ISOLATION OF PROTOPLASTS FROM SUSPENSION-CULTURE AND SUBSEQUENT SHOOTREGENERATION IN SUGARCANE

Cell suspension cultures with an ability to regenerate plants were ini tiated from 10 sugarcane clones tested. Cultures were initiated and ma intained in modified N6 liquid medium containing 2 mg/l 2,4-D, 500 mg/ l casein hydrolysate and 3% sucrose. Protoplasts isolated from these s uspensions were embedded in 1.2% agarose (modified KM8P medium) and cu ltured in modified KM8P medium with the addition of nurse culture cell s from suspension culture. After 5 to 8 weeks of culture, calluses wer e obtained in 5 clones. Calluses derived from protoplasts could not fo rm any organs in the following culture on R2 regeneration medium. Only , protoplast-derived calluses of NiF4 (cultivar) cultured for 2 weeks on PR4 medium regenerated green shoots on R9 medium. These shoots coul d not develop to plant, and died in the following culture.