In Vitro Callus Culture of Salvia Officinalis L. and the Effect of Some Amino Acids on Rosmarinic Acid Accumulation

This study investigated the in vitro callus formation ability of various explants obtained from the Salvia officinalis (common sage) and the production of rosmarinic acid in the resulting calli. Four types of explants, including young leaves (1-1.5 cm), nodes, and stem pieces, were cultured on MS medium containing 0.8 mg/l 2.4-D, 0.5 mg/l NAA, and 2.0 mg/l BAP to induce callus formation. The highest callus formation (100%) was achieved from young leaf explants. After 4 weeks, the formed calli were transferred to an MS medium containing 0.1 mg/l NAA+1.0 mg/l BAP, and in vitro rosmarinic acid production was enhanced by adding L-tyrosine (10 mg/l) and L-phenylalanine (10 mg/l) to the nutrient medium. The calli were cultured in these media for 1 and 2 months, and the production of rosmarinic acid in sage calli was analyzed using HPLC. The addition of amino acids to the medium significantly increased rosmarinic acid production in the calli. Furthermore, the results varied significantly with different amino acids and callus culture durations. Tyrosine was observed to be more effective in increasing rosmarinic acid production in sage calli. Moreover, calli cultured for 2 months in both tyrosine- and phenylalanine-supplemented media exhibited higher rosmarinic acid production compared to the control group cultured for only 1 month.