Vol. 9, Issue 9 (2020)

Author(s):
Md. Nizamuddin Ansari, Chandan Kr Singh, Nidhi Kumari and Md. Naseem

Abstract:
Medicinal plants are in demand since the beginning of human civilization and the plant products feature prominently in traditional therapeutics. Diabetes is one of the most common diseases in human population and more than 30% Indian population are suffering from this disease. There are large number of plants which are recommended for its treatment in herbal system of medicine. Vernonia divergens Benth commonly known as insulin plant is a potent sugar killer and is used as an excellent medicine for diabetes mellitus. This plant has restricted distribution in our locality and some people suffering from diabetes grow this plant in their courtyards. Due to ever increasing demand of this plant for officinal uses and to analyse biochemical constituents, it is desirable to develop a simple and efficient protocol for mass propagation and conservation to meet the growing need. Keeping in view its medicinal use, the in vitro studies of this plant were being undertaken to develop a protocol for mass propagation as well as to analyse its biochemical constituents. Regeneration of plantlets and callus differentiation were obtained using stem (node, internode) and shoot-tip of Vernonia divergens as explants. Sterilized explants taken from in vivo grown plant (about 4 years old) were cultured on MS (Murashige & Skoog, 1962) medium containing 0.8% agar, 3% sucrose and different combination and concentration of NAA/2, 4-D and Kinetin (Kn). Techniques were standardized for shoot regeneration directly from node and shoot-tip explants as well as shoots from callus. The development of shoots/multiple shoots was more frequent in culture, the best response was obtained on 3 mg/l Kn in nodal and shoot tip culture. Multiple shoots were obtained in nodal culture on 2 mgl-1 NAA + 3 mgl-1 Kn. Callus mediated shoot regeneration was frequent in culture and was achieved on 5 mgl-1 Kn and 2 mg/l-1 NAA on subculture. 2, 4-D alone or 2, 4-D + Kn resulted in callus differentiation from explants. Callus in general was greenish-white/white compact, hydrated and crystalline in appearance. Rooting of shoots (3-4 cm) was obtained on rooting medium (1⁄2 strength of MS salts) fortified with 1 mgl-1 NAA & 2 mgl-1 IBA. Ex-vitro rooting (dipping of shoots in IBA) technique was more promising in woody shoots and plantlets were successfully transferred to soil and survived well in nature. The success rates of transferred plantlets were satisfactory. Higher concentration (>5 mgl-1) of auxin and Kn was inhibitory for culture growth, nodal explants was superior to all other explants (internode, shoot-tip) with respect to shoot regeneration. Protocol was also developed for conservation of callus which survived till 2 years. In vitro developed plantlets were morphologically identical to parent plants. Works are in progress to develop protocol for isolation of active constituents and to determine the active compound which has ant diabetic properties.