An Efficient Micropropagation Protocol for the Endangered European Shrub February Daphne (Daphne mezereum L.) and Identification of Bacteria in Culture

Daphne mezereum of the Thymelaeaceae family is a medicinal shrub occurring naturally in Europe and under legal protection in Poland. In the present study, a protocol developed for mass propagation of February daphne from nodal explants is presented. Micropropagation is one of the in vitro techniques that allow the preservation of rare and valuable plants by developing efficient methods for their propagation. In the proliferation stage, explants were cultured in the Woody Plant Medium (WPM) with different cytokinins, and in the rooting stage on the semi-solid WPM medium with perlite, indole-3-butyric acid (IBA) in various concentrations was used. The maximum proliferation rate (five shoots per explant) was observed on the medium containing 4.14 mu M of meta-Topolin (mT). When the regenerated shoots were rooted in vitro in the presence of IBA in various concentrations, that of 19.68 mu M induced the highest number of roots per shoot (6.63) and the maximal root length (2.15 cm). It is also worth remembering that plants are often colonized by different groups of microorganisms, which also affect the diversity of the ecosystem. The endophytic bacteria inhabiting the D. mezereum shoots are Mycobacterium.