Protoplast Culture and Plant Regeneration from the Suspension Cells of Gynostemma pentaphyllum (Thumb) Mak.
绞股蓝悬浮细胞的原生质体再生植株

While the embryogenic callus induced from young stems of G. pentaphyllum on MS medium with Img/L 2,4-D and 0. 2mg/L KT were cultured in MS liquid medium on a rotary shaker, stable embryogenic suspensions were establidshed. A high yield of protoplasts could be released from the suspension cells incubated in a enzyme solution containing 2%cellulase RS, 0. 1% pectolyase Y-23, 0. 55mol/L mannital and CPW salt for 5 hours. These protoplasts were cultured in defferent medium with liquid and nurse culture method. Cell division was observes within 4 days and microcalli were formed within 4 weeks. After proliferating, protoplast-derived callus differentiated into embry-oid on MS solid medium with Img/L KT and 0. 5mg/L IAA. Stems and leaves were formed on MS medium with Img/L 6-BA and 0. 5mg/L IAA. Finally, complet plantlets were obtained on hormon free MS solid medium.