The biosynthesis of phospholipid and the composition of fatty acid were analyzed in mitochondria isolated from Chlorella ellipsoidea treated with carbon sources (glucose, sucrose, raffinose) during the culture. The growth of Chlorella and total lipid contents in mitochondria treated with various carbon sources was increased to compare with the control. When Chlorella mitochondria was treated with various carbon sources, four kinds of phospholipid were increased predominantly. The major fatty acids utilized for the biosynthesis of the phospholipid were analyzed linoleic acid (average 25.18%) and stearic acid (average 10.52%) in the control. But, it was shown that the major fatty acids in Chlorella mitochondria treated with glucose were stearic acid (average 30.93%), palmitic acid (average 17.47%) and stearic acid (average 20.31%), linoleic acid (average 16.68%) in sucrose treatment and oleic acid (average 17.17%), palmitic acid (average 15.64%) in raffinose treatment.