Micropropagation of Plantago media L. and the presence of phenolic compounds in organs of multiplied plants were investigated for the first time. Multiplication of plant material was achieved in shoot-tip cultures and via direct organogenesis on Murashige and Skoog (MS) medium with four variants of plant growth regulators (M1–M4). The best multiplication coefficient – 9.2 was obtained in seedling shoot-tip cultures on MS medium M3 with BA 0.2 mg/L and IAA 1.0 mg/L. Methanol extracts prepared separately from shoots and roots of in vitroderived plantlets were found to contain typical of the genus Plantago L. phenylethanoid glycosides as the only phenolics. Acteoside and plantamajoside were the major compounds – both known to possess a wide range of promising biological activities applicable for medicinal (therapeutic) and cosmetic uses. Martynoside, as a trace constituent, was also found for the first time in the studied species. The quantitative screening of the extracts by TLC video densitometric method showed a higher content of acteoside in shoots (range 62.43–93.03 mg/g, dry weight) and plantamajoside in roots (range 22.45–44.08 mg/g); the highest recorded values – 93.03 mg/g and 44.08 mg/g, respectively, were found in the organs obtained on MS medium M4 with BA 2.0 mg/L.
Abstract An efficient micropropagation protocol for production of genetically uniform clones of Eryngium campestre L. was developed. To determine the effect of nutritional and hormonal factors on shoot and root development and bioactive compounds production, three variants of media differing in the content of macro- and micronutrients, as well as plant growth regulators of various types and concentrations were tested. The highest regeneration (100%), with over 13 shoots per explant, was induced on Murashige and Skoog (MS) medium with 1.0 mg l−1 benzyladenine (BA) and 0.1 mg l−1 indole-3-acetic acid (IAA). The in vitro derived shoots multiplied through axillary bud formation were rooted and transferred to an experimental plot with 78% frequency of survival. Flow cytometry showed no variation in nuclear DNA between the seedlings and micropropagated plants. Preliminary thin layer chromatography (TLC) analysis indicated that phenolic acids, saponins, flavonoids and acetylenes were present in plant biomass. Ultra high performance liquid chromatography (UHPLC) analysis revealed that shoots and roots from in vitro derived plants and root cultures maintained the ability to produce rosmarinic acid (RA), rosmarinic acid hexoside (RA-HEX) and chlorogenic acid (CGA). The highest phenolic acid content was detected in roots of in vitro regenerated plants. The extract from those roots expressed the highest inhibitory effect against bacteria Staphylococcus aureus, as well as dermatophytes Trichophyton mentagrophytes and T. rubrum.