and phenylethyl acetate elicited significant EAG responses from antennae of female flies. Most of the volatile compounds in the extracts were identified by mass spectral and retention index comparisons with synthetic standards. In cage bioassays, lures with a blend of ethanol, acetic acid, and 2-phenylethanol in a ratio of 1:22:5 attracted six times more flies than any single EAG-active compound. This blend also attracted four times more flies than traps baited with overripe mango or unripe mango. However, in field trials, the blend was not as attractive as suggested by the laboratory bioassay.

Responses of Glossina morsitans morsitans to blends of electroantennographically active compounds in the odors of its preferred (buffalo and ox) and nonpreferred (waterbuck) hosts.

In a previous study, comparison of the behavior of teneral Glossina morsitans morsitans on waterbuck, Kobus defassa (a refractory host), and on two preferred hosts, buffalo, Syncerus caffer, and ox, Bos indicus, suggested the presence of allomones in the waterbuck odor. Examination of the volatile odors by coupled gas chromatography-electroantennographic detection showed that the antennal receptors of the flies detected constituents common to the three bovids (phenols and aldehydes), as well as a series of compounds specific to waterbuck, including C8-C13 methyl ketones, delta-octalactone, and phenols. In this study, behavioral respones of teneral G. m. morsitans to different blends of these compounds were evaluated in a choice wind tunnel. The flies' responses to known or putative attractant blends (the latter comprising EAG-active constituents common to all three animals and those common to buffalo and ox, excluding the known tseste attractants, 4-methylphenol and 3-n-propylphenol), and to putative repellent (the blend of EAG-active compounds specific to the waterbuck volatiles), were different. A major difference related to their initial and final behaviors. When a choice of attractant blends (known or putative) and clean air was presented, flies initially responded by flying upwind toward the odor source, but later moved downwind and rested on either side of the tunnel, with some preference for the side with the odor treatments. However, when presented with a choice of waterbuck-specific blend (putative repellent) and clean air, the flies' initial reaction appeared random; flies flew upwind on either side, but eventually settled down on the odorless side of the tunnel. Flies that flew up the odor plume showed an aversion behavior to the blend. The results lend further support to previous indications for the existence of a tsetse repellent blend in waterbuck body odor and additional attractive constituents in buffalo and ox body odors.

(Z,Z)-4,7-tridecadien-(S)-2-yl acetate: sex pheromone of Douglas-fir cone gall midge, Contarinia oregonensis.

Our objectives were to identify and field test the sex pheromone of female Douglas-fir cone gall midge, Contarinia oregonensis (Diptera: Ce-cidomyiidae). Coupled gas chromatographic-electroantennographic detection (GC-EAD) analyses of pheromone extract revealed a single compound (A) that elicited responses from male antennae. Hydrogenation of pheromone extract, followed by renewed GC-EAD analysis, revealed a new EAD-active compound with chromatographic characteristics identical to those of tridecan-2-yl acetate on five fused silica columns (DB-5, DB-210, DB-23, SP-1000, and Cyclodex-B). Syntheses, chromatography, and retention index calculations of all possible tridecen-2-yl acetates suggested that the candidate pheromone A was a tridecadien-2-yl acetate with nonconjugated double bonds. Synthetic candidate pheromone component (Z,Z)-4,7-tridecadien-2-yl acetate (Z4Z7) cochromatographed with A on all analytical columns and elicited comparable antennal activity. In GC-EAD analyses that separated the enantiomers (Z,Z)-4,7-tridecadien-(S)-2-yl acetate (2S-Z4Z7) and (Z,Z)-4,7-tridecadien-(R)-2-yl acetate (2R-Z4Z7) with baseline resolution, only 2S-Z4Z7 as a component in a racemic standard or in pheromone extract elicited antennal responses. In Douglas-fir seed orchards, sticky traps baited with 2S-Z4Z7 captured male C. oregonensis, whereas 2R-Z4Z7 was behaviorally benign. Comparable catches of males in traps baited with racemic Z4Z7 (50 microg) or virgin female C. oregonensis suggested that synthetic pheromone baits could be developed for monitoring C. oregonensis populations in commercial Douglas-fir seed orchards.

Evidence for a compound in Comstock-Kellog glands modulating premating behavior in male desert locust, Schistocerca gregaria.

The Comstock-Kellog glands in adult females of certain acridid species, including the desert locust, Schistocerca gregaria have been implicated as a source of volatiles that play a role in mating behavior. A dichloromethane extract of the glands was analyzed for metabolites by gas chromatography, coupled gas chromatography-electroantennographic detection, and mass spectrometry. Coupled gas chromatography-electroantennographic detector (GC-EAD) analysis revealed a component that elicited an electroantennogram response from the antenna of adult male S. gregaria. The compound was identified by GC-MS as pentanoic acid. The levels of the compound in the gland extract varied with age of female locust; it was present in detectable amounts only in 14- to 16-day-old females. In bioassays, pentanoic acid significantly stimulated pre-mating behavior in male desert locust. These results are discussed in relation to the biology of the locust.

Enantiospecific antennal response of bark beetles to spiroacetal (E)-conophthorin.

Optically pure synthetic enantiomers of (E)-conophthorin [(E)-7-methyl-1,6-dioxaspiro[4.5]decane], one of the volatiles affecting coniferophagous bark beetles, were tested on antennae of Ips typographus, I. duplicatus, I. subelongatus, Dendroctonus micans, and five Scolytus spp. by using combined gas chromatography-electroantennographic detection (GC-EAD). EAD dose-responses indicated that all three Ips species perceived only the naturally occurring and quantitatively dominant (5S,7S)-enantiomer, while its antipode, (5R,7R)-(E)-conophthorin was antennally inactive. Response thresholds for the Ips species were estimated as between 0.1 ng and 1 ng, or lower. The antennal responses of the Ips species caused by 100 ng of the (5R,7R)-enantiomer might be due to 1% impurity-(the active (5S,7S)-enantiomer) in the (5R,7R)-sample. At the 50-ng level, D. micans and five angiosperm Scolytus species (S. intracatus, S. mali, S. ratzeburgi, S. rugulosus, and S. scolytus) responded strongly to the (5S,7S)-enantiomer, while the (5R,7R)-enantiomer was antennally inactive. Currently updated knowledge on the natural occurrence, and electrophysiological and behavioral activity of (E)-conophthorin is summarized.

Pheromone components of the wheat stem sawfly: identification, electrophysiology, and field bioassay.

Volatiles collections and cuticular extracts of the wheat stem sawfly, Cephus cinctus Norton (Hymenoptera: Cephidae), were analyzed by coupled gas chromatographic-electroantennographic detection (GC-EAD) in order to detect possible pheromone components of this species. Volatiles collections from male and female sawflies contained the same 13 GC-EAD-active compounds that stimulated both male and female antennae. GC-EAD-active compounds were identified by mass spectrometry, and the identities were verified by acquiring or synthesizing authentic standards with identical GC retention times and mass spectra. 9-Acetyloxynonanal gave the strongest EAD response. Additional GC-EAD-active compounds included 13-acetyloxytridecanal, aldehydes with 9-16 carbon chain lengths, acids with 8-10 carbon chain lengths, and phenylacetic acid. The last was instantly detectable by human nose when several males were allowed to interact, but not from isolated insects. On a per-insect basis, grouped males overall released higher amounts of the GC-EAD-active compounds than grouped females. Notable quantitative differences between the sexes were relatively higher amounts of 9-acetyloxynonanal, phenylacetic acid, and tetradecanal from males and hexadecanal from females. In the absence of an absolute, qualitative difference between sexes, these relative differences might be important for sex recognition. Field bioassay demonstrated the attractiveness of 9-acetyloxynonanal for male and female C. cinctus. The major compounds of male and female cuticular extracts showed no GC-EAD activity, but ozonolysis of extracts gave dramatically increased amounts of GC-EAD-active material. Hence, double-bond-containing cuticular compounds are suggested as precursors of the aldehydic pheromone components.

2-Methyl-(Z)-7-octadecene: sex pheromone of allopatric Lymantria lucescens and L. serva.

Our objective was to identify the sex pheromone of Lymantria lucescens and Lymantria serva (Lepidoptera: Lymantriidae), whose larvae defoliate, respectively, Quercus spp. in temperate regions and Ficus spp. in the subtropics. Coupled gas chromatographic-electroantennographic (GC-EAD) detection analyses of pheromone gland extracts revealed one EAD active compound produced by female L. lucescens and by female L. serva. This was identified as 2-methyl-(Z)-7-octadecene (2me-Z7-18Hy) by retention index calculations on DB-5, DB-23, and DB-210 columns and by comparative GC-mass spectrometric (MS) and GC-EAD analyses of the insect-produced candidate pheromone and synthetic 2me-Z7-18Hy. In field experiments, traps baited with 2me-Z7-18Hy captured male L. lucescens near Toyota City, Japan, and male L. serva in Taipei, Taiwan. Allopatric distribution of L. lucescens and L. serva seems to allow both species to use the same sex pheromone without compromising its specificity.

Communication ecology of webbing clothes moth: 4. Identification of male- and female-produced pheromones

We investigated the hypothesis that aggregation signals produced by male webbing clothes moths (WCM), Tineola bisselliella (Hum.) (Lepidoptera: Tineidae), and close-range male attractant signals produced by females have a pheromonal basis, at least in part. Gas chromatographic-electroantennographic detection (GC-EAD) and GC-mass spectrometric analyses of bioactive methanolic extracts of male WCM disclosed three candidate pheromone components: hexadecanoic acid methyl ester (16:Ester), (Z)-9-hexadecenoic acid methyl ester (Z9—16:Ester), and octadecanoic acid methyl ester (18:Ester). In bioassay experiments in a large Plexiglas™ arena, a blend of synthetic 16:Ester plus Z9—16:Ester was attractive to male and virgin (but not mated) female WCM; the 18:Ester was inactive. GC-EAD analyses of pheromone gland extracts from female WCM revealed (E,Z)-2,13-octadecadienal (E2Z13—18:Ald) and (E,Z)-2,13-octadecadienol (E2Z13—18:OH) as candidate sex pheromone components. In arena bioassay experiments, 1—5 female equivalents of synthetic E2Z13—18:Ald (0.2 ng) and E2Z13—18:OH (0.1 ng) were more attractive to male WCM than were two virgin female WCM. We anticipate that the combination of aggregation and sex pheromones, male-produced sonic aggregation signals, and habitat-derived semiochemicals will be highly effective in attracting male and female WCM to commercial traps.

Volatile attractants for three Pteromalid parasitoids attacking concealed spruce bark beetles

The odour perceptive abilities, and preferences, of three bark beetle parasitoid species; Rhopalicus tutela (Walker), Roptrocerus mirus (Walker), and Roptrocerus xylophagorum (Ratzeburg) (Hymenoptera: Pteromalidae), were investigated to isolate and identify the essential compounds involved in host location. These parasitoids attack several economically important bark beetle species and oviposit preferentially on late larval stages concealed under the bark of conifers. Odours were collected from Norway spruce logs (Picea abies L. Karst.) containing Ips typographus L. (Coleoptera: Scolytidae) larvae. Biologically active compounds were isolated by coupled gas chromatographic-electroantennographic detection (GC-EAD), and identified by GC-mass spectrometry (GC-MS). Based on these analyses, four different synthetic baits were prepared and tested in a Y-tube walking bioassay. In the complex odour samples from spruce logs, only 16 compounds were EAD-active. The tested R. tutela and R. mirus females displayed similar trends in antennal activity to EAD-active compounds, responding mainly to oxygenated monoterpenes that indicate damaged conifers. Consequently, the synthetic baits were exclusively prepared with oxygenated monoterpenes. Parasitoid females (R. tutela and R. mirus) preferred spruce logs containing susceptible hosts over fresh logs, while male parasitoids (R. mirus) did not show any preference. However, when odours from fresh logs were mixed with synthetic baits (mimicking the odour composition of logs containing susceptible hosts), these combinations attracted female parasitoids (R. tutela, R. mirus, and R. xylophagorum). All synthetic baits seemed to be equally attractive to female parasitoids.

Olfactory responses of Ips duplicatus from inner Mongolia, China to nonhost leaf and bark volatiles.

Leaf and bark volatiles from nonhost angiosperm trees were tested on Ips duplicatus by gas chromatographic-electroantennographic detection (GC-EAD) and by pheromone-baited traps in Sweden and Inner Mongolia, China, respectively. GC-EAD analysis of the headspace volatiles from fresh bark chips of Betula pubescens revealed trans-conophthorin, two green leaf volatiles (GLVs): 1-hexanol and (Z)-3-hexen-1-ol, and two C8 alcohols: 3-octanol and 1-octen-3-ol, that consistently elicited antennal responses by I. duplicatus. The identification of these EAD-active compounds was confirmed in further GC-EAD recordings with synthetic mixtures. Antennal responses were also found to synthetic (E)-2-hexen-1-ol and linalool, which have been identified from the leaves of nonhost birch and aspen species. No antennal responses of I. duplicatus were found to hexanal, (E)-2-hexenal, and (Z)-3-hexyl acetates. In field trapping experiments, blends of EAD-active green leaf alcohols or C8 alcohols, or transconophthorin alone resulted in significant reductions (27-60%) in the number of I. duplicatus captured compared with pheromone-baited traps. The unsuitable host compound, verbenone (Vn), also significantly reduced trap catches by up to 60% in both experiments. The strongest disruptive effect resulted from the addition of the combination of green leaf alcohols, C8 alcohols, and verbenone to the pheromone trap, which caused an 84% reduction in trap catch. The blend of two green leaf aldehydes plus the acetate increased the trap catches in 1998 and had no negative or positive effects in 1999. Our results suggest that these nonhost volatiles (NHVs) are important olfactory signals used by I. duplicatus in host selection. They may have great significance in developing semiochemical-based management programs for I. duplicatus by reducing or stopping attacks on suitable hosts.