Evaluation of HSYA and Carthamin Accumulation in Safflower ( <scp> <i>Carthamus tinctorius</i> </scp> L.) Induced by Chemical Mutagens (EMS and SA) in M ₁ and M ₂ Generations

ABSTRACT Historically cultivated for medicinal and dyeing purposes, safflower ( Carthamus tinctorius L.) saw a shift towards widespread oilseed production after the 1920s due to the growing importance of oils. However, there is a renewed interest in its medicinal and dyeing properties today. Consequently, there is a critical need to develop new safflower lines with enhanced pigment content to meet the increasing industrial and pharmaceutical demand. This study aims to investigate the effect of chemical mutagens (ethyl methanesulfonate and sodium azide) on pigment (hydroxysafflor yellow A (HSYA) and carthamin) formation in safflower in M 1 and M 2 generations. The flower (intact and powder) colour values of HSYA and carthamin contents were analysed by comparing HSYA and carthamin values across generations. The correlation between HSYA and carthamin content and flower colour value was investigated by comparing HSYA and carthamin values across generations. The highest HSYA content was achieved in both generations with a 0.2% EMS treatment for 6 h, reaching 20.99 mg/g in the M 1 and 17.28 mg/g in the M 2 generation. The change in carthamin content of M 1 generation safflower flowers was not statistically significant. In contrast, the highest carthamin content in M 2 generation flowers was observed with SA 0.3% 4 h mutagen treatment (0.1 mg/g). Lightness (L) values were significantly positively correlated with HSYA concentration ( p &lt; 0.01). Increased HSYA content in safflower flowers is typically associated with higher L and b values, corresponding to a yellower and brighter appearance.