Microscopy
Protoplasts were isolated from cauliflower plants using anenzyme solution [10 mM CaCl2, 0.6 M mannitol, 10 mM MES,macerozyme (1%, w/v) and Cellulase Onozuka R-10 (2% w/v)]. Cellulases were sourced from Austratec Pty. Ltd. (Kilsyth, Australia).Leaf slices were incubated in the enzyme solution at 24◦C for 16 h. Protoplasts were pelleted at 60 × g then resuspended in W5 solution . Chloroplast phenotypes were observed in protoplasts, leaf samples or epidermal peels of leaves via a BH2Epi-fluorescence microscope (Olympus) using UV or bright field illumination. Images were captured with a MOTIC camera and soft-ware