Mature rhizomes of B. rotunda were purchased from herbal supplier at a local Market in Kuala Lumpur, Malaysia. The rhizomes were cleaned and rinsed. The cleaned rhizomes were then placed in an open container to allow shoots to sprout two to four cm in length. The sprouting buds were collected and washed with 20% (v/v) clorox for 15 min. Under aseptic conditions, shoots were surface sterilized with 0.5% (w/v) aqueous solution of mercuric chloride (HgCl 2) solution for five minutes and followed by three rinses in sterile distilled water. The shoot buds had their external leaves removed and trimmed down until the size ranged from 0.5 to 0.8 mm and used as explants (Figure 1A to C). They were then inoculated into 350 ml glass jar containing MS (Murashige and Skoog, 1962) medium supplemented with 30.0 g/l sucrose and 2.0 g/l gelrite for 30 days. The pH of the medium was adjusted to 5.7 prior to autoclaving at 121°C for 21 min. The cultures were maintained in a culture room with 16/8 h photoperiod (light/dark) at 25 ± 2°C. After 30 days, the aseptic buds of this species were transferred onto MS medium supplemented with 3.0 mg/l BAP and 0.5 mg/l NAA, the optimum medium was formulated for other Zingiberaceae species by Yusuf et al. (2007) for shoot multiplication for four (4) weeks (Figure 1D). The multiple shoots produced were separated and transferred into MS medium devoid of plant growth regulators for four (4) weeks and were used for all subsequent experiments.