Abstract
Three cotyledon-derived callus lines of Harpagophytum procumbens were established on agar-solidified Schenk and Hildebrandt (SH) medium supplemented with 0.2mgL−1 α-naphthaleneacetic acid (NAA) and 1mgL−1 6-benzylaminopurine (BAP) (CNB line), 0.5mgL−1 picloram (CP0.5 line) or 2mgL−1 picloram (CP2 line). They were maintained in three different culture vessels: culture tubes, Erlenmeyer flasks and Magenta vessels. Hormonal treatment and type of culture vessel influenced callus growth and morphology. The highest biomass (64- to 130-fold increase in fresh and dry weights within 4weeks) was achieved in Erlenmeyer flasks. The contents of major phenylethanoid and iridoid glycosides in the calli were determined by high performance liquid chromatography (HPLC). Harpagoside, harpagide, verbascoside and isoverbascoside were identified and the ability to produce these bioactive compounds sustained for over 3years. H. procumbens calli were also analyzed by colorimetric methods for total contents of phenolics, flavonoids and anthocyanins. The extracts of CNB callus had the highest antiradical activity against DPPH and ABTS and capacity to reduce metal ions (Ferric Reducing Antioxidant Power—FRAP). The same extracts were also characterized by the highest total content of phenolics and flavonoids.
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