Establishment of Cell Suspension Culture and Isolation of Protoplast of Pinus serotina

Callus was induced by embryos from mature seeds of Pinus serotina with MS medium contained 0.2 mg·L-1 of KT and 2.5 mg·L-1 of 2,4 D.After successive subculture and selection,six month old callus were transferred to a liquid medium with 0.2 mg·L-1 of KT and 0 5.0 mg·L-1 of 2,4 D and 200 mg·L-1 of LH for vibration culture,the suspension cultured cell lines were established with favorable dispersivity.A great amount of protoplasts were released from three month old cell suspension cultures,and protoplast yield and protoplast viability of suspension cells on linear phase are the highest under the enzyme combination cellulase R 10.2%+pectinase 0.25%?