On the basis of karyotypic analysis performed by conventional staining and Giemas C-banding technique, cytological relationship was inferred for 21 lines of Paeonia lactiflora Pal. cultivated in Korea. It was very difficult to infer their organized karyotypic classification system using the composition of somatic chromosomes involving sat-chromosomes, relative length of chromosomes, arm ratio and karyotypic formulae by conventional staining. From the distribution and number of Giemsa C-bands on the chromosomes b and c, 21 lines can be subclassified into 5 groups. It seems that the karyotypic polymorphism is observed in 21 lines of cultivated P. lactiflora because peony mainly propagates by outbreeding.