PLANT-REGENERATION FROM INDICA RICE PROTOPLASTS

We have standardized a protocol for efficient regeneration of rice plants from single protoplasts. Protoplasts were isolated enzymatically from embryogenic, rice suspension cultures of indica rice cv IR 50. The 45-day-old suspensions, subcultured at weekly intervals, were used for protoplast isolation. Protoplasts were isolated on the fourth day of subculture when cells were in active growth phase. Isolated protoplasts were purified and subjected to a heat shock prior to culture. They were then cultured by feeder layer technique. Protoplasts divided and formed microcalli in six weeks. These calli were nurtured to give fertile plants.