Flue gas desulfurization (FGD) gypsum, a coal combustion by-product, can be used to decrease water-extractable soil P, thereby lowering the potential for P export to surface waters. This decrease results from a conversion of loosely bound inorganic P (IP) which is readily desorbable to water, to less soluble Al- and Fe-bound IP and, to a lesser extent, calcium-bound IP pools. Although this conversion has little effect on predictors of plant-available soil P (e.g., Mehlich-3 P), little is known about the plant uptake of P over several growth cycles after high P soils are amended with FGD. In a greenhouse experiment, we measured P uptake by ryegrass (Lolium perenne) using a modified Stanford–Dement procedure (three growth cycles), and the extent to which IP was being removed from each soil IP fraction (Hedley fractionation), for three soils treated with FGD gypsum (equivalent to 22.4 Mg ha−1). Treatment with FGD decreased water extractable soil P 38 to 57%, but had little effect on Mehlich-3 soil P. During the first growth cycle, the shift from resin IP to less available Al, Fe, and Ca IP remained stable. Repeated growth cycles of ryegrass removed resin IP and thus, had a continued effect on lowering water-extractable P. After three growth cycles and harvests, ryegrass dry-matter production was not affected by FGD treatment (P > 0.05), although cumulative P uptake (20%) and P concentration of ryegrass tops (25%) were greater in FGD treated than untreated soils. Our results confirm that treatment of high P soils with FGD gypsum decreases water-extractable P by conversion to soil IP fractions that are stable with time, does not decrease plant production, and suggests that the potential for P export in surface runoff may be reduced for several years.