Shoot proliferation is a very important micropropagation phase, decisive for economic efficiency of this method for a given taxon. To obtain a high multiplication ratio and a good quality of microshoots a detailed propagation protocol must be developed for particular species or even cultivars. Rhododendron ‘Kazimierz Odnowiciel’ is a relatively new cultivar distinguished by large, beautiful flowers and high frost resistance so there is a need to develop an efficient method of its propagation to satisfy a growing demand for this plant. The aim of the experiment was to evaluate effects of cytokinins: meta-Topolin (mT), zeatin (ZEA), 6-benzyladenine (BA), thidiazuron (TDZ), 2-isopentenyladenine (2iP), or the combination of 2iP+ZEA on proliferation of shoots in R. ‘Kazimierz Odnowiciel’ cultured on Anderson’s medium (AN). Biochemical changes in plant material affected by cytokinins during this phase of micropropagation were determined and occurrence of genetical changes was followed using ISSR markers. TDZ, ZEA or the combination of ZEA+2iP resulted in 100% explant regeneration. On the medium with TDZ or ZEA over two new shoots per explant were produced but the highest proliferation was attained on the medium containing ZEA+2iP – over three shoots per explant. Microshoots developed in this treatment had also the highest contents of chlorophyll, carotenoids and soluble sugars as well as the highest catalase activity. Microshoots formed on the medium with zeatin showed the lowest polymorphism (below 4%) relative to a stock plant.