The effects of culture media and cytokinin types on micropropagation of mature Crataegus aronia L. were investigated. Using single-axillary bud explants, the growth of cultures on MS, WPM, DKW and NRM containing 4.44 lM benzyladenine (BA) plus 0.05 lM indole-3-butyric acid (IBA), and on NRM containing thidiazuron, metaTopolin (mT) or BA at 1.25, 2.5, 5.0 or 7.5 lM plus 0.05 lM IBA were compared. The culture medium had significant effects on shoot number and length. In comparison with MS, DKW and WPM, shoot production was greater on NRM (5.7 shoots per explant). Shoot production on MS, DKW and WPM (4.2, 4.2 and 4.1, respectively) were statistically similar to each other. Thidiazuron was detrimental to shoot formation and caused formation of rosette shoots and/or large callus to form on explants. In the presence of mT, only some of the explants developed into shoots. Benzyladenine was the only cytokinin that promoted both shoot proliferation and shoot elongation. Higher shoot numbers were obtained at 5.0 and 7.5 lM BA compared to lower concentrations of BA. Over 80% of microshoots rooted and rooted shoots were successfully acclimatized to ex vitro conditions.
Phytochemical investigation of Cephalaria tuteliana (Caprifoliaceae) endemic to Turkey was carried out for the first time. Thirteen compounds were isolated, including 2 triterpenoid sapogenins, 10 saponins, and 1 iridoid glycoside. This is the first report of the isolation of pomolic acid, tormentic acid, and 3- O-α-l-rhamnopyranosyl-(1→3)- β-d-glucopyranosyl hederagenin, not only from the Cephalaria genus, but also from the Caprifoliaceae family. The structures of the isolated compounds were elucidated by a combination of spectroscopy, including one- and two-dimensional nuclear magnetic resonance, and by comparison with the reported data in the literature. Two triterpene sapogenins, hederagenin and oleanoic acid, the latter being the most typical sapogenin of the oleanane type of triterpene saponins, were evaluated by the MTT method for their cytotoxic effect against A549, HeLa, PANC1, and SHSY5Y cancer cells, and the noncancerous HEK293 cell line. The cytotoxic activity of triterpene aglycones was discussed in detail and a structure-activity relationship was established. C. tuteliana may be considered as a useful source for developing new cytotoxic and immunotherapeutic agents due to its biologically active saponin content.
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