Abstract
Triterpenoid saponins, which are glycosidic derivatives of oleanolic acid, demonstrate numerous pharmacological properties. The hairy root cultures of marigold accumulate these phytoanticipins mainly in vacuoles, which may contain up to 40% of the triterpene glycosides synthesized in the cytoplasm. Dimethyl sulfoxide, Tween 20 (T20), Tween 80 (T80), and Triton X-100 (Tx100) were used as potential surfactants, allowing for an increase in the release of saponin into the culture medium. T20 at concentrations of 0.3–1.0% (v/v) caused a higher saponin content in the liquid medium, from 52 to 61 times for CH9, 15 to 22 times higher for CC16, and seven to twelve times higher for CH2, compared to the respective control cultures. DMSO was efficient toward the CC16 line, providing a 3–7 times higher saponin content for 0.5% and 1.0% (v/v) surfactant concentrations, respectively. The suitability of Triton X-100 for triterpenoid saponin could not be determined with the method used in this experiment, and there was a serious contamination of the analyzed samples. The ultrasound method accelerated surfactant action, and only for the CH9 line did it result in an increase in the secretion of glycosides to almost three-fold in the case of T80 and over two-fold in the case of T20. The conditions that were least harmful for the roots were: ultrasound, Tween 80, and T80 interacting with US. The permeabilized cultures, after 30 days of growth in the new medium, obtained fresh biomasses similar to the control or a reduction by the maximum of one-fifth for CH9. For the second line (CC16), the growth parameters were reduced twice. Polyoxoethylene sorbitan monolaurate was found to be the most powerful surfactant, and a proposed concentration and time of action allowed for culture viability only for the CC16 line. The young parts of the root tips generated a new culture, with the growth being reduced by 77% (FW) and 82% (DW).
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