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

Summary. 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.     

Two phylogenetically distinct species of sexually deceptive orchids mimic the sex pheromone of their single common pollinator, the cuckoo bumblebee Bombus vestalis

Sexually deceptive orchids of the genus Ophrys attract male insects for pollination. Pollinator attraction is achieved by mimicking sex pheromones of virgin females of their pollinators, mostly bee species. In earlier investigations, we showed that the phylogenetically distinct Ophrys species O. chestermanii and O. normanii on Sardinia attract their pollinator, males of the cuckoo bumblebee B. vestalis, with the same bouquets of relatively polar volatile compounds. In this investigation, we studied the sex pheromone of virgin females of B. vestalis with the aim of identifying male-attracting compounds and of comparing them with labellum extracts of the two orchids, which were found to release male-attracting compounds in earlier investigations (G?gler et al. 2009). In bioassays, shock-frozen females, cuticle extracts and polar fractions of cuticle extracts of virgin females stimulated mating behaviour in the males. Using gas chromatography coupled with electroantennography (GC-EAD) and gas chromatography coupled with mass spectrometry (GC-MS), we detected in polar fractions of cuticle extracts of B. vestalis females the same electrophysiologically active compounds as in labellum extracts of both orchid species, including aldehydes, esters, fatty acids and alcohols. Since statistical comparisons of the relative proportions of esters showed strong similarities between virgin females and orchids, our results support the hypotheses that this highly specialized Ophrys–pollinator relationship represents another case of chemical mimicry and that esters play a key role in male attraction.     

Is flower scent influencing host plant selection of leaf-galling sawflies (Hymenoptera, Tenthredinidae) on willows?

Though it is known that flower scent not only attracts pollinators but also herbivores, little is known about the importance of flower scent on the distribution of leaf herbivores among individuals within a plant species. In this study we determined the distribution of galls induced by the sawfly Pontania proxima (Serville 1823) (Hymenoptera, Tenthredinidae, Nematinae) on flowering and non-flowering representatives of several clones belonging to Salix fragilis and S. × rubens (Salicaceae). Further, amounts and composition of scent of flowering and non-flowering twigs were compared (dynamic headspace-gas chromatography–mass spectrometry, DHS-GC–MS), and a scent sample collected from flowering twigs of S. fragilis was tested by coupled gas chromatography and electroantennographic detection (GC-EAD) on the antennae of P. proxima females. The results show that the presence of flower catkins on plants led to a higher degree of allocation with galls, but the number of galls differed not between flowering and non-flowering plants. The DHS-GC–MS analyses revealed that the total amount of flower scent emitted per flowering twig is about 90 times higher than the scent emitted by a non-flowering twig. Further, several compounds were emitted only by flowering but not by non-flowering twigs. In the GC-EAD analyses, antennae consistently responded not only to green leaf volatiles, but also to compounds emitted only by the flowers (e.g. 1,4-dimethoxybenzene). These flower scent compounds are suggested to affect the host plant choice by attracting more sawflies from the distance to flowering plants compared to non-flowering plants. The EAD-active compounds emitted from vegetative plant parts are assumed to act as long-distance signals especially when flowers are absent on host plants, e.g. during the oviposition period of the second generation of P. proxima.     

Volatiles from the bark of trembling aspen, Populus tremuloides Michx. (Salicaceae) disrupt secondary attraction by the mountain pine beetle, Dendroctonus ponderosae Hopkins (Coleoptera: Scolytidae)

Summary. Coupled gas chromatographic-electroantennographic detection (GC-EAD) analysis of the Porapak Q-captured volatiles from the bark of trembling aspen, Populus tremuloides Michx., revealed four compounds that consistently elicited antennal responses by mountain pine beetles (MPBs), Dendroctonus ponderosae Hopkins. One of these, 1-hexanol, disrupted the capture of MPBs in multiple-funnel traps baited with the aggregation pheromones trans-verbenol and exo-brevicomin and the host kairomone myrcene, a blend of semiochemicals that mediates the secondary attraction response in which beetles mass attack and kill living pines. The other three EAD-active aspen bark volatiles, benzyl alcohol, benzaldehyde and nonanal, were inactive alone, but in binary and ternary combinations contributed to a disruptive effect in an additive and redundant manner when all four aspen bark volatiles were tested in all possible binary and ternary blends. The best ternary blend and the quarternary blend achieved ≥ 80% disruption. The quarternary blend enhanced the disruptive effect of the antiaggregation pheromone verbenone in traps, raising the disruptive effect to 98%, and also enhanced the inhibition of attack on attractant-baited lodgepole pines. This is the first demonstration of specific compounds from the bark of angiosperm trees that disrupt the secondary attraction response of sympatric coniferophagous bark beetles. The results support the hypothesis that such bark beetles are adapted to recognize and avoid non-host angiosperm trees by responding to a broad spectrum of volatiles that can act in various blends with equal effect. Received 27 October 1997; accepted 20 February 1998.     

Wind tunnel attraction of grapevine moth females, Lobesia Botrana, to natural and artificial grape odour

Summary. Host plant volatiles which attract insect herbivores for egg-laying are of principal interest with respect to insect ecology and evolution. Direct applications concern population monitoring and control through behavioural manipulation. Identification of behaviourally active plant secondary metabolites is essential also for plant breeding for insect resistance. Grapevine moth females Lobesia botrana are attracted by upwind flight to green grape berries Vitis vinifera. The headspace of grape berries was collected on air filters. A solvent extract of these filters, released from a sprayer, attracted females in the wind tunnel. The results demonstrate that volatile cues mediate attraction of grapevine moth females to grape berries, and that headspace collections capture the essence of this odour signal. The air filter extracts were examined by gas chromatography coupled with electroantennographic detection, and the compounds eliciting a consistent antennal response in L. botrana females were identified by mass spectrometry. The headspace collection apparatus was calibrated for collection efficiency of the active compounds. Their recovery rate ranged from 35 % for methyl salicylate to 83 % for (E,E)-α-farnesene. A synthetic ten-component blend was then formulated. The blend consisted of compounds eliciting an antennal response, formulated in a blend ratio corrected for differences in collection efficiency. Subsequent wind tunnel tests showed that female attraction to this synthetic ten-component blend was not significantly different from attraction to grape berries, or to headspace collections of the same berries. At a release rate of 35 ng/h of the most abundant compound (E)-β-caryophyllene, 20 % of the test females approached the source of sprayed headspace collections and the ten-component synthetic blend, respectively. In comparison, 100 g of green berries, releasing the main compound (E)-β-caryophyllene at a rate of ca. 4.7 ng/h, attracted 10 % of the females by upwind flight followed by source contact.     

Antennal responses to floral scents in the butterflies <Emphasis Type="Italic">Inachis io</Emphasis>, <Emphasis Type="Italic">Aglais urticae</Emphasis> (Nymphalidae), and <Emphasis Type="Italic">Gonepteryx rhamni</Emphasis> (Pieridae)

Summary. To better understand the biological role of floral scents for butterflies, electrophysiological responses to floral scents were investigated using combined gas chromatography and electroantennographic detection (GC-EAD). The antennal responses of three butterfly species, Aglais urticae L. (Nymphalidae), Inachis io L. (Nymphalidae), and Gonepteryx rhamni L. (Pieridae) to floral scent compounds from both natural and synthetic mixtures were examined. Floral scents were collected from the butterfly nectar plants Cirsium arvense (L.) (Asteraceae), and Buddleja davidii Franchet cv. (Loganicaeae) with dynamic head-space methods on Tenax-GR and eluted with pentane. These eluates, composed of natural floral scent blends, represent an array of compounds in their natural state. In the GC-EAD analyses eleven compounds were identified from C. arvense with the benzenoid compound phenylacetaldehyde in highest abundance. Seventeen compounds were identified from B. davidii with the irregular terpene oxoisophorone in highest abundance. Thirty-nine synthetic floral scent compounds were mixed in pentane, in equal amounts; about 35 ng were allowed to reach the antennae. The butterflies showed antennal responses to most of the floral scent compounds from both natural and synthetic blends except to the highly volatile monoterpene alkenes. Certain benzenoid compounds such as phenylacetaldehyde, monoterpenes such as linalool, and irregular terpenes such as oxoisophorone, were emitted in relatively large amounts from C. arvense and B. davidii, and elicited the strongest antennal responses. These compounds also elicited strong antennal responses when present in the synthetic scent blends. Thus, the butterflies seem to have many and /or sensitive antennal receptors for these compounds, which points to their biological importance. Moreover, these compounds are exclusively of floral scent origin. For B. davidii, which depends highly on butterflies for pollination, the exclusive floral scent compounds emitted in high abundance could be the result of an adaptive pressure to attract butterflies. Received 2 Septemter 2001; accepted 9 September 2002.     

Antennal electrophysiological responses of the giant swallowtail butterfly, Papilio cresphontes, to the essential oils of Zanthoxylum clava-herculis and related plants

We used the electroantennogram (EAG) technique to compare the antennal sensitivity of both sexes of the giant swallowtail butterfly, Papilio cresphontes to four doses (1, 10, 100, and 1,000 μg) of the leaf essential oils of Zanthoxylum clava-herculis and Ptelea trifoliata (key host plants) and Sassafras albidum (a marginal or non-host plant). The main hypothesis tested was that P. cresphontes will show greater olfactory sensitivity to volatiles of the key host plants than to volatiles of the marginal host plant, in particular at low doses. At the lower doses, extract of the key host plant, Z. clava-herculis elicited greater EAG responses in both sexes than extracts of the remaining two plants. At higher doses, however, extracts of P. trifoliata and S. albidum elicited greater EAG responses than extract of Z. clava-herculis. These results partly support our hypothesis and may suggest that Z. clava-herculis is a more preferred host plant of P. cresphontes than P. trifoliata. In general, female butterflies showed greater EAG responses than males to the three plant extracts at the higher doses. Preliminary coupled gas chromatography-electroantennogram (GC-EAD) tests revealed four components each from Z. clava-herculis and P. trifoliata (three peaks common to both extracts) and seven from S. albidum (one shared with Z. clava-herculis) which elicited GC-EAD activity in P. cresphontes females, but the peaks were un-indentified because most were detected in trace amounts. In addition, the chemical composition of the leaf essential oil of Z. clava-herculis was analyzed by GC–MS. The leaf essential oils consisted of 25 components, largely menthane monoterpenoids, dominated by limonene and 1,8-cineole, but neither of the two major components elicited significant GC-EAD response in P. cresphontes. These results are discussed in relation to host-plant selection in P. cresphontes.     

Behavioral and electrophysiological responses of the emerald ash borer, Agrilus planipennis, to induced volatiles of Manchurian ash, Fraxinus mandshurica

Summary. We investigated the volatile emissions of Manchurian ash seedlings, Fraxinus mandshurica, in response to feeding by the emerald ash borer, Agrilus planipennis, and to exogenous application of methyl jasmonate (MeJA). Feeding damage by adult A. planipennis and MeJA treatment increased volatile emissions compared to unexposed controls. Although the same compounds were emitted from plants damaged by beetles and treated with MeJA, quantitative differences were found in the amounts of emissions for individual compounds. Adult virgin female A. planipennis were similarly attracted to volatiles from plants damaged by beetles and those treated with MeJA in olfactometer bioassays; males did not respond significantly to the same volatiles. Coupled gas chromatographic-electroantennogram detection (GC-EAD) revealed at least 16 antennally-active compounds from F. mandshurica, including: hexanal, (E)-2-hexenal, (Z)-3-hexen-1-ol, 3-methyl-butylaldoxime, 2-methyl-butylaldoxime, (Z)-3-hexen-1-yl acetate, hexyl acetate, (E)-β-ocimene, linalool, 4,8-dimethyl-1,3,7-nonatriene, and E,E-α-farnesene. Electroantennogram (EAG) dose–response curves using synthetic compounds revealed that females had a stronger EAG response to linalool than males; and male responses were greater to: hexanal, (E)-2-hexenal, (Z)-3-hexen-1-ol, 3-methyl-butylaldoxime, 2-methyl-butylaldoxime, and hexyl acetate. These results suggest that females may use induced volatiles in long-range host finding, while their role for males is unclear. If attraction of females to these volatiles in an olfactometer is upheld by field experiments, host plant volatiles may find practical application in detection and monitoring of A. planipennis populations.     

Identification of polyenic hydrocarbons from the northern winter moth, Operophtera fagata, and development of a species specific lure for pheromone traps

Summary. In order to elucidate the composition of the female sex pheromone of the northern (beech) winter moth, Operophtera fagata Scharf. (Lepidoptera: Geometridae), ovipositor extracts of unmated, calling females were analysed by gas chromatography with simultaneous electroantennographic and flame ionization detection (GC-EAD/FID). Male antennal responses indicated three active components, two of which had distinct matching peaks in the FID trace. Using coupled gas chromatography- mass spectrometry (GC-MS), these two compounds were identified as (9Z)-nonadecene (9Z-19:Hy), and (6Z,9Z)-nonadecadiene (6Z9Z-19:Hy), respectively. The third component, present in very small amounts only, was identified as (1,3Z,6Z,9Z)-nonadecatetraene (1,3Z6Z9Z-19:Hy), known as the sex pheromone of the common winter moth, O. brumata. Field tests revealed that traps baited with 6Z9Z-19:Hy and 1,3Z6Z9Z-19:Hy caugth large numbers of male O. fagata. Both compounds were found to be essential for attraction of O. fagata. In addition, the diene prevented captures of co-occurring O. brumata. In contrast, 9Z-19:Hy neither influenced the attractiveness of the two-component mixture towards O. fagata nor contributed to bait specificity. A binary mixture of 6Z9Z-19:Hy and 1,3Z6Z9Z-19:Hy in a ratio of 10:1, applied to pieces of rubber tubing, constituted a highly attractive and species-specific bait for O. fagata, which can be used for monitoring of the flight of this defoliator pest of deciduous forests.     

Volatile attractants for three Pteromalid parasitoids attacking concealed spruce bark beetles

Summary. 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. Received 12 October 2000; accepted 18 January 2001     

Scent marking pheromones in lizards: cholesterol and long chain alcohols elicit avoidance and aggression in male Acanthodactylus boskianus (Squamata: Lacertidae)

Femoral gland secretions are believed to play an important role in chemical communication and social organization of lizards. In spite of this, few studies have investigated the chemical composition and the behavioural roles of these secretions. The lacertid lizard Acanthodactylus boskianus is a good example, having these well-developed glands in both sexes. We used GC–MS chemical analysis of gland secretions and y-maze choice test bioassays to investigate the ability of the lizards to detect and respond to different synthetic blends made from compounds identified in the gland secretions. Based upon the GC–MS quantification data, we selected representatives of the main chemical groups (steroids, alcohols, acids, alkanes) detected in the lizard secretions and used these in a behavioural bioassay against controls. Males showed significant avoidance behaviour for cholesterol and alcohol blends, combined with agonistic behaviour towards these stimuli. Females did not show any significant selection to particular odour combinations. The data support the hypotheses that lizards can potentially use femoral gland secretions in chemical odour trails and utilize scent to mark territories and potentially also to establish dominance hierarchies. Cholesterol and long chain alcohols are suggested as potential candidates functioning as scent marking pheromones in A. boskianus.     

Turning the game around: toxicity in a nudibranch-sponge predator–prey association

Escalation theory proposes enemy-related selection as the most relevant factor of natural selection among individual organisms. When hazardous to predators, prey might be considered enemies that influence predator evolution. Opisthobranch molluscs that prey on chemically defended prey are an interesting study case on this subject. Predation on chemically defended species paved the way for opisthobranchs to enter in an arms race, developing means to detoxify and/or excrete harmful compounds, which led to the sequestration of those compounds and their self-defensive use, an escalation of defenses. Here we aim to understand whether the opisthobranch predator is better protected than its chemically defended prey, using as predator–prey model, a nudibranch (Hypselodoris cantabrica) and the sponge it preys upon (Dysidea fragilis), and from which it obtains deterrent chemical compounds. Specimens of both species were collected on the Portuguese coast, and their crude extracts were analyzed and used in palatability tests. Nudibranchs revealed a higher natural concentration of crude extract, probably due to a progressive accumulation of the compounds. Both predator and prey extracts revealed similar mixtures of deterrent metabolites (furanosesquiterpenes). Palatability tests revealed a more effective deterrence in the nudibranch extracts because significant rejection rates were observed at lower concentrations than those necessary for the sponge extracts to have the same effect. We concluded that the predator is chemically better protected than its prey, which suggests that its acquisition of chemical defenses reveals a defensive escalation.     

Chemical mimicry in an incipient leaf-cutting ant social parasite

Some social parasites of insect societies are known to use brute force when usurping a host colony, but most use more subtle forms of chemical cheating either by expressing as few recognition cues as possible to avoid being recognized or by producing similar recognition cues to the host to achieve positive discrimination. The former “chemical insignificance” strategy represents a more general adaptive syndrome than the latter “chemical mimicry” strategy and is expected to be characteristic of early evolutionary stages of social parasitism. We tested this hypothesis by experimentally analyzing the efficiency by which Acromyrmex echinatior leaf-cutting ants recognize intruding workers of the incipient social parasite Acromyrmex insinuator. The results were consistent with the parasite being “chemically insignificant” and not with the “chemical mimicry” hypothesis. Gas chromatography–mass spectrometry analysis of cuticular hydrocarbon profiles showed that social parasite workers produce significantly fewer hydrocarbons overall and that their typical profiles have very low amounts of hydrocarbons in the “normal” C29–C35 range but large quantities of unusually heavy C43–C45 hydrocarbons. This suggests that the C29–C35 hydrocarbons are instrumental in normal host nestmate recognition and that the C43–C45 compounds, all of which are dienes and thus more fluid than the corresponding saturated compounds, may reinforce “chemical insignificance” by blurring any remaining variation in recognition cues.     

A survey of antennal responses by five species of coniferophagous bark beetles (Coleoptera: Scolytidae) to bark volatiles of six species of angiosperm trees

Summary. Using Porapak Q traps, we collected the bark volatiles of six angiosperm trees native to British Columbia: black cottonwood, Populus trichocarpa Torr. & A. Gray (Salicaceae), trembling aspen, P. tremuloides Michx. (Salicaceae), paper birch, Betula papyrifera Marsh. (Betulaceae), bigleaf maple, Acer macrophyllum Pursh (Aceraceae), red alder, Alnus rubra Bong. (Betulaceae), and Sitka alder, A. viridis ssp. sinuata (Regel) á. L?ve & D. L?ve (Betulaceae). Utilising coupled gas chromatographic-electroantennographic detection analysis, the captured volatiles were assayed for antennal responses in five species of coniferophagous bark beetles (Coleoptera: Scolytidae), sympatric with most or all of the angiosperm trees: the Douglas-fir beetle, Dendroctonus pseudotsugae Hopkins, the mountain pine beetle, D. ponderosae Hopkins, the spruce beetle, D. rufipennis (Kirby), the western balsam bark beetle, Dryocoetes confusus Swaine, and the pine engraver, Ips pini (Say). The identities of 25 antennally-active compounds were determined by coupled gas chromatographic-mass spectroscopic analysis, and co-chromatographic comparisons with authentic chemicals. The compounds identified were: hexanal, (E)-2-hexenal, (Z)-3-hexen-1-ol, 1-hexanol, heptanal, α-pinene, frontalin, benzaldehyde, β-pinene, 2-hydroxycyclohexanone, 3-carene, limonene, β-phellandrene, benzyl alcohol, (E)-ocimene, salicylaldehyde, conophthorin, guaiacol, nonanal, methyl salicylate, 4-allylanisole, decanal, thymol methyl ether, (E)-nerolidol, and dendrolasin. A number of these compounds are known semiochemicals that are active in the behaviour of other organisms, including bark beetles, suggesting a high degree of semiochemical parsimony. Antennally-active compounds ranged from seven in A. viridis to 17 in P. trichocarpa. The fewest number of compounds (9) were detected by I. pini and the largest number (24) were detected by D. pseudotsugae. Six compounds excited the antennae of all five species of bark beetles. The large number of antennally-active compounds detected in common by numerous bark beetles and present in common in numerous nonhost trees supports the hypothesis of olfaction-based recognition and avoidance of nonhost angiosperm trees during the process of host selection by coniferophagous bark beetles. Received 13 December 1999; accepted 14 March 2000     

Fight or flight? A geographic mosaic in host reaction and potency of a chemical weapon in the social parasite <Emphasis Type="Italic">Harpagoxenus sublaevis</Emphasis>

Ant social parasites use chemical warfare to facilitate host colony takeover, which is a critical but recurring step in their life cycle. Many slave-making ants use the secretion of the Dufour gland to manipulate host behaviour during parasitic nest foundation and slave raids. Harpagoxenus sublaevis applies this chemical weapon onto defending Leptothorax host workers, which elicits deadly fights amongst them. Host species are expected to evolve counter-adaptations against this behavioural manipulation and in this study we investigated the geographic structure of this co-evolving trait. We compared the effectiveness of the parasitic gland secretion from different H. sublaevis populations in host colonies from various sites and analysed the occurrence of local adaptation. The two host species L. muscorum and L. acervorum generally showed different responses to the parasites’ chemical weapon: L. acervorum attacked nestmates treated with Dufour gland secretion, while L. muscorum workers fled. Flight, instead of intraspecific fights, is an adaptive host reaction as it results in fewer host fatalities during raids. Beside interspecific host differences, we found a geographic mosaic of host resistance: parasites from a German population strongly manipulated the behaviour of both sympatric Leptothorax populations. Russian or Italian hosts instead did not react with intracolonial aggression, but fled when confronted with the gland secretion of their sympatric parasite. Not only variation in host resistance explains differences in the effectiveness of the parasitic gland secretion but also interpopulational differences in its chemical composition, which were revealed by gas chromatography and mass spectrometry.     

Antennal responses of the western pine beetle, <Emphasis Type="Italic">Dendroctonus brevicomis</Emphasis> (Coleoptera: Curculionidae), to stem volatiles of its primary host, <Emphasis Type="Italic">Pinus ponderosa</Emphasis>, and nine sympatric nonhost angiosperms and conifers

Summary. Stem volatile extracts from ten trees that are sympatric with the western pine beetle, Dendroctonus brevicomis LeConte (Coleoptera: Curculionidae) were assayed by gas chromatographic-electroantennographic detection analysis (GC-EAD). The extracts were from the primary host, ponderosa pine, Pinus ponderosa Dougl. ex Laws. (Pinaceae); two nonhost angiosperms, California black oak, Quercus kelloggii Newb. (Fagaceae), and quaking aspen, Populus tremuloides Michx. (Salicaceae); and seven nonhost conifers, white fir, Abies concolor (Gord. & Glend.) Lindl. ex Hildebr. (Pinaceae), incense cedar, Calocedrus decurrens (Torr.) Florin (Cupressaceae), Sierra lodgepole pine, P. contorta murrayana Grev. & Balf. (Pinaceae), Jeffrey pine, P. jeffreyi Grev. & Balf. (Pinaceae), sugar pine, P. lambertiana Dougl. (Pinaceae), Douglas-fir, Pseudotsuga menziesii (Mirb.) Franco (Pinaceae), and mountain hemlock, Tsuga mertensiana (Bong.) Carr. (Pinaceae). Sixty-four compounds were identified from the ten trees, 42 of which elicited antennal responses in D. brevicomis, usually in both sexes. In addition, several synthetic compounds, including a number of the antennally-active compounds from the extracted trees and some bark beetle pheromone components, elicited antennal responses in a manner similar to that observed with the extracts. Of the antennally-active compounds known to be present in trees sympatric with D. brevicomis, only geraniol was unique to its host. Four antennally-active compounds were found in the host and in other conifers; five compounds were found only in nonhost conifers; eight compounds were found in either or both of the nonhost angiosperms; eight compounds were found in either or both of the angiosperms and in nonhost conifers, but not in the host; and 19 were found in both the host and in angiosperms and/or nonhost conifers. Several bark beetle pheromone components were found in the stem volatile extracts. Conophthorin was identified from both nonhost angiosperms; exo-brevicomin was identified in A. concolor; verbenone was identified from a number of nonhost conifers; and chalcogran was identified from P. tremuloides. The number of nonhost volatile chemicals that D. brevicomis encounters and is capable of detecting, and the diversity of sources from which they emanate, highlight the complexity of the olfactory environment in which D. brevicomis forages. This provides a basis for further work related to chemically-mediated aspects of foraging in this insect and perhaps other coniferophagous bark beetles, and highlights the need to consider foraging context in the design and implementation of semiochemical-based management tactics for tree protection.     

GC-EAD responses to semiochemicals by eight beetles in the subcortical community associated with Monterey pine trees in coastal California: similarities and disparities across three trophic levels

Summary.  Antennae of six sympatric bark and ambrosia beetles (Scolytidae), Dendroctonus valens LeConte, Gnathotrichus retusus (LeConte), Hylastes tenuis Eichhoff, Ips mexicanus (Hopkins), Ips plastographus maritimus Lanier, and Pseudohylesinus sericeus (Mannerheim), and two scolytid predators, Enoclerus sphegeus (F.) (Cleridae) and Lascontonus tuberculatus Kraus (Colydiidae), were analyzed by gas chromatographic-electroantennographic detection (GC-EAD) for their responses to synthetic Ips spp. pheromone components, and host and nonhost volatiles. The beetles emerged from cut logs of pitch canker-infected Monterey pine trees, Pinus radiata D. Don. There were significant disparities in EAD response patterns to the hemiterpene and monoterpene alcohol pheromone components that are typically produced by Ips spp. Antennae of I. p. maritimus responded strongly to ( ± )-ipsdienol, ( ± )-ipsenol, amitinol, and lanierone; antennae of I. mexicanus responded strongly to (1S,2S)-(–)-cis-verbenol, with weaker responses to ( ± )-ipsdienol, ( ± )-ipsenol, and amitinol; antennae of H. tenuis responded to (1S, 2R)-(–)-trans-ver-benol, with less pronounced responses to (–)-cis-verbenol and 2-methyl-3-buten-2-ol; and antennae of D. valens, G. retusus, and P. sericeus generally responded to all Ips spp. pheromone components except 2-methyl-3-buten-2-ol (D. valens and G. retusus) and E-myrcenol (G. retusus and P. sericeus). Ips mexicanus responded only to the (–)-enantiomers of ipsenol and ipsdienol, whereas I. p. maritimus responded to (–)-ipsenol, but to both the (+)- and (–)-enantiomers of ipsdienol. The antennae of the two predaceous insects (E. sphegeus and L. tuberculatus) responded to a range of the Ips spp. pheromone components. Host monoterpenes elicited no antennal responses from E. sphegeus, G. retusus, H. tenuis, and I. mexicanus, but several monoterpenes elicited various levels of responses from D. valens and I. p. maritimus antennae. Interestingly, antennae of female D. valens responded to (–), but not (+)-limonene. α- and β-Pinene elicited weak responses from L. tuberculatus antennae. EAD responses to selected nonhost volatiles were almost identical among the six scolytid species, with trans-conophthorin eliciting the strongest response in most cases, followed by three C₆- alcohols and two C₈-alcohols. The antennal responses by most of these species to linalool or geranylacetone were very weak; (E)-2-hexenal, (Z)-3-hexenyl acetate, and benzyl alcohol elicited almost no response. The response pattern of P. sericeus to nonhost volatiles differed slightly from the rest of the scolytids: a strong response to linalool, weaker response to the C₈-alcohols. The two predaceous Coleoptera generally had weak, but detectable, responses to nonhost volatiles, except for a relatively strong response to trans-conophthorin by L. tuberculatus. No notable differences in EAD responses were observed between males and females of the two Ips spp. Our results provide an electrophysiological baseline for future efforts to identify attractive and repellent semiochemicals (aggregation pheromones, host kairomones, or nonhost interruptants) for this guild of scolytids and their key predators that are associated with moribund and pitch canker- infected P. radiata.     

Host-plant leaf surface compounds influencing oviposition in Delia antiqua

Summary. Delia antiqua (Diptera: Anthomyiidae) females lay eggs between the leaves of onion plants or in the soil around the base of the plants, then the maggots feed on the onion bulb and roots causing rapid secondary infection by fungi and bacteria. It is well known that the first sensory modality used by the onion fly is vision, therefore the shape (vertical narrow cylinders) and colour (yellow) of the plant play a crucial role in the recognition of a potential host plant. In the past it has been shown that n-dipropyl disulfide (Pr₂S₂), a typical component of onion volatiles, is an important chemical host plant cue. We extracted host leaf surface to verify if Pr₂S₂ is the major chemical oviposition stimulant and to determine if other as yet unknown substances may play a role in host-plant selection. We confirmed that the females laid more eggs around onion plants with leaves than when only the onion bulb was present and that the odour of chopped onion stimulates oviposition. Extraction of the surface of onion leaves revealed that only the apolar fraction contained substances that stimulate egg-laying in D. antiqua. GC-EAD analysis indicated that a minor constituent, Pr₂S₂, is perceived by the olfactory receptor on the antennae of the onion fly females. This confirmed the importance of Pr₂S₂ as oviposition stimulant. Contact with the polar fraction did not stimulate egg-laying behaviour in this Delia species. We discuss the oviposition strategy of D. antiqua in comparison with its closely related species, D. radicum, in which the oviposition behaviour is stimulated mainly through contact with the cabbage leaf surface and only partially by the host volatiles.     

Sex pheromone of <Emphasis Type="Italic">Ectinus aterrimus</Emphasis> (L<Emphasis Type="SmallCaps">inné</Emphasis>, 1761) (Coleoptera: Elateridae)

Summary.   Ectinus aterrimus (L.) is a fairly common European click beetle species which develops mainly in forests. In pheromone gland extracts of female E. aterrimus, examined using gas chromatography – mass spectrometry (GC-MS), one single compound was present. This was identified as 7-methyloctyl 9-methyldecanoate by comparison with a synthetic sample. Field trapping trials revealed a highly significant attraction of male E. aterrimus towards this ester. The structure of the compound differs remarkably from the pheromones of the closely related Agriotes spp., which exclusively use terpene esters.     

Chemosensory and behavioural responses of the turnip sawfly, Athalia rosae, to glucosinolates and isothiocyanates

Summary. The turnip sawfly Athalia rosae sequesters glucosinolates from its cruciferous host plants in the larval stage. Investigation of the chemosensory and behavioural responses of adult A. rosae to glucosinolates and their volatile hydrolysis products, isothiocyanates, revealed that females detect glucosinolates by contact chemoreception and isothiocyanates by antennal olfaction. In electroantennogram recordings, four isothiocyanates (allyl [2-propenyl] isothiocyanate, benzyl isothiocyanate, butyl isothiocyanate and iberverin [3-methylthiopropyl isothiocyanate]) were active at all doses presented, including the lowest (0.1 μg), whilst the threshold for detection of three others, iberin [3-methylsulphinylpropyl isothiocyanate], methyl isothiocyanate, and sulforaphane [4-methylsulphinylbutyl isothiocyanate], was higher, at between 1 and 10 μg (source concentration of volatiles). Allyl isothiocyanate attracted experienced females in a four-chambered olfactometer, whilst na?ve females showed no response. Allyl isothiocyanate also attracted mature females to baited yellow water traps in field trials, although immature females were repelled at high isothiocyanate concentrations. In laboratory behavioural bioassays the glucosinolates sinigrin (allyl [2-propenyl] glucosinolate) and sinalbin (p-hydroxybenzyl glucosinolate), stimulated ovipositor probing in mature female A. rosae to an extent comparable to hot-water extracts of their host plants. These responses show that glucosinolates and isothiocyanates play an important role in host finding and host recognition in A. rosae.