Nitric oxide (NO) production by Botrytis cinerea and the effect of externally supplied NO were studied during saprophytic growth and plant infection. Fluorescence analysis with 4,5-diaminofluorescein diacetate and electrochemical studies were conducted in vitro between 4 and 20 h of incubation and in planta between 15 and 75 h post-inoculation. The production of NO by B. cinerea in vitro was detected inside the germinating spores and mycelium and in the surrounding medium. In planta production of NO showed a large variation that was dependent on the host plant and developmental stage of the infection. The induced production of NO was detected from 16 h of in vitro incubation in response to externally added NO. The production of NO by B. cinerea is probably modulated to promote fungal colonization of the plant tissue. The production of NO which diffuses outside the fungal cells and the induction of NO production by exogenous NO open up the possibility of NO cross-talk between the fungus and the plant. Finally, the existence of an NO concentration threshold is proposed, which may increase or reduce the plant defence against necrotrophic fungal pathogens.