Vitro plantlet regeneration from Tinospora cordifolia (Willd.) Miers - a highly valuable medicinal plant and its chemo-profiling

Abstract

Tinospora cordifolia[(Willd.) Miers ex Hook. F. and Thom)] belongs to family Menispermaceae and is mostly found in tropical Indian subcontinent especially India, Sri Lanka and Bangladesh. It is an important medicinal plant species and considered as a rich source of alkaloids and terpenes which is used as a rejuvenating drug. The extract of the mature stem is useful in curing skin diseases and dry barks have anti-spasmodic, anti-pyretic, anti-allergic, anti-inflammatory and anti-leprotic properties. In the present investigation a viable in vitro regeneration protocol for mass multiplication of Tinospora cordifolia was standardized. Chemo-profiling of crude extract of Tinospora cordifolia was also investigated. The shoot buds were used for direct shoot induction and regeneration by using Murashige and Skoog (MS) medium supplemented with different concentrations of cytokinins BAP and Kinetin singly or in the combination. For auxiliary shoot proliferation and superior in shoot elongation, it was found that BAP was more effective than Kinetin among the cytokinins tested. An average multiplication rate of 6.3 shoots per explants was obtained with MS medium supplemented with 8.87 µM BAP alone. Small shootlets were transferred to MS medium supplemented with 8.87 µM BAP and 5.71 µM IAA for shoot elongation. The elongated shoots were rooted in half-strength MS medium supplemented with 2.22 µM BAP and 4.92 µM IBA. Rooted plantlets were successfully transferred to soil and established with 80% survival. The TLC profile was developed using hexane: ethyl acetate (25:5v/v) as a mobile phase and individual peak scanned at short wavelength 366nmto identify and separate the berberine in leaf, stem, and root of T. cordifolia In the antibacterial studies, it was found that Tinospora cordifolia has the most antibacterial properties against Bacillus subtilis as compare to other bacterial species. The methanolic extract of Tinospora cordifolia resulted a „zone of inhibition‟ of 18 mm at a concentration 80 mg/ml. Against Styphylococcus aureus, a 80 mg/ml methanolic extract worked best giving a „zone of inhibition‟ of 15 mm. The methanolic extracts of this species gave a „zone of inhibition‟ less than 1mm Mycococcus