Regulation of diacylglycerol acyltransferase in developing seeds of castor.

We have previously reported the cloning of castor diacylglycerol acyltransferase (RcDGAT) based on its homology to other plant type 1 diacylglycerol acyltransferases (DGATs). To elucidate the physiological role of the RcDGAT, we have investigated the regulation of RcDGAT expression in developing seeds of castor. The RcDGAT transcript appeared at 12 d after pollination (DAP), reached the highest level at 26 DAP, and declined rapidly after that. However, the RcDGAT protein started to accumulate at 26 DAP, reached its peak at 47 DAP, then remained at this high level until 54 DAP. The significant difference between the expression of mRNA and protein indicates that gene expression of RcDGAT in maturing castor seeds is controlled at the posttranscriptional level. We found that DGAT activity measured in microsomal membranes isolated from seed at different stages of development was parallel to RcDGAT protein level, suggesting DGAT activity is mainly a function of the level of RcDGAT protein. We monitored the triacylglycerol (TG) composition and content during seed development. Compared with the overall rate of TG accumulation, DGAT activity appeared coincidently with the onset of lipid accumulation at 26 DAP; the highest DGAT activity occurred during the rapid phase of lipid accumulation at 40 DAP; and a decline in DGAT activity coincided with a decline in the accumulation rate of TG after 40 DAP. The ricinoleate-containing TG content was very low (only about 7%) in oil extracted from seeds before 19 DAP; however, it increased up to about 77% of the oil at 26 DAP. The relative amount of triricinolein in oil at 26 DAP was 53 times higher than that at 19 DAP, and it was about 76% of the amount present in oil from mature castor seeds. The close correlation between profiles of RcDGAT activity and oil accumulation confirms the role of RcDGAT in castor oil biosynthesis.