Hypocotyls and cotyledons were transversely chopped in pre-plasmolysis solution (Table 1) into fine pieces
(0.05-1.0 mm thickness) in a 9 cm Petri plate. Protoplasts were isolated in a two step-stationary and gyration
digestion process. Following 1h pre-plasmolysis and stationary overnight digestion at 25°C in the enzyme
solution (Table 1) for cell wall degradation, they were incubated at 27°C on a rotary shaker maintained at 50 rpm
for 2h. The resultant mixture was pelleted and purified over sucrose solution (Table 1). All incubations were
carried in dark at 26±1°C.