Effects of various plant growth regulators upon callus induction potential using leaf explants of Clinacanthus nutans (Sabah Snake grass)

Sabah snake grass, also known as Clinacanthus nutans, is a medicinal plant which has been widely consumed by natives in the Asian countries to treat many ailments. At present, stem cutting is the only way this plant is propagated which in future could lead to extinction. Besides, variation in the active compounds had been reported in fresh samples of C. nutans. Not many in vitro works had been carried out to cultivate this plant. The aim of this study was to establish clean (in vitro) cultures of C. nutans and determine the optimal callus induction and proliferation protocol, using various combination of plant growth regulators at different concentrations, ranging from 0-2.0mg/l, using leaf-segments. Highest percentage (87.88%) of aseptic C. nutans shoot cultures were successfully obtained from nodal segments disinfected using in 50% home bleach Clorox® for 20 minutes, and with 30% Clorox® for 50 minutes. Best callus induction was obtained from explants cultured on Murashige and Skoog media supplemented with Picloram alone at 0.5mg/l. This protocol would be useful to minimize the destruction of wild grown C. nutans and effective callus propagation would be beneficial to growing resources in a big scale.