Abstract
Protoplasts were successfully isolated from mungbean leaves (Vigna radiata L. Wilczek) using an enzyme solution containing 4% Meicelase P-1, 1% Macerozyme R-10, 1% potassium dextran sulphate and 0.5 M mannitol. Approximately 4.1×107 protoplasts per gram fresh weight were released after digestion with an enzyme for 4 hours in the dark on a shaker. The cultured protoplasts started to divide 5 days after culture followed by sustained division, and a microcolony was produced on day 15 of culture. Microcalli appeared on the medium surface a month after culture and were large enough for transfer to a callus induction medium after 1.5 month of culture. Having transferred to the regeneration media, green and compact calli were developed in MS medium supplemented with NAA and BA. Globular-like calli appeared in the MS basal medium or in the mixture of MS salts and B5 vitamin supplemented with 2, 4-D. Root formation then occurred from this calli.
