*2.4. Rooting of Shootlets and Acclimation*

Individually harvested, the in vitro proliferating shootlets of *P. amboinicus* with 2–3 pairs of substantially developed leaves were transplanted to a half strength MS basal medium supplemented with 0, 0.25, 0.5, 1.0, and 2.0 μM of indole 3-butyric acid (IBA) solidified with 0.8% (*w*/*v*) agar-agar and having a pH of 5.7. Data on the frequency of shootlets producing roots and the number of roots per shootlets were recorded after 4 weeks of transfer in rooting media.

For ex vitro acclimation, well-rooted plantlets were gently washed under the laboratory running water to remove the medium and agar residues. Then, the plantlets were transferred to pots containing sterile planting soil substrates (Planta-GuardTM, Germany). Maintaining the plantlets at high humidity for the first few days following transplanting is a critical factor for the survival of micropropagated plants. To retain humidity, the ex vitro transplanted plants were covered with clear polybags, which were removed after 4 weeks and irrigated with half strength MS basal medium containing macro and micro-salts devoid of vitamins. After 1 month, the percent survival of acclimated plants was calculated as follows:

Total number of plants survived Total number of plants trasnferred <sup>×</sup> <sup>100</sup>
