Katalin Böröczky

Behavioral Evidence for a Contact Sex Pheromone Component of the Emerald Ash Borer, Agrilus Planipennis Fairmaire

The cuticular hydrocarbon profiles of emerald ash borers, Agrilus planipennis, were examined to determine if there are differences in these compounds between the sexes. We also assessed feral male EAB in the field for behavioral changes based on the application of a female-specific compound to dead, solvent-washed beetles. Males in the field spent significantly more time attempting copulation with dead, pinned female beetles coated with a three-beetle-equivalent dose of 3-methyltricosane than with solvent-washed beetles or those coated in 3-methyltricosane at lower concentrations. Males in the field spent the most time investigating pinned dead, unwashed female beetles. In the laboratory, sexually mature males were presented with one of several mixtures applied in hexane to filter paper disks or to the elytra of dead female beetles first washed in solvent. Male EAB also spent more time investigating dead beetles treated with solution applications that contained 3-methyltricosane than dead beetles and filter paper disks treated with male body wash or a straight-chain hydrocarbon not found on the cuticle of EAB.

Monoalkenes as Contact Sex Pheromone Components of the Woodwasp Sirex noctilio

A pheromone on the cuticle of females of the woodwasp Sirex noctilio, a recently introduced pest of pines in North America, induces conspecific males to attempt copulation. Dead females washed with hexane did not elicit copulation attempts from males, whereas reapplication of a female hexane body wash onto the cuticle of dead females elicited copulation attempts by 65% of males tested. Analysis of the hexane extract revealed saturated and unsaturated hydrocarbons as major components of the female cuticle. Behavior-guided fractionation of the female body wash led to the identification of three components, (Z)-7-heptacosene, (Z)-7-nonacosene, and (Z)-9-nonacosene, of the sex pheromone of S. noctilio that elicited copulatory responses from males.

Male-Produced Pheromone in the European Woodwasp, Sirex noctilio

A male-produced pheromone that attracts both males and females was identified for the European woodwasp, Sirex noctilio, a serious pest of pine trees. Males displayed excitatory behaviors when placed in groups, and were attracted to the odors from males that were 2-5-d-old, but not to odors from males that were 0-1-d-old. An unsaturated short-chain alcohol, (Z)-3-decen-1-ol, was discovered in samples collected on SuperQ filters over groups of males and identified by using micro-derivatization reactions and gas chromatography coupled with mass spectrometry (GC-MS). The compound was not detected in volatile samples from females. Gas chromatography coupled electroantennographic detection (GC-EAD) of antennae from males exposed to male headspace odors produced strong antennal responses to the main peak of (Z)-3-decen-1-ol, as well as to an unknown minor component that had a similar retention time. Antennae from both males and females responded to synthetic (Z)-3-decen-1-ol. Several different synthetic candidates for the GC-EAD active minor components were selected based on GC-MS and GC-EAD responses to male headspace collections. These synthetic compounds were tested for antennal activity using GC-EAD, and those that produced strong responses were blended with the major component and tested for male attraction in the Y-tube olfactometer at different concentrations and ratios. Males tested in the Y-tube olfactometer were attracted to a synthetic blend of (Z)-3-decen-1-ol and (Z)-4-decen-1-ol at a ratio of 100:1. Whereas the addition of some suspected minor compounds reduced attraction, the addition of a third compound found in male emanations that produced strong male antennal responses, (E,E)-2,4-decadienal (at a ratio of 100:1:1), resulted in attraction of both males (Y-tube and wind tunnel) and females (wind tunnel).

Volatile Profile Differences and the Associated Sirex noctilio Activity in Two Host Tree Species in the Northeastern United States

Sirex noctilio females are known to be attracted to stem sections of stressed pine trees for oviposition. The volatile profiles and attractiveness of Eastern white pine (Pinus strobus) and two chemotypes of Scots pine (P. sylvestris) were compared after stem injection with herbicide. In general, trap captures on herbicide-treated trees were higher than on controls. The high-carene chemotype of Scots pine captured the highest numbers of females, followed by the low-carene chemotype, and finally the Eastern white pine. Herbicide-treated trees of both species emitted larger quantities of volatiles than the controls. The herbicide treatment induced higher volatile emission rates in the Scots pine chemotypes than in white pine, although there was no difference between the two chemotypes. However, qualitative differences were found between the volatile profiles of the two species as well as between the two Scots pine chemotypes, which could account for the differential attractiveness of the species and chemotypes tested.

Differences in cuticular lipid composition of the antennae of Helicoverpa zea, Heliothis virescens, and Manduca sexta

Analyses of the hexane washes of antennae, forelegs and whole bodies of Helicoverpa zea, Heliothis virescens, and Manduca sexta revealed notable differences in the components of the cuticular coatings of each species. Most striking were the differences between the cuticular coatings of male and female antennae of both H. zea and H. virescens. Novel esters of short-chain acids (C2-C4) and long-chain secondary alcohols (C25-C32) were identified in the hexane washes of the male antenna and forelegs of H. zea and H. virescens. These compounds were found in only small amounts or were completely absent on the female antennae of both species. In H. zea, butyrates of 7- and 8-pentacosanol and 8- and 9-heptacosanol were found, whereas, in the foreleg extracts of H. virescens, acetates and propionates were detected in addition to butyrates. While cholesterol is a major component of antennal washes (10-15%), only traces were found in the foreleg extracts. Although the composition of the cuticular coating of M. sexta differed greatly from that of the other two species, the extractable coatings of the antennae of male and female M. sexta were nearly identical.

Identification of a queen and king recognition pheromone in the subterranean termite Reticulitermes flavipes

Significance Understanding the mechanisms that social insects use to communicate their individual status within the colony is vital to understanding the evolution of sociality. This study accomplishes this goal by identifying a royal-recognition pheromone in termites, as well as a king pheromone. Our behavioral assay defines royal-specific responses for one species of termites, which will foster future studies of termite behavior. This study also dates cuticular hydrocarbons as royal pheromones to the rise of termites ∼150 million years ago, suggesting that termites and social Hymenoptera convergently evolved the use of these ubiquitous compounds for communication. In conclusion, we have expanded our understanding of chemically mediated royal recognition in termites and helped to understand the evolution of insect societies. Chemical communication is fundamental to success in social insect colonies. Species-, colony-, and caste-specific blends of cuticular hydrocarbons (CHCs) and other chemicals have been well documented as pheromones, mediating important behavioral and physiological aspects of social insects. More specifically, royal pheromones used by queens (and kings in termites) enable workers to recognize and care for these vital individuals and maintain the reproductive division of labor. In termites, however, no royal-recognition pheromones have been identified to date. In the current study, solvent extracts of the subterranean termite Reticulitermes flavipes were analyzed to assess differences in cuticular compounds among castes. We identified a royal-specific hydrocarbon—heneicosane—and several previously unreported and highly royal enriched long-chain alkanes. When applied to glass dummies, heneicosane elicited worker behavioral responses identical to those elicited by live termite queens, including increased vibratory shaking and antennation. Further, the behavioral effects of heneicosane were amplified when presented with nestmate termite workers’ cuticular extracts, underscoring the importance of chemical context in termite royal recognition. Thus, heneicosane is a royal-recognition pheromone that is active in both queens and kings of R. flavipes. The use of heneicosane as a queen and king recognition pheromone by termites suggests that CHCs evolved as royal pheromones ∼150 million years ago, ∼50 million years before their first use as queen-recognition pheromones in social Hymenoptera. We therefore infer that termites and social Hymenoptera convergently evolved the use of these ubiquitous compounds in royal recognition.