Membrane lipid metabolism in Chlamydomonas reinhardtii 137+ and y-1: II. Cytochemical localization of acyltransferase activities.

We have cytochemically localized the acyltransferase activities in the alga Chlamydomonas reinhardtii. Glycerol 3-phosphate (G3P) acyltransferase and lysophosphatidate (LPA) acyltransferase activities were cytochemically assayed utilizing biochemically optimized reaction conditions (Jelsema et al., 1982). The cytochemical assays clearly demonstrate that in the wild-type cells (137+) and the y-1 mutant, both enzymes were present in the photosynthetic membranes, envelope, and pyrenoid tubules of the chloroplast. Activity was also localized to the Golgi apparatus and its associated vesicles. Additionally, LPA acyltransferase activity was found associated with the outer mitochondrial membranes. The cytochemical data from this study confirm the biochemical data obtained using purified chloroplast membranes (Jelsema et al., 1982) and establishes the presence of these glycerolipid-synthesizing enzymes in the photosynthetic membranes of the chloroplast. These findings support the earlier reports of the presence and activity of lipid-synthesizing enzymes in the chloroplast, and also is in agreement with the postulated role of the pyrenoid tubules in the synthesis of the thylakoid membranes. This report differs in that these lipid-synthesizing enzymes have been localized to the chloroplast photosynthetic membranes themselves as well as exhibiting significant levels of activity for both enzymes in the Golgi apparatus. During light-induced chloroplast biogenesis in the yellow mutant of C. reinhardtii (y-1), the photosynthetic membranes appear to be the primary site of acyltransferase activity, suggesting that in situ synthesis of the membrane lipids during this period of rapid membrane formation is the primary mode for the synthesis of the thylakoid lipids. That these intrinsic thylakoid enzyme activities are not exclusively a function of the growth phase of the cell, but are found in mature chloroplast of the 137+ cells as well, further supports the conclusion that the photosynthetic membranes of the chloroplast have the capacity for synthesis of their own membrane lipids.