Enhanced Bioproduction of Withaferin A from Suspension Cultures of Withania somnifera

By Chitturi D, Venisetty RK, Molmoori RK, Kokate CK and Apte SS


Abstract

Callus cultures of Withania somnifera from leaves were established on MS (Murashige and Skoog) media supplemented with Dicamba (2 mg/l), Kinetin (0.1 mg/l) and Sucrose (3% w/v). Suspension cultures were established and the growth and production kinetics were studied.  For growth kinetics MS media supplemented with Dicamba (2 mg/l), Kinetin (0.1 mg/l) and Sucrose (3% w/v) without agar was used which was found suitable for the initiation and maintenance of the suspension cultures cultures from the calli. Half B5 medium supplemented with 2,4-Dichlorophenylacetic acid(2,4-D) (1 mg/l), Kinetin (0.1 mg/l) and Sucrose (5% w/v) was employed.  Effect of various precursors and elicitors on suspension cultures was studied.  The addition of precursor’s sodium acetate (50 mg/l) (~10 folds), Mevalanolactone (50 mg/l) (~14 folds), Squalene (50 mg/l) in colloidal form (~23 folds) and Cholesterol (25 mg/l) in colloidal form (~30.5 folds) have shown significant increase in bioproduction of withaferin A. Among the various biotic and abiotic elicitors used Verticillium dahliae (5% w/v cells extract) (~10 folds) and copper sulphate (100µm/l) (~2.5 folds) have shown moderate increase in the bioproduction of withaferin A. The effective precursors and elicitors were studied for optimization of day of addition and found that maximum bioproduction of withaferin A was seen in the cultures when precursors and elicitors were added to 3 day old suspension cultures.

 

Key Words : Elicitors, Precursors, Suspension cultures, Withaferin A.

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