Calcium-calmodulin is required for abscisic acid-induced antioxidant defense and functions both upstream and downstream of H2O2 production in leaves of maize (Zea mays) plants

Using pharmacological and biochemical approaches, the role of calmodulin (CaM) and the relationship between CaM and hydrogen peroxide (H,0,) in abscisic acid (ABA)-induced antioxidant defense in leaves of maize (Zea mays) plants were investigated. Treatment with ABA or H2O2 led to significant increases in the concentration of cytosolic Ca2+ in the protoplasts of mesophyll cells and in the expression of the calmodulin 1 (CaM1) gene and the content of CaM in leaves of maize plants, and enhanced the expression of the antioxidant genes superoxide dismutase 4 (SOD4), cytosolic ascorbate peroxidase (cAPX), and glutathione reductase 1 (GR1) and the activities of the chloroplastic and cytosolic antioxidant enzymes. The up-regulation of the antioxidant enzymes was almost completely blocked by pretreatments with two CaM antagonists. Pretreatments with CaM antagonists almost completely inhibited ABA-induced H2O2 production throughout ABA treatment, but pretreatment with an inhibitor or scavenger of reactive oxygen species (ROS) did not affect the initial increase in the contents of CaM induced by ABA. Our results suggest that Ca2+-CaM is involved in ABA-induced antioxidant defense, and that cross-talk between Ca2+-CaM and H2O2 plays a pivotal role in ABA signaling.