Evidence for an essential role of carotenoids in the assembly of an active photosystem II

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Abstract
Dark-grown cells of mutant C-6D of the green alga Scenedesmus obliquus exhibit a high activity of photosystem I (PSI) but lack activity of photosystem II (PSII). These cells contain only the pigment-protein complex CPI, representing the reaction-center of PSI. Only chlorophyll a and precursors of carotenoids (lycopene, neurosporene, ξ-carotene, β-zeacarotene) could be detected in dark-grown cells by analysis using high-performance liquid chromatography. Activity of PSII and the corresponding pigment-protein complex, CPa, develop immediately upon transfer to light. Light-harvesting complexes and higher molecular forms of PSI are synthesized only in the later stages of light-induced chloroplast differentiation. During illumination the amounts of carotenoid precursors decrease and carotenes, xanthophylls and chlorophylls a and b are formed. β-Carotene and lutein are synthesized without a lag-phase. Their kinetics are similar to those of CPa formation and development of PSII activity. In contrast, all other xanthophylls are synthesized only after a lag-phase of about 30 min. Inhibition of the transformation of precursors into carotenoids by nicotine prevents the light-inducible development of PSII activity and CPa formation. During illumination under anaerobic conditions no xanthophylls are synthesized but high amounts of α- and β-carotene accumulate. Such cells exhibit no PSII activity and show only traces of CPa. After subsequent transfer to aerobic conditions the xanthophylls are synthesized and simultaneously active PSII units are formed. The results prove that carotenoids are essential components for the assembly of active PSII units. Strong evidence is given that lutein is the absolute necessary prerequisite for this process. Whether β-carotene is also an absolute necessary prerequisite for a functioning PSII unit cannot be deduced from our experiments.