Hiding In Plain Sight: Cuticular Compound Profile Matching Conceals A Larval Tortoise Beetle In Its Host Chemical Cloud

Larvae of tortoise beetles are postulated to have fecal shields as the main defensive strategy against predators. Such a device protects beetles both physically and chemically. In order to examine how larvae Chelymorpha reimoseri are protected against predatory ants, which frequently visit extrafloral nectaries in their host plant, the morning glory Ipomoea carnea, we conducted anti-predation bioassays with live 5th instars. In the field, larvae in contact with ants had survival between 40 and 73 %, independently of shield presence. In the laboratory, when exposed to Camponotus crassus, larvae with shields had significantly higher survival (85 %) than those without shields (64 %). In both scenarios, larval survival was significantly higher when compared with palatable Spodoptera frugiperda larvae, as the latter were all consumed. We also observed that when C. reimoseri larvae showed no movement, the ants walked on them without attacking. We hypothesized that if the larval integument has a pattern of cuticular compounds (CCs) similar to that of its host plant, larvae would be rendered chemically camouflaged. In the field and laboratory, the freeze-dried palatable larvae of S. frugiperda treated with CCs of 5th instar C. reimoseri and left on I. carnea leaves were significantly less removed by ants than controls without these compounds. We also found a similarity of approximately 50 % between the CCs in C. reimoseri larvae and I. carnea host leaves. Both findings provide evidence in support of the hypothesis that chemical camouflage plays an important role in larval defense, which is reported for the first time in an ectophagous leaf beetle larva. © 2014 Springer Science+Business Media New York.404341354Akino, T., Chemical and behavioral study on the phytomimetic giant geometer Biston robustum Butler (Lepidoptera: Geometridae) (2005) Appl Entomol Zool, 40, pp. 497-505Akino, T., Chemical strategies to deal with ants: a review of mimicry, camouflage, propaganda and phytomimesis by ants (Hymenoptera: Formicidae) and other arthropods (2008) Myrmecol News, 11, pp. 173-181Akino, T., Nakamura, K.-I., Wakamura, S., Diet induced chemical phyomimesis by twig-like caterpillars of Biston robustum Butler (Lepidoptera: Geometridae) (2004) Chemoecology, 14, pp. 165-174Austin, D., Huáman, Z., A synopsis of Ipomoea (Convolvulaceae) in the Americas (1996) Taxon, 45, pp. 3-38Bacher, S., Luder, S., Picky predators and the function of the faecal shield of a cassidine larva (2005) Funct Ecol, 19, pp. 263-272Bachmann, A.O., Cabrera, N., A catalog of the types of Chrysomelidae sensu lato (Insecta, Coleoptera, Polyphaga) deposited in the Museo Argentino de Ciencias Naturales, Buenos Aires (2010) Rev Mus Argent Cienc Nat, 12, pp. 57-80Borowiec, L., (1999) World Catalogue of Cassidinae (Coleoptera:Chrysomelidae), , Wroclaw: Biologica SilesiaeBottcher, A., Zolin, J.P., Nogueira-De-Sá, F., Trigo, J.R., Faecal shield chemical defence is not important in larvae of the tortoise beetle Chelymorpha reimoseri (Chrysomelidae: Cassidinae: Stolaini) (2009) Chemoecology, 19, pp. 63-66Budzikiewicz, H., Djerassi, C., Williams, D.H., (1967) Mass Spectrometry of Organic Compounds, , San Francisco: Holden-Day IncCarlson, D.A., Roan, C.-S., Yost, R.A., Hector, J., Dimethyl disulfide derivatives of long chain alkenes, alkadienes, and alkatrienes for gas chromatography/mass spectrometry (1989) Anal Chem, 61, pp. 1564-1571Carlson, D.A., Bernier, U.R., Sutton, B.D., Elution patterns from capillary GC for methyl-branched alkanes (1998) J Chem Ecol, 24, pp. 1845-1865Chaboo, C.S., Biology and phylogeny of the Cassidinae (tortoise and leaf-mining beetles) (Coleoptera: Chrysomelidae) (2007) Bull Am Mus Nat Hist, 305, pp. 1-250Davidson, D.W., Cook, S.C., Snelling, R.R., Chua, T.H., Explaining the abundance of ants in lowland tropical rainforest canopies (2003) Science, 300, pp. 969-972Eisner, T., Tassel, E., Carrel, J.E., Defensive use of 'fecal shield' by a beetle larva (1967) Science, 158, pp. 1471-1473Fernández, F., (2003) Introducción a las hormigas de la región Neotropical, , Instituto de Investigación de Recursos Biológicos Alexander von Humboldt, BogotáGomes, C.C.G., Trigo, J.R., Eiras, A.E., Sex pheromone of the American warble fly, Dermatobia hominis: the role of cuticular hydrocarbons (2008) J Chem Ecol, 34, pp. 636-646Guimarães, P.R., Raimundo, R.L.G., Bottcher, C., Silva, R.R., Trigo, J.R., Extrafloral nectaries as a deterrent mechanism against seed predators in the chemically defended weed Crotalaria pallida (Leguminosae) (2006) Austral Ecol, 31, pp. 776-782Haase, R., Seasonal growth of "algodão-bravo" (Ipomoea carnea spp. fistulosa) (1999) Pesq Agrop Brasileira, 34, pp. 159-163Haraguchi, M., Gorniak, S.L., Ikeda, K., Minami, Y., Kato, A., Watson, A.A., Nash, R.J., Asano, N., Alkaloidal components in the poisonous plant, Ipomoea carnea (Convolvulaceae) (2003) J Agric Food Chem, 51, pp. 4995-5000Heckman, C.W., (1998) The Pantanal of Poconé. Biota and Ecology in the Northern Section of the World's Largest Pristine Wetland, , Dordrecht: Kluwer Academic PublishersHölldobler, B., Wilson, E.O., (1990) The Ants, , Cambridge: Harvard University PressHoward, R.W., Cuticular hydrocarbons and chemical communication (1993) Insect Lipids: Chemistry, Biochemistry and Biology, pp. 179-226. , D. W. Stanley-Samuelson and D. R. Nelson (Eds.), Lincoln: University of Nebraska PressKrebs, C.J., (1999) Ecological Methodology, , Menlo Park: Addison-Wesley EducationalKusnezov, N., El género Camponotus en la Argentina (1951) Acta Zool Lilloana, XII, pp. 183-255Labandeira, C.C., The history of associations between plants and animals (2002) Plant-Animal Interactions. An Evolutionary Approach, pp. 26-74. , C. M. Herrera and O. Pellmyr (Eds.), Oxford: Blackwell ScienceMappes, J., Marples, N., Endler, J.A., The complex business of survival by aposematism (2005) Trends Ecol Evol, 20, pp. 598-603Mccullagh, P., Nelder, J.A., (1989) Generalized Linear Models, , 2nd edn., London: Chapman and HallMenéndez, R.D., Marrero, D., Más, R., Fernández, I., González, L., González, R.M., In vitro and in vivo study of octacosanol metabolism (2005) Arch Med Res, 36, pp. 113-119Müller, C., Hilker, M., Unexpected reactions of a generalist predator towards defensive devices of cassidine larvae (Coleoptera, Chrysomelidae) (1999) Oecologia, 118, pp. 166-172Müller, C., Hilker, M., Ecological relevance of fecal matter in Chrysomelidae (2004) New Contributions to the Biology of Chrysomelidae, pp. 693-705. , P. H. Jolivet, J. A. Santiago-Blay, and M. Schmitt (Eds.), The Hague: SPC Academic PublishersNelson, D.R., Sukkestad, D.R., Zaylskie, R.G., Mass spectra of methyl-branched hydrocarbons from eggs of the tobacco hornworm (1972) J Lipid Res, 13, pp. 413-421Nishida, R., Sequestration of defensive substances from plants by Lepidoptera (2002) Annu Rev Entomol, 47, pp. 57-92Nogueira-de-Sá, F., (2004) Defesas de larvas de Plagiometriona flavescens e Stolas areolata (Coleoptera: Chrysomelidae: Cassidinae) contra predadores, , Ph. D. Thesis. Instituto de Biologia, Universidade Estadual de Campinas, Campinas, BrasilNogueira-De-Sá, F., Trigo, J.R., Faecal shield of the tortoise beetle Plagiometriona aff. flavescens (Chrysomelidae: Cassidinae) as chemically mediated defence against predators (2005) J Trop Ecol, 21, pp. 189-194Opitz, S.E.W., Müller, C., Plant chemistry and insect sequestration (2009) Chemoecology, 19, pp. 117-154Piskorski, R., Trematerra, P., Dorn, S., Cuticular hydrocarbon profiles of codling moth larvae, Cydia pomonella (Lepidoptera: Tortricidae), reflect those of their host plant species (2010) Biol J Linn Soc, 101, pp. 376-384Portugal, A.H.A., Trigo, J.R., Similarity of cuticular lipids between a caterpillar and its host plant: a way to make prey undetectable for predatory ants? (2005) J Chem Ecol, 31, pp. 2551-2561Price, P.W., Denno, R.F., Eubanks, M.D., Finke, D.L., (2011) Insect Ecology: Behavior, Populations and Communities, , Cambridge: Cambridge University PressQuinn, G.P., Keough, M.J., (2002) Experimental Design and Data Analysis for Biologists, , Cambridge: Cambridge University PressRowe, C., Halpin, C., Why are warning displays multimodal? (2013) Behav Ecol Sociobiol, 67, pp. 1425-1439Ruxton, G.D., Non-visual crypsis: a review of the empirical evidence for camouflage to senses other than vision (2009) Phil Trans R Soc B, 364, pp. 549-557Ruxton, G.D., Sherratt, T.N., Speed, M.P., (2004) Avoiding Attack, , New York: Oxford University PressSilveira, H.C.P., Oliveira, P.S., Trigo, J.R., Attracting predators without falling prey: chemical camouflage protects honeydew-producing treehoppers from ant predation (2010) Am Nat, 175, pp. 261-268(2004) Statistica (data analysis software system), version 7, , www.statsoft.com, Statsoft, IncSteward, J.L., Keeler, K.H., Are there trade-offs among antiherbivore defenses in Ipomoea (Convolvulaceae)? (1988) Oikos, 53, pp. 79-86Świetojańska, J., The Immatures of Tortoise Beetles with Bibliographic Catalogue of All Taxa (Coleoptera: Chrysomelidae: Cassidinae) (2009) Polish Taxonomical Monographs, 16. , Wrocław: Biologica Silesiaevan den Dool, H., Kratz, P.D., A generalization of the retention index system including linear temperature programmed gas-liquid partition chromatography (1963) J Chromatogr, 11, pp. 463-471Vane-Wright, R.I., An unified classification of mimetic resemblances (1976) Biol J Linn Soc, 8, pp. 25-56Vasconcellos-Neto, J., Genetics of Chelymorpha cribraria, Cassidinae: Colour patterns and their ecological meaning (1988) Biology of Chrysomelidae, pp. 217-232. , P. H. Jolivet, E. Petitpierre, and T. H. Hsiao (Eds.), Dordrecht: Kluwer Academic PublishersVasconcellos-Neto, J., Jolivet, P.H., Une nouvelle stratégie de défense: la stratégie de défense annulaire (cycloalexie) chez quelquez larves de Chrysomélides brésiliens (1988) Bull Soc Entomol Fr, 92, pp. 291-299Vencl, F.V., Srygley, R.B., Enemy targeting, trade-offs, and the evolutionary assembly of a tortoise beetle defense arsenal (2013) Evol Ecol, 27, pp. 237-252Vencl, F.V., Nogueira-De-Sá, F., Allen, B.J., Windsor, D.M., Futuyma, D.J., Dietary specialization influences the efficacy of larval tortoise beetle shield defenses (2005) Oecologia, 145, pp. 404-414Vencl, F.V., Gómez, N.E., Ploss, K., Boland, W., The chlorophyll catabolite, pheophorbide a, confers predation resistance in a larval tortoise beetle shield defense (2009) J Chem Ecol, 35, pp. 281-288Vencl, F.V., Trillo, P.A., Geeta, R., Functional interactions among tortoise beetle larval defenses reveal trait suites and escalation (2011) Behav Ecol Sociobiol, 65, pp. 227-239Walters, D.W., (2011) Plant Defense: Warding off Attack by Pathogens, Herbivores, and Parasitic Plants, , Oxford: Blackwell Publishin