Abstract:Calli were induced and suspension cell lines were established from cotyledones of ramie (Boehmeria nivea). Protoplasts (2 × 10 6/g fr. wt) were isolated from suspension cell cultures in enzyme mixture solution containing 4. 5 % cellulase Onozuka R-10 and 0. 8 % Macerozyme R-10, 0.8 % hemicellulase. When cultivated on KM8p medium containing 2, 4-D 0.5 mg/L, KT 0.5 mg/L with alginate embedding method, they grew vigorously and produced microcalli within fifty days. After subcultured, the protoplast-derived ~alli produced shoots and roots on different differentiation media, then complete plants were formed. Protoplasts from cotyledones divided only several times.