Abstract:

A protocol for plant regeneration from protoplast of Musa AAB Silk cv. Guoshanxiang via somatic embryogenesis was developed. Viable protoplasts were isolated from embryogenic cell suspension (ECS) in a enzyme mixture of 3.5% cellulose R-10, 1% macerozyme R-10, 0.15% pectinase Y-23 and 0.41 mol/L manntitol, the yield was 3.1×107 protoplasts per mL packed cell volume (PCV) ECS. Liquid and feeder layer culture systems with medium 'A' and medium 'B' were used respectively for protoplast culture. In liquid culture system, medium 'B' was more efficient for inducing cell division and colony formation which was about 3-fold and 10-folod respectively, compared to that with medium 'A'. However, all protoplast-derived cell colonies obtained from liquid culture system could not develop further. In feeder layer culture system, there was no significant difference between medium 'A' and medium 'B' on cell division and colony formation of the cultured protoplasts. Protoplast-derived cell colonies from feeder layer culture system were then transferred onto embryo induction medium for somatic embryogenesis. Forty-five days after the cell colonies were transferred on embryo induction medium, 1550 mature embryos were obtained from 105 protoplasts. After another 30 d of culture, 7.8% of mature embryos germinated. Normal plantlets were obtained from MS basal medium supplemented with 0.1% activated charcoal and the plantlets were transferred into pots and grew well.

Key words: Musa AAB Silk cv. Guoshanxiang, protoplast, somatic embryogenesis, plant regeneration