ASIATICOSIDE AND ASIATIC ACID PRODUCTION IN SHOOT, CALLUS AND CELL SUSPENSION CULTURES OF CENTELLA ASIATICA

Abstract

Centella asiatica commonly known as gotu kola, Mandukaparni or Indian pennywort is a small herbaceous perennial medicinal plant belonging to Apiaceae family. Its active metabolites are triterpenoids which includes asiaticoside, asiatic acid, madecassoside, madecassic acid which has wide range of pharmaceutical activities like wound healing, memory enhancer, treatment of skin diseases etc. The present work is aimed at comparative estimation of Asiaticoside and Asiatic acid before and after the Methyl jasmonate (MEJA) elicitation in various cell cultures like shoot culture (SC), callus culture (CC) and cell suspension culture (CSC) of Centella asiatica . The in vitro shoot cultures were grown to study the effects of variations in the nitrogen, carbon, phosphorous, potassium, micronutrient (Manganese) and macronutrient (magnesium) concentrations in MS media and also elicitors like MEJA and malt extract on the shoot multiplication . The callus culture was established using leaf explants in the MS media supplemented with combination of plant hormones like BAP,NAA,2,4 D,IAA . CSC was established using callus as the explants and its growth curve was plotted . By HPLC analysis, maximum asiaticoside content in CSC (0.033mg/ml) followed by shoot and callus culture. When elicited with 100μM MEJA there was 39 fold increase in asiaticoside content( 0.146 mg/ml) in shoot and about 68 fold(0.2019 mg/ml) increase in CC. Asiatic acid content was highest in SC (0.653 mg/ml) followed by callus (0.0349 mg/ml) which was enhanced by 2 fold ( 0.0675 mg/ml) after MEJA elicitation. The antioxidant activity by DPPH assay showed maximum IC 50 value in CSC extract than shoot and callus extracts. The quantitative estimation of total phenolic, flavanoids and tannins indicates highest phytochemicals in SC when compared to CC and CSC.