Twenty cultivars of Matthiola incanagrowing in various areas with different percentages of double flowers were chromosomally investigated, using fixation and staining of root tip cells. The somatic chromosome number (2n = 2x = 14) in mitotic metaphase cells and quantitative karyotypic parameters were evaluated. Aneu-ploidy was observed in two cultivars with high percentage of double flowers. An extra chromosome was observed as a rod univalent or chain trivalent in PMCs at diakinesis stage in two cultivars. The satellites were found in four cultivars (mostly locating on short arms) and were differed in size (0.62 to 0.83 μm). Two chromosomal types (metacentric, sub-metacentric) formed six different karyotype formulas. The degree of karyotype asymmetry was studied using qualitative and quantitative methods. The most of stock cultivars were placed in 1A classes of Stebbins' karyotype asymmetry. Based on Ward’s method, the cultivars separated into three clusters. The results of PCA showed the first three principal components accounted for 95.50% ofthe total variability.