Figure 6.

Developing silique microsome GPAT activity. GPAT assays were done with microsomes isolated from whole developing siliques 5 to 7 and 9 to 11 d after flowering (DAF) from Col-0 and GPAT9 amiRNA knockdown lines 2 and 12. The assay utilized 3.55 nmol of [¹⁴C]G3P and 25 nmol of palmitoyl-CoA for 15 min at 24°C. A, Phosphor image of a thin-layer chromatography (TLC) plate indicating that [¹⁴C]LPA and [¹⁴C]PA are the major products of whole-silique microsome GPAT assays utilizing palmitoyl-CoA as an acyl donor. B and C, Quantification of total products from GPAT activity in whole-silique microsomes. Two representative GPAT assays from a total of six GPAT assays were performed with different batches of plants grown at different times. Total GPAT activity was dependent on growth conditions and stage of development. However, with each batch of plants or stage of development, the GPAT activity of amiRNA lines was always less than that of the wild-type control. An additional two representative GPAT assays are shown in Supplemental Figure S11. MAG, Monoacylglycerol.