Abstract
A method is described for isolating epidermal protoplasts from the primary leaves of barley (Hordeum vulgare L.). Epidermal protoplasts are lighter than mesophyll protoplasts because of their smaller ratio of cytoplasm to vacuole, and can be separated from the latter by density-gradient centrifugation after complete digestion of the leaves. We have started a basic characterization of the epidermal protoplast fraction in comparison with mesophyll protoplasts. Epidermal protoplasts had a mean diameter of 63.5 μm, whereas that of mesophyll protoplasts was 35.7 μm. Their respiratory oxygen consumption was not influenced by light. They contained acid hydrolases and cytoplasmic enzymes in relative activities different from those of mesophyll protoplasts. Their polypeptide pattern as judged from two-dimensional separations was, in principle, similar to that of mesophyll cells after elimination of the plastids from the latter by the preparation of vacuoplasts. However, in addition, a considerable number of epidermis-specific polypeptides were observed. Isolated epidermal protoplasts were viable and efficiently incorporated [35S]methionine into newly synthesized proteins. The results show that epidermal protoplasts are suitable for the investigation of the physiological and molecular properties of epidermal cells in leaves.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
Abbreviations
- SDS-PAGE:
-
sodium dodecyl sulfate-polyacrylamide gel electrophoresis
References
Boller, T., Kende, H. (1979) Hydrolytic enzymes in the central vacuole of plant cells. Plant Physiol. 63, 1123–1132
Delieu, T., Walker, D.A. (1974) An improved cathode for the measurement of photosynthetic oxygen evolution by isolated chloroplasts. New Phytol. 71, 201–225
Dietz, K.-J., Bogorad, L. (1987) Plastid development in Pisum sativum leaves during greening. 1. A comparison of plastid polypeptide composition and in organello translation characteristics. Plant Physiol. 85, 808–815
Dietz, K.-J., Kaiser, G., Martinoia, E. (1988) Characterization of vacuolar polypeptides of barley mesophyll cells by two-dimensional gel electrophoresis and by their affinity to lectins. Planta 176, 362–367
Dietz, K.-J., Schramm, M., Lang, B., Lanzl-Schramm, A., Dürr, C., Martinoia, E. (1992) Characterization of the epidermis from barley primary leaves. II. The role of the epidermis in ion compartmentation. Planta 187, 431–437
Gorton, H.L., Williams, W.E., Binns, M.E., Gemmell, C.N., Leheny, E.A., Shepard, A.C. (1989) Circadian stomatal rhythms in epidermal peels from Vicia faba. Plant Physiol. 90, 1329–1334
Hampp, R., Outlaw, W.H., Tarczynski, M.C. (1982) Profile of basic carbon pathways in guard cells and other leaf cells of Vicia faba L. Plant Physiol. 70, 1582–1585
Kaiser, G., Heber, U. (1983) Photosynthesis of leaf cell protoplasts and permeability of the plasmalemma to some solutes. Planta 157, 462–470
Kaiser, G., Martinoia, E., Wiemken, A. (1982) Rapid appearance of photosynthetic products in the vacuoles isolated from barley mesophyll protoplasts by a new fast method. Z. Pflanzenphysiol. 107, 103–113
Knuth, M.E., Keith, B., Clark, C., Garcia-Martinez, J.L., Rappaport, L. (1983) Stabilization and transport capacity at cowpea and barley vacuoles. Plant Cell Physiol. 24, 423–432
Kolattukudy, P.E. (1977) Biosynthesis and degradation of lipid polymers. In: Lipid and lipid polymers in higher plants, pp. 271–299, Tevini, M., Lichtenthaler, H.K., eds. Springer, Berlin Heidelberg New York
Kolattukudy, P.E. (1980) Cutin, suberin, and waxes. In: The biochemistry of plants, pp. 571–645, Stumpf, P.K., Conn, E.E., eds. Academic Press, New York London Toronto
Mimura, T., Dietz, K.-J., Kaiser, W.M., Schramm, M.J., Kaiser, G., Heber, U. (1990) Phosphate transport across biomembranes and cytosolic ion homeostasis in barley leaves. Planta 180, 139–146
Pfanz, H., Dietz, K.-J. (1987) A fluorescence method for the determination of the apoplastic proton concentration in intact leaf tissue. J. Plant Physiol. 129, 41–48
Outlaw, W.H., Manchester, J., DiCamelli, C.A., Randall, D.D., Rapp, B., Veith, G.M. (1979) Photosynthetic carbon reduction pathway is absent in chloroplasts of Vicia Faba guard cells. Proc. Natl. Acad. Sci. USA 76, 6371–6375
Outlaw, W.H., Tarczynski, M.C., Miller, W.I. (1984) Histological compartmentation of phosphate in Vicia faba. L. leaflet. Plant Physiol. 74, 430–433
Outlaw, W.H., Springer, S.A., Tarczynski, M.C. (1985) Histochemical technique: a general method for quantitative enzyme assays of single cell ‘extracts’ with a time resolution of seconds and reading precision of femtomoles. Plant Physiol. 77, 659–666
Rother, Th., Acker, G., Scheibe, R. (1988) Immunogold localization of chloroplast protein in spinach leaf mesophyll, epidermis, and guard cells. Bot. Acta 101, 311–320
van Steveninck, R.F.M., van Steveninck, M.E., Stelzer, R., Läuchli, A. (1982) Studies on the distribution of Na and Cl in two species of lupin (Lupinus luteus and Lupinus angustifolius) differing in salt tolerance. Physiol. Plant. 56, 465–473
Zeiger, E., Hepler, P. (1977) Light and stomatal function: blue light stimulates swelling of guard cell protoplasts. Science 196, 887–889
Author information
Authors and Affiliations
Additional information
We are grateful to Professor U. Heber (Lehrstuhl Botanik 1, Würzburg) for his continuous support. This work was supported by the DFG and the University of Würzburg within the Sonderforschungsbereich 176.
Rights and permissions
About this article
Cite this article
Dietz, K.J., Schramm, M., Betz, M. et al. Characterization of the epidermis from barley primary leaves. Planta 187, 425–430 (1992). https://doi.org/10.1007/BF00199959
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00199959