Callus Induction and Cell Suspension Culture Establishment of Salvia sahendica for The Production of Medicinal Triterpenoids Betulinic, Oleanolic and Ursolic Acid

Betulinic, oleanolic and ursolic acid are highly valuable pentacyclic triterpenoids because of their wide spectrum of biological activities such as anti-inflammatory, hepatoprotective, antitumor, anti-HIV, antimicrobial, antifungal, anti-ulcer, gastroprotective, hypoglycemic, and antihyperlipidemic. Salvia sahendica Boiss. & Buhse is an aromatic, endemic perennial herb which grows in the restricted regions of Northwest of Iran. In the present study, callus induction and cell suspension culture establishment of the plant for the production of triterpenoids were studied. Callus induction was conducted from in vitro-grown leaf explant cultured on Murashige and Skoog (MS) medium supplemented with different concentrations of 2,4-D (0.5, 1.0, 1.5 and 2.0 mg l−1) and BA (0.5, 1.0 and 1.5 mg l−1) base on factorial complete random design experiment. Maximum callus induction (100%) was obtained in MS medium supplemented with 1 mg l−1 2,4-D and 0.5 mg l−1 BA. The study of cell growth curve showed that the highest fresh weight (13.2g) and dry weight (2.2g) obtained after 4 week of culture. High content of ursolic acid (173.81 mg/100g DW), betulinic acid (162.2 mg/100g DW) and oleanolic acid (174.32 81 mg/100g DW) were obtained at third week of culture. Results showed that the content of betulinic acid and ursolic acid at third week of culture was 10.4 and 1.5-fold than the aerial part of wild growing plant. These results can be considered for scaling-up of triterpenoids production in bioreactors