Abstract
Male orchid bees (Euglossini) collect fragrances from flowers and other natural sources, a behaviour that has shaped the euglossine pollination syndrome. Males store such chemicals in hind leg pouches and later expose them during courtship display. In the present study, we show that complex bouquets of two sympatric species of Eulaema, E. meriana and E. bombiformis, are chemically distinct. When exposed during bioassays at display perches individual hind leg extracts rapidly and consistently attracted other males of the correct species, even if derived from males of disparate localities (French Guiana and Panama). Conspecific males as well as females of E. bombiformis arrived at natural perch sites only from downwind, and two copulations were observed. Our findings demonstrate that acquired odours mediate exclusive attraction within species and support the idea that such fragrances are pheromone analogues. Their role in acquiring matings and during male–male interaction is discussed.
Similar content being viewed by others
References
Ackerman JD, Montalvo AM (1985) Longevity of euglossine bees. Biotropica 17:79–81
Adams RP (2001) Identification of essential oil components by gas chromatography/quadrupole mass spectroscopy. Allured Publishing, Carol Stream, USA
Ayasse M, Paxton RJ, Tengo J (2001) Mating behavior and chemical communication in the order Hymenoptera. Annu Rev Entomol 46:31–78
Bembé B (2004) Functional morphology in male euglossine bees and their ability to spray fragrances (Hymenoptera, Apidae, Euglossini). Apidologie 35:283–291
Clarke KR, Green RH (1988) Statistical design and analysis for a ‘biological effects’ study. Mar Ecol Prog Ser 46:213–226
Clarke KR, Gorley RN (2001) PRIMER v5: user manual/tutorial. Primer-E, Plymouth
Clarke KR, Warwick RM (2001) Change in marine communities: an approach to statistical analysis and interpretation, 2nd edn. Natural Environment Research Council, UK
Dressler RL (1982) Biology of the orchid bees (Euglossini). Ann Rev Ecol Syst 13:373–394
Eberhard WG (1997) Graverobbing by male Eulaema sebrai bees (Hymenoptera: Apidae). J Kans Entomol Soc 70:66
Eltz T, Lunau K (2005) Antennal response to fragrance compounds in male orchid bees. Chemoecology 15:135–138
Eltz T, Whitten WM, Roubik DW, Linsenmair KE (1999) Fragrance collection, storage, and accumulation by individual male orchid bees. J Chem Ecol 25:157–176
Eltz T, Roubik DW, Whitten WM (2003) Fragrances, male display and mating behaviour of Euglossa hemichlora—a flight cage experiment. Phys Entomol 28:251–260
Eltz T, Roubik DW, Lunau K (2005a) Experience-dependent choices ensure species-specific fragrance accumulation in male orchid bees. Behav Ecol Sociobiol 59:149–156
Eltz T, Sager A, Lunau K (2005b) Juggling with volatiles: exposure of perfumes by displaying male orchid bees. J Comp Physiol A 191:575–581
Eltz T, Ayasse M, Lunau K (2006) Species-specific antennal response to tibial fragrances in male orchid bees. J Chem Ecol 32:71–79
Free JB (1987) Pheromones of social bees. Chapman and Hall, London
Gerlach G, Schill R (1991) Composition of orchid scents attracting euglossine bees. Bot Acta 104:379–391
Goulson D (2003) Bumblebees—behavior and ecology. Oxford University Press
Hills HG, Williams NH, Dodson CH (1972) Floral fragrances and isolating mechanisms in the genus Catasetum (Orchidaceae). Biotropica 4:61–76
Kimsey LS (1980) The behaviour of male orchid bees (Apidae, Hymenoptera, Insecta) and the question of leks. Anim Behav 28:996–1004
Kimsey LS (1984) The behavioural and structural aspects of grooming and related activities in euglossine bees (Hymenoptera: Apidae). J Zool 204:541–550
Kindl J, Hovorka O, Urbanova K, Valterova I (1999) Scent marking in male premating behavior of Bombus confusus. J Chem Ecol 25:1489–1500
Kullenberg B (1973) Field experiments with chemical sexual attractants in Aculeate Hymenoptera males II. Zoon Suppl 1:31–42
Legendre P, Legendre L (1998) Numerical ecology, 2nd edn. Elsevier, Amsterdam
O’Neill KM, Evans EJ, Bjostad LB (1991) Territorial behaviour in males of three North American species of bumblebees (Hymenoptera: Apidae, Bombus). Can J Zool 69:604–613
Ramírez S, Dressler RL, Ospina M (2002) Abejas euglosinas (Hymenoptera: Apidae) de la región Neotropical: listado de especies con notas sobre su biología. Biota Colombiana 3:7–118
Roelofs WL (1995) The chemistry of sex attraction. In: Meinwald J (ed) Chemical ecology. National Academy, Washington, DC, pp 103–118
Roubik DW (1998) Grave-robbing by male Eulaema (Hymenoptera, Apidae): implications for euglossine biology. J Kans Entomol Soc 71:188–191
Roubik DW, Hanson PE (2004) Orchid bees of tropical America: biology and field guide. Instituto Nacional de Biodiversidad(INBio), Heredia, Costa Rica
Schemske DW, Lande R (1984) Fragrance collection and territorial display by male orchid bees. Anim Behav 32:935–937
Stern DL (1991) Male territoriality and alternative male behaviors in the euglossine bee, Eulaema meriana (Hymenoptera: Apidae).J Kans Entomol Soc 64:421–437
Stern DL, Dudley R (1991) Wing buzzing by male orchid bees, Eulaema meriana (Hymenoptera: Apidae). J Kans Entomol Soc 64:88–94
Vogel S (1966) Parfümsammelnde Bienen als Bestäuber von Orchidaceen ond Gloxinia. Österr Botan Zeit 113:302–361
Whitten WM, Williams NH (1992) Floral fragrances of Stanhopea (Orchidaceae). Lindleyana 7:130–153
Whitten WM, Williams NH, Armbruster WS, Battiste MA, Strekowski L, Lindquist N (1986) Carvone oxide: an example of convergent evolution in euglossine pollinated plants. Syst Bot 11:222–228
Whitten WM, Hills HG, Williams NH (1988) Occurrence of ipsdienol in floral fragrances. Phytochemistry 27:2759–2760
Whitten WM, Young AM, Williams NH (1989) Function of glandular secretions in fragrance collection by male euglossine bees. J Chem Ecol 15:1285–1295
Whitten WM, Young AM, Stern DL (1993) Nonfloral sources of chemicals that attract male euglossine bees (Apidae: Euglossini). J Chem Ecol 19:3017–3027
Williams NH (1982) The biology of orchids and euglossine bees. In: Arditti J (ed) Orchid biology: reviews and perspectives. Cornell University Press, Ithaca, NY, pp 119–171
Williams NH, Whitten WM (1983) Orchid floral fragrances and male euglossine bees: methods and advances in the last sesquidecade. Biol Bull 164:355–395
Acknowledgements
We thank Oris Acevedo and the entire BCI staff for continuous support during field seasons. The chemical-analytical part of the study was substantiated by Roman Kaiser who analysed reference samples and identified critical fragrance compounds. His help is gratefully acknowledged, as are the valuable discussions with Klaus Lunau and the members of the Sensory Ecology seminar in Düsseldorf. The study was sponsored by the Deutsche Forschungsgemeinschaft (EL 249/2-1)
Author information
Authors and Affiliations
Corresponding author
Additional information
Communicated by R.F.A. Moritz
Appendix
Appendix
Incidence and relative abundance of chemical compounds detected in hexane extracts of hind legs of Eulaema meriana and E. bombiformis. Abundances are the average of a given compound’s (peak) contribution to the individual GC/MS ion chromatogram (in % of the total ion current). Compounds are indexed as exogenous fragrances (F), labial gland lipids also found in head extracts (H), and other high molecular weight non-fragrance compounds (NF).
RT (min) | Compound | Number of individuals with compound | Average rel. abundance (% of total ion current) | ||||
---|---|---|---|---|---|---|---|
Class | E. bombiformis N=7 | E. meriana N=41 | Both N=48 | E. bombiformis | E. meriana | ||
5.32 | Thujene, alpha- | F | 0 | 4 | 4 | 0.00 | 0.01 |
5.52 | Pinene, alpha- | F | 0 | 14 | 14 | 0.00 | 0.04 |
6.24 | Benzaldehyde | F | 0 | 8 | 8 | 0.00 | 0.07 |
6.56 | Sabinene | F | 0 | 34 | 34 | 0.00 | 0.73 |
6.83 | Pinene, beta- | F | 0 | 11 | 11 | 0.00 | 0.05 |
7.07 | Myrcene | F | 4 | 4 | 8 | 0.11 | 0.01 |
7.47 | Menthene, 3-para- ? | F | 0 | 6 | 6 | 0.00 | 0.02 |
7.75 | cyclic monoterpene | F | 0 | 2 | 2 | 0.00 | 0.01 |
8.02 | Terpinene, alpha- | F | 0 | 2 | 2 | 0.00 | 0.00 |
8.29 | Cymene, para- (ortho- ?) | F | 0 | 27 | 27 | 0.00 | 0.32 |
8.46 | Limonene | F | 0 | 10 | 10 | 0.00 | 0.04 |
8.51 | Phellandrene, beta | F | 0 | 3 | 3 | 0.00 | 0.01 |
8.54 | Benzyl alcohol | F | 5 | 7 | 12 | 0.50 | 0.51 |
8.58 | Cineole, 1,8- | F | 0 | 27 | 27 | 0.00 | 0.59 |
8.66 | Ocimene, (Z)-beta- | F | 0 | 2 | 2 | 0.00 | 0.02 |
9.06 | Ocimene, (E)-beta- | F | 7 | 0 | 7 | 1.55 | 0.00 |
9.50 | Terpinene, gamma- | F | 0 | 22 | 22 | 0.00 | 0.23 |
9.93 | Sabinene hydrate, cis- (rel. to OH) | F | 0 | 21 | 21 | 0.00 | 0.68 |
9.99 | ? | F | 0 | 17 | 17 | 0.00 | 0.84 |
10.53 | Terpinolene | F | 0 | 7 | 7 | 0.00 | 0.03 |
11.10 | Sabinene hydrate, trans-? | F | 0 | 12 | 12 | 0.00 | 0.05 |
11.13 | ? | F | 1 | 0 | 1 | 0.04 | 0.00 |
11.15 | ? | F | 0 | 13 | 13 | 0.00 | 0.07 |
11.35 | ? | F | 0 | 11 | 11 | 0.00 | 0.02 |
11.58 | Phenylethyl alcohol, 2- | F | 5 | 0 | 5 | 2.98 | 0.00 |
12.71 | Limonene oxide, trans- ? | F | 1 | 15 | 16 | 0.03 | 0.32 |
12.86 | Ipsdienol | F | 7 | 0 | 7 | 3.65 | 0.00 |
13.71 | Benzyl acetate | F | 5 | 8 | 13 | 2.74 | 0.99 |
14.58 | Terpinen-4-ol | F | 0 | 40 | 40 | 0.00 | 4.80 |
14.95 | Methyl salicylate | F | 0 | 22 | 22 | 0.00 | 0.27 |
15.17 | Terpineol, alpha- | F | 0 | 2 | 2 | 0.00 | 0.01 |
15.36 | Dihydro carvone, cis- ? | F | 0 | 1 | 1 | 0.00 | 0.00 |
15.60 | Dihydro carvone, trans- | F | 0 | 9 | 9 | 0.00 | 0.08 |
16.25 | aromatic compound | F | 0 | 1 | 1 | 0.00 | 0.01 |
16.93 | ? | F | 1 | 0 | 1 | 0.18 | 0.00 |
17.31 | Carvone | F | 3 | 27 | 30 | 0.27 | 0.73 |
17.65 | ? | F | 1 | 0 | 1 | 0.22 | 0.00 |
17.71 | Phenylethyl acetate, 2- | F | 4 | 0 | 4 | 4.27 | 0.00 |
17.90 | ? | F | 1 | 0 | 1 | 2.44 | 0.00 |
18.00 | Carvone epoxide, cis- | F | 5 | 38 | 4 | 3.47 | 12.92 |
18.49 | Geranial | F | 1 | 0 | 1 | 0.09 | 0.00 |
18.82 | Carvone oxide, trans- | F | 0 | 1 | 1 | 0.00 | 0.00 |
18.97 | ? | F | 0 | 8 | 8 | 0.00 | 0.12 |
19.35 | Indole | F | 3 | 0 | 3 | 0.22 | 0.00 |
20.12 | ? | F | 0 | 5 | 5 | 0.00 | 0.01 |
20.99 | ? | F | 2 | 4 | 6 | 0.09 | 0.04 |
21.16 | trans-(trans-Carveol) epoxide? | F | 0 | 12 | 12 | 0.00 | 0.23 |
21.26 | ? | F | 0 | 1 | 1 | 0.00 | 0.01 |
21.47 | ? | F | 0 | 1 | 1 | 0.00 | 0.00 |
21.58 | ? | F | 0 | 1 | 1 | 0.00 | 0.00 |
21.76 | ? | F | 1 | 0 | 1 | 0.04 | 0.00 |
22.22 | Eugenol | F | 0 | 1 | 1 | 0.00 | 0.00 |
23.06 | ? | F | 7 | 0 | 7 | 0.64 | 0.00 |
23.21 | Copaene, alpha- | F | 1 | 0 | 1 | 0.05 | 0.00 |
23.27 | Geranyl acetate | F | 2 | 0 | 2 | 0.11 | 0.00 |
23.58 | Methyl cinnatamte, (E)- | F | 0 | 8 | 8 | 0.00 | 0.07 |
23.74 | Cubebene, beta- | F | 0 | 1 | 1 | 0.00 | 0.01 |
23.75 | ? | F | 0 | 2 | 2 | 0.00 | 0.04 |
23.89 | ? | F | 0 | 3 | 3 | 0.00 | 0.04 |
24.05 | ? | F | 0 | 2 | 2 | 0.00 | 0.01 |
24.75 | ? | F | 0 | 7 | 7 | 0.00 | 0.03 |
24.95 | ? | F | 0 | 5 | 5 | 0.00 | 0.04 |
24.96 | ? | F | 0 | 1 | 1 | 0.00 | 0.01 |
24.99 | Caryophyllene, (Z)- | F | 0 | 1 | 1 | 0.00 | 0.01 |
25.02 | ? | F | 0 | 2 | 2 | 0.00 | 0.02 |
25.67 | Bergamotene, (E-)-alpha- | F | 0 | 2 | 2 | 0.00 | 0.02 |
25.71 | ? | F | 1 | 0 | 1 | 0.04 | 0.00 |
25.72 | ? | F | 1 | 4 | 5 | 0.05 | 0.02 |
26.52 | ? | F | 0 | 1 | 1 | 0.00 | 0.00 |
26.55 | Farnesene, (E)-beta- | F | 0 | 1 | 1 | 0.00 | 0.01 |
26.61 | ? | F | 1 | 0 | 1 | 0.08 | 0.00 |
26.70 | ? | F | 0 | 1 | 1 | 0.00 | 0.01 |
27.51 | ? | F | 0 | 1 | 1 | 0.00 | 0.00 |
27 53 | Dodecanol | H | 0 | 1 | 1 | 0 00 | 0 00 |
27.72 | Germacrene D | F | 1 | 3 | 4 | 0.25 | 0.11 |
27.83 | aff. Farnesene | F | 0 | 1 | 1 | 0.00 | 0.01 |
28.25 | Bicyclogermacrene ? | F | 0 | 2 | 2 | 0.00 | 0.07 |
28.48 | Bulnesene, alpha- ? | F | 2 | 0 | 2 | 0.07 | 0.00 |
28.55 | ? | F | 0 | 1 | 1 | 0.00 | 0.01 |
28.78 | Farnesene | F | 0 | 4 | 4 | 0.00 | 0.02 |
29.00 | ? | F | 0 | 1 | 1 | 0.00 | 0.01 |
30.39 | ? | F | 7 | 20 | 27 | 2.86 | 0.23 |
31.10 | nerolidol, (E)- | F | 0 | 1 | 1 | 0.00 | 0.00 |
31.65 | Spathulenol ? | F | 0 | 1 | 1 | 0.00 | 0.00 |
32.90 | Dodecyl acetate | H | 2 | 12 | 14 | 0.19 | 0.21 |
33.25 | Farnesene epoxide, (E)-alpha- | F | 0 | 6 | 6 | 0.00 | 0.03 |
33.28 | ? | F | 0 | 1 | 1 | 0.00 | 0.01 |
33.31 | ? | F | 0 | 3 | 3 | 0.00 | 0.04 |
33.31 | Tetradecanal (=Myristylaldehyde) | H | 1 | 5 | 6 | 0.35 | 0.01 |
34.38 | ? | F | 7 | 0 | 7 | 2.94 | 0.00 |
34.56 | Methoxynaphtol | F | 7 | 0 | 7 | 1.55 | 0.00 |
35.28 | 2-Propenoic acid, 3-[4-Methoxyphenyl]- | F | 0 | 3 | 3 | 0.00 | 0.01 |
35.65 | Tetradecanol | H | 1 | 1 | 2 | 0.06 | 0.00 |
35.98 | ? | F | 0 | 2 | 2 | 0.00 | 0.01 |
36.48 | ? | F | 0 | 1 | 1 | 0.00 | 0.00 |
36.68 | Bergamotol, (Z)-alpha-, trans- | F | 0 | 15 | 15 | 0.00 | 0.13 |
38.87 | Benzyl benzoate | F | 0 | 6 | 6 | 0.00 | 0.04 |
38.89 | ? | F | 0 | 2 | 2 | 0.00 | 0.01 |
39.24 | Tetradecanoic acid | H | 0 | 1 | 1 | 0.00 | 0.00 |
40.62 | Tetradecyl acetate | H | 3 | 19 | 2 | 6.66 | 1.16 |
41.08 | Hexadecanal | F | 1 | 0 | 1 | 0.15 | 0.00 |
41.50 | ? | F | 0 | 1 | 1 | 0.00 | 0.01 |
41.67 | Hexahydrofarnesyl acetone | F | 0 | 10 | 10 | 0.00 | 0.04 |
42.70 | Benzyl salicylate | F | 0 | 1 | 1 | 0.00 | 0.00 |
43.21 | Hexadecanol | H | 1 | 4 | 5 | 0.10 | 0.02 |
43.41 | ? | F | 0 | 1 | 1 | 0.00 | 0.00 |
45.10 | Cyclohexadecanolid ? | NF | 0 | 1 | 1 | 0.00 | 0.01 |
45.70 | aromatic compound | F | 0 | 1 | 1 | 0.00 | 0.00 |
45.97 | Hexadecanoic acid? | H | 0 | 2 | 2 | 0.00 | 0.03 |
47.47 | Hexadecyl acetate | H | 3 | 22 | 25 | 4.10 | 1.84 |
47.53 | ? | NF | 0 | 2 | 2 | 0.00 | 0.01 |
47.86 | ? | NF | 2 | 5 | 7 | 0.06 | 0.02 |
47.91 | ? | F | 0 | 6 | 6 | 0.00 | 0.03 |
48.36 | ? | F | 0 | 1 | 1 | 0.00 | 0.04 |
48.37 | Falcarinol, (Z)- | F | 0 | 3 | 3 | 0.00 | 0.05 |
49.38 | ? | H | 0 | 2 | 2 | 0.00 | 0.02 |
49.50 | ? | H | 1 | 36 | 37 | 0.02 | 0.46 |
49.91 | ? | H | 1 | 2 | 3 | 0.02 | 0.01 |
50.44 | Heneicosane | H | 1 | 1 | 2 | 0.34 | 0.02 |
50.62 | ? | F | 0 | 1 | 1 | 0.00 | 0.03 |
51.82 | ? | H | 1 | 6 | 7 | 0.50 | 0.37 |
53.06 | ? | F | 1 | 1 | 2 | 0.24 | 0.03 |
53.76 | Octadecanol acetate | H | 3 | 8 | 11 | 1.07 | 0.48 |
55.58 | Tricosene | H | 6 | 40 | 46 | 23.83 | 20.49 |
55.59 | ? | H | 1 | 2 | 3 | 7.75 | 1.00 |
56.47 | Tricosane | H | 6 | 37 | 43 | 2.71 | 2.62 |
58.86 | Octadecanoic acid, butyl ester ? | NF | 1 | 1 | 2 | 0.20 | 0.03 |
61.12 | ? | NF | 0 | 1 | 1 | 0.00 | 0.04 |
61.20 | Pentacosene | H | 1 | 34 | 35 | 0.17 | 2.28 |
62.06 | Pentacosane | H | 7 | 39 | 46 | 3.75 | 3.16 |
64.05 | ? | NF | 0 | 7 | 7 | 0.00 | 0.11 |
65.96 | ? | H | 1 | 12 | 13 | 0.89 | 0.42 |
66.45 | ? | H | 0 | 1 | 1 | 0.00 | 0.25 |
66.56 | ? | H | 3 | 40 | 43 | 3.97 | 6.23 |
66.91 | ? | H | 1 | 0 | 1 | 0.19 | 0.00 |
67.21 | Heptacosane | H | 2 | 14 | 16 | 0.62 | 0.36 |
69.07 | ? | H | 1 | 9 | 10 | 0.18 | 0.18 |
70.77 | Nonacosadiene ? | H | 1 | 39 | 40 | 1.51 | 6.20 |
71.03 | ? | NF | 1 | 10 | 11 | 0.08 | 0.19 |
71.19 | ? | H | 5 | 40 | 45 | 6.84 | 10.06 |
71.95 | Nonacosane | NF | 2 | 14 | 16 | 0.28 | 0.30 |
72.64 | ? | NF | 0 | 2 | 2 | 0.00 | 0.04 |
73.19 | ? | NF | 0 | 3 | 3 | 0.00 | 0.03 |
74.89 | ? | H | 1 | 38 | 39 | 0.43 | 4.30 |
75.38 | Untriacontadien ? | NF | 0 | 3 | 3 | 0.00 | 0.07 |
75.75 | ? | H | 3 | 19 | 22 | 1.09 | 1.29 |
76.25 | Hentriacontane | H | 1 | 7 | 8 | 0.10 | 0.13 |
79.49 | Tritriacontadiene ? | NF | 0 | 2 | 2 | 0.00 | 0.12 |
79.51 | ? | NF | 0 | 14 | 14 | 0.00 | 0.92 |
80.67 | Phytosterole ? | NF | 0 | 6 | 6 | 0.00 | 0.18 |
81.31 | similar beta-Amyrin | NF | 0 | 32 | 32 | 0.00 | 2.50 |
81.59 | similar alpha-Amyrin | NF | 0 | 3 | 3 | 0.00 | 0.17 |
81.62 | Phytosterole | NF | 0 | 13 | 13 | 0.00 | 0.61 |
82 39 | similar alpha-Amyrin | NF | 0 | 38 | 38 | 0 00 | 4 84 |
Rights and permissions
About this article
Cite this article
Zimmermann, Y., Roubik, D.W. & Eltz, T. Species-specific attraction to pheromonal analogues in orchid bees. Behav Ecol Sociobiol 60, 833–843 (2006). https://doi.org/10.1007/s00265-006-0227-8
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00265-006-0227-8