Journal of Chemical Ecology

, Volume 29, Issue 5, pp 1099–1116 | Cite as

Antioxidants in the Midgut Fluids of a Tannin-Tolerant and a Tannin-Sensitive Caterpillar: Effects of Seasonal Changes in Tree Leaves

  • Raymond V. BarbehennEmail author
  • Ann C. Walker
  • Farhan Uddin


The seasonal decline in foliar nutritional quality in deciduous trees also effects the availability of essential micronutrients, such as ascorbate and α-tocopherol, to herbivorous insects. This study first examined whether there are consistent patterns of seasonal change in antioxidant concentrations in deciduous tree leaves. α-Tocopherol concentrations increased substantially through time in late summer in sugar maple (Acer saccharum), red oak (Quercus rubra), and trembling aspen (Populus tremuloides). However, seasonal change in the concentrations of other antioxidants differed between each species: P. tremuloides had higher levels of ascorbate and glutathione in the spring, Q. rubra had higher levels of glutathione but lower levels of ascorbate in the spring, and A. saccharum had lower levels of both ascorbate and glutathione in the spring. To test the hypothesis that tannin-tolerant caterpillars maintain higher concentrations of antioxidants in their midgut fluids than do tannin-sensitive species, we measured antioxidants in Orgyialeucostigma (a spring- and summer-feeding, tannin-tolerant species) and Malacosoma disstria (a spring-feeding, tannin-sensitive species) that were fed tree leaves in the spring and summer. The midgut fluids of O. leucostigma larvae generally had higher concentrations of antioxidants in the summer than did those of M. disstria, and were significantly higher overall. The results of this study are consistent with the hypothesis that higher concentrations of antioxidants form an important component of the defenses of herbivores that feed on mature, phenol-rich tree leaves. Some limitations of the interpretation of total antioxidant capacity are also discussed.

Ascorbic acid α-tocopherol glutathione Lepidoptera herbivore antioxidant tree leaves 


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. Anderson, J. V., Chevone, B. I., and Hess, J. L. 1991. Seasonal variation in the antioxidant system of eastern white pine needles. Plant Physiol. 98:501–508.Google Scholar
  2. Appel, H. M. and Maines, L. W. 1995. The influence of host plant on gut conditions of gypsy moth (Lymantria dispar) caterpillars. J. Insect Physiol. 41:241–246.Google Scholar
  3. Appel, H. M., Govenor, H. L., D'Ascenzo, M. D., Siska, E., and Schultz, J. C. 2001. Limitations of folin assays of foliar phenolics in ecological studies. J. Chem. Ecol. 27:761–778.Google Scholar
  4. Aucoin, R. R., Fields, P., Lewis, M. A., Philogene, B. J. R., and Arnnason, J. T. 1990. The protective effect of antioxidants to a phototoxin-sensitive herbivore, Manduca sexta. J. Chem. Ecol. 16:2913–2924.Google Scholar
  5. Ayres, M. P. and MacLean, S. F., Jr. 1987. Development of birch leaves and the growth energetics of Epirrita autumnata (Geometridae). Ecology 68:558–568.Google Scholar
  6. Baker, W. L. 1972. Eastern Forest Insects. USDA miscellaneous publication no. 1175. Washington, DC.Google Scholar
  7. Barbehenn, R. V. 2003. Antioxidants in grasshoppers: higher levels defend the midgut tissues of a polyphagous species than a graminivorous species. J. Chem. Ecol. 29:665–684.Google Scholar
  8. Barbehenn, R. V., Bumgarner, S. L., Roosen, E. F., and Martin, M. M. 2001. Antioxidant defenses in caterpillars: Role of the ascorbate-recycling system in the midgut lumen. J. Insect Physiol. 47:349-357. (Erratum 47:1095)Google Scholar
  9. Barbehenn, R. V. and Martin, M. M. 1992. The protective role of the peritrophic membrane in the tannin-tolerant larvae of Orgyia leucostigma (Lepidoptera). J. Insect Physiol. 12:973–980.Google Scholar
  10. Barbehenn, R. V. and Martin, M. M. 1994. Tannin sensitivity in larvae of Malacosoma disstria (Lepidoptera): Roles of the peritrophic envelope and midgut oxidation. J. Chem. Ecol. 20:1985–2001.Google Scholar
  11. Barbehenn, R. V., Martin, M. M., and Hagerman, A. E. 1996. Reassessment of the roles of the peritrophic envelope and hydrolysis in protecting polyphagous grasshoppers from ingested hydrolyzable tannins. J. Chem. Ecol. 22:1901–1919.Google Scholar
  12. Barbehenn, R. V., Poopat, U., and Spencer, B. 2003. Semiquinone and ascorbyl radicals in the gut fluids of caterpillars measured with EPR spectrometry. Insect Biochem. Mol. Biol. 33:125–130.Google Scholar
  13. Bi, J. L. and Felton, G. W. 1995. Foliar oxidative stress and insect herbivory: Primary compounds, secondary metabolites, and reactive oxygen species as components of induced resistance. J. Chem. Ecol. 21:1511-1530Google Scholar
  14. Cilliers, J. J. L. and Singleton, V. L. 1989. Nonenzymic autoxidative phenolic browning reactions in a caffeic acid model system. J. Agric. Food Chem. 37:390–396.Google Scholar
  15. Dadd, R. H. 1973. Insect nutrition: current developments and metabolic implications. Annu. Rev. Entomol. 18:381–420.Google Scholar
  16. Dash, J. A. and Jenness, R. 1985. Ascorbate content of foliage of eucalypts and conifers by some Australian and North American mammals. Experientia 41:952–955.Google Scholar
  17. Feeny, P. P. 1968. Effect of oak leaf tannins on larval growth of the winter moth Operophtera brumata. J. Insect Physiol. 14:805–817.Google Scholar
  18. Feeny, P. P. 1970. Seasonal changes in oak leaf tannins and nutrients as a cause of spring feeding by winter moth caterpillars. Ecology 51:565–581.Google Scholar
  19. Felton, G. W., Donato, K. K., Del Vecchio, R. J., and Duffey, S. S. 1989. Activation of plant polyphenol oxidases by insect feeding damage reduces the nutritive quality of foliage. J. Chem. Ecol. 15:2667–2694.Google Scholar
  20. Felton, G. W. and Duffey, S. S. 1992. Ascorbate oxidation reduction in Helicoverpa zea as a scavenging system against dietary oxidants. Arch. Insect Biochem. Physiol. 19:27–37.Google Scholar
  21. Felton, G. W. and Summers, C. B. 1993. Potential role of ascorbate oxidase as a plant defense protein against insect herbivory. J. Chem. Ecol. 19:1553–1568.Google Scholar
  22. Hagerman, A. E. 1988. Extraction of tannin from fresh and preserved leaves. J. Chem. Ecol. 14:453–461.Google Scholar
  23. Hagerman, A. E., Riedl, K. M., Jones, G. A., Sovik, K. N., Ritchard, N. T., Hartzfeld, P. W., and Riechel, T. L. 1998. High molecular weight plant polyphenolics (tannins) as biological antioxidants. J. Agric. Food Chem. 46:1887–1892.Google Scholar
  24. Halliwell, B. and Gutteridge, J. M. C. 1999. Free Radicals in Biology and Medicine. Oxford University Press, Oxford, England.Google Scholar
  25. Hess, J. L. 1993. Vitamin E, α-tocopherol, pp. 111-134, in R. G. Alscher and J. L. Hess (Eds.). Antioxidants in Higher Plants. CRC Press, Boca Raton, Florida.Google Scholar
  26. Hunter, A. F. and Lechowicz, M. J. 1992. Foliage quality changes during canopy development of some northern hardwood trees. Oecologia 89:316–323.Google Scholar
  27. Johnson, K. S. and Felton, G. W. 1996. Physiological and dietary influences on midgut redox conditions in generalist lepidopteran larvae. J. Insect Physiol. 42:191–198.Google Scholar
  28. Johnson, K. S. and Felton, G. W. 2001. Plant phenolics as dietary antioxidants for herbivorous insects: A test with genetically modified tobacco. J. Chem. Ecol. 27:2579–2597.Google Scholar
  29. Karowe, D. N. 1989. Differential effect of tannic acid on two tree-feeding Lepidoptera: Implications for theories of plant-herbivore chemistry. Oecologia 80:507–512.Google Scholar
  30. Kramer, K. and Seib, P. A. 1982. Ascorbic acid and the growth and development of insects, pp. 275-291, in P. A. Seib and B. M. Tolbert (Eds.). Ascorbic Acid: Chemistry, Metabolism and Uses. ACS, Washington, DC.Google Scholar
  31. Kunert, K. J. and Ederer, M. 1985. Leaf aging and lipid peroxidation: The role of the antioxidants vitamin C and E. Physiol. Plant. 65:85–88.Google Scholar
  32. Lindroth, R. L., Barman, M., and Weisbrod, A. W. 1991. Nutritional deficiencies in the gypsy moth, Lymantria dispar: Effects on larval performance and detoxication enzyme activities. J. Insect Physiol. 37:45–52.Google Scholar
  33. Lindroth, R. L., Hsia, M. T. S., and Scriber, J. M. 1987. Seasonal patterns in the phytochemistry of three Populus species. Biochem. Syst. Ecol. 15:681–686.Google Scholar
  34. Lindroth, R. L. and Weiss, A. P. 1994. Effects of ascorbic acid deficiencies on larvae of Lymantria dispar (Lepidoptera: Lymantriidae). Great Lakes Entomol. 27:169–174.Google Scholar
  35. Luwe, M. 1996. Antioxidants in the apoplast and symplast of beech (Fagus sylvatica L.) leaves: Seasonal variations and responses to changing ozone concentrations in air. Plant Cell Environ. 19:321–328.Google Scholar
  36. Marabottini, R., Schraml, C., Paolacci, A. R., Sorgona, A., Raschi, A., Rennenberg, H., and Badiani, M. 2001. Foliar antioxidant status of adult Mediterranean oak species (Quercus ilex L. and Q. pubescens Willd.) exposed to permanent CO2-enrichment and to seasonal water stress. Environ. Pollut. 113:413–423.Google Scholar
  37. Maufette, Y. and Oechel, W. C. 1989. Seasonal variation in leaf chemistry of the coast live oak Quercus agrifolia and implications for the California oak moth Phryganidia californica. Oecologia 79:439–445.Google Scholar
  38. Meister, A. 1992. On the antioxidant effects of ascorbic acid and glutathione. Biochem. Pharmacol. 44:1905–1915.Google Scholar
  39. Navon, A. 1978. Effects of dietary ascorbic acid on larvae of the Egyptian cotton leafworm, Spodoptera littoralis. J. Insect Physiol. 24:39–44.Google Scholar
  40. Nicol, R. W., Arnason, J. T., Helson, B., and Abou-Zaid, M. M. 1997. Effect of host and nonhost trees on the growth and development of the forest tent caterpillar, Malacosoma disstria (Lepidoptera: Lasiocampidae). Can. Entomol. 129:991–999.Google Scholar
  41. Panzuto, M., Lorenzetti, F., Mauffette, Y., and Albert, P. J. 2001. Perception of aspen and sun/shade sugar maple leaf soluble extracts by larvae of Malacosoma disstria. J. Chem. Ecol. 27:1963–1978.Google Scholar
  42. Pardini, R. S. 1995. Toxicity of oxygen from naturally occurring redox-active pro-oxidants. Arch. Insect Biochem. Physiol. 29:101–118.Google Scholar
  43. Pedersen, J. A. 2000. Distribution and taxonomic implications of some phenolics in the family Lamiaceae determined by ESR spectroscopy. Biochem. Syst. Ecol. 28:229–253.Google Scholar
  44. Pietta, P., Simonetti, P., and Mauri, P. 1998. Antioxidant activity of selected medicinal plants. J. Agric. Food Chem. 46:4487–4490.Google Scholar
  45. Pratt, D. E. 1992. Natural antioxidants from plant material, pp. 54-71, in M.-T. Huang, C-T. Ho, and C. Y. Lee (Eds.). Phenolic Compounds in Food and their Effects on Health. II. Antioxidants and Cancer Prevention (ACS Symposium Series 507). American Chemical Soc., Washington, DC.Google Scholar
  46. Prior, R. L., Cao G., Martin, A., Sofic, E., McEwen, J., O'Brien, C., Lischner, N., Ehlenfeldt, M., Kalt, W., Krewer, G., and Mainland C. M. 1998. Antioxidant capacity as influenced by total phenolic and anthocyanin content, maturity, and variety of Vaccinium species. J. Agric. Food Chem. 46:2686–2693.Google Scholar
  47. Re, R., Pellegrini, N., Proteggente, A., Pannala, A., Yang, M., and Rice-Evans, C. 1999. Antioxidant activity applying an improved ABTS radical cation decolorization assay. Free Rad. Biol. Med. 26:1231–1237.Google Scholar
  48. Ricklefs, R. E. and Matthew, K. K. 1982. Chemical characteristics of the foliage of some deciduous trees in southeastern Ontario. Can. J. Bot. 60:2037–2045.Google Scholar
  49. Rossiter, M., Schultz, J. C., and Baldwin, I. T. 1988. Relationship among defoliation, red oak phenolics, and gypsy moth growth and reproduction. Ecology 69:267–277.Google Scholar
  50. Roth, S., Lindroth, R., and Montgomery, M. 1994. Effects of foliar phenolics and ascorbate on performance of the gypsy moth (Lymantria dispar). Biochem. Syst. Ecol. 22:341–351.Google Scholar
  51. SAS Institute. 2000. The SAS System for Windows, Version 8e. SAS Institute, Cary, North Carolina.Google Scholar
  52. Schroeder, L. A. 1986. Changes in tree leaf quality and growth performance of lepidopteran larvae. Ecology 67:1628–1636.Google Scholar
  53. Schultz, J. C., Nothnagle, P. J., and Baldwin, I. T. 1982. Seasonal and individual variation in leaf quality of two northern hardwood tree species. Am. J. Bot. 69:753–759.Google Scholar
  54. Schupp, R. and Rennenberg, H. 1988. Diurnal changes in the glutathione content of spruce needles (Picea abies L.). Plant Sci. 57:113–117.Google Scholar
  55. Schwanz, P., Kimball, B. A., Idso, S. B., Hendrix, D. L., and Polle, A., 1996a. Antioxidants in sun and shade leaves of sour orange trees (Citrus aurantium) after long-term acclimation to elevated CO2. J. Exp. Bot. 47:1941–1950.Google Scholar
  56. Schwanz, P., Picon, C., Vivin, P., Dreyer, E., Guehl, J. M., and Polle, A., 1996b. Responses of antioxidative systems to drought stress in pedunculate oak and maritime pine as modulated by elevated CO2. Plant Physiol. 110:393–402.Google Scholar
  57. Scriber, J. M. 1977. Limiting effects of low leaf-water content on the nitrogen utilization, energy budget, and larval growth of Hyalophora cecropia (Lepidoptera: Saturniidae). Oecologia 28:269–287.Google Scholar
  58. Sokal, R. R. and Rohlf, F. J. 1981. Biometry, 2 Edition. Freeman, San Francisco, California.Google Scholar
  59. Stehr, F. W. and Cook, E. F. 1968. A Revision of the Genus Malacosoma Hübner in North America (Lepidoptera: Lasiocampidae): Systematics, Biology, Immatures, Parasites. Smithsonian Institution Press, Washington, DC.Google Scholar
  60. Strohm, M., Jouanin, L., Kunert, K. J., Pruvost, C., Polle, A., Foyer, C. H., and Rennenberg, H. 1995. Regulation of glutathione synthesis in leaves of transgenic poplar (Populus tremula X P. alba) overexpressing glutathione synthetase. Plant J. 7:141–145.Google Scholar
  61. Strube, M., Haenen, G. R. M. M., Van Den Berg, H., and Bast, A. 1997. Pitfalls in a method for assessment of total antioxidant capacity. Free Rad. Res. 26:515–521.Google Scholar
  62. Summers, C. B. and Felton, G. W. 1994. Prooxidant effects of phenolic acids on the generalist herbivore Helicoverpa zea (Lepidoptera: Noctuiidae): Potential mode of action for phenolic compounds in plant anti-herbivore chemistry. Insect Biochem. Mol. Biol. 24:943–953.Google Scholar
  63. Swain, T. 1979. Tannins and lignins, pp. 657-682, in G. A. Rosenthal and D. H. Janzen (Eds.). Herbivores: Their Interaction with Secondary Plant Metabolites. Academic Press, New YorksGoogle Scholar
  64. Thiboldeaux, R. L., Lindroth, R. L., and Tracy, J. W. 1998. Effects of juglone (5-hydroxy-1,4-naphthoquinone) on midgut morphology and glutathione status in Saturniid moth larvae. Comp. Biochem. Physiol. 120:481–487.Google Scholar
  65. Timmerman, S. E., Zangerl, A. R., and Berenbaum, M. R. 1999. Ascorbic and uric acid responses to xanthotoxin ingestion in a generalist and a specialist caterpillar. Arch. Insect Biochem. Physiol. 42:26–36.Google Scholar
  66. Vanderzant, E. S., Pool, M. C., and Richardson, C. D. 1962. The role of ascorbic acid in the nutrition of three cotton insects. J. Insect Physiol. 8:287–297.Google Scholar
  67. Velioglu, Y. S., Mazza, G., Gao, L., and Oomah, B. D. 1998. Antioxidant activity and total phenolics in selected fruits, vegetables and grain products. J. Agric. Food Chem. 46:4113–4117.Google Scholar

Copyright information

© Plenum Publishing Corporation 2003

Authors and Affiliations

  • Raymond V. Barbehenn
    • 1
    Email author
  • Ann C. Walker
    • 1
  • Farhan Uddin
    • 1
  1. 1.Departments of Ecology and Evolutionary Biology and Molecular, Cellular and Developmental BiologyUniversity of MichiganAnn ArborUSA

Personalised recommendations