Vol. 4, Issue 12 (2016)
A novel and efficient method for the enhanced production of multiple shoots and camptothecin from Ophiorrhiza trichocarpos blume through subculture passages in media of alternating strength
Author(s): Sibi CV, Renjith R, Roja Gopalakrishnan, Ravichandran P, Satheeshkumar K
Abstract: Ophiorrhiza trichocarpos Blume is a herbaceous medicinal plant reported for its high value medicinal compound, camptothecin. The current study describes for the first time an in-vitro method using nodal segments for the mass propagation of O. trichocarpos. Initiation of culture using nodal segment was obtained in quarter strength Murashige and Skoog (MS) solid media containing 0.5 mgL-1 6-Benzylaminopurine (BAP) and 0.05 mgL-1 Indole 3-acetic acid (IAA) and multiplication was achieved by frequent subcultures using the same hormonal combination. We report that alternating the salt strength of media for subcultures can enhance shoot multiplication as compared to constant salt strength. Maximum shoot proliferation (58.40 ± 3.63) and biomass production (1.22±0.102 g) was obtained in third subculture, thereafter a consistency was achieved. High performance thin layer chromatography (HPTLC) estimation of camptothecin content (0.0426±0.0012 mg/g dwt) was highest in second subculture followed by a reduction in further subcultures. A significant increase in camptothecin was observed in in-vitro shoots (0.0426±0.0012 mg/g dwt) as compared to wild plants cultivated in poly-house (0.0283±0.0031mg/g dwt).