The glycerolipid composition of pea (Pisum sativum L.) root plastids and their capacity to synthesize glycerolipids from [UL-'4C]glycerol-3-phosphate were determined. Pea root plastids primarily consist of monogalactosyldiacylglycerol, triacylglycerol, phosphatidylcholine, digalactosyldiacylglycerol, and diacylglycerol. Maximum rates of total glycerolipid biosynthesis were obtained in the presence of 2.4 mM glycerol-3-phosphate, 15 mM KHCO,, 0.2 mM sodium-acetate, 0.5 mM each of NADH and NADPH, 0.05 mM coenzyme A, 2 mM MgCI,, 1 mM ATP, 0.1 M Bis-Tris propane (pH 7.5), and 0.31 M sorbitol. Clycerolipid biosynthesis was completely dependent on exogenously supplied ATP, coenzyme A, and a divalent cation, whereas the remaining cofactors improved their activity from 1.3-to 2.4-fold. Radioactivity from glycerol-3-phosphate was recovered predominantly in phosphatidic acid, phosphatidylglycerol, diacylglycerol, and triacylglycerol with lesser amounts in phosphatidylcholine and monoacylglycerol. The proportions of the various radiolabeled lipids that accumulated were dependent on the pH and the concentration of ATP and glycerol-3-phosphate. The data presented indicate that pea root plastids can synthesize almost all of their component glycerolipids and that glycerolipid biosynthesis is tightly coupled to de novo fatty acid biosynthesis. pH and the availability of ATP may have important roles in the regulation of lipid biosynthesis at the levels of phosphatidic acid phosphatase and in the reactions that are involved in phosphatidylglycerol and triacylglycerol biosynthesis.Plastids play a central role in plant lipid metabolism. As the only subcellular site for de novo fatty acid biosynthesis, plastids provide the entire plant cell with fatty acids for the synthesis of membrane and storage lipids (Stumpf, 1984;Ohlrogge et al., 1993). Plastids are also actively engaged in their own membrane glycerolipid biosynthesis. Most of the information about the glycerolipid biosynthetic capacities of plastids is derived from studies that have been conducted with photosynthetically active chloroplasts (Roughan and Slack, 1982). Chloroplasts contain and synthesize approximately 75% of the total leaf lipid and are thus the most important sites of glycerolipid biosynthesis in leaves (Harwood, 1980 Slack, 1982). In contrast, little information is available about lipid metabolism in nonphotosynthetic plastids. The available information suggests that nonphotosynthetic plastids may be similar to chloroplasts in terms of their autonomy for fatty acid and glycerolipid biosynthesis and the types of lipids comprising their membranes. However, the actual lipid composition of each plastid varies from one type to another, depending on the physiology or function of the tissues from which the plastids are isolated as well as differences in the regulation of lipid metabolism in these plastids (Sparace and KleppingerSparace, 1993).A few studies have described glycerolipid biosynthesis in nonphotosynthetic plastids from such tissues as cauliflower buds an...