Document Type : Research Paper

Authors

• R. Jafari Hajati ¹
• V. Payamnoor ¹
• K. Ghasemi Bezdi ¹
• N. Ahmadian Chashmi ²

¹ Faculty of Forest Sciences, Gorgan University of Agricultural Sciences and Natural Resources, Gorgan, Iran

² Department of Biology, Faculty of Basic Sciences, University of Mazandaran, Babolsar, Iran

Abstract

This study aimed to enhance the production of betulin and betulinic acid using suspension cultures of birch (Betula pendula Roth) and elicitation of the cell cultures by methyl jasmonate (MeJA) and salicylic acid (SA). To do this, at the first step, the cell growth curve was investigated in a 16-day period. Then, two elicitors, namely, MeJA (at final concentration of 0, 50, 100, 150 and 200 µM) and SA (at final concentration of 0, 100, 200, 300 and 400 µM) were separately supplemented to 8-day-old cell cultures and the cells were harvested 1, 2, 3, 5 and 7 days after elicitations. Fresh weight (FW), dry weight (DW) and cell viability were measured. In addition, betulin and betulinic acid content were analyzed using HPLC. The results showed the significant effects of different concentrations of SA and MeJA on metabolites content and FW and DW. Maximum amount of betulin was observed about 4-fold (2.5 mg g^-1 DW) higher than the control treatment by addition of 100 µM SA, two days after elicitation. Moreover, betulinic acid content was enhanced about 5 mg g^-1 DW, 4.5-fold compared to control, one day after addition of 200 µM SA. Furthermore, the high accumulation of betulin (2.3 mg g^-1 DW) was obtained in the elicited cell by 50 µM MeJA, seven days after elicitation. Also, the maximum amount of betulinic acid, about 3 mg g^-1 DW, was observed in the cells elicited by 100 µM MeJA, two days after elicitation. Overall, the effect of SA on the production of betulin and betulinic acid was significantly more than the effect of MeJA.

Keywords

• betulin
• Betulinic acid
• Elicitor
• HPLC