| Abstract [eng] |
Calendula officinalis L. is a medicinal herb, used in traditional medicine to treat various eye, skin and other health disorders. Various biologically active compounds can be found in the extracts of this plant, which provide anti-inflammatory and antiseptic properties. Due to extreme changes in temperature and humidity, pollution, plants accumulate metabolites, which are used in the manufacturing industry, unevenly. In vitro cultivation of callus cultures is a promising technology that allows to evaluate the synthesis of biologically active compounds in controlled environment and to improve their accumulation by using different growth regulators at various concentrations in the nutrient medium. The aim of the study was to investigate the phytochemical, antioxidant and antibacterial activity of Calendula officinalis L. in in vitro callus cultures and in vivo extracts. The studies used in vitro callus cultures of leaves, stems and roots of Calendula officinalis, grown in MS nutrient medium, supplemented with 0.5 mg/L thidiazuron (TDZ), 0.1 mg/L 3-indolylacetic acid (IAA), 0.5 mg/L L-tryptophan and 0.5 mg/L melatonin, as well as in vivo leaves and stems. The concentrations of phytochemical compounds and antioxidant activity were assessed spectrophotometrically, for which DPPH, ABTS, FRAP methods were applied and reducing properties were measured. Antibacterial activity was assessed against E. coli and B. subtilis by measuring the zone of inhibition. The results of the study show that certain combinations of growth regulators in the nutrient medium may affect the synthesis of biologically active compounds in callus cultures. The highest concentrations of phenolic compounds, phenolic acids, flavonoids, malondialdehyde, chlorophyll a and b were determined in in vivo leaf extracts. Extracts of in vitro callus cultures grown in medium supplemented with only TDZ and IAA phytohormones revealed higher anthocyanin concentrations, superoxide dismutase activity and reducing properties. Extracts of root callus cultures grown in medium supplemented with phytohormones and melatonin (0.5 mg/L) showed the highest L-proline concentration - 511.48 mol/g. The highest antioxidant activity, according to DPPH and FRAP methods, was assessed in extracts of root callus cultures, grown in medium with TDZ, IAA and L-tryptophan (0.5 mg/). ABTS method study showed that the antioxidant activity of almost all extracts reached 99%. Antibacterial studies revealed significant inhibition of E.coli and B.subtilis activity when extracts of in vitro callus cultures were used. Against E. coli, the largest inhibition zone was determined to be 1.57 cm, using extracts from leaf callus cultures grown in MS medium with phytohormones and melatonin (0.5 mg/L). Against B. subtilis, the greatest inhibitory effect was observed using in vivo stem extracts, but extracts from root callus cultures grown in medium, supplemented with phytohormones and melatonin (0.5 mg/L), also had significant inhibition. |
