This study investigated the levels of c-linolenic (GLA, 18:3n-6) and stearidonic acid (SDA, in various parts of the borage plant (Borago officinalis L.) to elucidate D6-desaturase activity. Lipid class and fatty acid (FA) composition during germination of the seeds and FA composition of various borage parts were determined as well as FA compositions of neutral lipids (NL), polar lipids (PL), glycolipids (GL) and monogalactosyl diacylglycerols (MGDG) of borage leaves. When seeds were germinated for 12 days in the dark then exposed to light for 8 h, an overall decrease in oil content was seen with a significant increase in PL from 0.2 to 39.1%. An increase in SDA (from 0.2 to 0.6 g/100 g oil) indicated minor FA synthesis during germination and therefore, D6-desaturase activity with the most marked increase after light exposure. The FA compositions of developing and mature seeds were similar, suggesting oil/FA synthesis takes place at the initial stage of seed development. Among all the borage parts, the leaves had the highest amounts of a-linolenic acid (ALA, 18:3n-3) (36.2%), indicating D15-desaturase activity, and SDA (25.2%), indicating D6-desaturase activity. In leaves, the GL and especially, MGDG fractions had the highest amounts of SDA (31.8 and 39.8%, respectively), indicating that D6-desaturase is most active in chloroplasts. Leaves and developing seeds appear to be the major sources of D6-desaturase in borage, associated with different organelles in the different tissues.