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  • 1
    ISSN: 1432-2048
    Keywords: Amino acid (cryoprotective media) ; Chlorophyll fluorescence ; Freezing damage ; Photosynthesis (electron transport, photophos-phorylation) ; Spinacia (freezing damage) ; Thyllakoid, isolated
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Chloroplast thylakoid membranes were isolated from leaves of unhardened and cold-acclimated spinach (Spinacia oleracea L.). For freezethaw treatment, the membranes were suspended in complex media composed to simulate the solute concentrations in the chloroplast stroma in the unhardened and hardened states of the leaves. In particular, high concentrations of amino acids were applied for simulating the hardened state. After frost treatment, photosynthetic activities and chlorophyll fluorescence parameters of the thylakoids were tested to determine the degree of freezing damage. The results revealed a pattern of freezing injury similar to that observed upon frost treatment of thylakoids in situ. A major manifestation of damage was the inhibition of photosynthetic electron transport. Uncoupling of photophosphorylation, which is the dominating effect of freezing of thylakoids suspended in binary solutions (e.g., containing one sugar and one inorganic salt), was also visible but less pronounced in the complex media. Thylakoids obtained from cold-acclimated leaves did not exhibit an increased frost tolerance in vitro, as compared with thylakoids from unhardened plants. The results, furthermore, indicated a strong protective effect of free amino acids at the concentrations and composition found in chloroplasts of hardened leaves. The presence of inorganic salts in the complex media slightly stabilized rather than damaged the membranes during freezing. It is concluded that inactivation of thylakoids in situ may be understood as the destabilizing action of the combined solutes surrounding the thylakoids, occurring when solute concentration is raised due to freezing of water.
    Type of Medium: Electronic Resource
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  • 2
    Electronic Resource
    Electronic Resource
    Springer
    Planta 138 (1978), S. 73-78 
    ISSN: 1432-2048
    Keywords: Chlorophyll ; Chloroplasts ; Fluorescence ; Magnesium transport ; Spinacia ; Uncouplers
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Uncoupling concentrations (about 1 μmol l-1) of desaspidin or carbonyl cyanide-4-trifluoromethoxyphenyl hydrazone reverse the slow light-induced, Mg2+-dependent quenching of fluorescence of chlorophyll a in isolated (intact and broken) spinach chloroplasts. Likewise, uncoupling inhibits the light-induced increase of the Mg2+ concentration in the stroma of intact chloroplasts, as determined with Eriochrome Blue SE. Addition of higher amounts of the uncouplers to the chloroplasts leads to a slow, light-dependent fluorescence lowering which appears to be promoted by high light intensities and is not reversed in the dark. The reversal of the fluorescence quenching by uncoupling is interpreted to reflect exchange of protons for Mg2+ ions at negative sites of the inner thylakoid face, caused by the collapse of the proton gradient across the membrane. The secondary fluorescence lowering caused by high levels of the uncouplers and high light intensities is suggested to be related to an inhibition of non-cyclic photosynthetic electron transport.
    Type of Medium: Electronic Resource
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