The aim of the study was to examine the possibility of propagating in vitro four of the most common cultivars in Tuscany (central Italy): Terom, Violetto di Toscana, Chiusure and Empolese. The first three belong to the “Violetti” group, while cv Empolese belongs to the “Romaneschi” group. Explants were cultured on an induction medium (IM), which is a modified MS medium consisting of nitrate concentrations reduced by one quarter, 0.8 mg L−1 6-benzylaminopurine (BA) and 0.2 mg L−1 3-indole butyric acid (IBA). Explants were then transferred to a proliferation medium (PM) consisting of the same basal medium together with 0.03 mg L−1 BA and 0.05 mg L−1 gibberellic acid (GA3). A rooting double-phase was then established. The pre-rooting medium (PRM), consisting of a basal MS medium with half strength nitrate concentrations, 0.5 mg L−1 indole-3-acetic acid (IAA) and 1 mg L−1 paclobutrazol (PBZ) was used for two weeks. Over the next four weeks, a rooting medium (MR) was used, consisting of a basal MS medium with 2 mg L−1 β-cyclodextrin and 2 mg L−1 α-naphthaleneacetic acid sodium salt (NAA). The cv Empolese provided the highest number of proliferated explants and rooted plantlets using the method described.
Curcuma longa L. (turmeric) is one of the most important spice and safe food additives. Its main constituents, curcuminoids, showed anti-inflammatory, antitumor and antioxidant effects. In the present work, an in vitro propagation method was developed to achieve selected plant organs with quantified curcuminoid content. In vitro plants were obtained from sprouting buds as primary explants. The major curcuminoid constituents, such as curcumin (CUR), demethoxycurcumin (DEM), and bis-demethoxycurcumin (bis-DEM) were examined in different organs by LC-DAD-ESI-MS. A significant production of curcumin (more than 260 µg g-1 fresh weight) was obtained from in vitro microrhizomes, especially grown in a Murashige and Skoog medium (MS) supplemented with kinetin (0.1 mg L-1), -naphthaleneacetic acid (NAA, 1 mg L-1), sucrose (6%), agar (5%) and activated charcoal (0.1%). The analyzed microrhizomes showed reduced amounts of DEM and bis-DEM in comparison with CUR levels. In addition a shoot culture line was suitable to biosynthesize curcuminoids, in a ratio very similar to that identified in the fresh rhizomes of parent plants. This study represents the first direct quantification of curcuminoids in turmeric in vitro shoots and microrhizomes to be used in dietary supplements.
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