The pH medium was adjusted to 5.6 with 0.1N NaOH or HCl before the addition of 0.8% agar (Agar-Agar). Seeds were maintained at room temperature (25±1 ◦C) under 16/8 h photoperiod for 14 days. Then, after germination, new emerged hypocotyl buds were excised and used for shoot induction.