Keng Hong Tan

Methyl Eugenol: Its Occurrence, Distribution, and Role in Nature, Especially in Relation to Insect Behavior and Pollination

Abstract This review discusses the occurrence and distribution (within a plant) of methyl eugenol in different plant species (> 450) from 80 families spanning many plant orders, as well as various roles this chemical plays in nature, especially in the interactions between tephritid fruit flies and plants.

Accumulation ofDendrobium superbum (orchidaceae) fragrance in the rectal glands by males of the melon fly,Dacus cucurbitae.

Abstract4-(4-Hydroxyphenyl)-2-butanone was characterized from flowers of the orchidDendrobium superbum as a specific attractant factor for the male melon fly,Dacus cucurbitae. The male flies compulsively licked the flower surface and sequestered the compound in significant quantities in their rectal glands. The compound was detected within 6 hr after ingestion and was retained for more than six days in the rectal gland sacs.

Comparative sensitivity to and consumption of methyl eugenol in three Bactrocera dorsalis (Diptera: Tephritidae) complex sibling species

Summary. Sensitivities to methyl eugenol of three sibling species in the Bactrocera dorsalis complex were compared. The degree of species sensitivity to methyl eugenol, i.e. B. dorsalis > B. papayae > B. carambolae (in decreasing order), was concomitant with the species age-related response to methyl eugenol as previously reported. The ability to consume methyl eugenol by the three sibling species showed similar trend - the average ME consumption per male was 0.70 ml for B. dorsalis, 0.58 ml B. papayae and 0.18 μl B. carambolae. Results obtained were discussed in relation to area-wide control of fruit fly.

Pharmacophagy of methyl eugenol by males enhances sexual selection of Bactrocera carambolae.

After pharmacophagy of methyl eugenol (ME), males of Bactrocera carambolae (Diptera: Tephritidae) produced (E)-coniferyl alcohol (CF) along with its endogenously synthesized pheromonal compounds. CF was shown to be released into the air by the ME-fed males only during the courtship period at dusk and attracted significantly more males and females than the ME-deprived males in wind tunnel assays. However, earlier onset of sexual attraction and a higher mating success were observed only in the wind tunnel and field cage assays on the third day posttreatment of ME. Field cage observations on the male-to-male interaction indicated that the ME-deprived males did not exhibit aggregation behavior, but that ME feeding promoted aggregation behavior in B. carambolae. Field cage observations revealed that the ME-deprived males were not only attracted to the ME-fed males, but also appeared to feed on their anal secretions. The secretions were subsequently confirmed to contain CF along with endogenously produced pheromonal compounds. Results obtained for B. carambolae were compared to those previously obtained from its sibling species, Bactrocera dorsalis, and are discussed in light of species advancement in fruit fly–plant relationships.

Floral phenylpropanoid cocktail and architecture of Bulbophyllum vinaceum orchid in attracting fruit flies for pollination.

It is widely believed that most orchid flowers attract insects by using deception or chemical rewards in the form of nectar. Flowers of Bulbophyllum vinaceum produce a large array of phenylpropanoids that lure tephritid fruit fly males and also act as floral reward, which the flies subsequently convert to pheromone components. The major floral volatile components identified are methyl eugenol (ME), trans-coniferyl alcohol (CF), 2-allyl-4,5-dimethoxphenol (DMP), and trans-3,4-dimethoxycinnamyl acetate, whereas the minor components are eugenol, euasarone, trans-3,4-dimethoxy cinnamyl alcohol, and cis-coniferyl alcohol. Among the various floral parts, the lip (which is held in a closed position up against the sexual organs) has the highest concentration of the major compounds. An attracted male fly normally lands on one of the petals before climbing up onto and forcing the “spring loaded” floral lip into the open position, hence exposing the floral sexual organs. The architecture and location of chemical attractants of the lip compel the fly to align itself along the lip’s longitudinal axis in a precise manner. As the fly laps up the compounds and moves towards the base of the lip, it passes the point of imbalance causing the lip to spring back to its normal closed position. The fly is catapulted headfirst into the column cavity, and its dorsum strikes the protruding sticky base of the hamulus and adheres to it. The momentum of the fly and the structural morphology of the long stiff hamulus act to pry out the pollinia from its anther cover. Hence, the pollinarium (pollinia + hamulus) is detached from the flower and adhered to the fly’s dorsum. In this unique mutualistic association, both species receive direct reproductive benefits—the flower’s pollinarium is transported for cross pollination, and the fly is offered a bouquet of phenylpropanoids (synomone) that it consumes, converts, and/or sequesters as sex pheromonal components, thus enhancing sexual attraction and mating success.

Phenylpropanoids in the fragrance of the fruit fly orchid, Bulbophyllum cheiri, and their relationship to the pollinator, Bactrocera papayae

Flowers of a southeast Asian orchid, Bulbophyllum cheiri, attract males of Bactrocera papayae and other fruit fly species sensitive to methyl eugenol for pollination. In addition to the major attractive component methyl eugenol, several other related phenylpropanoids were identified as minor components from the floral extracts. 2-Allyl-4,5-dimethoxyphenol, known as an element of sex and aggregation pheromone of B. papayae males was detected as a significant volatile ingredient together with its O-methyl ether (euasarone). The attractancy of each floral component and efficiency of incorporation of these phenylpropanoids into the rectal pheromone glands were examined in B. papayae males.

Evidence of Natural Hybridization Between Two Sympatric Sibling Species of Bactrocera dorsalis Complex Based on Pheromone Analysis

Bactrocera carambolae and B. papayae are major fruit fly pests and sympatric sibling species of the B. dorsalis complex. They possess distinct differences in male pheromonal components. In the 1990’s, wild Bactrocera fruit flies with morphological traits intermediate between those of B. carambolae and B. papayae were often captured in traps baited with methyl eugenol (ME). Chemical analyses of rectal glands of ME-fed males revealed that the laboratory F1, F2, and backcross hybrids possessed ME-derived sex pheromonal components ranging from that typical of B. papayae to that of B. carambolae without any specific trend, which included a combination of pheromonal components from both parental species within an individual hybrid. ME-fed hybrids without any ME-derived pheromonal components were also detected. Further chemical analysis of rectal glands from wild Bactrocera males, after ME feeding in the laboratory, showed a combination of pheromonal components similar to that found in the ME-fed, laboratory-bred hybrids. These findings present circumstantial evidence for the occurrence of a natural hybrid of the two Bactrocera species.

Comparison of methyl eugenol metabolites, mitochondrial COI, and rDNA sequences of Bactrocera philippinensis (Diptera: Tephritidae) with those of three other major pest species within the dorsalis complex

Males of the oriental fruit fly, Bactrocera dorsalis (Hendel) and some of its sibling species have strong affinity for methyl eugenol (ME). Methyl eugenol ingested by male flies is biotransformed in the crop to two ME metabolites that eventually accumulate in the rectal gland, which is known to serve as a reservoir for B. dorsalis sex pheromones. When fed with ME, males of laboratory and wild B. philippinensis Drew and Hancock selectively accumulated two metabolites, 2-allyl-4,5-dimethoxyphenol and (E)-coniferyl alcohol, in the rectal gland, as was seen for B. dorsalis sensu stricto, B. invadens Drew, Tsuruta and White, and B. papayae Drew and Hancock. Phylogenetic analysis of COI and rDNA sequence data of these four taxa also revealed a close relationship among B. philippinensis, B. dorsalis s.s., B. invadens, and B. papayae (all four are members of the dorsalis species complex). This result corroborates pheromone analysis. The usefulness of pheromonal analysis as a chemotaxonomy tool to complement molecular and other analysis in differentiation of closely related sibling species within the Bactrocera dorsalis complex, for which use of morphological characters had been inadequate, is highlighted.

Allomonal and Hepatotoxic Effects Following Methyl Eugenol Consumption in Bactrocera papayae Male Against Gekko monarchus

Methyl eugenol (ME), is converted into two major phenylpropanoids, 2-allyl-4,5-dimethoxyphenol and trans-coniferyl alcohol, following consumption by the male fruit fly Bactrocera papayae. Chemical analysis of wild male B. papayae rectal glands, where the compounds are sequestered, revealed the presence of ME metabolites in varying quantities. These phenylpropanoids are shown to be involved in the fruit fly defense both in no-choice and choice feeding tests against the Malayan spiny gecko, Gekko monarchus. After being acclimatized to feeding on fruit flies, geckos consumed significantly fewer ME-fed male flies than controls that consumed all the ME-deprived male flies offered throughout a two-week period. Diagnosis of dissected livers from geckos that consumed ME-fed male flies revealed various abnormalities. These included discoloration and hardening of liver tissue, whitening of the gallbladder, or presence of tumorlike growths in all geckos that consumed ME-fed male flies. Control geckos fed on ME-deprived male flies had healthy livers. When given an alternative prey, geckos preferred to eat untreated house flies, Musca domestica to avoid preying on ME-fed fruit flie

Temporal accumulation of phenylpropanoids in male fruit flies, Bactrocera dorsalis and B. carambolae (Diptera: Tephritidae) following methyl eugenol consumption

Summary.Males of dacine tephritids, Bactrocera carambolae and B. dorsalis are strongly attracted to, and compulsively feed on methyl eugenol (ME), a potent attractant for many Bactrocera species. While ME was shown to be biotransformed into phenylpropanoids, 2-allyl-4,5-dimethoxyphenol and (E)-coniferyl alcohol, and temporarily stored in the rectal gland of male B. dorsalis prior to release during courtship at dusk, B. carambolae male produces only the latter compound along with its de novo synthesized pheromone components. Both species were also shown to have different age-related response, sensitivity and consumption levels of ME. Here, we monitored and compared temporal changes in the accumulation profiles of these phenylpropanoids by the two sibling species, with male rectal glands being individually excised at different time intervals from 15 minutes to 20 days after initial ME feeding and analysed quantitatively. Results are discussed in light of plant-fruit fly co-evolution relationship.