International Journal of Agriculture, Environment and Biotechnology

Ophiorrhiza mungos var. angustifolia is a herbaceous medicinal plant belonging to the family Rubiaceae. It is a rich source of potent anticancer compound camptothecin (CPT). Massive uprooting of the species from its natural habitat for the extraction of CPT along with poor seed viability has led to the destruction of its population. At this context, the present study was proposed to develop in vitro protocol from leaf explant for the propagation and production of CPT from the species. In vitro multiple shoot cultures maintained in half strength Murashige and Skoog medium with 3% sucrose and 0.8% agar supplemented with 8.88 μM Benzyl Amino purine (BA) were served as the explant source. Multiple shoots were induced on leaf explant cultures on half strength MS solid medium fortified with different concentrations of (4.44–22.19 μM BA, 4.67–23.23 μM Kinetin, 0.49–2.45 μM 2 isopentanyladinine, 4.57–22.85 μM Zeatin) either individually or in a combination. Medium supplemented with 13.31 μM BA showed the best result with 52.33±3.06 shoots/explant. The synergic effect of BA with other cytokinins also favoured shoot induction in a higher rate but they were very minute in nature. The shoot buds initiated were repeatedly sub cultured and multiple shoot cultures were developed. The CPT content of thus produced multiple shoots were checked (0.13 ± 0.1 mg/g DW) and found comparable with that of the field grown plants (0.14 ± 0.1 mg/g DW). The authenticity of CPT was checked with HPLC analysis. Protocol developed in the present study can be used for the regeneration of adventitious shoots from leaf explant of O. mungos var. angustifolia.