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)

Optimizing the potential utilization of bioreactors for the mass propagation of Indonesian Dendrobium varieties

“…Plantlets of E. ava grew in TIS (5 min/4 h), resulting in 1.5 shoots per explant, 4.4 roots per plantlet, 29.4 mm shoot height, 4.4 leaves per shoot (Kunakhonnuruk et al, 2019). The best multiple-shoots of Mokara Orchid were conducted in TIS (1 min/2 h) with 14.8 shoots per cluster, 2.6 cm height shoot, 2.1 cm diameter shoots clusters, and 0,0 number of dead shoots per cluster (Minh, 2022). TIS (5 min/6 h) on D. nobile produced 349.23 g L -1 fresh and 54.48 g L -1 dry weight (B.…”

“…In addition, using a temporary immersion bioreactor system (TIS-bioreactor) provides an opportunity to overcome the browning problem due to continuous immersion in the airlift bioreactor. Reducing browning explants and improving EC proliferation under a temporary immersion system (TIS) bioreactor were successfully carried out by applying 1 min/4 h dried and immersion frequencies for C. forbesii Lindl (Ekmekçigil et al, 2019); 5 min/4 h for Epipactis ava Seidenf (Kunakhonnuruk et al, 2019); 5 min/6 h for Dendrobium nobile Lindl (Zhang et al, 2020); and 1 min/2 h for Mochara orchid (Minh, 2022). Banana extract (BE) noted had optimal effect on regeneration and preparation of well rooted shoots (Nurfadilah et al, 2018;Sidhu and Zafar, 2018;Novita et al, 2022) with 50 g L -1 for Cymbidium pendulum (Kaur and Bhutani, 2012), 100 ml L -1 for Dendrobium sp.…”