Abstract
Changes in the weight and in the chlorophyll, free amino-acid and protein content of developing and senescing, vegetative and reproductive organs of Pisum sativum L. (cv. Burpeeana) were measured, and the proteolytic activity in extracts from the senescing leaf and the subtended pod was followed in relation to these changes. Protein content decreased in the ageing leaf and pod while it increased in the developing cotyledon. The proteolytic activity of the leaf did not increase as the leaf protein content decreased. In contrast, proteolytic activity in the subtended pod increased while the protein level decreased. The proteolytic activity in the extracts from the ageing organs was greater than the rates of protein loss. The proteolytic activity of leaf and pod extracts was greater on protein prepared from the respective organ than on non-physiological substrates. Proteolysis was increased by 2-mercaptoethanol and ethylenediaminetetraacetate but was not influenced by addition of ATP to the reaction mixture. The pH optimum was at 5.0. Free amino acids did not accumulate in the senescing leaf or pod when protein was degraded in each organ. It is suggested that these amino acids were quickly metabolized in situ or translocated to sink areas in the plant, especially to the developing seeds.
Similar content being viewed by others
References
Anderson, J.W., Rowan, K.S.: Activity of peptidase in tobacco leaf tissue in relation to senescence. Biochem. J. 97, 741–746 (1965)
Anderson, J.W., Rowan, K.S.: The effect of kinetin on senescence in tobacco leaf tissue after harvest. In: Biochemistry and physiology of plant growth substances, pp. 1437–1446, Wightman, F., Setterfield, G., eds. Ottawa: Runge Press 1968
Arnon, D.I.: Copper enzymes in isolated chloroplasts: polyphenoloxidase in Beta vulgaris. Plant Physiol. 24, 1–15 (1949)
Basha, S.M.M., Beevers, L.: The development of proteolytic activity and protein degradation during the germination of Pisum sativum L. Planta (Berl.) 124, 77–87 (1975)
Basha, S.M.M., Beevers, L.: Glycoprotein metabolism in the cotyledon of Pisum sativum L. during development and germination. Plant Physiol. 57, 93–98 (1976)
Beevers, L.: Growth regulator control of senescence in leaf discs of nasturtium (Tropaeolum majus). In: Biochemistry and physiology of plant growth substances, pp. 1417–1435, Wightman, F. and Setterfield, G., eds Ottawa: Runge Press 1968
Beevers, L.: Senescence: In: Experimental plant physiology, pp. 69–74, San Pietro, A., ed. St. Louis: Mosby 1974
Beevers, L.: Senescence. In: Plant Biochemistry, 3rd edn., pp. 771–794, Bonner, J., Varner, J.E., eds., New York: Academic Press 1976
Beevers, L., Poulson, R.: “Protein synthesis in cotyledons of Pisum sativum. I. Changes in cell free amino acid incorporation capacity during seed development and maturation. Plant Physiol. 49, 476–682 (1972)
Beevers, L., Storey, R.: Glutamate synthetase in developing cotyledons of Pisum sativum. Plant Physiol. 57, 862–866 (1976)
Bidwell, R.G.S., Barr, R.A., Steward, F.C.: Protein synthesis and turn-over in cultured plant tissue: source of carbon for synthesis and the fate of protein breakdown products. Nature (Lond.) 203, 367–373 (1964)
Brady, C.S.: The leaf protease of Trifolium repens. Biochem. J. 78, 631–640 (1961)
Dalling, M.J., Boland, G., Wilson, J.H.: Relation between acid proteinase activity and redistribution of nitrogen during grain development in wheat. Aust. J. Plant Physiol. 3, 721–730 (1977)
Davies, D.R.: Restructuring the pea plant. Sci. Prog. (Oxf.) 64, 201–214 (1977)
Etlinger, J.D., Goldberg, A.L.: A soluble ATP-dependent proteolytic system responsible for the degradation of abnormal proteins in reticulocytes. Proc. Natl. Acad. Sci. USA 74, 54–58 (1977)
Feller, U.K., Soong, T., Hageman, R.H.: Leaf proteolytic activities and senescence during grain development of field-grown corn (Zea mays L.). Plant Physiol. 59, 290–294 (1977)
Flinn, A.M., Pate, J.S.: Biochemical and physiological changes during maturation of fruit of the field pea (Pisum arvense L.) Ann. Bot. 32, 479–495 (1968)
Flinn, A.M., Pate, J.S.: A quantitative study of carbon transfer from pod and subtending leaf to the ripening seeds of the field pea (Pisum arvense L.). J. Exp. Bot. 21, 71–82 (1970)
Frith, G.J.T., Bruce, D.G., Dalling, M.J.: Distribution of acid protease activity in wheat seedlings. Plant Cell Physiol. 16, 1085–1091 (1975)
Goldberg, A.L., St. John, A.C.: Intracellular protein degradation in mammalian and bacterial cells. Pt. 2. Ann. Rev. Biochem. 45, 747–803 (1976)
Hall, J.M., Baker, D.A.: The chemical composition of Ricinus phloem exudate. Planta (Berl.) 106, 131–141 (1972)
Harvey, B.M.R., Oaks, A.: Characteristics of an acid protease from maize endosperm. Plant Physiol. 53, 449–452 (1974)
Hopkinson, J.M.: Studies on the expansion of leaf surface. VI. Senescence and usefulness of old leaves. J. Exp. Bot. 17, 762–770 (1966)
Huffaker, R.C., Peterson, L.W.: Protein turnover in plants and its possible means of regulation. Ann. Rev. Plant Physiol. 25, 363–392 (1974)
James, W.O.: Plant Respiration. London: Oxford Univ. Press 1953
Kipps, A.E., Boulter, D.: Origins of the amino acids in pods and seeds of Vicia faba L. New Phytol. 73, 675–684 (1974)
Lewis, O.A.M., Pate, J.S.: The significance of transpirationally derived nitrogen in protein synthesis in fruiting plants of pea (Pisum sativum L.). J. Exp. Bot. 24, 596–606 (1973)
Linck, A.S., Sudia, T.W.: Translocation of labeled photosynthetate from bloom node leaf to the fruit of Pisum sativum. Experientia 18, 69–70 (1962)
Lovell, P.H., Lovell, P.S.: Fixation of CO2 and export of photosynthate by the carpel in Pisum sativum. Physiol. Plant. 23, 316–322 (1970)
Lowry, O.H., Rosebrough, N.J., Farr, A.L.: Randall, R.J.: Protein measurement with the Folin phenol reagent. J. Biol. Chem. 193, 265–275 (1951)
McKee, H.S., Robertson, R.N., Lee, J.B.: Physiology of pea fruits. I. The developing fruit. Aust. J. Biol. Sci. 8, 136–163 (1955)
Millerd, A.: Biochemistry of legume seed proteins. Ann. Rev. Plant Physiol. 25, 53–72 (1975)
Muntz, K., Schalldach, I., Manteuffel, R.: The function of the pod for protein storage in seeds of Vicia faba L. Biochem. Physiol. Pflanzen 170, 465–477 (1976)
Oaks, A., Bidwell, R.G.S.: Compartmentation of intermediary metabolites. Ann. Rev. Plant Physiol. 21, 43–66 (1970)
Pate, J.S.: Physiological aspects of nitrogen metabolism with special reference to the legume (Pisum arvense). Symp. Long Ashton Res. Sta. Bristol. pp. 219–240 (1968)
Pate, J.S.: Pea. In: Crop physiology, some case histories, pp. 191–224, Evans, L.T., ed. Cambridge: University Press 1975
Pate, J.S., Flinn, A.M.: Carbon and nitrogen transfer from vegetative organs to ripening seeds of field pea (Pisum arvense L.). J. Exp. Bot. 24, 1090–1099 (1973)
Pate, J.S., Sharkey, P.M., Lewis, O.A.M.: Phloem bleeding from legume fruits — a technique for study of fruit nutrition. Planta (Berl.) 120, 229–243 (1974)
Pate, J.S., Sharkey, P.J., Lewis, O.A.M.: Xylem to phloem transfer in fruiting shoots of legumes studied by a phloem bleeding technique. Planta (Berl.) 122, 11–26 (1975)
Peterson, L.W., Huffaker, R.C.: Loss of ribulose-1,5-diphosphate carboxylase and increase in proteolytic activity during senescence of detached primary barley leaves. Plant Physiol. 55, 1009–1015 (1975)
Raache, I.D.: Protein synthesis in ripening pea seeds. III. Study of the pods. Biochem. J. 66, 113–118 (1957)
Racusen, D., Foote, M.: Protein turnover rate in bean leaf discs. Plant Physiol. 37, 640–647 (1962)
Racusen, D., Foote, M.: An endopeptidase of bean leaves. Canad. J. Bot. 48, 1017–1021 (1970)
Ryan, C.A.: Proteolytic enzymes and their inhibitors in plants. Ann. Rev. Plant Physiol. 24, 173–196 (1973)
Sakaro, S., Wildman, S.: Crystalline Fraction I protein, preparation in large yield. Science 176, 1145–1146 (1972)
Singh, N.: Proteolytic activity of leaf extracts. J. Sci. Food Agr. 13, 332 (1962)
Spencer, P.W., Spencer, R.D.: Globulin-specific proteolytic activity in germinating pumpkin seeds as detected by a fluorescence assay method. Plant Physiol. 54, 925–930 (1974)
Tracey, M.V.: Leaf proteases of tobacco and other plants. Biochem. J. 42, 281–287 (1948)
Van Die, J., Tammes, P.M.L.: Phloem exudation from monocotyledons axes. In: Encyclopedia of plant physiology, N.S., vol. I, transport in plants, Pt. I, pp. 196–222, Zimmermann, M.H., Milburn, J.A., eds. Berlin-Heidelberg-New York: Springer 1975
Wallace, W., Pate, J.S.: Nitrate assimilation in higher plants with special reference to the cocklebur (Xanthium pennsylvanicum Wallr). Ann. Bot. 31, 213–288 (1967)
Woolhouse, H.W.: The nature of senescence in plants. Symp. Soc. Exp. Biol. 21, 179–213 (1967)
Yemm, E.W., Cocking, E.C.: The determination of amino acids with ninhydrin. Analyst 80, 209–213 (1955)
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Storey, R., Beevers, L. Proteolytic activity in relationship to senescence and cotyledonary development in Pisum sativum L.. Planta 137, 37–44 (1977). https://doi.org/10.1007/BF00394432
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00394432