Comparison of phenylpropanoid volatiles in male rectal pheromone gland after methyl eugenol consumption, and molecular phylogenetic relationship of four global pest fruit fly species: Bactrocera invadens, B. dorsalis, B. correcta and B. zonata

Males of many tephritid fruit fly species of the genus Bactrocera show a very strong affinity to methyl eugenol (ME). An attracted male compulsively ingests ME, which is then biotransformed before its metabolites are accumulated into the rectal gland. The glandular organ is known to serve as a reservoir for sex pheromone in some species. Upon ME-feeding, males of the oriental fruit fly, Bactrocera dorsalis, selectively accumulated two metabolites, 2-allyl-4,5-dimethoxyphenol (DMP) and (E)-coniferyl alcohol (E-CF), in the rectal pheromone gland. We compared the profiles of phenylpropanoid metabolites accumulated by three other species of very high economic and quarantine importance-Bactrocera invadens, Bactrocera zonata and Bactrocera correcta, with that of B. dorsalis. Males of each species were fed artificially on ME and the metabolites stored in the rectal glands were examined by means of chromatography and spectroscopy. Similar to B. dorsalis, males of laboratory-raised B. invadens accumulated DMP and E-CF, in almost equal quantities, in the rectal sac. The sum of DMP and E-CF increased gradually with time after ME consumption and reached as high as 150 mu g/male 2 days post ME-feeding. Wild males of B. invadens captured in Kenya also possessed both the compounds in varying quantities. In contrast, males of B. zonata accumulated DMP and (Z)-coniferyl alcohol (Z-CF) in an approximate ratio of 1:1; whereas B. correcta is known to convert ME to (Z)-3,4-dimethoxycinnamyl alcohol (Z-DMC) and Z-CF also in an approximately 1:1 ratio. Thus, there are three types of binary combinations of rectal phenylpropanoid volatiles (i.e. DMP + E-CF; DMP + Z-CF; Z-CF + Z-DMC) utilized among the four Bactrocera species. Such differences in phenylpropanoid ingredients may play a critical role in differentiating these species if encountered in the natural habitat. In this context, the two putative sibling species-B. invadens and B. dorsalis, possess the identical subset of rectal volatiles (DMP and E-CF) in a similar proportion. Furthermore, the phylogenetic analyses of the four Bactrocera species by comparing nucleotide sequences in the mitochondrial genes showed that B. invadens clearly belonged to the same clade as B. dorsalis species. Therefore, we consider the two as the same biological species, and certainly not distinct.