Cold stress is one of the most important causes of reduced crop production and result in a lot of damages every year. In order to investigate the effect of gamma aminobutyric acid (GABA) spray on physiological and biochemical traits and improvement of cold tolerance, a study was conducted on Bidaneh Sefid and Hosseini cultivars factorial in a completely randomized design at the Urmia University. In this experiment, GABA was sprayed at four concentrations (0, 15, 30 and 60 mM) during 10-15 leaf stage in two cultivars (Bidaneh Sefid and Hosseini). The treated seedlings were then placed in a growth chamber at 4 °C and sampling of young and fully developed leaves was performed at defined times (0, 3, 6, 12 and 24 hours) and biochemical parameters such as total phenol, total soluble carbohydrate, antioxidant capacity, lipid peroxidation, proline content and activity of enzymes such as catalase, guaiacol peroxidase and ascorbate peroxidase were investigated. The results showed that the most positive impact against deleterious effects of cold was observed in 60 mM GABA at 24 hours after cold treatment compared to other treatments in both cultivars. The results also showed that the highest total soluble carbohydrate, total phenol, and proline content and the lowest lipid peroxidation were observed at 12 and 24 hours after cold treatment in Hosseini cultivar. The use of this compound increases plant signaling and prevents severe damage by removing free radicals during stress.