Drought-induced H2O2 accumulation in subsidiary cells is involved in regulatory signaling of stomatal closure in maize leaves

Increasing H2O2 levels in guard cells in response to environmental stimuli are recently considered a general messenger involved in the signaling cascade for the induction of stomatal closure. But little is known as to whether subsidiary cells participate in the H2O2-mediated stomatal closure of grass plants. In the present study, 2-week-old seedlings of maize (Zea mays) were exposed to different degrees of soil water deficit for 3 weeks. The effects of soil water contents on leaf ABA and H2O2 levels and stomatal aperture were investigated using physiological, biochemical, and histochemical approaches. The results showed that even under well-watered conditions, significant amounts of H2O2 were observed in guard cells, whereas H2O2 concentrations in the subsidiary cells were negligible. Decreasing soil water contents led to a significant increase in leaf ABA levels associated with significantly enhanced O-2 (-) and H2O2 contents, consistent with reduced degrees of stomatal conductance and aperture. The significant increase in H2O2 appeared in both guard cells and subsidiary cells of the stomatal complex, and H2O2 levels increased with decreasing soil water contents. Drought-induced increase in the activity of antioxidative enzymes could not counteract the significant increase in H2O2 levels in guard cells and subsidiary cells. These results indicate that subsidiary cells participate in H2O2-mediated stomatal closure, and drought-induced H2O2 accumulation in subsidiary cells is involved in the signaling cascade regulating stomatal aperture of grass plants such as maize.