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Reactive oxygen species (ROS) originate as a natural byproduct in standard metabolism of oxygen activities. The principal sites of ROS generation in the cell are apoplast, mitochondria, chloroplasts, and peroxisomes. These ROS can induce cellular injuries by proteins oxidation, lipid peroxidation, and DNA damage, which finally may result in plant cellular death. Under regular circumstances, there is a steadiness between generation and elimination of ROS, but this balance is hampered by different biotic and abiotic stress factors such as exposure to heavy metals, high and low-light conditions, pathogens, insects and temperature extremes, resulting in a high generation of ROS which should be counteracted by the antioxidant machinery in cells. The antioxidant system of defense is composed by two groups: (i) Enzymatic antioxidants such as superoxide dismutase (SOD), catalase (CAT), ascorbate peroxidase (APX), general peroxidases (PRX) (e.g. guaiacol peroxidase GPX), glutathione reductase (GR), monodehydroascorbate reductase (MDHAR), and dehydroascorbate reductase (DHAR); (ii) Non-enzymatic antioxidants such as ascorbic acid (AA), reduced glutathione (GSH), α-tocopherol, carotenoids, plastoquinone/ubiquinone and flavonoids. These two groups of metabolites and enzymes work together with the main aim of ROS scavenging, but also in determining plant signaling, immune response, and plant growth and development. Finally, the molecular genetics of ROS genes and related metabolic pathways are briefly outlined, including gene isoforms, cellular localization, detection methods used and interactions amongst them. This information is crucial in better understanding and designing procedures for plants´stress tolerance; leading to a better management of agricultural plants under challenging and changing climatic conditions and food security.