Selection of double-flowered plants at the seedling stage is one of the main purposes of stock breeding programs. Eight stock cultivars of Matthiola incana L. named 'Nobel', 'Cinderella', 'Pacific', 'Avalanche', 'Midblue', 'Lavender', 'Goddess' and 'Esfahan', with different percentage of double-flowered plants were used for examining the relationship with three morphological types of cotyledons. The results of a chi-square test indicated that in heartshaped (HC) and cup-shaped cotyledon (CC) populations, the number of plants with double flowers was much more than that of single flowers and CC seedlings rarely produced single flowers. Therefore, increasing the number of CC seedlings can improve the percentage of double flowers. The highest and lowest numbers of CC seedling were observed in high double and single flower cultivars, respectively. Single flower cultivars showed the maximum count of dumbbell-shaped cotyledons. Chromosome pairing of these cultivars was evaluated using the squash technique. Aneuploid cells were found in 'Nobel' and 'Goddess' cultivars, which showed the highest percentage of CC seedling. Based on morphological measurements, the highest value of inflorescence size was observed in the seedlings with cup-shaped cotyledons. The authors of the present research wish to acknowledge Mr. Momeni, the head of the cytology laboratory at Aburaihan College for his kindness in providing the required facilities and equipment for chromosome counting. Special thanks also go to Tehran University authorities and Sabzparvar Co. for financially supporting of this study.