Protoplast studies in cotton (Gossypium Spp.)

Different explants like tender leaves, cotyledonary leaves, hypocotyl and hypocotyl derived callus of G. hirsutum cv. Abadhita and G. herbaceum cv. Jayadhar were used for protoplast isolation. Protoplasts were isolated in the enzyme mixture consisting of 5 per cent cellulase, 5 per cent pectinase, 7.5 per cent mannitol and inorganic salts (pH-5.S). Cotton protoplasts originating from different explants varied in size and shape. The variation in size was more evident in callus derived protoplasts. Callus derived protoplast yield was highest followed by tender leaves and ieast in hypocotyl segments. Protoplast viability test based on Evan's blue staining indicated that per cent of nonviable protoplasts was more from hypocotyl explants followed by cotyledonary leaf and least in callus. Protoplasts derived from different explants did not show any cell division but callus derived protoplasts cultured on MS liquid medium +2.5 g/l glucose+0.1 mg/l NAA developed cell wall and microcolonies after 12–15 days of incubation.