2.2. Callus induction
Fresh leaves and rhizomes of H. pinifolia were washed thor-oughly and sequentially with native seawater and sterilizedseawater (32‰). The explants were dissected into pieces 5 mmin size and surface sterilized with ethanol (70%, v/v) for 3 min,sodium hypochlorite solution (0.6%, v/v) for 30 s, followed by thor-ough washing with sterile seawater for 4–5 min. The explants weresoaked in antibacterial solution composed of Nystatin (120 mg l−1),Kanamycin (60 mg l−1) and Gentamycin (60 mg l−1) with continu-ous mild agitation for 24 h.