Interaction between the signaling molecules hydrogen sulfide and hydrogen peroxide and their role in vacuolar H+ -ATPase regulation in cadmium-stressed cucumber roots.

Abstract

Vacuolar H+ -ATPase (V-ATPase; EC 3.6.3.14) is the main enzyme responsible for generating a proton gradient across the tonoplast. Under cadmium (Cd) stress conditions, V-ATPase activity is inhibited. In the present work, hydrogen sulfide (H2 S) and hydrogen peroxide (H2 O2 ) cross-talk was analyzed in cucumber (Cucumis sativus L.) seedlings exposed to Cd to explain the role of both signaling molecules in the control of V-ATPase. V-ATPase activity and gene expression as well as H2 S and H2 O2 content and endogenous production were determined in roots of plants treated with 100μM CdCl2 and different inhibitors or scavengers. It was found that H2 S donor improved photosynthetic parameters in Cd-stressed cucumber seedlings. Cd-induced stimulation of H2 S level was correlated with the increased activities of the H2 S-generating desulfhydrases. Increased H2 O2 and lowered H2 S contents in roots were able to reduce V-ATPase activities similar to Cd. H2 O2 and H2 S-induced modulations in V-ATPase activities were not closely related to the transcript level of encoding genes, suggesting posttranslational modifications of enzyme protein. On the other hand, exogenous H2 O2 raised H2 S content in root tissues independently from the desulfhydrase activity. Although treatment of control plants with H2 S significantly stimulated NADPH oxidase activity and gene expression, H2 S did not affect H2 O2 accumulation in roots exposed to Cd. The results suggest the existence of two pathways of H2 S generation in Cd-stressed cucumber roots. One involves desulfhydrase activity, as was previously demonstrated in different plant species. The other, the desulfhydrase-independent pathway induced by H2 O2 /NADPH oxidase, may protect V-ATPase from inhibition by Cd.