PLANT CELL BIOTECHNOLOGY AND MOLECULAR BIOLOGY

The present work describes an effective, high-frequency multiple shoot induction protocol by using a cotyledonary node. The seeds of Cryptocarya stocksii were surface sterilized with HgCl₂ (0.1 % w/v) for 7 min and washed 3 times with sterile water. Seeds were cultured on Murashige and Skoog (MS) medium containing 0.5 mg/L Thidiazuron (TDZ) for 20 days. Cotyledonary nodes were excised from 20 days old seedlings were used as explants for multiple shoot induction. Cotyledonary node were pre-inoculated for 20 days on MS medium containing 1 mg/L TDZ, 0.5 mg/L 1-Naphthaleneacetic acid (NAA), 0.5 mg/L Gibberellic acid (GA₃) and 0.2% Activated charcoal (AC) for multiple shoot bud induction. Thereafter, they were transferred to the secondary medium for shoot elongation using MS medium containing 3 mg/L 6-Benzylaminopurine (BAP), 0.5 mg /L NAA and 1 mg /L Kinetin (KN). Those shoots with 4 to 5 leaves were transferred to root induction MS medium containing 3 mg/L indole 3-butyric acid (IBA). Plantlets with well-developed roots were transferred to pots containing sterile sand and maintained in the growth room.