In vitro production of Swertiamarin from Swertia chirata Using Liquid Culture

Abstract

newline ABSTRACT newlineA protocol for in vitro production of swertiamarin from swertia chirata using liquid culture was developed. The sterilized explants (leaves) cultured on MS medium supplemented with 2,4-D (2mg/l) alone, 2,4-D (1mg/l) + BAP (0.5mg/l) + TDZ (0.5mg/l), 2,4-D (2.5mg/l) + BAP (0.7mg/l) and BAP (3mg/l + NAA (1.5mg/l) gave best results with respect to their percentage response for callus induction. The callus was further multiplied on medium supplemented with 2,4-D (1 mg/l) + IAA (0.5mg/l). The shoot regeneration was obtained from callus on MS medium supplemented with Kn (2 mg/l) + IAA (1.5 mg/l) + GA3 (0.2 mg/l). The same medium was found to be the best for in vitro shoot multiplication. newlineCell suspension, proliferation and shoot multiplication was optimized in liquid culture medium. Different combinations and concentrations of phytohormones were tested. Out of which MS+ Kn (0.3mg/l) + BAP (0.4mg/l) + NAA (0.3mg/l) + TDZ (0.1mg/l) with 83% survival rate found to be the best for cell suspension and proliferation. For shoot multiplication in liquid culture, MS medium supplemented with Kn (1mg/l) + BAP (2mg/l) + GA3 (1.5mg/l) with an average number of 17.5 shoots per explant was found to be the best. newlineConcentration of swertiamarin in lab callus and field grown plants were identified through HPLC. Field grown plants had much higher concentration (2.91%) than lab callus (0.67%). Elicitor (methyl jasmonate) was used to enhance the concentration of swertiamarin in lab callus with different concentrations in liquid culture and semi-solid medium. Finally, with elicitor concentration (50and#956;g) for 16 days of incubation in liquid culture the concentration of swertiamarin was found to be the best (4.30%) than the field grown plants.