Jocelyn G. Millar

Advances in insect chemical ecology

Preface 1. Phytochemical diversity of insect defenses in tropical and temperate plant families John T. Arason, Gabriel Guillet and Tony Durst 2. Recruitment of predators and parasitoids by herbivore-injured plants Ted C. J. Turlings and Felix Wackers 3. Chemical ecology of astigmatid mites Yasumasa Kuwahara 4. Semiochemistry of spiders Stefan Schulz 5. Why do flowers smell? The chemical ecology of fragrance-driven pollination Robert A. Raguso 6. Sex pheromones of cockroaches Cesar Gemeno and Coby Schal 7. A quest for alkaloids: curious relationship between tiger moths and plants containing pyrrolizidine alkaloids William E. Conner and Susan J. Weller 8. Structure of the pheromone communication channel in moths Ring T. Carde and Kenneth F. Haynes Index.

Specialized cells tag sexual and species identity in Drosophila melanogaster.

Social interactions depend on individuals recognizing each other, and in this context many organisms use chemical signals to indicate species and sex. Cuticular hydrocarbon signals are used by insects, including Drosophila melanogaster, to distinguish conspecific individuals from others. These chemicals also contribute to intraspecific courtship and mating interactions. However, the possibility that sex and species identification are linked by common chemical signalling mechanisms has not been formally tested. Here we provide direct evidence that a single compound is used to communicate female identity among D. melanogaster, and to define a reproductive isolation barrier between D. melanogaster and sibling species. A transgenic manipulation eliminated cuticular hydrocarbons by ablating the oenocytes, specialized cells required for the expression of these chemical signals. The resulting oenocyte-less (oe-) females elicited the normal repertoire of courtship behaviours from males, but were actually preferred over wild-type females by courting males. In addition, wild-type males attempted to copulate with oe- males. Thus, flies lacking hydrocarbons are a sexual hyperstimulus. Treatment of virgin females with the aversive male pheromone cis-vaccenyl acetate (cVA) significantly delayed mating of oe- females compared to wild-type females. This difference was eliminated when oe- females were treated with a blend of cVA and the female aphrodisiac (7Z,11Z)-heptacosadiene (7,11-HD), showing that female aphrodisiac compounds can attenuate the effects of male aversive pheromones. 7,11-HD also was shown to have a crucial role in heterospecific encounters. Specifically, the species barrier was lost because males of other Drosophila species courted oe- D. melanogaster females, and D. simulans males consistently mated with them. Treatment of oe- females with 7,11-HD restored the species barrier, showing that a single compound can confer species identity. These results identify a common mechanism for sexual and species recognition regulated by cuticular hydrocarbons.

Conserved class of queen pheromones stops social insect workers from reproducing

Long Live the Queen Eusociality is often considered to have arisen, at least in part, due to the inclusive fitness that workers gain through helping their queen sister to raise her offspring. Van Oystaeyen et al. (p. 287; see the Perspective by Chapuisat) characterized the sterility-inducing queen pheromone across three distantly related eusocial hymenopterans (a wasp, a bumblebee, and a desert ant) and synthesized data across 69 other species. Queen pheromones appear to be remarkably conserved, which suggests that reproductive manipulation has ancient roots. Social insect queens use an ancient, evolutionarily conserved class of pheromones to prevent worker reproduction. [Also see Perspective by Chapuisat] A major evolutionary transition to eusociality with reproductive division of labor between queens and workers has arisen independently at least 10 times in the ants, bees, and wasps. Pheromones produced by queens are thought to play a key role in regulating this complex social system, but their evolutionary history remains unknown. Here, we identify the first sterility-inducing queen pheromones in a wasp, bumblebee, and desert ant and synthesize existing data on compounds that characterize female fecundity in 64 species of social insects. Our results show that queen pheromones are strikingly conserved across at least three independent origins of eusociality, with wasps, ants, and some bees all appearing to use nonvolatile, saturated hydrocarbons to advertise fecundity and/or suppress worker reproduction. These results suggest that queen pheromones evolved from conserved signals of solitary ancestors.

Identification of volatile synomones, induced by Nezara viridula feeding and oviposition on bean spp., that attract the egg parasitoid Trissolcus basalis.

Bean plants (Vicia faba L. and Phaseolus vulgaris L.) damaged by feeding activity of Nezara viridula (L.) (Heteroptera: Pentatomidae), and onto which an egg mass had been laid, produced volatiles that attracted the egg parasitoid Trissolcus basalis (Wollaston) (Hymenoptera: Scelionidae). Extracts of volatiles of broad bean and French bean plants induced by adults of N. viridula as a result of their feeding activity, oviposition activity, and feeding and oviposition activity combined were analyzed by gas chromatography–mass spectrometry (GC–MS), and tested in Y-tube olfactometer bioassays as attractants for T. basalis females. In extracts from undamaged leguminous plants, green-leaf volatiles were absent or scarcely detected, and monoterpenes and sesquiterpenes were present at trace levels. No significant differences were detected in the profiles of volatiles of undamaged plants, and undamaged plants on which bugs were allowed only to lay eggs. In contrast, feeding and oviposition by adults of N. viridula induced in both leguminous plants a significant increase in terpenoids such as linalool, (E)-β-caryophyllene, (E,E)-4,8,12-trimethyl-1,3,7,11-tridecatetraene, and (3E)-4,8-dimethyl-1,3,7-nonatriene, which was induced only in French bean plants. Quantitative comparisons revealed increased levels of (E)-β-caryophyllene in extracts from feeding-damaged plants with N. viridula egg masses compared to feeding-damaged plants without egg masses. In Y-tube olfactometer bioassays, T. basalis females were attracted by extracts of both leguminous plants only when N. viridula adults were allowed to feed and oviposit upon them. Fractionation of extracts of volatiles from broad bean plants with N. viridula feeding damage and egg masses yielded two fractions, but only the fraction containing (E)-β-caryophyllene was attractive to the egg parasitoid. These findings indicate that N. viridula feeding and oviposition induce leguminous plants to produce blends of volatiles that are characterized by increased amounts of (E)-β-caryophyllene, and these blends attract female T. basalis. The role of (E)-β-caryophyllene as a potential synomone for T. basalis is discussed.

Male-Produced Aggregation Pheromone of the Cerambycid Beetle Neoclytus acuminatus acuminatus

This is the first fully verified report of an aggregation pheromone produced by a cerambycid beetle species. Field bioassays with adultNeoclytus acuminatus acuminatus (F.) (Coleoptera: Cerambycidae) revealed that males produce a pheromone that attracts both sexes. Extracts of odors from males contained a single major male-specific compound, (2S,3S)-hexanediol. Field trials determined that both sexes were attracted by the racemic blend of (2S,3S)- and (2R,3R)-hexanediols and that activity was similar to enantiomerically enriched (2S,3S)-hexanediol (e.e. 80.2%). However, the blend of all four 2,3-hexanediol stereoisomers attracted few beetles, indicating inhibition by one or both of the (2R*,3S*)-stereoisomers. Females of the cerambycidCurius dentatus Newman were attracted to traps baited with the four component blend, suggesting that a male-produced sex pheromone for this species may contain (2R,3S)-hexanediol and/or (2S,3R)-hexanediol. The pheromone ofN. a. acuminatus, and presumed pheromone ofC. dentatus, bear structural similarities to those produced by males of six other species in the Cerambycinae (straight chains of 6, 8, or 10 carbons with hydroxyl or carbonyl groups at C2 and C3). It is likely that males of other species in this large subfamily produce pheromones that are variations on this structural motif.

Development and Optimization of Methods for Using Sex Pheromone for Monitoring the Mealybug Planococcus ficus (Homoptera: Pseudococcidae) in California Vineyards

Abstract The sex pheromone of the vine mealybug Planococcus ficus Signoret has been identified as a single component, lavandulyl senecioate. Racemic lavandulyl senecioate was as attractive to male mealybugs as the insect-produced (S)-enantiomer, indicating that the unnatural enantiomer is not inhibitory. Lavandulol, which also was found in extracts from virgin females, antagonized attraction of males at higher doses. Rubber septum lures loaded with 10- to 1,000-μg doses of the pheromone were equally attractive, and lures loaded with 100 μg of racemic pheromone remained attractive for at least 12 wk under field conditions. Delta traps were more effective than double-sided sticky cards and minimized captures of nontarget insects. Pheromone-baited traps had an effective range of at least 50 m. Comparison of visual sampling methods and sampling of males with pheromone-baited traps revealed that trap catches were significantly correlated with the results from visual sampling methods, and with economic damage.

Treating Panel Traps with a Fluoropolymer Enhances their Efficiency in Capturing Cerambycid Beetles

ABSTRACT The most effective traps for capturing cerambycids and other saproxylic beetles are intercept designs such as funnel traps and cross-vane panel traps. We have observed that adult cerambycids of many species often alight and walk upon panel traps, and few are actually captured. In an effort to improve trap capture and retention, researchers have treated intercept traps with Rain-X, a polysiloxane formulation that renders surfaces more slippery. Here, we summarize experiments that compared the efficacies of Rain-X and Fluon, a PTFE fluoropolymer dispersion, as surface treatments for panel traps that are deployed to capture cerambycid beetles, using untreated traps as controls. Fluon-treated traps captured on average >14× the total number of beetles, and many more cerambycid species, than were captured by Rain-X-treated or control traps. Beetles captured by Fluon-treated traps ranged in body length by 350%. They could not walk on vertical panels treated with Fluon but easily walked on those treated with Rain-X and on untreated traps. Moreover, a single Fluon treatment remained effective for the entire field season, even in inclement weather. We conclude that treating panel traps with Fluon greatly improves their efficiency in capturing cerambycid beetles. This increased efficacy will be particularly important when traps are deployed to detect very low-density populations, such as incursions of exotic species, or remnant communities of rare and endangered species. The influence of Fluon on trap efficiency may vary with product formulation and its source and also with climatic conditions.

Water relations of host trees and resistance to the phloem-boring beetle Phoracantha semipunctata F. (Coleoptera: Cerambycidae).

Abstract Environmental stresses, particularly water deficit, predispose eucalypt trees to attack by the eucalyptus longhorned borer, Phoracantha semipunctata F. (Coleoptera: Cerambycidae). Our experiments with potted eucalypts revealed that reduced tree water potential was associated with lower resistance to colonization by neonate P. semipunctata, but the linear relationship between water potential and colonization success was reversed at higher larval densities. There was no indication that the bark exudate “kino” served to defend trees from borer attack. Larvae were not able to colonize the cambium of eucalypt logs with high bark moisture, and survival was low under high moisture conditions in artificial hosts composed of pure cellulose. In trees and cut logs with moist bark, larvae failed to reach the cambium, feeding instead in poorer-quality tissues just beneath the bark surface. Our findings suggest that variation in resistance of eucalypts to attack by the borer is associated with moisture content of the bark.

Pheromone-Based Mating Disruption of Planococcus ficus (Hemiptera: Pseudococcidae) in California Vineyards

Experiments were conducted to test a mating disruption program for the mealybug Planococcus ficus (Signoret) (Hemiptera: Pseudococcidae) in California vineyards. The sprayable, microencapsulated formulation of the racemic sex pheromone lavandulyl senecioate was applied with an air-blast sprayer, using three and four applications in 2003 and 2004, respectively. Mating disruption was combined with an application of buprofezin (2004) in June. Compared with a no-pheromone control, there were significantly lower season-long trap catches of adult males, season-long mealybug densities (2003 only), and crop damage in mating disruption plots. The amount of mealybug reduction and mechanisms that resulted in lower crop damage in mating disruption plots is discussed. In samples taken during the growing season (April to September), mealybug density was only 12.0 +/- 15.6 and 31.1 +/- 11.6% lower in the mating disruption plots than in control plots in 2003 and 2004, respectively. In the mating disruption treatment, mealybug egg production was significantly lower (2003 only), as were the proportion of ovisacs and crawlers produced. There was no treatment impact on percentage of parasitism. Mealybug density influenced treatment impact. In 2004, vines were categorized as having low, medium, or high mealybug densities during a preapplication survey. After treatment application, mealybug density was reduced by 86.3 +/- 6.3% on vines in the low mealybug density category, but it was unchanged on vines in the high density category. Another factor that reduced treatment impact was the relatively short effective lifetime of the sprayable formulation.

Role of Contact Pheromones in Mate Recognition in Xylotrechus colonus

Adult male and female rustic borers, Xylotrechus colonus F. (Coleoptera: Cerambycidae), aggregate on cut logs and fallen trees that are the hosts of their larvae. Our studies show that male X. colonus actively search for females, and only respond to them after contacting them with their antennae. Stripping cuticular hydrocarbons from females with solvent rendered them unattractive to males, suggesting that males did not recognize females by mechanoreception alone. Reapplying solvent extract to washed females restored their attractiveness to males, confirming the role of cuticular hydrocarbons in mate recognition. Female cuticular hydrocarbon extracts contain n-pentacosane, 9-methylpentacosane, and 3-methylpentacosane, components that were either absent or present in very small amounts on males. We demonstrate that the contact pheromone is a blend of these three cuticular hydrocarbons.