Changes in the carotenoid composition of chloroplast membranes from Chlamydomonas reinhardtii double mutants with alterations of various sites in photosystem II

The variable fluorescence and polypeptide and carotenoid compositions of the chlorophyll b-deficient mutant C-48 of the unicellular green alga Chlamydomonas reinhardtii and its double mutants without chlorophyll b and with inactive photosystem II were compared with those of the wild-type algal cells. Studying variable fluorescence demonstrated the alterations at the donor side (AC-121), the acceptor side (AC-234) or immediately in the photosystem II reaction centre (AC-184, AC-864). Gel electrophoresis showed that the absence of chlorophyll b in all mutants was due to the lack of 26, 28 and 31 kDa polypeptides in the light-harvesting chlorophyll a/b-protein complex II (LHC II). As a result of the second mutation, the chlorophyll a-protein complex of photosystem II did not form in chloroplast membranes. The disassembly of this complex in the mutants AC-121, AC-234 and AC-864 was related to the deficiency of both polypeptides of the reaction centre (30 and 32 kDa) and polypeptides of the water-oxidizing system (18, 23 and 34 kDa). Besides the loss of these polypeptides, the contents of polypeptides with molecular masses of 47 and 51 kDa decreased in the double mutant AC-184. Substantial changes were revealed in the carotenoid composition of the double mutants. We observed the considerable accumulation of carotenes that accompanied alterations in the donor (mutant AC-121) or acceptor (mutant AC-234) sides of PS II. In the first case, beta-carotene predominantly accumulated (87%); in the second case, it was alpha-carotene (52%). Alterations in the PS II reaction centre (mutants AC-184, AC-864) caused accumulation of xanthophylls, mainly lutein (38-41%). We suppose that alterations in different parts of the PS II chloroplast membrane lead to substantial changes in the carotenoid composition.