Downy mildew disease caused by Bremia lactucae causes one of the most devastating diseases of lettuce worldwide. An obligat parasite, B.lactucae's sporulation in unnatural environments and storage is more difficult than necrotrophic pathogens and some special methods are needed to be developed. For this aim, disease samples were collected during 2016 lettuce growing seasons. The all samples were stored at -20 °C and -80 °C. Small leaf parts showing symptoms from the samples were dissected and spores were scraped by various methods. The spore suspensions were obtained using three methods (keep at room illumination+room temperature, room illumination+stored in ice, darkness+room temperature). In order to determine the suitable inoculation method, seedling, and detached leaf tests (blotter, floating, waxed slide test and water agar methods) were performed. A series of petri plates and seedlings were kept at 13±1 °C in the climate chamber and another series of assay were incubated at 18-21 °C until sporulation.

Among the inoculation methods used in the present study, the best results were obtained from waxed slide and MS+Agar methods. Both incubation temperatures were found to be appropriate and there was no significant difference between them.