[en] Males of polyandrous species have to overcome sperm competition. They should select their mate based on the reproductive status of the female to increase their own fitness. Because the sexual behavior of lady beetles relies on semiochemicals, with cuticular hydrocarbons (CHCs) being used for mate recognition, we developed and tested two hypotheses. First, we hypothesized that the cuticular hydrocarbon profile qualitatively and quantitatively differs between virgin and mated Harmonia axyridis females, regardless of the color morph. Second, we hypothesized that males discriminate virgin and mated females, preferring copulating with virgin females, rather than previously mated ones, to avoid sperm competition and subsequently increase their fitness. CHCs were solvent-extracted before being quantified and identified by gas chromatography. We found no qualitative differences between mated and unmated females irrespective of the morph; however, quantitative differences were detected. Specifically, the CHC profiles of mated females presented higher concentrations of alkenes, including 9-pentacosene, 9-heptacosene, and 9-hentriacontene. During dual-choice behavioral assays, males equally copulated with virgin and mated females. Our results suggest that there is no CHC-based discrimination strategy in virgin males of H. axyridis between virgin and once-mated females. We discuss alternative strategies that might be used in this lady beetle species.
Andersson, J., Borg-Karlson, A.K. & Wiklund, C. (2003): Antiaphrodisiacs in pierid butterflies: theme with variation! – Journal of Chemical Ecology 29: 1489−1499.
Arnqvist, G. & Nilsson, T. (2000): The evolution of polyandry: Multiple mating and female fitness in insects. – Animal Behaviour 60: 145−164.
Awad, M., Piálek, L., Krejčí, A., Laugier, G. & Nedvěd, O. (2017): Paternity following multiple mating in ladybird Harmonia axyridis. – Biocontrol 62: 297−307.
Bro-Jørgensen, J. (2010): Dynamics of multiple signalling systems: animal communication in a world in flux – Trends in Ecology & Evolution 25: 292−300.
Birkhead, T. & Hunter, F.M. (1990): Mechanisms of sperm competition. – Trends in Ecology & Evolution 5: 164−194.
Carazo, P., Sanchez, E., Font, E. et al. (2004): Chemosensory cues allow male Tenebrio molitor beetles to assess the reproductive status of potential mates. – Animal Behaviour 68: 123−129.
Chaudhary, D.D., Mishra, G. & Omkar A (2016): Last male wins the egg fertilization fight: A case study in ladybird, Menochilus sexmaculatus. – Behavioural Process 131: 1−8.
Cuvillier-Hot, V., Cobb, M., Malosse, C. et al. (2001): Sex, age and ovarian activity effect cuticular hydrocarbons in Diacamma ceylonense a queenless ant. – Journal of Insect Physiology 47: 485−493.
Daly, M. (1978): The cost of mating. – American Naturalist 112: 771−774.
De Jong, P.W., Brakefield, P.M. & Geerinck, B.P. (1998): The effect of female mating history on sperm precedence in the two-spot ladybird, Adalia bipunctata (Coleoptera, Coccinellidae). – Behavioural Ecology 9: 559−565.
Dillwith, J.W., Adams, T.S. & Blomquist, G.J. (1983): Correlation of housefly sex-pheromone production with ovarian development. – Journal of Insect Physiology 29: 377−386.
Everaerts, C., Farine, J.P., Cobb, M. et al. (2010): Drosophila cuticular hydrocarbons revisited: Mating status alters cuticular profiles. – PLoS ONE 5: 1−12.
Fassotte, B., Fischer, C., Durieux, D. et al. (2014): First evidence of a volatile sex pheromone in lady beetles. – PLoS ONE 9 (12): e115011.
Fassotte, B., Francis, F. & Verheggen, F.J. (2016): The scent of love: how important are semiochemicals in the sexual behavior of lady beetles? – Journal of Pest Science 89: 347−358.
Friberg, U. (2006): Male perception of female mating status: its effect on copulation duration, sperm defence and female fitness. – Animal Behaviour 72: 1259−1268.
Gilbert, L.E. (1976): Postmating female odour in Heliconius butterflies: a male-contributed antiaph-rodisiac? – Science 193: 419−420.
Haddrill, R., Shuker, M. & Majerus, E.N. (2007): Temporal effects of multiple mating on components of fitness in the two-spot ladybird, Adalia bipunctata (Coleoptera: Coccinellidae). – European Journal of Entomology 104: 393−398.
Happ, G.M. (1969): Multiple sex pheromones of the mealworm beetle, Tenebrio molitor L. – Nature 222: 180−181.
Hemptinne, J.L., Dixon, F.G. & Lognay, G. (1996): Searching behaviour and mate recognition by males of the two-spot ladybird beetle, Adalia bipunctata. – Ecological Entomology 21: 165−170.
Hodek, I. & Ceryngier, P. (2000): Sexual activity in Coccinellidae (Coleoptera): a review. – European Journal of Entomology 97: 449−456.
Howard, R.W. & Blomquist, G.J. (2005): Ecological, behavioral, and biochemical aspects of insect hydrocarbons. – Annual Review of Entomology 50: 371−393.
Ingleby, F.C. (2015): Insect cuticular hydrocarbons as dynamic traits in sexual communication. – Insects 6: 732−742.
Johansson, B.G. & Jones, T.M. (2007): The role of chemical communication in mate choice. – Biological Reviews 82: 265−289.
Koch, R.L. (2003): The multicolored Asian lady beetle, Harmonia axyridis, a review of its biology, uses in biological control, and non-target impacts. – Journal of Insect Science 3: 1−16.
Laugier, G.J.M., Le Moguéde, G., Tayeh, A. et al. (2013): Increase in male reproductive success and female reproductive investment in invasive populations of the Harlequin Ladybird Harmonia axyridis. – PlosOne 8 (10): e77083.
Lockey, K.H. (1988): Lipids of the insect cuticle: origin, composition and function. – Comparative Biochemistry and Physiology Part B. – Comparative Biochemistry 89: 595−645.
Mallet, G., Dimitriades, C. & Ucciani, E. (1985): Apport des techniques de dérivatisation à la sépara-tion des esters d’acides gras par chromatographie gazeuse capillaire. – Revue française des corps gras 11–12: 439–446.
McCornack, B.P., Koch, R.L. & Ragsdale, D.W. (2007): A simple method for in-field sex determination of the multicolored Asian lady beetle Harmonia axyridis. – Journal of Insect Science 7 (10): 1−12.
Nedvěd, O. & Honěk, A. (2012): Life history and development. – In: Hodek, I., van Emden, H.F. & Honěk, A. (eds): Ecology and Behaviour of the Ladybird Beetles (Coccinellidae). – Wiley-Blackwell, 54–109, Oxford, UK.
Obata, S. (1987): Mating behavior and sperm transfer in the Ladybird Beetle, Harmonia axyridis Pallas (Coleoptera: Coccinellidae). – Applied Entomology and Zoology 22: 434–442.
Omkar & Mishra, G. (2005): Mating in aphidophagous ladybirds: Costs and benefits. – Journal of Applied Entomology 129: 432−436.
Omkar & Pervez, A. (2005): Mating behavior of an aphidophagous ladybird beetle, Propylea dissecta (Mulsant). – Insect Science 12: 37−44.
Omkar A & Sahu, J. (2012): Costs and benefits of reproduction in predaceous ladybird: Effect of multiple matings on reproduction and offspring development. – Journal of Asia-Pacific Entomology 15: 219−224.
Omkar, Singh, S.K. & Mishra, G. (2010): Multiple matings affect the reproductive performance of aphidophagous ladybird beetle, Coelophora saucia (Coleoptera: Coccinellidae). – European Journal of Entomology 107: 177−182.
Omkar & Srivastava, S. (2002): The reproductive behaviour of an aphidophagous ladybeetle, Coccinella septempunctata (Coleoptera: Coccinellidae). – European Journal of Entomology 99: 465−470.
Osawa, N. (1994): The occurrence of multiple mating in a wild population of the ladybird beetle Harmonia axyridis Pallas (Coleoptera: Coccinellidae). – Journal Ethology 12: 63−66.
Pattanayak, R., Mishra, G., Omkar et al. (2014): Does the volatile hydrocarbon profile differ between the sexes: A case study on five aphidophagous ladybirds. – Archives of insect biochemistry and physiology 87: 105−125.
R Development Core Team (2008): R: A language and environment for statistical computing. – R Foundation for Statistical Computing, Vienna, Austria, ISBN 3-900051-07-0, http://www.R-project.org.
Romano, D., Donati, E., Canale, A., Messing, R.H., Benelli, G. & Stefanini, C. (2016): Lateralized courtship in a parasitic wasp. – Laterality 21: 243−254.
Romano, D., Benelli, G., Donati, E., Remorini, D., Canale, A. & Stefanini, C. (2017): Multiple cues produced by a robotic fish modulate aggressive behaviour in siamese fighting fishes. – Scientific Reports 7 (1): 4667.
Romano, D., Benelli, G., Stefanini, C., Desneux, N., Ramirez-Romero, R., Canale, A. & Lucchi, A. (2018): Behavioral asymmetries in the mealybug parasitoid anagyrus sp. near pseudococci: Does lateralized antennal tapping predict male mating success? – Journal of Pest Science 91: 341−349.
Scott, D. (1986): Sexual mimicry regulates the attractiveness of mated Drosophila melanogaster females. – Proceedings of the National Academy of Sciences of the United States of America 83: 8429−8433.
Soares, A.O., Borges, I., Borges, P.A.V. et al. (2008): Harmonia axyridis: What will stop the invader? – Biocontrol 53: 127–145.
Srivastava, S. & Omkar (2005): Short-and long-term benefits of promiscuity in the seven-spotted ladybird Coccinella septempunctata (Coleoptera: Coccinellidae). – International Journal of Tropical Insect Science 25: 176−181.
Thomas, M.L. (2011): Detection of female mating status using chemical signals and cues. – Biological Review 86: 1−13.
Thomas, M.L. & Simmons, L.W. (2007): Male crickets adjust the viability of their sperm in response to female mating status. – The American Naturalist 170: 190−195.
Thomas, M.L. & Simmons, L.W. (2009): Male-derived cuticular hydrocarbons signal sperm competition intensity and affect ejaculate expenditure in crickets. – Proceedings of the Royal Society B: Biological Sciences 276: 383−388.
Ueno, H. (1996): Estimate of multiple insemination in a natural population of Harmonia axyridis (Coleoptera: Coccinellidae). – Applied Entomology and Zoology 31: 621−623.
Ueno, H., Sato, Y. & Tsuchida, K. (1998): Colour-associated mating success in a polymorphic ladybird beetle, Harmonia axyridis. – Functional Ecology 12: 7573−761.
Wang, S., Michaud, J.P., Zhang, R.Z. et al. (2009): Seasonal cycles of assortative mating and reproductive behaviour in polymorphic populations of Harmonia axyridis in China. – Ecological Entomology 34: 483–494.
Wenninger, E.J., Stelinski, L.L. & Hall, D.G. (2008): Behavioral evidence for a female-produced sex attractant in Diaphorina citri. – Entomologia experimentalis et applicata 128: 450−459.
Zimmerman, D.W. & Zumbo, B.D. (1993): Relative Power of the Wilcoxon Test, the Friedman Test, and Repeated-Measures ANOVA on Ranks. – Journal of Experimental Education 62: 75−86.
