Biotechnological production of antitumoral phytochemicals in endemic Centaurea kilaea plant

This study focuses on Centaurea kilaea, a significant species widely used in traditional medicine and renowned for its notable biological activities. Particularly, its secondary metabolites with antitumor and antioxidant properties hold immense potential for pharmaceutical applications. The primary objective of this research is to optimize secondary metabolite production using callus and cell suspension cultures derived from C. kilaea, thereby investigating its biotechnological production potential and developing sustainable production methods. For callus induction, various hormone combinations and concentrations were tested, and the best results were obtained using a medium containing 1 mg/L 2,4-D. Under these conditions, approximately 5-fold increase in callus biomass was observed over a 12-week period. To produce secondary metabolites, Cell Suspension Cultures (CSC) were established, targeting the production of antitumor phenolic compounds. Analyses were conducted using media containing 1 mg/L BAP and 1 mg/L 2,4-D, with the highest metabolite production observed in the medium containing 1 mg/L 2,4-D. Analysis of callus and cell suspension cultures identified several target antitumor compounds, including 2,5-dihydroxybenzoic acid, ascorbic acid, caffeic acid, chlorogenic acid, eupatorine, fumaric acid, hispidulin 7-glucoside, indole-3-acetic acid, jaceosidin, lithospermic acid, p-coumaric acid, protocatechuic acid, pyrocatechol, salicylic acid, syringic acid, and vanillic acid. Furthermore, it was observed that ascorbic acid, caffeic acid, fumaric acid, indole-3-acetic Acid, p-coumaric acid, and salicylic acid were produced at higher levels in culture media compared to the natural plant. Conversely, compounds such as 2,5-dihydroxybenzoic acid, chlorogenic acid, eupatorine, jaceosidin, lithospermic acid, protocatechuic acid, pyrocatechol, syringic acid, and vanillic acid were found at higher concentrations in the natural plant than in the culture media. Considering these findings, optimizing the conditions for the production of antitumor compounds from C. kilaea can facilitate a more efficient exploration of its biotechnological potential. Future studies will play a crucial role in developing sustainable and high-yield production methods, paving the way for innovative pharmaceutical applications.