The effect of photoperiod on ent-kaurene biosynthesis was determined in the long-day (LD) plants spinach (Spinacia oleracea 1.) and Agrostemma githago L . Further metabolism of ent-kaurene was blocked by application of the growth retardant tetcyclacis, and ent-kaurene accumulation was measured by isotopic dilution using gas chromatography-selected ion monitoring (CC-SIM) (E.Physiol 96: 1099-1104). In spinach, the rate of ent-kaurene accumulation in shoots grown under LD conditions was 3 times higher than in shoots grown under short-day (SD) conditions. ent-Kaurene also accumulated in fully expanded leaves, but at a lower rate than in shoots (15 and 55 pmol g-' dry weight h-', respectively). In Agrostemma, ent-kaurene accumulated at a rate 2.5 times higher in plants grown under LD conditions than in those grown under SD conditions. In spinach, enhanced ent-kaurene accumulation was detectable after 1 long day, and with exposure to additional long days, the rate of ent-kaurene accumulation increased further.Conversely, when plants were exposed to LD conditions and then returned to SD conditions, the rate of ent-kaurene accumulation decreased. Following tetcyclacis application, ent-kaurene accumulation was observed in all parts of spinach that were analyzed, but there were large quantitative differences between organs of different ages. As the leaves matured, enf-kaurene biosynthesis declined. Petioles accumulated more ent-kaurene than the corresponding leaf blades. It is concluded that stimulation of entkaurene biosynthesis by LD conditions leads to a higher rate of gibberellin biosynthesis, which is essential for stem elongation in rosette plants.There is considerable evidence that the rate of GA biosynthesis in LD rosette plants is higher under LD conditions, when stem elongation and flower formation take place, than during vegetative growth in the rosette stage under SD conditions. For example, the GA content increased severalfold after the transfer of Silene plants from SD to LD conditions