Taking the leaves of Salanum sisymbriifolium and S. torvum as protoplast fusion materials，this paper studied the enzymatic hydrolysate concentration，enzymatic hydrolysis time and electrofusion parameters．The results showed that the optimal concentration was 0.3%（m/V）cellulase R-10+0.1%（m/V）macerozyme R-10．Under this condition，the maximum number of single cells could be observed by microscope．The optimal enzymolysis time were 14 hours for S. sisymbriifolium and 13 hours for S. torvum，which the protoplasts yield was 14.06×10⁶ protoplasts per gram and 15.92×10⁶ protoplasts per gram，respectively．The optimum electric fusion parameters of field strength was 80 V·cm^-1，for 10 s action time．The D.C. square pulses was 1 250 V·cm^-1 for 45 μs．Just once the best fusion effect could be observed by microscope．