A novel biotechnological source of secondary metabolites: Phylogenetic and biochemical analysis of callus and cell suspension cultures of Cirsium helenioides (L.) Hill
摘要
Cirsium helenioides (L.) Hill is a species with promising potential as a source of plant secondary metabolites. In the present study, callus and cell suspension cultures of C. helenioides were successfully established and optimized in vitro, and their capacity for secondary metabolite accumulation was systematically evaluated. Species identity was confirmed using nuclear ITS and chloroplast rbcL markers, combined with phylogenetic analysis. Callus cultures maintained under dark conditions presented the highest levels of metabolite accumulation, although the maximal values were dependent on the explant origin and phytohormonal composition of the culture medium. The highest total flavonoid content was detected in cotyledon-derived callus cultures in darkness on Murashige and Skoog medium supplemented with 1.0 mg L−1 2,4-D and 0.5 mg L−1 BAP, reaching 5.94 ± 0.15 mg g−1 DW. The maximum total phenolic content was observed in leaf derived callus cultures grown in darkness on MS medium supplemented with 2.0 mg L−1 NAA and 0.5 mg L−1 BAP, with a value of 27.20 ± 0.56 mg GAE g−1 DW. The highest polysaccharide accumulation was recorded in root derived callus cultures in darkness on MS medium supplemented with 1.0 mg L−1 2,4-D and 0.5 mg L−1 BAP, reaching 76.2 ± 1.23 mg g−1 DW. In all the cases, the metabolite concentrations in the callus cultures were several-fold greater than those detected in the intact in vitro grown plants, whereas the concentrations in the suspension cultures were lower than those in the solid medium callus. Overall, the results demonstrated that C. helenioides callus cultures represent an efficient and controllable in vitro system for the accumulation of phenolic compounds and polysaccharides, highlighting their potential for further biotechnological optimization and application.