Small pasque flower is a perennial species widely distributed in Central and Eastern Europe; the occurrence is rare, however. Due to environmental changes natural habitats of small pasque flower populations are endangered. Current populations consist often predominantly of old plants, indicating that the germination and the development of young plants is faulty. So far, causes that lie behind are unclear. In vitro propagation could be a strategy for preservation and multiplication of small pasque flower. Plants in vitro could easily be multiplied on RV medium (MS + 0.009 mg L-1 + 1.15 mg L-1 BAP). For rooting a higher amount of auxin is needed. It was found that on ½ MS salts + MS vitamins + 1.5 mg L-1 IAA almost all explants developed roots. However, the in vitro roots are thickening, fragile and without adventitious roots. Up to 83 % of plants could be transferred to soil. After a period in the greenhouse plants grew up in an open field. The plants were evaluated morphologically for two growing seasons. They looked phenotypically very uniform. Flow cytometry did not show any changes in the ploidy. Therefore, using the proposed protocol true to type, small pasque flower plants suitable for replanting in the natural habitat are produced.