Application of Gamma Aminobutyric Acid Treatment on Frost Damage and Antioxidant Properties of Cavendish Banana (Musa acuminata cv. Cavendish) during Cold Storage

Introduction Fresh fruits and vegetables are physiologically active and perishable after harvest. Continued metabolic processes such as transpiration or respiration may significantly affect their quality and thus shorten their useful life. Since keeping at low temperatures and without freezing for a long time effectively reduces the physiological activity of the products, it can be used as a strategy to maintain the quality of the products and increase their life after harvesting. Among the most economically important tropical fruits, ripe green bananas are very sensitive to cold and when stored below the threshold temperature, they show all the symptoms of frost damage. While banana cultivars, maturity stage, and ripening all influence cold sensitivity, there's a growing interest in extending the shelf life of produce using natural, plant-friendly compounds. Gamma-aminobutyric acid (GABA), a naturally occurring four-carbon, non-protein amino acid found in plants, animals, and bacteria, is a promising candidate in this area.Abiotic stresses such as cold, heat, drought, ultraviolet rays and low oxygen can cause the accumulation of GABA in plants. Generally, the purpose of this research was to investigate the effect of gamma-aminobutyric acid treatment to improve freezing and antioxidant properties of Cavendish banana at 5 degrees Celsius for 24 days in 90% relative humidity.   Materials and Methods Cavendish banana fruits (Musa acuminata cv. Cavendish) at the time of commercial maturity (ripe green) were obtained from a banana garden in Kerman and immediately transferred to the horticultural science laboratory of Shahid Bahoner University, Kerman. Healthy and uniform fruits were selected in terms of size, shape, color, and degree of ripening, and after washing with water and drying them, frost tolerance, malondialdehyde, and antioxidants were measured for zero day. GABA (Sigma-Aldrich, USA) required after weighing was dissolved in water and prepared in two concentrations of 2.5 and 5 mM. The fruits were divided into three groups of 54 and each repetition included 18 fruits. The first and second groups were immersed in GABA solution of 2.5 mM and 5 mM for 5 minutes, respectively. The third group was immersed in distilled water for 5 minutes and was used as a control (Khaliq et al., 2023). Each treatment was repeated three times. Then, all the fruits were dried in the air for one hour and kept for 24 days at 5 degrees Celsius and relative humidity of 85-90%. Biochemical observations were measured on days 0, 4, 8, 12, 16, 20 and 24 of storage.     Results and Discussion The results of this research showed that frost damage gradually increased during the storage period and the control fruits showed significantly more frost damage symptoms than the fruits treated with GABA. GABA treatments of 2.5 and 5 mM at the end of the storage period reduced the amount of frost damage by 55.64 and 69.95%, respectively, compared to control fruits. As shown in Figure 1b, MDA content as an index of membrane lipid peroxidation in the control and GABA-treated fruit showed an upward trend, which was associated with the destruction of banana fruit membrane under cold stress. Compared to control, banana fruits treated with GABA showed lower MDA accumulation during the entire storage period at 4 degrees Celsius. On the last day of storage, GABA treatment with a concentration of 2.5 mM and 5 mM reduced the amount of MDA in banana fruits by 30.99% and 59.80%, respectively, compared to the control. Post-harvest treatment with GABA reduced frostbite, ion leakage and MDA levels in banana fruits, thereby maintaining fruit quality during low temperature storage. GABA treatment increased the activity of catalase (CAT), ascorbate peroxidase (APX), peroxidase (POD) and superoxide dismutase (SOD) enzymes in banana fruit compared to the control under low temperature storage. The activity of antioxidant enzymes CAT, APX, POD and SOD increased significantly until the 20th day of storage at low temperature, especially in the 5 mM GABA treatment compared to the other two treatments, and then decreased slightly at the end of the storage period. An increase in the concentration of oxygen free radicals, including hydrogen peroxide, leads to an increase in catalase enzyme activity. Catalase enzyme is often present in the peroxisome and causes the decomposition of hydrogen peroxide into water and oxygen. The specific activity of catalase enzyme increased during cold storage, especially in GABA treatments, so it seems that this enzyme is an efficient scavenger for removing hydrogen peroxide and thus causes better protection of cells against peroxidation. In the ascorbate-glutathione cycle, the ascorbate peroxidase enzyme reduces the amount of hydrogen peroxide by using ascorbate as an electron donor. In the present study, the activity of ascorbate peroxidase enzyme in GABA treatment was significantly higher than the control, which indicates the importance of the role of ascorbate peroxidase in plant tissues against oxidative damage. Guaiacol peroxidase enzyme is another antioxidant enzyme that decomposes hydrogen peroxide into water and oxygen. Peroxidase enzyme plays a role in the oxidation of precursors of phenolic compounds, lignin production, and removal of free radicals. The activity of peroxidase enzyme showed a similar trend in all three treatments, although its activity in GABA treatments was more than the control. Therefore, this enzyme effectively eliminated free radicals in banana fruits. In confirmation of these findings, it was reported in research that the activity of peroxidase enzyme increased in fir cuttings during the cold period. In research, post-harvest treatment of GABA with a concentration of 5 mM reduced frostbite and increased the activity of antioxidant enzymes such as CAT, APX, POD and SOD in peach fruits.   Conclusion The results of this research showed that the applied post-harvest treatments reduced the signs of frostbite and preserved the antioxidant properties of banana fruits. Among the treatments, 5 mM concentration of GABA was the most effective treatment in the storage period. Therefore, GABA treatment can be used as a practical solution to reduce frostbite and preserve the antioxidant properties of Cavendish bananas during long-term storage.