| Abstract [eng] |
Medicinal plants are a valuable source of biologically active compounds that are widely used in pharmaceuticals, food additives, cosmetics, and nanoparticle synthesis. Secondary metabolites have complex chemical structures and pharmacological properties. The widespread use of these biologically active compounds in many industrial sectors creates a need for increased production of secondary metabolites. Biotechnological tissue culture techniques are used to preserve plants and improve metabolite biosynthesis and modify synthesis pathways. The aim of the study was to determine and compare the variation and biological activity of phytochemical compounds produced in in vitro callus cultures of Calendula grown in media with different phytohormones and additives and in vivo extracts. The study was carried out using cultures of marigold leaf, stem and root callus grown in MS media supplemented with different cytokinins (0.5 mg/l BAP or 0.5 mg/l TDZ) and auxins (0.5 mg/l NAA or 0.1 mg/l IAA). To obtain higher concentrations of phytochemical compounds and stronger antioxidant and antibacterial effects, the nutrient media were also supplemented with 10 mg/l ZnO and 1.25 g/l L-phenylalanine and L-tryptophan. The antioxidant activity of Calendula was evaluated by DPPH, FRAP and reducing power assays. The antioxidant enzyme activity of Calendula callus culture extracts was also evaluated. No significant effect of the different phytohormones was detected, but the addition of 10 mg/l ZnO and 1.25 g/l amino acids promoted a higher antioxidant effect and antioxidant enzyme activity of Calendula cultures. Studies have shown that higher concentrations of chlorophyll a and b, carotenoids, anthocyanins, and L-proline were obtained in the extracts of callus cultures grown with 0.5 mg/l BAP and 0.5 mg/l NAA phytohormones. Higher concentrations of phenolic compounds and phenolic acids were detected in the extracts of callus cultures grown on media containing 0.1 mg/l IAA and 0.5 mg/l TDZ. It was also found that calamus cultures grown with 0.5 mg/l BAP and 0.5 mg/l NAA phytohormones were subjected to higher oxidative stress. In vivo extracts showed higher concentrations of phenolic compounds, phenolic acids, flavonoids and anthocyanins, as well as ascorbate peroxidase activity, compared to in vitro callus culture extracts. The highest inhibitory effect against Escherichia coli was found in the extract of leaf callus cultures grown with 0.5 mg/l BAP and 0.5 mg/l NAA phytohormones (0.107±0.008 cm). |
