An efficient protocol for production of rosmarinic acid in Salvia nemorosa L.

Rosmarinic acid (RA), as the major secondary metabolite of Salvia species, is a valuable bioactive compound for pharmaceutical applications. Toward rosmarinic acid production, Murashige and Skoog (MS) medium supplemented with 0.5 mg L −1 2,4-D and 2 mg L −1 BA or 1 mg L −1 2,4-D and 1 mg L −1 BA showed the highest percentage of callus induction (100%). In this research, MS medium supplemented with various concentrations of sucrose, phenylalanine, salicylic acid (SA), and methyl jasmonate (MeJA) were evaluated using cell suspension cultures of Salvia nemorosa L. Sucrose treatment (4.5%) increased cell biomass (20.58 g L −1 ) and RA content (17.1 mg g −1 DW). Phenylalanine had no significant effect on cell biomass; however, phenylalanine led to a higher accumulation of RA (16.2 mg g −1 DW) compared to non-treated cells. MeJA caused a slight decrease in cell growth. Maximum RA content (14.2 mg g −1 DW) observed in cells treated with 200 μM MeJA, 48 h after elicitation. SA (0.5 μM) treatment showed an impressive effect on RA production (26.4 mg g −1 DW) with an increase of 8 times compared to the control. Phenylalanine ammonia-lyase and RA synthase expression were positively correlated with RA biosynthesis after induction with SA. Results showed that SA might be considered as an economical elicitor compound for large-scale production of RA.