Abstract
In vitro tissue culture of hard woody, endangered, medicinal plant Coscinium fenestratum is most challenging to plant tissue culturists. In the present study, petiole and leaf explants of Coscinium fenestratum were induced to form callus when cultured on vermicompost extract media along with coelomic fluid. Suspension medium was developed using vermicompost extract and coelomic fluid in 3:1 ratio. Phytochemical analysis of the alkaloid berberine was confirmed from callus, suspension cell culture and suspension medium by Thin Layer Chromatography and High Performance Liquid Chromatography. Vermicompost and its extracts with coelomic fluid have shown maximum (100 per cent) response of callus induction. Callus mass enlarged with increasing concentration of coelomic fluid and callus growth was assessed from the biomass. Incubation of culture tubes in dark supported callus development significantly. The Rf value of 0.36 confirmed the presence of berberine by Thin Layer Chromatography. Qualitative analysis confirmed the presence of alkaloid berberine with the retention time of 2.8 minutes similar to that of standard reference sample from Sigma chemicals, USA. The suspension medium turned deep yellow because of the release of the alkaloid. Vermicompost and its extracts along with coelomic fluid have shown the economical approach for micropropagation of economically and medicinally important plants.
Highlights
Coscinium fenestratum Colebr. (Menispermaceae) is widely spread across the Western Ghats (India) and Sri Lanka
Protocol was developed for obtaining berberine-producing callus and cell suspension cultures established from the petiole segments of C. fenestratum
Collection of Medicinal Plant Sample (Explant) Stem cuttings and fresh leaves of C. fenestratum authenticated in June 2012 by taxonomist, Dr Rajanna, University of Agricultural Sciences (UAS), GKVK, Bangalore were collected
Summary
Coscinium fenestratum Colebr. (Menispermaceae) is widely spread across the Western Ghats (India) and Sri Lanka. (Menispermaceae) is widely spread across the Western Ghats (India) and Sri Lanka. It is commonly known as tree turmeric. The stem portions contained many alkaloids like berberine, hentriacontane, ceryl alcohol, sitosterol, oleic acid, palmitic acid and saponin with some resinous material [2]. Antidiabetic activity of C. fenestratum has been reported [4]. Preliminary studies on micropropagation of C. fenestratum were reported [8]. Protocol was developed for obtaining berberine-producing callus and cell suspension cultures established from the petiole segments of C. fenestratum. With 4 mg/L of NAA, highest yield of alkaloid berberine was acquired and 2 mg/L of 2,4-D yielded the best cell growth [9]. Intercellular berberine and berberine recovered from the suspension media were studied [10]
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