, Volume 29, Issue 1, pp 25–34 | Cite as

Phenylpropanoid sex pheromone component in hemolymph of male Carambola fruit fly, Bactrocera carambolae (Diptera: Tephritidae)

  • Wei-Wei Hiap
  • Suk-Ling Wee
  • Keng-Hong Tan
  • Alvin Kah-Wei HeeEmail author
Original Article


Males of the Carambola fruit fly, Bactrocera carambolae Drew & Hancock (Diptera: Tephritidae) are strongly attracted to, and feed on methyl eugenol (ME) that exists as a plant secondary compound in over 480 plant species worldwide. Upon feeding on this highly potent attractant, the males convert ME into a phenylpropanoid, (E)-coniferyl alcohol (ECF), that is stored in the rectal gland prior to its release as a sex pheromone component during calling and courtship. Here, using a series of chemical and behavioural assays, we provide evidence for the presence of ECF in the hemolymph and suggest the latter’s involvement in transport of ECF to the male rectal gland following consumption of ME. The greatest concentration of ECF was detected in the hemolymph at 3 h after feeding on ME and subsequently decreased, whereas accumulation of ECF in the rectal gland reached a maximum at 2 days post-feeding. Using male flies as biodetectors, fractions of 1.5–9.2 kDa from fractionated hemolymph of ME-fed males were found to be attractive and contained ECF as sex pheromone. In addition, the significant increase in the total concentration of protein in hemolymph from ME-fed males compared with that of ME-deprived males suggests a direct protein carrier involvement in hemolymph transport of the sex pheromone in B. carambolae. All these results are further discussed in comparison with previous results obtained from its sibling species, the Oriental fruit fly—B. dorsalis.


Bactrocera carambolae Methyl eugenol (E)-coniferyl alcohol Hemolymph Rectal gland Sex pheromone 



Carambola fruit fly


Oriental fruit fly


(E)-coniferyl alcohol


Methyl eugenol


Male annihilation technique


Sterile insect technique




Days after emergence


Internal standard


Bioactive fractions



This research was funded by the Fundamental Research Grant Scheme (FRGS No.: 01-11-09-715FR). We also thank Dr R. Nishida, Professor Emeritus in Kyoto University for providing us (E)-coniferyl alcohol.

Author contributions

AKWH and KHT conceived and designed the methodology; WWH collected the data; WWH, SLW, AKWH and KHT analyzed the data; WWH and AKWH led the writing of the manuscript. All authors contributed critically to the drafts and approved the final draft for publication.

Compliance with ethical standards

Conflict of interest

The authors declare that there is no conflict of interest.


  1. Aketarawong N, Isasawin S, Sojikul P, Thanaphum S (2015) Gene flow and genetic structure of Bactrocera carambolae (Diptera, Tephritidae) among geographical differences and sister species, B. dorsalis, inferred from microsatellite DNA data. ZooKeys 540:239–272CrossRefGoogle Scholar
  2. Allwood AJ, Chinajariyawong A, Drew RAI, Hameck EL, Hancock DL, Hengsawad JC, Jipanin M, Konkrong C, Kritsaneepaiboon S, Leong CTS, Vijaysegaran S (1999) Host plant records for fruit flies (Diptera: Tephritidae) in South East Asia. Raffles B Zool 7:1–92Google Scholar
  3. Barclay HJ, McInnis D, Hendrichs J (2014) Modeling the area-wide integration of male annihilation and the simultaneous release of methyl eugenol-exposed Bactrocera spp. sterile males. Ann Entomol Soc Am 107:97–112CrossRefGoogle Scholar
  4. Bradford MM (1976) A rapid and sensitive method for quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254CrossRefGoogle Scholar
  5. Clarke AR, Armstrong KF, Carmichael AE, Milne JR, Raghu S, Roderick GK, Yeates DK (2005) Invasive phytophagous pests arising through a recent tropical evolutionary radiation: the Bactrocera dorsalis complex of fruit flies. Annu Rev Entomol 50:293–319CrossRefGoogle Scholar
  6. Drew RAI, Romig MC (1997) Overview–Tephritidae in the Pacific and South-East Asia. In: Allwood AJ, Drew RAI (eds) Management of fruit flies in the pacific. A regional symposium, ACIAR proceedings, vol 76, pp 46–53Google Scholar
  7. Fan Y, Schal C, Vargo EL, Bagnères AG (2004) Characterization of termite lipophorin and its involvement in hydrocarbon transport. J Insect Physiol 31:609–620CrossRefGoogle Scholar
  8. Gu X, Quilici D, Juarez P, Blomquist GJ, Schal C (1995) Biosynthesis of hydrocarbons and contact sex pheromone and their transport by lipophorin in females of the German cockroach (Blattella germanica). J Insect Physiol 41:257–267CrossRefGoogle Scholar
  9. Hee AKW, Tan KH (1998) Attraction of female and male Bactrocera papayae to conspecific males fed with methyl eugenol and attraction of females to male sex pheromone components. J Chem Ecol 24:753–764CrossRefGoogle Scholar
  10. Hee AKW, Tan KH (2004) Male sex pheromonal components derived from methyl eugenol in the hemolymph of the fruit fly Bactrocera papayae. J Chem Ecol 30:2127–2137CrossRefGoogle Scholar
  11. Hee AKW, Tan KH (2005) Bioactive fractions containing methyl eugenol-derived sex pheromonal components in haemolymph of the male fruit fly Bactrocera dorsalis (Diptera: Tephritidae). Bull Entomol Res 95:615–619CrossRefGoogle Scholar
  12. Hee AKW, Tan KH (2006) Transport of methyl eugenol-derived sex pheromonal components in the male fruit fly, Bactrocera dorsalis. Comp Biochem Physiol C Toxicol Pharmacol 143:422–428CrossRefGoogle Scholar
  13. Iwaizumi R (2004) Species and host record of the Bactrocera dorsalis complex (Diptera: Tephritidae detected by the plant quarantine of Japan. Appl Entomol Zool 39:327–333CrossRefGoogle Scholar
  14. Khoo CCH, Tan KH (2005) Rectal gland of Bactrocera papayae: Ultrastructure, anatomy, and sequestration of autofluorescent compounds upon methyl eugenol consumption by the male fruit fly. Microsc Res Tech 67:219–226CrossRefGoogle Scholar
  15. Lim CL, Tan ST, Tan KH (1998) Enzymatic assay of a mixed-function monooxygenase in Bactrocera papayae. In: Fifth international symposium on fruit flies of economic importance, Penang, Malaysia, 1–5 June, 1998, p 139Google Scholar
  16. Marchioro CA (2016) Global potential distribution of Bactrocera carambolae and the risks for fruit production in Brazil. PLoS One 11:e0166142CrossRefGoogle Scholar
  17. Myers SW, Cancio-Martinez E, Hallman GJ, Fontenot EA, Vreysen MJ (2016) Relative tolerance of six Bactrocera (Diptera: Tephritidae) species to phytosanitary cold treatment. J Econ Entomol 109:2341–2347CrossRefGoogle Scholar
  18. Nishida R, Tan KH, Serit M, Lajis NH, Sukari AM, Takahashi T, Fukami H (1988a) Accumulation of phenylpropanoids in the rectal glands of males of the Oriental fruit fly, Dacus dorsalis. Cell Mol Life Sci 44:534–536CrossRefGoogle Scholar
  19. Nishida R, Tan KH, Fukami H (1988b) Cis-3,4-dimethoxycinnamyl alcohol from the rectal glands of male Oriental fruit fly, Dacus dorsalis. Chem Express 3:207–210Google Scholar
  20. Opitz SE, Müller C (2009) Plant chemistry and insect sequestration. Chemoecology 19:117–154CrossRefGoogle Scholar
  21. Perkins MV, Fletcher MT, Kitching W, Drew RAI, Moore CJ (1990) Chemical studies of rectal gland secretions of some species of Bactrocera dorsalis complex of fruit flies (Diptera: Tephritidae). J Chem Ecol 16:2475–2487CrossRefGoogle Scholar
  22. San Jose M, Doorenweerd C, Leblanc L, Barr N, Geib S, Rubinoff D (2018) Tracking the origins of fly invasions; using mitochondrial haplotype diversity to identify potential source populations in two genetically intertwined fruit fly species (Bactrocera carambolae and Bactrocera dorsalis [Diptera: Tephritidae]). J Econ Entomol. Google Scholar
  23. Schal C, Sevala V, Cardé RT (1998) Novel and highly specific transport of a volatile sex pheromone by hemolymph lipophorin in moths. Naturwissenschaften 85:339–342CrossRefGoogle Scholar
  24. Schuler MA (1996) The role of cytochrome P450 monooxygenases in plant-insect interactions. Plant Physiol 112:1411–1419CrossRefGoogle Scholar
  25. Schutze MK, Aketarawong N, Amornsak W, Armstrong KF, Augustinos A, Barr N, Bo W, Bourtzis K, Boykin LM, Ca´ceres C, Cameron SL, Chapman TA, Chinvinijkul S, Chomicˇ A, De Meyer M, Drosopoulou ED, Englezou A, Ekesi S, Gariou-Papalexiou A, Hailstones D, Haymer D, Hee AKW, Hendrichs J, Hasanuzzaman M, Jessup A, Khamis FM, Krosch MN, Leblanc L, Mahmood K, Malacrida AR, Mavragani-Tsipidou P, McInnis DO, Mwatawala M, Nishida R, Ono H, Reyes J, Rubinoff DR, San Jose M, Shelly TE, Srikachar S, Tan KH, Thanaphum S, Ul Haq I, Vijaysegaran S, Wee SL, Yesmin F, Zacharopoulou A, Clarke AR (2015) Synonymization of key pest species within the Bactrocera dorsalis complex (Diptera: Tephritidae): taxonomic changes based on 20 years of integrative morphological, genetic, behavioural, and chemoecological data. Syst Entomol 40:456–471CrossRefGoogle Scholar
  26. Sevala VL, Bachmann JA, Schal C (1997) Lipophorin: a hemolymph juvenile hormone binding protein in the German cockroach, Blattella germanica. Insect Biochem Mol Biol 31:663–670CrossRefGoogle Scholar
  27. Shapiro JP, Law JH, Wells MA (1988) Lipid transport in insects. Annu Rev Entomol 33:297–318CrossRefGoogle Scholar
  28. Silva TL, Lima AL, Sousa MSM, Jesus-Barros CR, Bariani A, Pereira JF, Adaime R (2016) Potential of Amazonian isolates of Metarhizium to control immatures of Bactrocera carambolae (Diptera: Tephritidae). Fla Entomol 99:788–789CrossRefGoogle Scholar
  29. Steiner LF, Hart WG, Harris EJ, Cunningham RT, Ohinata K, Kamakahi DC (1970) Eradication of the Oriental fruit fly from the Mariana Islands by the methods of male annihilation and sterile insect release. J Econ Entomol 63:131–135CrossRefGoogle Scholar
  30. Strauss AS, Peters S, Boland W, Burse A (2013) ABC transporter functions as a pacemaker for sequestration of plant glucosides in leaf beetles. eLife 2, e01096CrossRefGoogle Scholar
  31. Tan KH (1985) Estimation of native populations of male Dacus spp. by using Jolly’s stochastic method using a new designed attractant trap in a village ecosystem. J Plant Prot Trop 2:87–95Google Scholar
  32. Tan KH, Nishida R (1996) Sex pheromone and mating competition after methyl eugenol consumption in the Bactrocera dorsalis complex. In: McPheron B, Steck GJ (eds) Fruit fly pest. Lucie Press, Florida, pp 147–153Google Scholar
  33. Tan KH, Nishida R (2012) Methyl eugenol: its occurrence, distribution, and role in nature, especially in relation to insect behavior and pollination. J Insect Sci 12(1):1–60CrossRefGoogle Scholar
  34. Tan KH, Nishida R, Jang EB, Shelly TE (2014) Pheromones, male lures, and trapping of tephritid fruit flies. In: Shelly TE, Epsky N, Jang EB, Reyes-Flores J, Vargas RI (eds) Trapping and the detection, control, and regulation of tephritid fruit flies. Springer, Dondrecht, pp 15–74Google Scholar
  35. Vijaysegaran S (1997) Fruit fly research and development in tropical Asia. In: Allwood AJ, Drew RAI (eds) Management of fruit flies in the Pacific. A regional symposium, ACIAR proceedings, vol 76, pp 21–29Google Scholar
  36. Wee SL, Tan KH (2000) Sexual maturity and intraspecific mating success of two sibling species of the Bactrocera dorsalis complex. Entomol Exp Appl 94:133–139CrossRefGoogle Scholar
  37. Wee SL, Tan KH (2005) Evidence of natural hybridization between two sympatric sibling species of Bactrocera dorsalis complex based on pheromone analysis. J Chem Ecol 31:845–858CrossRefGoogle Scholar
  38. Wee SL, Tan KH (2007) Temporal accumulation of phenylpropanoids in male fruit flies, Bactrocera dorsalis and B. carambolae (Diptera: Tephritidae) following methyl eugenol consumption. Chemoecology 17:81–85CrossRefGoogle Scholar
  39. Wee SL, Hee AKW, Tan KH (2002) Comparative sensitivity to and consumption of methyl eugenol in three Bactrocera dorsalis (Diptera: Tephritidae) complex sibling species. Chemoecology 12:193–197CrossRefGoogle Scholar
  40. Wee SL, Tan KH, Nishida R (2007) Pharmacophagy of methyl eugenol by males enhances sexual selection of Bactrocera carambolae.. J Chem Ecol 33:1279–1282CrossRefGoogle Scholar
  41. Westerlund SA, Hoffmann KH (2004) Rapid quantification of juvenile hormones and their metabolites in insect haemolymph by liquid chromatography–mass spectrometry (LC–MS). Anal Bioanal Chem 379:540–543CrossRefGoogle Scholar
  42. Wyatt G, Pan ML (1978) Insect plasma proteins. Annu Rev Biochem 47:779–817CrossRefGoogle Scholar
  43. Xu HX, Zheng XS, Yang YJ, Tian JC, Lu YH, Tan KH, Heong KL, Lu ZX (2015) Methyl eugenol bioactivities as a new potential botanical insecticide against major insect pests and their natural enemies on rice (Oryza sativa). Crop Prot 72:144–149CrossRefGoogle Scholar
  44. Yagi KJ, Tobe SS (2001) The radiochemical assay for juvenile hormone biosynthesis in insects: problems and solutions. J Insect Physiol 47:1227–1234CrossRefGoogle Scholar

Copyright information

© Springer Nature Switzerland AG 2018

Authors and Affiliations

  1. 1.Laboratory of Entomology and Chemical Ecology, Department of Biology, Faculty of ScienceUniversiti Putra Malaysia, UPMSerdangMalaysia
  2. 2.School of Environmental and Natural Resource SciencesUniversiti Kebangsaan MalaysiaBangiMalaysia
  3. 3.Centre of Insect Systematics, Faculty of Science & TechnologyUniversiti Kebangsaan MalaysiaBangiMalaysia
  4. 4.Mobula ResearchTanjong BungahMalaysia

Personalised recommendations