Hydrilla is one of the most serious aquatic weed problems in the United States, and fluridone is the United States Environment Protection Agency (USEPA)-approved herbicide that provides relatively long-term systemic control. Mature (6-wk-old) and young (freshly planted from 10-cm apical shoot apices) hydrilla were grown in 540 L fiberglass vaults under short- (natural 8 to 10 hr light/14 to 16 hr dark photoperiod) or long- (artificially extended 16 h light/8 h dark photoperiod) day greenhouse conditions. Fluridone treatments of 0, 1, 5, and 10 µg L−1 were applied after 2 wk and maintained within each population and photoperiodic regime throughout the study. Short days promoted subterranean turion formation, but this effect was reduced by long days and 5 and 10 µg L−1 fluridone. Fluridone caused a reduction in chlorophyll and carotenoid levels but the effect on anthocyanin content was variable. Short days caused elevated anthocyanin, and this effect was diminished by fluridone. Fluridone reduced the abscisic acid content of mature apical stems and was higher under short days in younger plants. These studies provide further evidence that fluridone can be used as a fall herbicide application to reduce turion production.