Abstract
Metabolite concentrations in subcellular compartments from mature barley (Hordeum vulgare L. cv. Apex) leaves after 9 h of illumination and 5 h of darkness were determined by nonaqueous fractionation and by the stereological evaluation of cellular and subcellular volumes from light and electron micrographs. Twenty one-day-old primary leaves of barley with a total leaf volume of 902 μL per mg chlorophyll were found to be composed of 27% epidermis, 42% mesophyll cells, 6% veins, 4.5% apoplast and 23% gas space. While in epidermal cells 99% of the volume was occupied by the vacuole, mesophyll cells with an average volume of 31.3 pL consisted of 23 pL (73%) vacuole, 4.6 pL (19%) chloroplasts, 2.06 pL (6,7%) cytosol (including smaller organelles and vesicles), 0.34 pL (1%) mitochondria and 107 fL (0.34%) nucleus. The differences between leaves harvested after 9 h of illumination and after 5 h of darkness were in the size of the stromal compartment and the starch grains therein. Subcellular metabolite concentrations were calculated from the compartmental volumes and metabolite contents of the compartments as determined by nonaqueous fractionation. The amino-acid concentrations in stroma and cytosol were rather similar after 9 h of illumination and 5 h of darkness. In contrast, the vacuolar amino-acid concentrations were about one order of magnitude lower than the stroma and cytosol values, and there was a slight increase in concentration after 5 h of darkness.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
Abbreviations
- Chl:
-
chlorophyll
References
Arnon, D.J. (1949) Copper enzymes in isolated chloroplasts. Polyphenoloxidase in Beta vulgaris. Plant Physiol. 24, 1–15
Delesse, M.A. (1847) Procédé mecanique pour determiner la composition des roches. C. R. Acad. Sci. (Paris) 25, 544
Dietz, K.-J., Jäger, R., Kaiser, G., Martinoia, E. (1990) Amino acid transport across the tonoplast of vacuoles isolated from barley mesophyll protoplasts. Plant Physiol. 92, 123–129
Gerhardt, R., Heldt, H.W. (1984) Measurement of subcellular metabolite levels in leaves by fractionation of freeze-stopped material in nonaqueous media. Plant Physiol. 75, 542–547
Gerhardt, R., Stitt, M., Heldt, H.W. (1987) Subcellular metabolite levels in spinach leaves. Plant Physiol. 83, 399–407
Gimmler, H., Schäfer, G., Kraminer, H., Heber, U. (1974) Amino acid permeability of the chloroplast envelope as measured by light scattering, volumetry and amino acid uptake. Planta 120, 47–61
Glauert, A.M. (1974) Fixation, dehydration and embedding of biological specimens. In: Practical methods in electron microscopy, vol. 3, pp. 1–201, Glauert, A.M., ed. North-Holland Publ. Comp., Amsterdam Oxford
Goerlach, J., Willms-Hoff, I. (1992) Glycine uptake into barley mesophyll vacuoles is regulated but not energized by ATP. Plant Physiol. 99, 134–139
Heldt, H.W. (1976) Metabolite transport in intact spinach chloroplasts. In: The intact chloroplast, pp. 215–234, Barber, J., ed. Elsevier, Amsterdam
Heldt, H.W., Sauer, F. (1971) The inner membrane of the chloroplast envelope as the site of specific metabolite transport. Biochim. Biophys. Acta 234, 83–91
Homeyer, U., Schulz, G., (1988) Transport of phenylalanine into vacuoles isolated from barley mesophyll protoplasts. Planta 176, 378–382
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
Kubinova, L. (1989) Stereological analysis of the leaf of barley. Acta Stereol 8, 19–26
Kushida, H. (1966) Block staining with lead acetate. J. Electron Microsc. 15, 90–91
Martinoia, E., Heck, U., Wiemken, A. (1980) Vacuoles as storage compartments for nitrate in barley leaves. Nature 289, 292–294
Martinoia, E., Thume, M., Vogt, E., Rentsch, D., Dietz, K.-J. (1991) Transport of arginine and aspartic acids into isolated barley mesophyll vacuoles. Plant Physiol. 97, 664–650
Miller, A.J., Smith, S.J. (1992) The mechanism of nitrate transport across the tonoplast of barley root cells. Planta 187, 554–557
Randall, P.J., Bouma, D. (1973) Zinc deficiency, carbonic anhydrase and photosynthesis in leaves of spinach. Plant Physiol. 52, 229–239
Riens, B., Heldt, H.W. (1992) Decrease of nitrate-reductase activity in spinach leaves during a light-dark transition. Plant Physiol. 98, 573–577
Riens, B., Lohaus, G., Heineke, D., Heldt, H.W. (1991) Amino acid and sucrose content determined in the cytosolic, chloroplastic and vacuolar compartment and in the phloem sap of spinach leaves. Plant Physiol. 97, 227–233
Schumaker, K.S., Sze, H. (1987) Decrease of pH gradients in tonoplast vesicles by NO3 − and Cl−: evidence for H+-coupled anion transport. Plant Physiol. 83, 490–496
Speer, M., Kaiser, W.M. (1991) Ion relations of symplastic and apoplastic space in leaves from Spinacia oleracea L. and Pisum sativum L. under salinity. Plant Physiol. 97, 990–997
Spurr, A.R. (1969) A low viscosity epoxy resin embedding medium for electron microscopy. J. Ultrastruct. Res. 26, 31
Steer, M.W. (1981) Understanding cell ultrastructure. Cambridge University Press, Cambridge UK
Steingröver, E., Ratering, P., Siesling, J. (1986) Daily changes in uptake, reduction and storage of nitrate in spinach grown at low light intensity. Physiol. Plant. 66, 550–556
Stitt, M., Gerhardt, R., Kürzel, B., Heldt, H.W. (1983) A role for fructose 2,6-bisphosphate in the regulation of sucrose synthesis in spinach leaves. Plant Physiol. 72, 1139–1141
Wagner, W., Keller, F., Wiemken, A. (1983) Fructan metabolism in cereals: induction in leaves and compartmentation in protoplasts and vacuoles. Z. Pflanzenphysiol. 112, 359–372
Weibel, E.R., Bolender, R.P. (1973) Stereological techniques for electon microscopic morphometry. In: Principles and techniques of electron microscopy, Biological applications, vol. 3, pp. 237–296, M.A. Hayat, ed. Van Nostrand Reinhold Company, New York
Wilson, P.D., Wheeler, K.P. (1973) Permeability of phospholipid vesicles to amino acids. Biochem. Soc. Transact. 1, 369–372
Winter, H., Lohaus, G., Heldt, H.W. (1992) Phloem transport of amino acids and sucrose depending on the corresponding metabolite levels in the leaves of barley. Plant Physiol. 99, 996–1004
Wolfe, I., Steponkus, P.L. (1983) Mechanical properties of the plasma membrane of isolated protoplasts. Plant Physiol. 71, 276–285
Woo, K.C., Flügge, U.I., Heldt, H.W. (1987) A two-translocator model for the transport of 2-oxoglutarate and glutamate in chloroplasts during ammonia assimilation in light. Plant Physiol. 84, 624–632
Yu, J., Woo, K.C., (1988) glutamine transport and the role of the glutamine translocator in chloroplasts. Plant Physiol. 88, 1048–1054
Author information
Authors and Affiliations
Additional information
The authors are grateful to Prof. M.W. Steer (Department of Botany, University College, Dublin, Ireland) for a critical appraisal of the manuscript. This work was supported by the Deutsche Forschungsgemeinschaft.
Rights and permissions
About this article
Cite this article
Winter, H., Robinson, D.G. & Heldt, H.W. Subcellular volumes and metabolite concentrations in barley leaves. Planta 191, 180–190 (1993). https://doi.org/10.1007/BF00199748
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00199748