Labelling of glycerolipids in the cotyledons of developing oilseeds by [1-14C]acetate and [2-3H]glycerol

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Abstract
3-sn-Phosphatidylcholine was identified as the major lipid in cotyledons from the developing seeds of soya bean [Glycine max], linseed [Linum usitatissimum] and safflower [Carthamus tinctorious] when tissue was steamed before lipid extraction. The proportion of oleate in this lipid decreased markedly and that of the poly-unsaturated C18 fatty acids increased when detached developing cotyledons were incubated for up to 3 h. Similar but less pronounced changes occurred in diacylglycerol, which had a fatty acid composition resembling that of the 3-sn-phosphatidylcholine from cotyledons of the same species. [1-14C]Acetate supplied to detached cotyledons was incorporated into the acyl moieties of mainly 3-sn-phosphatidylcholine, 1,2-diacylglycerol and triacylglycerol. Initially label was predominantly in oleate, but subsequently entered at accelerating rates the linoleoyl moieties of the above lipids in soya-bean and safflower cotyledons and the linoleoyl and linolenyl moieties of these lipids in linseed cotyledons. In pulse-chase experiments label was rapidly lost from the oleate of 3-sn-phosphatidylcholine and accumulated in the linoleoyl and linolenoyl moieties of this phospholipid and of the di- and tri-acylglycerols. [2-3H]Glycerol was incorporated into the glycerol moieties of 3-sn-phosphatidylcholine and di- and tri-acylglycerols of developing linseed and soya-bean cotyledons. The label entered the phospholipid and diacylglycerol at rates essentially linear with time from the moment the substrate was supplied, and entered the triacylglycerol at an accelerating rate. With linseed cotyledons the labeled glycerol was incorporated initially into species of 3-sn-phosphatidylcholine and diacylglycerol that contained oleate, but accumulated with time in more highly unsaturated species. In pulse-chase experiments with linseed cotyledons, label was lost from both 3-sn-phosphatidylcholine and diacylglycerol, preferentially from the dioleoyl species, and accumulated in triacylglycerol, mainly in species containing 2 molecules of linolenate. The results suggest a rapid turnover of 3-sn-phosphatidylcholine during triacylglycerol accumulation in developing oilseeds, and are consistent with the operation of a biosynthetic route whereby oleate initially esterified to the phospholipid is first desaturated, then polyunsaturated fatty acids are transferred to triacylglycerol via diacylglycerol. The possible role of oleoyl phosphatidylcholine as a substrate for oleate desaturation is discussed.