Potentilla alba L. is a valuable medicinal plant widely used in folk and traditional medicine and particularly promising in complex treatment of thyroid pathology. Natural resources of this species are insufficient due to ever-growing use in contemporary medicine. Comprehensive investigations of different P. alba populations are essential for the successful extension of P. alba plantings. Aiming for a better understanding of karyotype structure, chromosome behaviour in meiosis and developing new diagnostic characters, we performed molecular cytogenetic characterization and leaf structure and ultrastructure analyses of two introduced P. alba samples originating from different habitats. Based on chromosome morphology, distribution of 45S/5S rDNA and DAPI-banding patterns, all chromosomes in the karyotypes were identified and the P. alba chromosomal idiogram was constructed. Our findings confirmed P. alba karyotype stability and also revealed several diagnostic characters of this species: the features of cells of upper and lower leaf epidermis, the presence of calcium oxalate druses and three types of leaf indumentum, essential for evaluation of genetic diversity in different populations, validation of raw materials and further selection progress. The meiotic abnormalities were detected probably related to low pollen activity and indicated the advantages of vegetative propagation in the development of a P. alba plantation system. © 2018, Springer Science+Business Media B.V., part of Springer Nature.