Protoplasts were washed twice further and resuspended in one of 2 culture media; KMP8 medium [4] (0.2 mg/l 2,4-D , 0.5 mg/l zeatin, 1 mg/l NAA, 750 mg/l casein hydrolysate, 250 mg/l sucrose, 100 mg/l glucose or L1D2 medium [3] at densities of 1-6x105 protoplasts/ml. Protoplast viability and assessment of the presence of cell wall were performed as mentioned by Power et al.