A protocol for protoplast isolation in mandarins

The factors affecting protoplast isolation were studied in two mandarins namely, kinnow (Citrus nobilis Lour × Citrus deliciosa Tenora) and Local Sangtra (Citrus reticulata Blanco). In kinnow, the best yield (3.1 × 10⁵ protoplasts/ml) was obtained from epicotyl segments using enzyme mixture containing cellulase (2.5%) and macerozyme (0.5%). In Local Sangtra, the best yield (2.9 × 10⁵ protoplasts/ml) was obtained from leaf, using enzyme mixture containing cellulase (2.5%) and macerozyme (0.5%). The optimum time of pre-treatment in all the explants was 1 hour. The best temperature and duration of incubation was 28°C and 16 h respectively, both for kinnow and Local Sangtra.