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Metabolism and Enzymology

Pathway for the Synthesis of Triacylglycerols from Monogalactosyldiacylglycerols in Ozone-Fumigated Spinach Leaves

Division of Environmental Biology, The National Institute for Environmental Studies, Onogawa, Tsukuba, Ibaraki 305, Japan, Department of Biology, University of Tokyo, Komaba, Meguro-ku, Tokyo 153, Japan

When the upper leaf surface of spinach (Spinacia oleracea L.) plants was treated with [1-¹⁴C]acetate and grown for 2 days, ¹⁴C was effectively incorporated into acyl moieties of leaf lipids in ratios approximately their composition by mass. Fumigation of the plants with ozone (0.5 microliter per liter) caused a redistribution of ¹⁴C among lipid classes, i.e. a marked increase of ¹⁴C content in triacylglycerol (TG) and 1,2-diacylglycerol (1,2-DG) and a decrease of label in monogalactosyldiacylglycerol (MGDG) without affecting ¹⁴C distribution in leaf fatty acids. Label in both TG and 1,2-DG was found predominantly in their polyene molecular species. Since MGDG consists of similar polyene molecular species, the results indicate the synthesis of TG from MGDG via 1,2-DG. Label was also accumulated in tri- and tetragalactosyldiacylglycerol, products of galactolipid:galactolipid galactosyltransferase (GGGT). Moreover, there was a close relation between increases in the amounts of TG and the oligogalactolipids in ozonetreated leaves. These results indicate that MGDG was converted to 1,2-DG by GGGT and then to TG. In intact chloroplasts isolated from ozone-treated leaves, there was an enhanced production of free fatty acid (FFA), which was diminished by the addition of coenzyme A (CoA) and ATP, indicating that ozone stimulated the hydrolysis of MGDG to liberate FFA, which was in turn converted to acyl-CoA. The final step of TG synthesis, acylation of 1,2-DG with acyl-CoA, was confirmed by feeding with [1-¹⁴C]linolenic acid in leaf discs excised from ozone-fumigated leaves; ¹⁴C was effectively incorporated into TG but not into 1,2-DG. These results demonstrate the synthesis of TG from 1,2-DG and FFA which were liberated from MGDG in ozone-fumigated spinach leaves.

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