Establishment of callus tissue and effect of growth regulators on enhanced sterol production in Cissus quadrangularis L

Abstract

The present investigation was carried out for isolation, identification and enhancement of phytosterol from in vitro tissue of Cissus quadrangularis L. Aim of this investigation was to develop an efficient protocol for enhancement of phytosterol from in vitro cultured callus tissue of Cissus quadrangularis (L.), an important bone healing plant. The production of callus tissue was achieved from mature nodal segment by culturing on MS medium supplemented with auxins and cytokinins. Identification of the steroidal substances from these tissues was done by various chromatographic techniques such as-Colour reaction, TLC, comparing IR and HPLC chromatogram. Enhancement of the steroidal substances from these tissues was achieved by the addition of various concentrations of plant growth regulators (IAA, NAA and 2, 4-D). Best results for callus induction were achieved on MS medium supplemented with 3% sucrose, NAA (2.0 mg L-1) and BAP (0.5 mgL-1). β-sitosterol and stigmasterol were identified by above mentioned techniques and enhanced production of these sterols up to 10-12 fold was achieved with IAA (5 ppm) and 6-8 fold with 2, 4-D (5 ppm) in callus tissue. Hence, these tissue culture and enhanced steroidal content can be used after scale-up procedure, on a large scale for commercial production of valuable steroidal compounds. © 2011 Asian Network for Scientific Information.