Abstract
Tannic acid and quebracho precipitate many times their weight of the abundant leaf protein, ribulose-1,5-bisphosphate carboxylase/oxygenase (RuBPC). The use of this protein in protein precipitation assays for tannin content is described. Extracts of mature foliage of pin, bur, and black oak precipitate 2.01, 0.69, and 0.09 mg RuBPC/ mg (dry wt) of leaf powder extracted, respectively, at pH 6.1. From these measurements it can be calculated that all three of these oak species have sufficient tannins to precipitate all of the RuBPC present in their foliage. At mildly alkaline pHs, however (pH ⩾ 7.5), RuBPC is not precipitated by tannins. Since RuBPCis the most abundant protein present in photosynthetic tissues, often constituting as much as 50% of the soluble proteins and 25% of the total proteins in leaf tissue, the interactions of this protein with tannins are highly relevant to an evaluation of the role of tannins as antiherbivore, digestibility-reducing substances. Our measurements provide no basis for arguing that differences in tannin levels in different species reflect differences in the digestibility of leaf proteins or that tannins have any effect whatsoever upon the digestibility of leaf protein under conditions which normally prevail in most insects' guts. These findings emphasize the need to test more of the assumptions underlying contemporary interpretations of the importance of tannins in plant herbivore interactions.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Akazawa, T. 1970. The structure and function of fraction-I protein.Progr. Phytochem. 2:107–141.
Bate-Smith, E.C. 1973. Haemanalysis of tannins: The concept of relative astringency.Phytochemistry 12:907–912.
Bate-Smith, E.C., andMetcalfe, C.R. 1957. The nature and distribution of tannins in dicotyledonous plants.J. Linn. Soc., London, Bot. 55:669–705.
Becker, P., andMartin, J.S., 1982. Protein-precipitating capacity of tannins inShorea (Dipterocarpaceae) seedling leaves.J. Chem. Ecol. 8:1353–1367.
Berenbaum, M. 1980. Adaptive significance of midgut pH in larval Lepidoptera.Am. Nat. 115:138–146.
Bernays, E.A. 1981. Plant tannins and insect herbivores: an appraisal.Ecol. Entomol. 6:353–360.
Bradford, M. 1976. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding.Anal. Biochem. 72:248–254.
Calderon, P., Van Buren, J., andRobinson, W.B. 1968. Factors influencing the formation of precipitates and hazes by gelatin and condensed and hydrolyzable tannins.J. Agric. Food Chem. 16:479–482.
Dow, J.A.T., Gupta, B.L., andHall, T.A. 1981. Microprobe analysis of Na, K, Cl, P, S, Ca, Mg and H2O infrozen-hydrated sections of anterior caecae of the locust,Schistocerca gregaria.J. Insect Physiol. 27:629–639.
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.
Feeny, P. 1976. Plant apparency and chemical defense.Recent. Adv. Phytochem. 10:1–40.
Gartlan, J.S., McKey, D.B., Waterman, P.G., Mbi, C.N. andStruhsakers, T.T. 1980. A comparative study of the phytochemistry of two African rain forests.Biochem. Syst. Ecol. 8:401–422.
Giordana, B., andSacchi, F. 1978. Cellular ionic concentrations in the midgut of two larvae of Lepidoptera in vivo and in vitro.Comp. Biochem. Physiol. 59A:17–20.
Goldstein, J.L., andSwain, T. 1965. The inhibition of enzymes by tannins.Phytochemistry 4:185–192.
Griffiths, D. W. 1979, The inhibition of digestive enzyme by extracts of field bean (Vicia faba).J. Sci. Food Agric. 30:458–462.
Hagerman, A.E., andButler, L.G. 1978. Protein precipitation method for the quantitative determination of tannins.J. Agric. Food Chem. 26:809–812.
Hagerman, A.E., andButler, L.G. 1980. Determination of protein in tannin-protein precipitates.J. Agric. Food Chem. 28:944–947.
Hagerman, A.E., andButler, L.G. 1981. The specificity of proanthocyanidin-protein interactions.J. Biol. Chem. 256:4494–4497.
Haslam, E. 1974. Polyphenol-protein interactions.Biochem. J. 139:285–288.
Jensen, R.G., andBahr, J.T. 1977. Ribulose-1,5-bisphosphate carboxylase-oxygenase.Annu. Rev. Plant Physiol. 28:379–400.
Lawson, D.L.,Merritt, R.W.,Klug, M. J., andMartin, J.S. 1982. The utilization of late season foliage by the orange striped oakworm,Anisota senatoria, (J.E. Smith) (Cither-oniidae).Entomol. Exp. Appl. In press.
Lyttleton, J.W. 1973. Proteins and nucleic acids.Chem. Biochem. Herb. 1:63–103.
Mandels, M., andReese, E.T. 1963. Inhibition of cellulases and β-glucosidases, pp. 115–157,in E.T. Reese (ed.). Advances in Enzymic Hydrolysis of Cellulose and Related Materials. Pergamon Press, New York.
Martin, J.S., andMartin, M.M. 1982. Tannin assays in ecological studies: Lack of correlation between phenolics, proanthocyanidins and protein-precipitating constituents in mature foliage of six oak species.Oecologia 54:205–211.
Rhoades, D. F., andCates, R.G. 1976. Toward a general theory of plant antiherbivore chemistry.Recent Adv. Phytochem. 10:168–213.
Schultz, J.C., Baldwin, I.T., andNothnagle, P. J. 1981. Hemoglobin as a binding substrate in the quantitative analysis of plant tannins.J. Agric. Food Chem. 29:823–826.
Singer, E.J., Eggman, L., Campbell, J.M., andWildman, S.G. 1952. The proteins of green leaves. IV. A high molecular weight protein comprising a large part of the cytoplasmic protein.J. Biol. Chem. 197:233–239.
Sokal, R.R., andRohlf, F.J. 1969. Biometry, W.H. Freeman, San Francisco.
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 York.
Van Buren, J.P., andRobinson, W.B. 1969. Formation of complexes between protein and tannic acid.J. Agric. Food Chem. 17:772–777.
Van Sumere, C.F., Albrecht, J., Dedonder, A., DePooter, H. andPe, I. 1975. Plant protein and phenolics, pp. 211–264,in J.B. Harborne and C.F. Van Sumere (eds.). The Chemistry and Biochemistry of Plant Proteins. Academic Press, New York.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Martin, J.S., Martin, M.M. Tannin assays in ecological studies Precipitation of ribulose-1,5-bisphosphate carboxylase/oxygenase by tannic acid, quebracho, and oak foliage extracts. J Chem Ecol 9, 285–294 (1983). https://doi.org/10.1007/BF00988046
Received:
Revised:
Issue Date:
DOI: https://doi.org/10.1007/BF00988046