The method of isolating and transiently transforming oil palm leaf protoplasts was established. Protoplasts were isolated from leaves of oil palm with 30 g/L cellulase and 8 g/L macerozyme for 3.5 h,and collected by centrifugating at 2 000 r/min for 5 min at 4℃. The dual plasmid transient coexpression system was optimized with green fluorescent protein (GFP) and red fluorescent protein (RFP). The best transformation efficiency was the obtained when the plasmids were mixed with oil palm protoplasts with the mass ratio of 8∶1,heat shocked at 45℃ for 20 min after placed on ice for 30 min,and incubated at room temperature under the dark condition for 30 min with equal amount of PEG/Mg2+ (200 g/L PEG 4000 and 100 g/L MgCl2). It will provide a novel method for further identifying the function of the genes from oil palm.