In vitro Callus Induction and Rosmarinic Acid Quantification in Callus Culture of Satureja khuzistanica Jamzad (Lamiaceae)

Abstract

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 mg l-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.