Abstract

OPTIMIZATION OF BIOMASS YIELD AND ASIATICOSIDE ACCUMULATION IN THE CALLUS CULTURES FROM THE LEAVES OF CENTELLA ASIATICA (L). URBAN

Netala Vasudeva Reddy, Sukhendu Bikash Ghosh, Kalla Chandramouli,Tartte Vijaya*, Bobbu Pushpalatha, Dandu Anitha, Duggina Pragathi

ABSTRACT

Due to increasing evidences of potential medicinal and clinical applications of Asiaticoside, an active phytoconstituent of Centella asiatica leaves, it is an essential to produce this bioactive compound with optimal yield which can be achieved through invitro callus cultures from the young leaves. Callus growth, biomass yield and in vitro asiaticoside accumulation in callus cultures from young leaf explants of Centella asiatica was studied by the application of different plant growth regulators in the culture media. Callus induction was observed from young leaf explants on MS medium with different concentrations of 2,4-dichlorophenoxy aceticacid (2,4-D) and α- napthalene aceticacid (NAA) supplied singly and in combination with different concentrations of kinetin, 6-benzylaminopurine. Optimum callus (84%) was developed on murashig-skoog (MS) medium with α- napthalene aceticacid (2.0 mg/l) and kinetin (1.0 mg/l) which induced whitish green and compact callus and was found best for the optimal biomass yield (Fresh weight 1.58 g/20mL; Dry weight 110 mg /g FW) as determined by growth curve analysis and gave highest asiaticoside content(0.94 mg/gm dry weight) which was quantified by High- Performance Liquid Chromatography (HPLC) on C18 reverse phase column using mobile phase water:acetonitrile (20:80) with the flow rate of 1 mL/min and read at 220 nm with PDA detector. Based on the callus growth curve, we also reported the suitable growth time point for callus harvesting in terms of biomass accumulation and the highest biomass yield was observed on 35th day after inoculation.

Keywords: Centella asiatica, asiaticoside, callus cultures, growth curve, biomass yield, High-performance liquid chromatography.