This research aimed to investigate the effect of different concentrations and treatment durations of Ethyl Methanesulfonate (EMS) on the morphophysiological traits and genetic variations of spinach (Spinacia oleracea L.) ‘Viroflay’ cultivar. The experiment was conducted during the years 2022-2023 in the horticultural greenhouse of the University of Kurdistan, using a factorial experiment in a completely randomized design. Spinach seeds were treated with four concentrations of EMS: 0 (control), 0.25%, 0.5%, and 1%, and exposed for two treatment times: 75 and 150 minutes. After thorough seed washing, cultivation was carried out under uniform greenhouse conditions. Morphological traits, including plant height, shoot fresh weight, and shoot dry weight, were measured. Furthermore, several phytochemical indices were evaluated, such as flavonoid content, Vitamin C, antioxidant capacity, nitrate, oxalic acid, and mineral elements (sodium and potassium). To assess genetic variations, ISSR and SCOT molecular markers were employed to analyze DNA banding patterns.

The results indicated that increasing EMS concentration significantly reduced plant height, decreasing from 10.96 cm in the control treatment to 8.66 cm at a 1% concentration. Conversely, low EMS concentrations had a positive effect on biomass accumulation. The highest shoot fresh weight (15.56 g) and shoot dry weight (1.76 g) were observed at the 0.25% EMS concentration. Phytochemical indices were also affected by EMS; for instance, the highest ascorbic acid content (709.17 ppm) was observed at the 1% concentration. Additionally, the highest flavonoid content (13.17 mg/g FW) and antioxidant capacity (37.44 µmol/g FW) were recorded at the 0.25% EMS concentration. Molecular analysis results revealed that EMS treatment altered DNA banding patterns, leading to the appearance of new bands and the disappearance of some existing ones, suggesting an increase in genetic diversity. Overall, the findings of this study suggest that controlled application of EMS at low concentrations can be an effective method for inducing beneficial mutations and enhancing genetic diversity in the ‘Viroflay’ spinach cultivar.This research aimed to investigate the effect of different concentrations and treatment durations of Ethyl Methanesulfonate (EMS) on the morphophysiological traits and genetic variations of spinach (Spinacia oleracea L.) ‘Viroflay’ cultivar. The experiment was conducted during the years 2022-2023 in the horticultural greenhouse of the University of Kurdistan, using a factorial experiment in a completely randomized design. Spinach seeds were treated with four concentrations of EMS: 0 (control), 0.25%, 0.5%, and 1%, and exposed for two treatment times: 75 and 150 minutes. After thorough seed washing, cultivation was carried out under uniform greenhouse conditions. Morphological traits, including plant height, shoot fresh weight, and shoot dry weight, were measured. Furthermore, several phytochemical indices were evaluated, such as flavonoid content, Vitamin C, antioxidant capacity, nitrate, oxalic acid, and mineral elements (sodium and potassium). To assess genetic variations, ISSR and SCOT molecular markers were employed to analyze DNA banding patterns.

The results indicated that increasing EMS concentration significantly reduced plant height, decreasing from 10.96 cm in the control treatment to 8.66 cm at a 1% concentration. Conversely, low EMS concentrations had a positive effect on biomass accumulation. The highest shoot fresh weight (15.56 g) and shoot dry weight (1.76 g) were observed at the 0.25% EMS concentration. Phytochemical indices were also affected by EMS; for instance, the highest ascorbic acid content (709.17 ppm) was observed at the 1% concentration. Additionally, the highest flavonoid content (13.17 mg/g FW) and antioxidant capacity (37.44 µmol/g FW) were recorded at the 0.25% EMS concentration. Molecular analysis results revealed that EMS treatment altered DNA banding patterns, leading to the appearance of new bands and the disappearance of some existing ones, suggesting an increase in genetic diversity. Overall, the findings of this study suggest that controlled application of EMS at low concentrations can be an effective method for inducing beneficial mutations and enhancing genetic diversity in the ‘Viroflay’ spinach cultivar.

Additionally, the highest flavonoid content (13.17 mg/g FW) and antioxidant capacity (37.44 µmol/g FW) were recorded at the 0.25% EMS concentration. Molecular analysis results revealed that EMS treatment altered DNA banding patterns, leading to the appearance of new bands and the disappearance of some existing ones, suggesting an increase in genetic diversity. Overall, the findings of this study suggest that controlled application of EMS at low concentrations can be an effective method for inducing beneficial mutations and enhancing genetic diversity in the ‘Viroflay’ spinach cultivar.