Journal of Chemical Ecology

, Volume 36, Issue 12, pp 1335–1345 | Cite as

Leaf and Floral Parts Feeding by Orange Tip Butterfly Larvae Depends on Larval Position but Not on Glucosinolate Profile or Nitrogen Level

  • Niels AgerbirkEmail author
  • Frances S. Chew
  • Carl Erik Olsen
  • Kirsten Jørgensen


In an attempt to identify chemical signals governing the general flower and silique feeding behavior of larvae of the orange tip butterfly, Anthocharis cardamines (L.), we investigated feeding behavior and chemistry of two major host plants: Cardamine pratensis L. and Alliaria petiolata (Bieb.) Cavara & Grande (garlic mustard). Larvae reportedly feed mainly on flowers and siliques rather than leaves in nature, and did so when observed on the original host plants. Behavioral experiments, using detached A. petiolata branches, however, showed that larvae readily accepted leaves and only the final instar showed a tendency for directed movement towards floral parts. To search for semiochemicals that control plant part preference and to assess possible nutritional consequences of floral parts feeding, we determined glucosinolate profiles and total nitrogen levels of floral parts and leaves. There was only moderate difference between glucosinolate profiles of leaves and floral parts within each of two host plant species. In contrast, the profiles of floral parts differed significantly between them. A. petiolata was dominated by 2-propenyl glucosinolate, while C. pratensis was dominated by aromatic glucosinolates and branched aliphatic glucosinolates, with considerable variation among populations. Nitrogen levels tended to be higher in floral parts than in leaves in A. petiolata, but not in C. pratensis, so floral feeding could not generally be attributed to higher N content. With the exception of a tendency of last instar larvae (L5) to move to the apex and ingest flowers and upper stem, we did not find either a plant chemistry basis or larval acceptance/rejection behavior that could explain the usual feeding of floral parts by orange tip larvae of all instars. However, by artificial manipulation of vertical larval position on host plants, we found that the frequency of leaf vs. flower feeding during 24 hr depended significantly on the initial larval position. Hence, we suggest that the placement of eggs on floral parts by ovipositing female butterflies is a major explanation of orange tip feeding habits previously known from field observations.

Key Words

Larval feeding behavior and preference Glucosinolate profile Total nitrogen Leaves Flowers Siliques 



We thank two anonymous reviewers for helpful comments and suggestions to an earlier version of the text, Claus T. Ekström and J. Michael Reed for statistical advice, Birgitte B. Rasmussen for skillful glucosinolate analysis, and the laboratory of Jeffrey Dukes for determination of total nitrogen and carbon. This research was financially supported by Torben og Alice Frimodts Fond to NA and the Arabis Fund to FC.


  1. Agerbirk, N., and Jørgensen, K. 2008. Aurora-larvens korte liv i en vindueskarm. Lepidoptera IX, 176–179 (in Danish).Google Scholar
  2. Agerbirk, N., Olsen, C.E., Bibby, B. M., Frandsen, H. O., Brown, L. D., Nielsen, J. K., and Renwick, J. A. A., 2003. A saponin correlated with variable resistance of Barbarea vulgaris to the diamond back moth Plutella xylostella. J. Chem. Ecol. 29, 1417–1433.CrossRefPubMedGoogle Scholar
  3. Agerbirk, N., Müller, C., Olsen, C. E., and Chew, F. S. 2006. A common pathway for metabolism of 4-hydroxybenzylglucosinolate in Pieris and Anthocaris (Lepidoptera: Pieridae). Biochem. Syst. Ecol. 34, 189–198.CrossRefGoogle Scholar
  4. Agerbirk, N., Olsen, C. E., Topbjerg, H. B., and Sørensen, J. C. 2007. Host plant dependent metabolism of 4-hydroxybenzylglucosinolate in Pieris rapae: Substrate specificity and effects of genetic modification and plant nitrile hydratase. Insect Biochem. Mol. Biol. 37, 1119–1130.CrossRefPubMedGoogle Scholar
  5. Agerbirk, N., Warwick, S., Hansen, P. R., and Olsen, C. E. 2008. Sinapis phylogeny and evolution of glucosinolates and specific nitrile degrading enzymes. Phytochemistry 69, 2937–2949.CrossRefPubMedGoogle Scholar
  6. Agerbirk, N., Olsen, C. E., Chew, F. S, and Ørgaard, M. 2010a. Variable glucosinolate profiles of Cardamine pratensis (Brassicaceae) with equal chromosome numbers. J. Agric. Food Chem. 58, 4693–4700.CrossRefPubMedGoogle Scholar
  7. Agerbirk, N., Olsen, C. E., Poulsen, E., Jacobsen, N., and Hansen, P. R. 2010b. Complex metabolism of aromatic glucosinolates in Pieris rapae caterpillars involving nitrile formation, hydroxylation, demethylation, sulfation, and host plant dependent carboxylic acid formation. Insect Biochem. Mol. Biol. 40, 126–137. CrossRefPubMedGoogle Scholar
  8. Arvanitis, L., Wiklund, C., and Ehrlén, J. 2007. Butterfly seed predation: effects of landscape characteristics, plant ploidy level and population structure. Oecologia 152, 275–285.CrossRefPubMedGoogle Scholar
  9. Arvanitis, L., Wiklund, C., and Ehrlén, J. 2008. Plant ploidy level influences selection by butterfly seed predators. Oikos 117, 1020–1025.CrossRefGoogle Scholar
  10. Badenes-pérez, F. R., Reichelt, M., Gershenzon, J., and Heckel, D. G. 2010. Phylloplane location of glucosinolates in Barbarea spp. and misleading assessment of host suitability by a specialist herbivore. New Phytologist. doi: 10.1111/j.1469-8137.2010.03486.x.
  11. Bandeili, B., Muller, C. 2010. Folivori versus florivory - adaptiveness of flower feeding. Naturwissenschaften 97, 79–88.CrossRefPubMedGoogle Scholar
  12. Bidart-bouzat, M. G., and Kliebenstein, D. J. 2008. Differential levels of insect herbivory in the field associated with genotypic variation in glucosinolates in Arabidopsis thaliana. J. Chem. Ecol. 34, 1026–1037.CrossRefPubMedGoogle Scholar
  13. Brown, P. D., Tokuhisa, J. G., Reichelt, M., Gershenzon, J. 2003. Variation of glucosinolate accumulation among different organs and developmental stages of Arabidopsis thaliana. Phytochemistry 62, 471–481.CrossRefPubMedGoogle Scholar
  14. Courtney, S. P. 1981. Coevolution of Pierid butterflies and their cruciferous foodplants III. Anthocharis cardamines (L.) survival, development and oviposition on different host plants. Oecologia 51, 91–96.CrossRefGoogle Scholar
  15. Courtney, S. P., and Chew, F. C. 1987. Coexistence and host use by a large community of Pierid butterflies: habitat is the templet. Oecologia 71, 210–220.CrossRefGoogle Scholar
  16. De Vos, M., Kriksunov, K., and Jander, G. 2008. Indole-3-acetonitrile production from indole glucosinolates deters oviposition by Pieris rapae (white cabbage butterfly). Plant Physiol. 146, 916–926.CrossRefPubMedGoogle Scholar
  17. Dempster, J. P. 1997. The role of larval food resources and adult movement in the population dynamics of the orange-tip butterfly (Anthocharis cardamines). Oecologia 111, 549–556.CrossRefGoogle Scholar
  18. Giamoustaris A., and Mithen, R. 1995. The effect of modifying the glucosinolate content of leaves of oilseed rape (Brassica napus ssp. oleifera) on its interaction with specialist and generalist pests. Ann. Appl. Biol. 126, 347–363.CrossRefGoogle Scholar
  19. Gols, R., Wagenaar, R., Bukovinszky, T., Van Dam, N. M., Dicke, M., Bullock, J. M., and Harvey, J. A. 2008. Genetic variation in defense chemistry in wild cabbages affect herbivores and their endoparasitoids. Ecology 89, 1616–1626.CrossRefPubMedGoogle Scholar
  20. Griffiths, D. W., Deighton, N., Birch, A. N. E., Patrian, B., Baur, R., and Städtler, E. 2001. Identification of glucosinolates on the leaf surface of plants from the cruciferae and other closely related plants. Phytochemistry 57, 693–700.CrossRefPubMedGoogle Scholar
  21. Hopkins, R. J., Van Dam, N. M., and Van Loon, J. J. A. 2009. Role of glucosinolates in insect-plant relationships and multitrophic interactions. Annu. Rev. Entomol. 54, 57–83.CrossRefPubMedGoogle Scholar
  22. Huang, X. P., and Renwick, J. A. A. 1994. Relative activities of glucosinolates as oviposition stimulants for Pieris rapae and P. napi oleracea. J. Chem. Ecol. 20, 1025–1037CrossRefGoogle Scholar
  23. Larsen, L. M., Nielsen, J. K., and Sørensen, H. 1992. Host plant recognition in monophageous weevils: Specialization of Ceutorhyncus inaffectatus to glucosinolates from its host plant Hesperis matronalis. Entomol. Exp. Appl. 64, 49–55.CrossRefGoogle Scholar
  24. Li, Q., Eigenbrode, S. D., Stringham, G. R., and Thiagarajah, M. R. 2000. Feeding and growth of Plutella xylostella and Spodoptera eridania on Brassica juncea with varying glucosinolate concentrations and myrosinase activities. J. Chem. Ecol. 26, 2401–2419.CrossRefGoogle Scholar
  25. Mattson, W. J. 1980. Herbivory in relation to plant nitrogen-content. Annu. Rev. Ecol. Syst. 11, 119–161.CrossRefGoogle Scholar
  26. Miles, C. I., Del Campo, M. L., and Renwick, J. A. A. 2005. Behavioral and chemosensory responses to a host recognition cue by larvae of Pieris rapae. J. Comp. Physiol. A 191, 147–155.CrossRefGoogle Scholar
  27. Nielsen, J. K., Nagao, T., Okabe, H., Shinoda, T. 2010. Resistance in the plant, Barbarea vulgaris, and counter-adaptations in flea beetles mediated by saponins. J. Chem. Ecol. 36, 277–285.CrossRefPubMedGoogle Scholar
  28. Nomakuchi, S., Masumoto, T., Sawada, K., Sunahara, T., Itakura, N., and Suzuki, N. 2001. Possible age-dependent variation in egg-loaded host selectivity of the pierid butterfly, Anthocharis scolymus (Lepidoptera: Pieridae): A field observation. J. Insect Behav. 14, 451–458.CrossRefGoogle Scholar
  29. Reifenrath, K., Städler, E. 2009. Glucosinolates on the leaf surface perceived by insect herbivores: review of ambiguous results and new investigations. Phytochem. Rev. 8, 207–225.CrossRefGoogle Scholar
  30. Reifenrath, L., Riederer, M., and Müller, C. 2005. Leaf surface wax layers of Brassicaceae lack feeding stimulants for Phaedon cochleariae. Entomol. Exp. Appl. 115, 41–50.CrossRefGoogle Scholar
  31. Renwick, J. A. A., and Chew, F.S. 1994. Oviposition in Lepidoptera. Annu. Rev. Entomol. 39, 377–400.CrossRefGoogle Scholar
  32. Rice, W. R. 1988. A new probability model for determining exact p-values for 2 × 2 contingency tables when comparing binomial proportions. Biometrics 44, 1–22.CrossRefGoogle Scholar
  33. Rodman, J. E., and Chew, F. S. 1980. Phytochemical correlates of herbivory in a community of native and naturalized Cruciferae. Biochem. Syst. Ecol. 8:43–50CrossRefGoogle Scholar
  34. Schoonhoven, L. M., Van Loon, J. J. A., Dicke, M., 2005. Insect-Plant Biology, 2nd edn., Oxford University Press.Google Scholar
  35. Shapiro, A. M. 1981. The pierid red-egg syndrome. Am. Nat. 117, 276–294.CrossRefGoogle Scholar
  36. Smallegange, R. C., Van Loon, J. J. A., Blatt, S. E., Harvey, J.A., Agerbirk, N., and Dicke, M. 2007. Flower vs. leaf feeding by Pieris brassicae: glucosinolate-rich tissues are preferred and sustain higher growth rate. J. Chem. Ecol. 33, 1831–1844.CrossRefPubMedGoogle Scholar
  37. Strauss, S. Y., Irwin, R. E., and Lambrix, V. M. 2004. Optimal defence theory and flower petal colour predict variation in the secondary chemistry of wild radish. J. Ecol. 92, 132–141.CrossRefGoogle Scholar
  38. Sun, J. Y., Sønderby, I. E., Halkier, B. A., Jander, G., De Vos, M. 2009. Non-volatile intact indole glucosinolates are host recognition cues for ovipositing Plutella xylostella. J. Chem. Ecol. 35, 1427–1436.CrossRefGoogle Scholar
  39. Theunissen, J., Den Ouden, H., and Wit, A. K. H. 1985. Feeding capacity of caterpillars on cabbage, a factor in crop loss assessment. Entomol. Exp. Appl. 39, 255–260.Google Scholar
  40. Traw, M. B., and Feeny, P. 2008. Glucosinolates and trichomes track tissue value in two sympatric mustards. Ecology 89, 763–772.CrossRefPubMedGoogle Scholar
  41. Van Loon, J. J. A., Blaakmeer, A., Griepink, F. C., Van Beek, T. A., Schoonhoven, L.M., and De Groot, A. 1992. Leaf surface compound from Brassica oleracea (Cruciferae) induces oviposition by Pieris brassicae (Lepidoptera: Pieridae). Chemoecology 3, 39–44.CrossRefGoogle Scholar
  42. Vergara, F., Svatoš, A., Schneider, B., Reichelt, M, Gershenzon, J., and Wittstock, U. 2006. Glycine conjugates in a lepidopteran insect herbivore – the metabolism of benzylglucosinolate in the cabbage white butterfly, Pieris rapae. Chem. Bio. Chem 7, 1982–1989.Google Scholar
  43. Wiklund, C., and Åhrberg, C. 1978. Host plants, nectar source plants, and habitat selection of males and females of Anthocharis cardamines (Lepidoptera). Oikos 31, 169–183.CrossRefGoogle Scholar
  44. Wiklund, C., and Friberg, M. 2009. The evolutionary ecology of generalisation: among-year variation in host plant use and offspring survival in a butterfly. Ecology 90, 3406–3417.CrossRefPubMedGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2010

Authors and Affiliations

  • Niels Agerbirk
    • 1
    Email author
  • Frances S. Chew
    • 2
  • Carl Erik Olsen
    • 1
  • Kirsten Jørgensen
    • 1
  1. 1.Faculty of Life SciencesUniversity of CopenhagenFrederiksbergDenmark
  2. 2.Department of BiologyTufts UniversityMedfordUSA

Personalised recommendations