In the present study, an efficient protocol has been developed for callus induction and production of RA in callus culture of Satureja khuzistanica for the first time. In-vitro callus induction was achieved from young shoot tip explants cultured on MS and B5 media supplemented with different concentrations of IBA (0.1, 1.0, 2.0 and 5.0 mgL(-1)) solely or in combination with cytokinins BAP and KIN (1.0, 2.0 and 5.0 mgL(-1)). B5 medium supplemented with 1.0 mgL(-1) IBA plus 5.0 mgL(-1) BAP and MS medium fortified with 2.0 mgL(-1) IBA and 2.0 mgL(-1) BAP were the most favorable media for callus formation with the highest induction rate (96%). Maximum growth index (2.89 and 2.63) and maximum callus biomass (2.34 and 2.33 g fresh weight) were obtained from the callus cultured on B5 medium supplemented with 1.0 mgL(-1) IBA plus 5.0 mgL(-1) BAP and MS medium fortified with 1.0 mgL(-1) IBA plus 1.0 mgL(-1) KIN, respectively. Determination and quantification of RA in cultured calli were performed by HPLC UV/MS analysis. Calli induced from the plant and maintained on supplements of IBA and BAP in the absence of light produced RA 7.5% based on dry weight (DW). No differentiation was observed in any callus during the course of this study.
Keywords: Callus culture; HPLC UV/MS; Lamiaceae; Rosmarinic acid; Satureja sp.