Isolation of Protoplasts from Suspension Culture and Subsequent Shoot Regeneration in Sugarcane
Cell suspension cultures with an ability to regenerate plants were initiated from 10 sugarcane clones tested. Cultures were initiated and maintained in modified N6 liquid medium containing 2 mg/l 2,4-D, 500 mg/l casein hydrolysate and 3% sucrose. Protoplasts isolated from these suspensions were embedded in 1.2% agarose (modified KM8P medium) and cultured in modified KM8P medium with the addition of nurse culture cells from suspension culture. After 5 to 8 weeks of culture, calluses were obtained in 5 clones. Calluses derived from protoplasts could not form any organs in the following culture on R2 regeneration medium. Only, protoplastderived calluses of NiF4 (cultivar) cultured for 2 weeks on PR4 medium regenerated green shoots on R9 medium. There shoots could not develop to plant, and died in the following culture.
Moriyuki SHODA Hiroshi NAKANO