*4.2. Chloroplast DNA Isolation and Extraction*

Plants were kept in the dark for 48 h before harvesting to reduce starch accumulation. Fresh leaves (15–25 g) were collected and used for chloroplast isolation with discontinuous sucrose gradient [56]. Purified chloroplasts were lysed with a detergent and proteins eliminated by proteinase K and phenol/chloroform treatments following the procedure described by [57].
