Suspension culture of Taxus chinensis was used as a model plant cell system to study the effects of pulsed electric field (PEF) on cell growth and the biosynthesis of secondary metabolites. Different exposure time of PEF had different effects on Taxus chinensis cells in different cell growth phases. Significant intracellular enhanced accumulation of taxuyunnanine C (Tc, a bioactive secondary metabolite) was observed by exposing the cells in early exponential growth phase to a 30 min PEF. The Tc content increased by 30% after exposure to PEF, without any loss of dry cell weight compared with that of the control. Moreover, combination of PEF with sucrose feeding showed a great synergistic accumulative effect on secondary metabolite production.