Wendell L. Roelofs

Evolution of moth sex pheromones via ancestral genes

Mate finding in most moth species involves long-distance signaling via female-emitted sex pheromones. There is a great diversity of pheromone structures used throughout the Lepidoptera, even among closely related species. The conundrum is how signal divergence has occurred. With strong normalizing selection pressure on blend composition and response preferences, it is improbable that shifts to pheromones of diverse structures occur through adaptive changes in small steps. Here, we present data supporting the hypothesis that a major shift in the pheromone of an Ostrinia species occurred by activation of a nonfunctional desaturase gene transcript present in the pheromone gland. We also demonstrate the existence of rare males that respond to the new pheromone blend. Their presence would allow for asymmetric tracking of male response to the new blend and, thus, evolution of an Ostrinia species with structurally different sex pheromone components.

Sex Attractant of the Codling Moth: Characterization with Electroantennogram Technique

trans-8, trans-10-Dodecadien-1-ol is a sex attractant of the codling moth, Laspeyresia pomonella. Antennal responses (electroantennograms) to a series of monounsaturated compounds were used in determining the location and configuration of the two double bonds. The synthetic compound is very attractive to male codling moths in the field.

Fruit odor discrimination and sympatric host race formation in Rhagoletis

Rhagoletis pomonella is a model for incipient sympatric speciation (divergence without geographic isolation) by host-plant shifts. Here, we show that historically derived apple- and ancestral hawthorn-infesting host races of the fly use fruit odor as a key olfactory cue to help distinguish between their respective plants. In flight-tunnel assays and field tests, apple and hawthorn flies preferentially oriented to, and were captured with, chemical blends of their natal fruit volatiles. Because R. pomonella rendezvous on or near the unabscised fruit of their hosts to mate, the behavioral preference for apple vs. hawthorn fruit odor translates directly into premating reproductive isolation between the fly races. We have therefore identified a key and recently evolved (<150 years) mechanism responsible for host choice in R. pomonella bearing directly on sympatric host race formation and speciation.

Molecular genetics and evolution of pheromone biosynthesis in Lepidoptera

A great diversity of pheromone structures are used by moth species (Insecta: Lepidoptera) for long-distance mating signals. The signal/response channel seems to be narrow for each species, and a major conundrum is how signal divergence has occurred in the face of strong selection pressures against small changes in the signal. Observations of various closely related and morphologically similar species that use pheromone components biosynthesized by different enzymes and biosynthetic routes underscore the question as to how major jumps in the biosynthetic routes could have evolved with a mate recognition system that is based on responses to a specific blend of chemicals. Research on the desaturases used in the pheromone biosynthetic pathway for various moth species has revealed that one way to make a major shift in the pheromone blend is by activation of a different desaturase from mRNA that already exists in the pheromone gland. Data will be presented to support the hypothesis that this process was used in the evolution of the Asian corn borer, Ostrinia furnacalis species. In that context, moth sex-pheromone desaturase genes seem to be evolving under a birth-and-death process. According to this model of multigene family evolution, some genes are maintained in the genome for long periods of time, whereas others become deleted or lose their functionality, and new genes are created through gene duplication. This mode of evolution seems to play a role in moth speciation, as exemplified by the case of the Asian corn borer and European corn borer, Ostrinia nubilalis species.

Sex pheromone perception: Electroantennogram responses of the red-banded leaf roller moth

Abstract Electroantennograms (EAG) were recorded for various compounds which can modify the attractancy of the red-banded leaf roller moth sex pheromone. Inhibitors and synergists elicit definite antennal responses in accordance with a proposed protein receptor model for pheromone perception. The sex pheromone produced the highest amplitude EAG responses of all monounsaturated positional isomers, but no correlation could be made between EAG shape and behavioural activity.

Biosynthesis of lepidopteran pheromones

Abstract Research on lepidopteran sex pheromone component biosynthetic pathways has revealed general systems that may have significance in understanding the evolution of these moth mating communication signals. Studies with the redbanded leafroller moth, cabbage looper moth, and the domestic silkworm showed that they all possess a unique delta-11 unsaturated acid precursor. Radiolabeled precursor acids were used to show that various combinations of limited beta-oxidation chain-shortening or chain-elongation steps with the desaturase enzyme could produce most of the pheromone components identified for noctuid, pyralid, and tortricid moths. Evolution of the delta-11 desaturase enzyme from the ubiquitous delta-9 desaturase enzyme was suggested by finding primitive species that use the intermediate delta-10 desaturase enzyme. It is suggested that pheromone components of other primitive species are produced by using only the chain-shortening steps on available oleate, linoleate, and linolenate. Pheromone componets of some more advanced species appear to be produced by chain elongation of these available acids, with subsequent reductive decarboxylation to hydrocarbon.

Two sex pheromone components of the tobacco budworm moth, Heliothisvirescens

Abstract Two compounds were isolated from female Heliothis virescens (Lepidoptera: Noctuidae) extracts and identified as cis -9-tetradecenal and cis -ll-hexadecenal. Together they elicit intense male H . virescnes response in laboratory tests and have attracted males in the field. Although cis -ll-hexadecenal is an H . zea sex pheromone, no evidence was obtained for cis -9-tetradecenal in H . zea .

Sex pheromone of the european corn borer,Ostrinia nubilalis (Lepidoptera: Pyralidae), in New York

Abstractteans-11-Tetradecenyl acetate (96%) andcis-11-tetradecenyl acetate (4%) are pheromone components of the European corn borer,Ostrinia nubilalis (Hübner), from New York. This isomeric blend extracted from the female abdominal tips is very similar to the most attractive synthetic mixture in the field in New York. AnO. nubilalis strain from London, Ontario, was found to produce 97%cis-11-tetradecenyl acetate and 3%trans-11-tetradecenyl acetate.

Male moth sensitivity to multicomponent pheromones: Critical role of female-released blend in determining the functional role of components and active space of the pheromone.

In the present study male redbanded leafroller (Argyrotaenia velutinana), cabbage looper (Trichoplusia ni), and Oriental fruit moths, (Grapholita molesta), were tested in a flight tunnel to (1) the major pheromone component, (2) theZ/E pheromone component mixtures for Oriental fruit moth and redbanded leafroller, (3) and the female-released blends, over a series of dosages. Experiments were designed to test the hypothesis that male response downwind of a female is initiated by the major component and that minor components function only to elicit behaviors close to the female during close-range approach and courtship. The results did not support this hypothesis, but rather showed that males initiated upwind flight in significantly higher percentages to the complete blends of components, at all dosages, compared to single components or partial blends. Addition of minor components also signficantly enhanced male perception of the major component at lower dosages, resulting in completed flights to dosages of the major component that alone did not elicit any upwind flight. Our results support the concept that minor components function to enhance male sensitivity to the pheromone, and the specificity of the signal. Our results also support the hypothesis that the active space of the pheromone is a function of the upper and lower concentration thresholds for the blend of components, and not simply for the major component.

Identification of new sex pheromone components inTrichoplusia ni, predicted from biosynthetic precursors.

In addition to the previously identified components (Z)-7-dodecenyl acetate and dodecyl acetate, sex pheromone glands ofTrichoplusia ni release (Z)-5-dodecenyl acetate, 11-dodecenyl acetate, (Z)-7-tetradecenyl acetate, and (Z)-9-tetradecenyl acetate. Bioassays in a flight tunnel showed that a synthetic blend of these six compounds elicited complete flights to the source from 95% of the males tested and elicited hairpenciling responses at the end of the flights from 88% of the males tested. This blend was not significantly different from intact pheromone glands, which elicited complete flights to the source from 98% of the males tested and hairpenciling responses from 91% of the males tested. In contrast, the previously identified two-component blend elicited significantly fewer complete flights to the source (33%) and did not elicit hairpenciling responses from any of the males tested. The search for additional sex pheromone components was prompted by our previous identification of unusual fatty acyl moieties in the gland that seemed to be possible biosynthetic intermediates.