Summary
The effect of freezing, desiccation and various electrolytes on photophosphorylation, electron transport and some enzyme reactions of isolated spinach chloroplasts has been investigated. Freezing of broken chloroplasts took place at-25°C for 3 hrs; desiccation was performed at +2°C in vacuo over CaCl2 for 3 hrs. The influence of various electrolytes during freezing or drying or during incubation of thylakoids or stroma enzymes for 3 hrs at +2°C in electrolyte solutions was determined. After treatment, the activities of a number of enzymes and enzyme systems were measured under normal conditions, e. g. in the absence of elevated electrolyte levels in a reaction medium which contained only the substrates and cofactors which are necessary for the respective enzyme reactions.
Only photophosphorylation and electron transport were affected by freezing, desiccation and high concentrations of electrolytes; various soluble enzymes investigated here were not inactivated under the same conditions. In general, mild dehydration and lower concentrations of electrolytes resulted in an irreversible inactivation of ATP synthesis but did not impair ferricyanide reduction. With increasing dehydration or at higher concentrations of electrolytes the Hill reaction was also inhibited. In a certain range of dehydration and electrolyte concentration uncoupling of photophosphorylation from electron transport took place. Sugar protects the sensitive structures against the deleterious effect of both dehydration and high concentration of electrolytes.
Various electrolytes affected thylakoid membranes differently. Inactivation of the membranes increased with increasing ion radius and decreasing hydration envelope of univalent or divalent cations. Divalent cations were more destructive than univalent cations. Anions did not follow these rules. Within a group of similar anions (halides or organic anions) effectivity decreased with increasing hydration envelope. On a molar basis, polyvalent anions were less effective than univalent anions. Inactivation by anions followed Hofmeister's series in seversed order. However, exceptions were observed and it appears that various ions affect the membrane in a specific manner.
Inactivation of photophosphorylation and electron transport due to freezing or desiccation is identical to that due to high concentrations of electrolytes. This suggests that during dehydration due to freezing or drying the concentration of electrolytes in the remaining solution is responsible for the inactivation of the sensitive membranes.
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Literatur
Arnon, D. I.: Copper enzymes in isolated chloroplasts. Polyphenoloxidase in Beta vulgaris. Plant Physiol. 24, 1–15 (1949).
Asada, K., R. Deura, and Z. Kasai: Effect of sulfate ions on photophosphorylation by spinach chloroplasts. Plant and Cell Physiol. 9, 143–146 (1968).
Asahina, É: Freezing injury in egg cells of the sea urchin. In: É. Asahina, Cellular injury and resistance in freezing organisms. Proc. Intern. Conf. Low Temp. Sci., Sapporo 1966, vol. II, p. 211–229 (1967).
Baldry, C. W., W. Cockburn, and D. A. Walker: Inhibition, by sulphate, of the oxygen evolution associated with photosynthetic carbon assimilation. Biochim. biophys. Acta (Amst.) 153, 467–483 (1968).
Bergmeyer, H. U.: Methoden der enzymatischen Analyse. Weinheim: Verlag Chemie 1962.
Duane, W. C., and D. W. Krogmann: Chloroplast storage with retention of photosynthetic activities. Biochim. biophys. Acta (Amst.) 71, 195–196 (1963).
Heber, U.: Ursachen der Frostresistenz bei Winterweizen. III. Mitt. Die Bedeutung von Proteinen für die Frostresistenz. Planta (Berl.) 54, 34–67 (1959).
—: Freezing injury in relation to loss of enzyme activities and protection against freezing. Cryobiology 5, 188–201 (1968).
Heber, U., and R. Ernst: A biochemical approach to the problem of frost injury and frost hardiness. In: É. Asahina, Cellular injury and resistance in freezing organisms. Proc. Intern. Conf. Low Temp. Sci., Sapporo 1966, vol. II, p. 63–77 (1967).
—, and K. A. Santarius: Loss of adenosine triphosphate synthesis caused by freezing and its relationship to frost hardiness problems. Plant Physiol. 39, 712–719 (1964).
Hofmeister, F.: Zur Lehre von der Wirkung der Salze. Naunyn-Schmiedebergs Arch. exp. Path. Pharmak. 24, 247–260 (1888).
Huggins, C. E.: Preservation of organized tissues by freezing. Fed. Proc., Suppl. 24/2, S190-S195 (1965).
Izawa, S., and N. E. Good: Effect of salts and electron transport on the conformation of isolated chloroplasts. I. Light-scattering and volume changes. Plant Physiol. 41, 533–543 (1966).
Jagendorf, A. T., and M. Avron: Cofactors and rates of photosynthetic phosphorylation by spinach chloroplasts. J. biol. Chem. 231, 277–290 (1958).
—, and M. Smith: Uncoupling phosphorylation in spinach chloroplasts by absence of cations. Plant Physiol. 37, 135–141 (1962).
Lovelock, J. E.: The haemolysis of human red blood-cells by freezing and thawing. Biochim. biophys. Acta (Amst.) 10, 414–426 (1953a).
—: The mechanism of the protective action of glycerol against haemolysis by freezing and thawing. Biochim. biophys. Acta (Amst.) 11, 28–36 (1953b).
—: The denaturation of lipid-protein complexes as a cause of damage by freezing. Proc. roy. Soc. B 147, 427–433 (1957).
Luck, W.: Zur Assoziation des Wassers. II. Ber. Bunsenges. physik. Chemie 69, 69–76 (1965).
Meryman, H. T.: Modified model for the mechanism of freezing injury in erythrocytes. Nature (Lond.) 218, 333–336 (1968).
Nishida, K., and K. Koshi: On the volume-changes of isolated spinach chloroplasts caused by electrolytes and sugars. Physiol. Plant. 846–854 (1964).
Partmann, W.: Physikalische und biochemische Veränderungen von Lebensmitteln beim Gefrieren und bei der Gefrierlagerung. In: Probleme der Ernährung durch Gefrierkost. Wiss. Veröff. Dtsch. Ges. für Ernährung, Bd. 12, S. 12–32. Darmstadt: D. Steinkopff 1964.
Santarius, K. A.: Protective mechanisms in frost-hardy plants. In: J. Hawthorne and E. J. Rolfe, Low temperature biology of foodstuffs, p. 135–151. Oxford: Pergamon Press 1968.
—, u. R. Ernst: Das Verhalten von Hill-Reaktion und Photophosphorylierung isolierter Chloroplasten in Abhängigkeit vom Wassergehalt. I. Wasserentzug mittels konzentrierter Lösungen. Planta (Berl.) 73, 91–108 (1967).
—, u. U. Heber: Das Verhalten von Hill-Reaktion und Photophosphorylierung isolierter Chloroplasten in Abhängigkeit vom Wassergehalt. II. Wasserentzug über CaCl2. Planta (Berl.) 73, 109–137 (1967).
Stein, W. D.: The movement of molecules across cell membranes, p. 91. New York and London: Academic Press 1967.
Ullrich, H., u. U. Heber: Über das Denaturieren pflanzlicher Eiweiße durch Ausfrieren und seine Verhinderung. Planta (Berl.) 51, 399–413 (1958).
Vambutas, V. K., and E. Racker: Partial resolution of the enzymes catalyzing photophosphorylation. I. Stimulation of photophosphorylation by a preparation of a latent, Ca++-dependent adenosine triphosphatase from chloroplasts. J. biol. Chem. 240, 2660–2667 (1965).
Webb, S. J.: Bound water in biological integrity, p. 32. Springfield (Ill.): Ch. C. Thomas 1965.
Williams, R. J., and H. T. Meryman: A calorimetric method for measuring ice in frozen solutions. Cryobiology 1, 317–323 (1965).
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Santarius, K.A. Der Einfluß von Elektrolyten auf Chloroplasten beim Gefrieren und Trocknen. Planta 89, 23–46 (1969). https://doi.org/10.1007/BF00386494
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DOI: https://doi.org/10.1007/BF00386494