Micropropagation of Mokara Orchid by Temporary Immersion System Technique

Abstract

Based young leaves of Mokara Leuen Berger Gold were used as cultured materials. Callus was initiated and increase biomass on medium of semisolid and liquid cultures: MS medium supplemented with CW (30%), sucrose (30 g/l), 2.4D (1 mg/l). Callus was used as materials for initiation and biomass propagation on medium: (1) MS medium supplemented with NAA (1 mg/l), B1 (5 mg/l), Adenin sulfate (10 mg/l), peptone (1 g/l), CW (10%), sucrose (30 g/l) (2) MS medium supplemented with 2.4D (1 mg/l), CW (30%), Adenin sulfate (10 mg/l), peptone (1 g/l), CW (10%), sucrose (30 g/l). Semisolid medium for callus cell regeneration and to induce multiple shoots were: MS medium supplemented with BA (0.5 mg/l), B1 (5 mg/l), Adenin sulfate (10 mg/l), peptone (1 g/l), CW (10%), sucrose (20 g/l). Multiple-shoots were propagated on MS medium supplemented with BA (0.5 mg/l), B1 (5 mg/l), CW (10%), sucrose (20 g/l). Propagation of multiple-shoots in TIS on MS medium supplemented with BA (0.5 mg/l), B1 (5 mg/l), CW (10%), sucrose (20 g/l). Interval culture time was optimum for 2 hours floating and 1 minute rising. Plantlets were induced roots on MS medium supplemented with NAA (1 mg/l), B1 (5 mg/l), CW (10%), sucrose (20 g/l). Experiments on callus formation and callus growth of mokara orchids both on semi-solid medium (agar) and liquid medium to create callus suspension, the best medium selected was: MS supplemented with 2.4D (1 mg/l), CW (30%), sucrose (30 g/l)