A Parallel Study of Pigment Bleaching and Cytochrome Breakdown during Aging of Thylakoid Membranes

Aging of freshly isolated thylakoid membranes from spinach leaves(Spinacia oleracea L.) leads to dramatic alterations in boththe cytochrome (b_{559 (HP)} and f) composition and pigment (chlorophyllsa and b and ß-carotene) content. These changes occurat a faster rate under anaerobic conditions or after heatingthylakoid membranes, and in light as well as in darkness. Inaddition, when thylakoid membranes are heated at 78°C for8 min, or incubated in the presence of an emulsion of linoleicacid, a huge decrease in both cytochrome (particularly cyt.b_{559 (HP)}) and pigment contents occur. Whatever the experimentalconditions, cytochrome b_{559 (HP)} destruction occurs as soonthe aging process starts. Conversely, pigment bleaching is detectableafter an initial lag phase of about 60–70 min. Then, thetwo processes (cytochrome breakdown and bleaching of pigments)appear to take place in parallel. The addition of salicylhydroxamicacid or 8-hydroxy-quinoline, two radical scavenger components,to the aging medium strongly reduces the rate and extent ofcytochrome breakdown and pigment bleaching. On the basis of these results, a tentative scheme accountingfor the bleaching of pigments and the breakdown of cytochromesduring aging in vitro of thylakoid membranes is proposed. Itis suggested that these changes are mediated via a non-enzymaticmechanism in which free radicals could be implicated. The possiblerole of free radicals inducing ultrastructural changes at thelevel of chloroplast membranes in senescent leaves is also considered. (Received October 11, 1985; Accepted January 24, 1986)