Mitigating the toxicity of reactive oxygen species induced by cadmium via restoring citrate valve and improving the stability of enzyme structure in rice

The mechanism of reactive oxygen species (ROS) burst in rice cells induced by cadmium (Cd) stress remains poorly understood. The present study shows that the burst of superoxide anions (O2·-) and hydrogen peroxide (H2O2) in roots and shoots led by Cd stress was attributed to the disturbance of citrate (CA) valve and the damage of antioxidant enzyme structure in the rice seedlings. Cd accumulation in cells altered the molecular structure of superoxide dismutase (SOD), catalase (CAT) and peroxidase (POD) through attacking glutamate (Glu) and other residues, leading to the significant reduction of their activities in clearing O2·- and decomposing H2O2. Citrate supplementation obviously increased the activity of antioxidant enzymes and decreased ∼20-30% of O2·- and H2O2 contents in roots and shoots. Meanwhile, the synthesis of metabolites/ligands such as CA, α-ketoglutarate (α-KG) and Glu as well as the activities of related enzymes in CA valve were remarkably improved. The activities of antioxidant enzymes were protected by CA through forming stable hydrogen-bonds between CA and antioxidant enzymes, and forming the stable chelates between ligands and Cd. These findings indicate that exogenous CA mitigated the toxicity of ROS under Cd stress by the ways of restoring CA valve function to reduce the production of ROS, and improving the stability of enzyme structure to enhance antioxidant enzymes activity.