A STUDY ON THE ISOLATION OF PROTOPLASTS FROM THE CALLUS OF LILIUM LONGIFLORUM OVERIG

Lilium longiflorum Overig is a Lilaceous plant grown for ornamental as well as certain other general purposes. The research presented here focuses on the method of protoplast isolation from the In vitro grown callus of Lilium longiflorum Overig. Series of experiments were conducted in order to optimize the conditions. Four different amounts of callus and a range of incubation time were used to achieve maximum number of viable protoplasts. However, the calli used were treated with Enzyme solution containing 0.5% (w/v) Macerozyme, 2% (w/v) Cellulase and 0.1% (w/v) Pectinase in combination, to obtain maximum number of viable protoplasts through the process of cell -wall digestion. Furthermore, an osmoticum of 20% (w/v) sucrose and a washing solution consisting of 0.1% (w/v) MES and 0.5M Mannitol were added to suspend a ring of protoplast at the interphase of both the solutions. This ring of protoplast was then isolated and tested for viability and yield. Consequently, amongst the various sets of experiment, callus of 1.5 grams and incubation time of 4 hours was found an ideal approach to obtain maximum number of viable protoplasts.