Micropropagation in Verbisina enceloides– The Medicinal Herb

Callus formation was best observed on MS medium from young leaves, appearing ceramist yellow in colour having soft appearance. Small shoots regenerated transferred to MS medium with BAP alone which gave sufficient shoot proliferation in three weeks. Callus was also successfully established on MS medium supplemented with BAP in combination with auxins. It was whitish brown in colour, nodular, compact, and nonchlorophyllous and friable in nature which soon degenerated without differentiation. This gave emergence to the fact that may be plant contains enough amount of auxins. Interestingly IAA content was found maximum (5 µg·g^–1) in the middle portion of the leaf segment and was least in basal part (3 µg·g^–1) of leaf segment. Shoot proliferation was achieved directly from the nodal explants inoculated on MS medium supplemented with BAP (0.5 mg·L^–1). Other explants did not give anticipated results. Other cytokinins (Kn, TDZ) also gave good results but were not as efficient as BAP alone. Different carbon sources were tried amongst glucose, fructose, maltose, and sucrose used at different concentrations and only sucrose (3%) gave the maximum number of shoots and shoot length. Various nitrogen sources were also tried such as arginine, adenine sulphate, tyrosine, and glycine; here, glycine gave the maximum shoot proliferation with maximum number of shoots. Rooting was induced in the in vitro raised plantlets using two approaches, by adding auxins to the MS medium and by using dip pulse treatment (where micro plant cut ends were dipped for varying time period in pre-autoclaved auxins), rooting was best observed in dip pulse treatment with IBA(0.5 mg·L^–1) for 15 min.