Abstract
The goal of this study was to establish an efficient protocol for the large-scale propagation of Mertensia maritima (L.) Gray, and evaluate the carotenoid, fatty acid, and tocopherol contents in the leaves of in vitro regenerated shoots. Surface-disinfected node and shoot tip explants were placed on semisolid Murashige and Skoog (MS) medium with 0-16 µM N6-benzyladenine (BA), kinetin, (KN), and thidiazuron (TDZ) alone, or in combination with, 1 or 2 µM α-naphthaleneacetic acid (NAA). Of the three different cytokinins employed, TDZ elicited the best results for axillary shoot proliferation. A maximum frequency of shoot initiation above 84%, with a mean of 8.9 and 4.8 shoots per node and shoot tip, respectively, was achieved on the culture medium supplemented with 4 µM TDZ. A combination of TDZ + NAA significantly increased the percentage of multiple shoot formation and number of shoots per explant. The best shoot induction response occurred on MS medium with 4 µM TDZ and 1 µM NAA. On this medium, the node (93.8%) and shoot tip (95.9%) explants produced an average of 17.7 and 8.6 shoots, respectively. The highest root induction frequency (97.4%) and number of roots per shoot (25.4), as well as the greatest root length (4.2 cm), were obtained on half-strength MS medium supplemented with 4 µM indole-3-butyric acid (IBA). The presence of six carotenoids and α-tocopherol in the leaf tissues of M. maritima was confirmed by HPLC. Gas chromatography-mass spectrometry analysis confirmed the presence of 10 fatty acids, including γ-linolenic acid and stearidonic acid in the leaf tissues of M. maritima. All-E-lutein (18.49 μg g-1 fresh weight, FW), α-tocopherol (3.82 μg g-1 FW) and α-linolenic acid (30.37%) were found to be the significant compounds in M. maritima. For the first time, a successful protocol has been established for the mass propagation of M. maritima with promising prospects for harnessing its bioactive reserves.