Effect of carbon source and elicitors on biomass production and accumulation of friedelin and epifriedelanol in hairy roots of hemp (Cannabis sativa L.)

Friedelin and epifriedelanol are pentacyclic triterpenoids that preferentially accumulate in the roots of hemp (Cannabis sativa L.) and are valued for their antidiabetic, hypolipidemic, antioxidant, liver protective, anti-ulcer, anti-inflammatory, antimicrobial, anticancer, and antisenescence properties. The aim of the present study was to investigate the influence of media, carbon sources, and elicitation on the production of C. sativa hairy root biomass and their metabolites. The highest biomass (9.45 +/- 0.00 g/100 mL flask fresh weight (FW) from the hairy root culture was obtained in MS liquid medium supplemented with 3% sucrose after 28 days. The highest levels of epifriedelanol (3.79-fold) and friedelin (3.25-fold) were found at the end of the exponential phase. Based on the carbon sources experiment, the presence of 3% sucrose provided the highest accumulation of epifriedelanol (0.930 +/- 0.013 mg/g DW) and friedelin (0.574 +/- 0.024 mg/g DW) in the hairy root culture after 28th days. The effects of methyl jasmonate (MJ) and salicylic acid (SA) on the enhancement of friedelin and epifriedelanol in C. sativa hairy root cultures were investigated. Between the two elicitors tested, SA at 100 mu M showed the highest production of epifriedelanol (5.018 +/- 0.35 mg/g DW) and SA at 50 mu M showed maximum content of friedelin 1.56 +/- 0.34 mg/g DW in hairy root culture. These represented 5.22- and 2.88-fold increase over the control (0.96 +/- 0.01 mg/g DW and 0.54 +/- 0.03 mg/g DW) after 96 h of treatment, respectively. The maximum accumulations of epifriedelanol (3.59 +/- 0.12 mg/g DW) and friedelin (1.31 +/- 0.01 mg/g DW) were observed in the treatment with MJ (100 mu M) after 24 h of exposure and were 3.73- and 2.44-fold higher than the control, respectively. The result of this study indicates that hairy root culture of C. sativa enhanced friedelin and epifriedelanol contents, which can be used for commercial production in pharmaceutical companies.