Opposing Effects of Host Monoterpenes on Responses by Two Sympatric Species of Bark Beetles to Their Aggregation Pheromones

The effects of host tree monoterpenes on primary and secondary attraction of the bark beetles, Ips pini and Ips grandicollis, and their associated phloeophagous insects were investigated. Monoterpenes alone were not attractive to I. pini. However, monoterpenes mediated the attraction of I. pini to its aggregation pheromones. With the exception of 3–carene, the effect of monoterpenes on I. pini's response to its pheromone, ipsdienol plus lanierone, was inhibitory. In contrast, (–)--pinene both attracted I. grandicollis and enhanced the attraction of I. grandicollis to its pheromone, ipsenol. No monoterpene inhibited the response of I. grandicollis to its pheromone. The inhibitory effect of host monoterpenes on I. pini response to its aggregation pheromone differs from previous work, in which monoterpenes either synergized responses or had no effect. In addition to possible geographic differences, the concentrations used in our study simulated trees that had begun to respond to attack, whereas previous studies deployed lower concentrations, which simulated constitutive phloem from unattacked trees. These results support the view that trees that undergo induced responses to bark beetles sometimes inhibit attraction of additional beetles, despite the beetles' production of aggregation pheromones. Neither species displayed cross-attraction to the pheromone of the other. The red turpentine beetle, Dendroctonus valens, showed weak and consistent attraction to (+)--pinene and in some cases to (–)--pinene. Attraction to (–)--pinene was usually enhanced by Ips spp. pheromones. The absence of strong attraction to (+)--pinene and partial attraction to (–)--pinene suggest that the effects of different stereoisomers of -pinene on D. valens vary throughout its geographical range. Hylastes porculus was also attracted to some monoterpenes, particularly (–)--pinene. An additional 10 species of phloeophagous insects were caught in response to monoterpenes and/or pheromones, including the pine root weevils, Hylobius pales and Pachylobius picivorus, and the longhorned beetle, Monochamus carolinensis.     

Monoterpenes and Epicuticular Waxes Help Female Autumn Gum Moth Differentiate Between Waxy and Glossy Eucalyptus and Leaves of Different Ages

The autumn gum moth, Mnesampela privata, is a native Australian species whose preferred host, Eucalyptus globulus (Myrtaceae), is an aromatic evergreen tree that has long-lived waxy leaves during the juvenile phase of growth. We compared the behavioral and antennal responses of female moths to whole leaves (new and old) and samples of foliar chemicals (from new and old leaves) from a typical E. globulus subsp. pseudoglobulus with responses to a glossy, half-sibling E. g. subsp. pseudoglobulus putative hybrid (the result of natural cross-pollination). We also studied larval survival and development on leaves from the same trees. In laboratory binary-choice assays, female M. privata laid more eggs on waxy leaves than on glossy leaves thereby confirming the nonpreference for the glossy tree that was observed in the field. Analyses of the monoterpenes and waxes of both trees revealed that they had comparable suites of monoterpenes and total oil contents but different suites of epicuticular waxes. Headspace extracts differed in the intensity of component monoterpenes. Gas chromatographic analyses with electroantennographic detection showed different patterns of monoterpene detection. Leaves of the glossy tree had a less diverse array of epicuticular waxes than those of the waxy tree. Electroantennographic screening of responses to wax extracts from leaves (new and old) from either tree revealed positive dose-dependent responses of female antennae to waxes from new leaves only. Binary-choice assays also revealed a strong preference by ovipositing females for new, compared to old, leaves whether they were from the waxy or the glossy tree. However, new leaves from either tree could be manipulated (by physical abrasion of epicuticular waxes) so that females would lay almost no eggs on them. Larval survival did not differ between groups reared on leaves from both trees (new and old). Over 70% of all larvae survived to pupation. However, larvae reared on leaves from the glossy tree took longer to pupate than those reared on leaves from the waxy tree. Also, larvae reared on new leaves from either tree did not perform as well as those reared on old leaves. Monoterpene and wax cues are suggested as helping female M. privata locate preferred hosts in native forests.     

Gallery initiation byTomicus piniperda (Coleoptera: Scolytidae) on scots pine trees baited with host volatiles

In a trapping experiment, wood chips from freshly cut Scots pine trees attracted flying adults of the bark beetleTomicus piniperda (L.) Healthy Scots pine trees, which usually are not attacked byT. piniperda, were baited with chips from freshly cut trees. In other experiments trees were baited with a mixture of (–)--pinene, (+)-3-carene, and terpinolene; the individual monoterpenes; or with ethanol alone. All baited treatments were attacked byT. piniperda. Most of the attacks resulted in short egg galleries and in larval galleries which were only a few millimeters in length. In trees from which a new generation of beetles emerged, net reproduction was well below 1.0. Unbaited control trees remained unattacked or received a few isolated attacks.     

Correspondence of soldier defense secretion mixtures with cuticular hydrocarbon phenotypes for chemotaxonomy of the termite genus Reticulitermes in North America

Soldier defense secretions from samples of Reticulitermes collected in California, Nevada, Arizona, New Mexico, and Georgia were characterized and correlated with cuticular hydrocarbon phenotypes. Twenty-seven cuticular hydrocarbon phenotypes have been defined, and soldier defense secretion (SDS) phenotypes have been described for 25 of these. Forty-five terpenoid compounds were found, including monoterpenes, sesquiterpenes, and a few diterpenes. The monoterpenes include (–)--pinene, (–)--pinene, (–)-camphene, myrcene, (Z)- and (E)-ocimene, and (–)-limonene. The major sesquiterpenes produced are (+)--cadinene, (+)--cadinene aldehyde, (–)-germacrene A, germacrene B, -himachalene, and -bisabolene. Some SDS phenotypes pair with more than one cuticular hydrocarbon phenotype; however, with two exceptions, each hydrocarbon phenotype is associated with only one SDS phenotype. These chemical characterizations lend support to the conclusion that there are numerous undescribed species of Reticulitermes in North America.     

Electroantennogram responses ofHyles lineata (Sphingidae: Lepidoptera) to volatile compounds fromClarkia breweri (Onagraceae) and other moth-pollinated flowers

Electroantennograms (EAGs) from field-collectedHyles lineata moths were recorded in response to 10 individual floral volatiles identified fromClarkia breweri (Onagraceae), to 22 scent compounds produced by other moth-pollinated flowers and to eight ubiquitous green leaf volatiles. Females' EAGs were generally 1.5- to 2-fold greater than those observed for male moths. Female:male EAG rank orders were significantly correlated, but marked differences in order were observed for some compounds (e.g., benzyl alcohol, cinnamic aldehyde, geraniol, and linalool). Linalool, benzyl acetate, methyl salicylate, and pyranoid linalool oxide elicited the largest EAG responses (–1.2 to –0.8 mV) among scent compounds fromC. breweri. EAG responses were significantly lower for monoterpenes as a pooled compound class than for aromatic esters, alcohols and aldehydes, fatty acid derivatives, N-bearing compounds and oxygenated terpenoids. EAG responses to structurally related scent compounds were not significantly different in most cases. Both male and femaleH. lineata were sensitive to mostC. breweri scent compounds at 10^–2 to 10^–4 µg/µl doses, and rank order in potency varied with the dose/concentration tested.H. lineata's olfactory sensitivity to diverse volatile compounds across a range of doses/concentrations suggests that a broad array of volatiles could function as floral attractants for foraging hawkmoths.     

Chemical inhibition of fire-prone grasses by fire-sensitive shrub,Conradina canescens

In an investigation of potential chemical activity of fire-sensitive shrubs in Florida's sand pine scrub community, bioassays of foliar washes ofConradina canescens showed significant inhibitory activity on three native grasses that are known to fuel frequent surface fires; inhibition was concentrated seasonally in spring and summer. Application of runoff fromConradina leaves to one of the grasses caused a 50% reduction in growth over a 20-week period. Isolation of the biologically active fractions from the fresh leaves ofC. canescens yielded numerous monoterpenes, a number of which were identified from a GC-MS reference library and/or MS comparison to authentic compounds: 11 from the diethyl ether extract, 11 from steam distillation, and four from the foliar leaf wash. Numerous other monoterpenes present in the extractions were unknown. The terpenoid fraction completely inhibited seed germination of one of the native grasses and of lettuce. Saturated aqueous solutions of nine of the monoterpenes inhibited germination and radicle growth of two native grasses. SEM views of the leaf surfaces ofConradina reveal secretory trichomes that appear to be the source of the monoterpenes as well as the triterpene, ursolic acid. The biological activity ofC. canescens as a fire-sensitive component of the scrub community is reviewed in light of the chemical evidence.     

Pheromone production by axenically rearedDendroctonus ponderosae andIps paraconfusus (Coleoptera: Scolytidae)

Mountain pine beetles,Dendroctonus ponderosae Hopkins, and California five-spined ips,Ips paraconfusus Lanier, were reared axenically from surface-sterilized eggs on aseptic pine phloem. After 24 hr in host logs, axenip femaleD. ponderosae and maleI. paraconfusus produced the aggregation pheromones,trans-verbenol (D. ponderosae), and ipsenol and ipsdienol (I. paraconfusus). Emergent, axenically reared maleD. ponderosae contained normal amounts of the pheromoneexo-brevicomin. Axenic femaleD. ponderosae treated with juvenile hormone or exposed to vapors of -pinene, produced the pheromonetrans-verbenol. By 25–35 days after eclosion, axenic females exposed to -pinene vapors produced over six times as muchtrans-verbenol as wild females, suggesting that while microorganisms in wild females may producetrans-verbenol, they may also inhibit production of the pheromone or use it as a substrate.Research supported in part by Natural Sciences and Engineering Research Council, Canada, Operating Grants A3881 and A3706, and H.R. MacMillan Family Fund Fellowships to J.E. Conn and D.W.A. Hunt.     

Odor Perception in the Bark Beetle Parasitoid Roptrocerus xylophagorum Exposed to Host Associated Volatiles

Y-tube olfactometer bioassays and combined gas chromatography–electroantennographic detector (GC-EAD) analyses were performed to investigate the specific odors utilized as host location cues by the bark beetle parasitoid, Roptrocerus xylophagorum, originating from the southeastern United States. R. xylophagorum parasitizes several economically important holarctic bark beetle species and females oviposit preferentially on late larval stages. Both female and male parasitoids were tested with volatiles derived from host infestations of either Dendroctonus frontalis, the southern pine beetle, or Ips grandicollis, the southern pine engraver. Tested volatiles were steam distillates from the bark of loblolly pine, Pinus taeda, infested with larvae of the respective bark beetle species. Combined gas chromatography–mass spectrometry (GC-MS) was employed for identification and quantification of the compounds in the steam distillates. To confirm the EAD activity of identified compounds, GC-EAD analyses were repeated with a synthetic blend composed predominantly of compounds in the crude extracts that had revealed apparent electrophysiological activity. In Y-olfactometer bioassays, female parasitoids were attracted to both of the above-mentioned distillates. Male parasitoids were tested with one of the distillates but failed to respond. In GC-EAD analyses, the sexes displayed similar relative sensitivities to the components of the blends. Males exhibited generally higher amplitudes of response to the tested compounds than females. Monoterpene hydrocarbons associated with the constitutive resin of the host tree did not elicit significant EAD responses. Compounds known to be associated specifically with the host–tree complex, such as certain oxygenated monoterpenes, generated the greatest EAD responses. Female parasitoids were attracted by a synthetic blend composed of several of the EAD active oxygenated monoterpenes.     

Allelopathic potential of menthofuran monoterpenes fromCalamintha ashei

A reversed-phase HPLC analysis was used to separate and quantify five menthofuran monoterpenes inCalamintha ashei leaf soaks and washes. (+)-Evodone and desacetylcalaminthone were the major constituents of both soaks and washes. Concentrations of (+)-evodone and desacetylcalaminthone were as high as 0.66 and 0.74 mM, respectively, in leaf soaks. The highest concentration of monoterpenes in leaf washes obtained by misting was 0.021 mM. Aqueous solubilities of the menthofurans were determined to exceed concentrations required for growth inhibition. Bioassays of individualCalamintha monoterpenes demonstrated effects on germination as low as 0.05 mM for (+)-evodone. An equimolar mixture of desacetylcalaminthone and (+)-evodone reducedRudbeckia hirta germination by 17% at a combined concentration of 0.025 mM forLeptochloa dubia. Confirmation of allelopathic effects byCalamintha ashei will require long-term bioassays ofCalamintha menthofurans on the growth of native sandhill species under conditions comparable to the harsh environment of the Florida scrub.