Fig. 2

Isolation of protoplasts from rice calli induced from mature seeds. a Donor tissue for protoplast isolation were translucent, pale yellow, and nodular calli propagated on OsCIM2 medium. b To degrade the tissue cell walls, digestion solution was added. c A visual indication of successful enzymatic digestion was a milky appearance of the solution after three hours of gentle shaking. d Large particles and spent tissue were removed from the protoplast solution via cell strainer filtration. e Protoplasts formed a visible band, marked by a yellow bracket, after centrifugation through a 0.55 M sucrose cushion. f Protoplasts derived from rice calli. Healthy cells are round and colorless. Dead cells and debris are stained by Evans Blue. Bar = 50 µm