Abstract
We have compared the movement of a series of fluorescent tracers of increasing molecular weight injected into the cytoplasm in the epidermal cells of leaves of Egeria densa Planch. In general, the tracers showed major movement into three cellular compartments: first, to the cytoplasm of adjacent cells; secondly, from the cytoplasm, to the vacuole (irreversible); and thirdly, from the cytoplasm to the nucleus (reversible). No visible accumulation in chloroplasts or mitochondria, or loss across the plasmalemma was observed. No evidence for metabolic breakdown was found in extracts from injected leaves. The time course of accumulation of the dye in the three major compartments (cytoplasm, nucleus, vacuole) was monitored using fluorescence microscopy. The rate measurements and the quantified geometry of the cells were used to generate a model of compartmentation during intercellular transport. Permeability coefficients were calculated and related to the molecular sizes of the tracers. The coefficients for the tonoplast and nuclear envelope were independent of the molecular sizes of the tracers, and were in the range 2.4·10−6–4.1· 10−6 cm·s−1 for the tonoplast, and 2.6·10−5-9.4.10−5 cm· s−1 for the nuclear envelope. For intercellular movement, permeabilities were strongly dependent on molecular size, and ranged from 1.1·10−4 cm·s−1 for 6-carboxyfluorescein (376 daltons (Da)) to 9·10−9 cm·s−1 for fluorescein leucyldiglutamylleucine (874 Da). Thus, the differences in cell-to-cell movement of these tracers are based upon their differing ability to cross the intercellular walls, not upon differences in their intracellular compartmentation.
Similar content being viewed by others
Abbreviations
- 6COOHF:
-
6-Carboxyfluorescein
- Da:
-
daltons
- DAPI:
-
4′,6-diamidino-2-phenylindole
- F:
-
fluorescein-isothiocyanate isomer I
- FGlu:
-
fluorescein glutamic acid
- F(Glu)2 :
-
fluorescein glutamyl-glutamic acid
- F(Gly)6 :
-
fluorescein hexaglycine
- FLGGL:
-
fluorescein leucyl diglutamyl-leucine
References
Beffanga, N., Romani, G. (1988) Effects of two plasmalemma AT-Pase inhibitors on H+ extrusion and intracellular pH in Elodea densa leaves. J. Exp. Bot. 39, 1033–1043
Boller, T., Alibert, G. (1983) Photosynthesis in protoplasts from Metilotus alba: distribution of products between vacuole and cytosol. Z. Pflanzenphysiol. 110, 231–238
Boller, T., Vögeli, V. (1984) Vacuolar localisation of ethylene-induced chitinase in bean leaves. Plant Physiol. 74, 442–444
Boller, T., Wiemken, A. (1986) Dynamics of vacuolar compartmentation. Annu. Rev. Plant Physiol. 37, 137–164
Caveney, S. (1985) The role of gap junctions in development. Annu. Rev. Physiol. 47, 319–335
Deus-Neumann, B., Zenk, M.H. (1984) A highly selective alkaloid uptake system in vacuoles of higher plants. Planta 162, 250–260
Erwee, M.G., Goodwin, P.B. (1985) Symplast domains in extrastelar tissues of Egeria densa Planch. Planta 163, 9–19
Felle, H., Berthl, A. (1986) Light-induced cytoplasmic pH changes and their interrelation to the activity of the electrogenic proton pump in Riccia fluitans. Biochim. Biophys. Acta 848, 176–182
Flügge, U.I., Benz, R. (1984) Pore-forming activity in the outer membrane of the chloroplast envelope. FEBS Lett. 169, 85–89
Geiger, D.R., Ploeger, B.J., Fox, T.C., Fondy, B.R. (1983) Sources of sucrose translocated from illuminated sugar beet source leaves. Plant Physiol. 72, 964–970
Goodwin, P.B. (1981) Short distance transport in the plant symplast. Abstr. XIIIth Int. Bot. Congr. Sydney, p 35
Goodwin, P.B. (1983) Molecular size limits for movement in the symplast of the Elodea leaf. Planta 157, 124–130
Goodwin, P.B., Erwee, M.G. (1985) Intercellular transport studied by micro-injection methods. In: Botanical microscopy 1985, pp. 335–358, Robards, A.W., ed. Oxford University Press, Oxford
Goodwin, P.B., Erwee, M.G. (1990) Transport across the tonoplast in relation to cell communication in plants. In: Transport across membranes of plants and fungi, pp. 303–306. Beilby, M.J., Walker, N.A., Smith, J.R., eds. Sydney University Press, Sydney
Goodwin, P.B., Shepherd, V. (1987) Compartmentation of fluorescent probes injected into the epidermal cells of Egeria densa leaves. Abstr. XIV Int. Bot. Cong. Berlin, p. 212
Oparka, K.J., Prior, D.A.M. (1988) Movement of lucifer yellow CH in potato storage tissues: a comparison of symplastic and apoplastic transport. Planta 176, 533–540
Paine, P.L., Moore, L.C., Horowitz, S.B. (1975) Nuclear envelope permeability. Nature 254, 109–114
Raven, J.A. (1984) Vacuoles: storage, flotation, and amplification of surface/volume ratio of cytoplasm. In: Energetics and transport in aquatic plants (Marine Biological Laboratory Lectures in Biology) 4, 411–470
Raven, J.A. (1987) The role of vacuoles. New Phytol. 106, 357–422
Serpersu, E.H., Kinosita, K., Tsong, T.Y. (1985) Reversible and irreversible modification of erythrocyte membrane permeability by electric field. Biochim. Biophys. Acta 812, 779–785
Taylor, D.L., Wang, Y.L. (1980) Fluorescently labelled molecules as probes of the structure and function of living cells. Nature 284, 405–410
Terry, B.R., Robards, A.W. (1987) Hydrodynamic radius alone governs the mobility of molecules through plasmodesmata. Planta 171, 145–157
Tucker, E.B. (1982) Translocation in the staminal hairs of Setcreasea purpurea. I. A study of cell ultrastructure and cell-to-cell passage of molecular probes. Protoplasma 113, 193–201
Tucker, E.B., Spanswick, R.M. (1985) Translocation in the staminal hairs of Setcreasea purpurea. II. Kinetics of intercellular transport. Protoplasma 128, 167–172
Vogel, A.I. (1966) A textbook of quantitative inorganic analysis including elementary instrumental analysis, 3rd edn. Longmans, Green & Co., London
Walker, N.A., Bostrom, T.E. (1983) Intercellular movement of chloride in Chara — a test of models for chloride influx. In: Ion transport in plants, pp. 447–458, Anderson, W.P., ed. Academic Press, London
Weiner, H., Burnell, J.N., Woodrow, I.E., Heldt, H.W., Hatch, M.D. (1988) Metabolite diffusion into bundle sheath cells from C4 plants. Relation to C4 photosynthesis and plasmodesmatal function. Plant Physiol. 88, 815–822
Zimmerman, A.L., Rose, B. (1985) Permeability properties of cellto-cell channels: kinetics of fluorescent tracer diffusion through a cell junction. J. Membr. Biol. 84, 269–283
Author information
Authors and Affiliations
Additional information
This work was supported by the Australian Research Grant Scheme. The assistance of Professor B.E.S. Gunning (Australian National University, Canberra, ACT) in providing facilities for making the photometer measurements is gratefully acknowledged.
Rights and permissions
About this article
Cite this article
Goodwin, P.B., Shepherd, V. & Erwee, M.G. Compartmentation of fluorescent tracers injected into the epidermal cells of Egeria densa leaves. Planta 181, 129–136 (1990). https://doi.org/10.1007/BF00202335
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00202335