*Department of Botany, University of Delhi, India.
Tissue culture lab., Central Sericultural Research and Training Institute, Srirampura, Mysore-570008
Callus tissue was obtained from hypocotyl and cotyledon explants of Morus alba L. by culturing on Murashige and Skoog medium supplemented with 1 mg/l, 2, 4-dichlorophenoxy acetic acid and 0.5mg/l benzylamino purine. Protoplasts were isolated from freshly grown callus using enzyme mixture of 2% Onozuka cellulase R-10 and 1% macerozyme R-10. Twenty one media combinations were tested to induce division in cultured protoplasts. Wall regeneration as observed under UV flourescence microscope occured in many cases. Protoplasts cultured in thin liquid layer of MS medium containing sucrose (3%), mannitol (0.5M) coconut water (20%) 2.4-D(l mg/l) and BAP (0.5 mg/l) exhibited division whereas the system did not respond to microdrops and solid medium plating techniques. Protoplasts were divided further in the same medium forming multicellular microcolonies after the addition of fresh medium of reduced osmotic strength to the previous suspension.
