توليد و نگهداري كشت سلولي گياه steud and Hochst Digitalis nervosa و بررسي متابوليت هاي ثانويه آن

Cell culture edtablishment and Secondary metabolites of Digitalis nervosa Steud & Hochst

Background and purpose: Digitalis nervosa is a wild species growing in Tran and the major cardenolode of this species is lanatoside A. D nervosa cell culture was estabilished and its secondary metabolites were studied. Materials and methods : Seeds of D. nervosa were surface sterilized in 70% ethanol or 5% Sodiume hypochlorite, then healthy seedlings were produced. The jemol, Hypocoty l root tip of seedling were transferred onto the Murshige and Skoog (MS) culture media containing different concentrations of plant growth rgulators. The Phytochemical studies on callus and plant were carried out using Thin layer chromatography (YLC). Total Cardiac glaycosides in plant and callus were determined by spectrophotometric method. Results: Sodium hypochlirite had less negative effect on the seedling production comparing with Ethanol. The best callus intiation and estabilishment was observed in media containing K (0.5mg/1), 2,4-D (0.5 mg/1), NAA( 1 mg/l) as plant growth regulators. Callus formed from the root part of seedies revealed the presence of saponins, tannins, flavonoids, steroids and Cardiac glycosides in both callus and plant extracts. The amount of cardiac glysosides in plant was 358 mg/100 gr. Dry w. This amount varied from 29.68 to 15.90 mg/100 gr. Dry. wt in the 7 th Generation of callus. Conclusions: The ratio between auxin and cytokinin is one of the important factors to callus production. The 3: 1 ratio demonstrated the best callus growth on D. Nervosa cell culture. The callus culture of D. nervosa showed the ability of producing cardiac glycosides howere, this ability reduced by repeated subculturing.