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This study is aimed at the responses of grapevine adventitious root explants to zinc (Zn 2+ ) excess. Within 24 h Zn 2+  induced oxidative burst in concentration-dependent manner. The time course analysis revealed biphasic response in superoxide (O 2–· ) production. Hydrogen peroxide (H2O2) accumulation rose gradually within 24 h. Enhanced Zn 2+  concentrations did not induce progression of cell death. Immediately upon Zn 2+  addition to the perfusion solution, root epidermal cells exhibited hyperpolarization of their electrical membrane potential (E M ), that was transient and independent of Zn 2+  concentration. The subsequent, transient depolarization of EM was concentration-dependent and its magnitude increased with increasing Zn 2+  concentration. After 24 h the EM in treated roots recovered and its values were identical with those of control roots. Membrane permeability of root cells increased in the roots treated with 5 mM Zn 2+  within 24 h while the lower concentrations did not show any impact on membrane permeability. Differences in protein expression pattern identified by proteomic approach involving antibody microarray expression profiling revealed Zn 2+ -induced upregulation of apoptosis-related protein dolichyl-diphosphooligosaccharide-protein glycosyltransferase subunit (DAD1), extracellular signal-regulated kinase 1/2 (ERK1/2), some antioxidant enzymes and structural proteins in the roots. Moreover, the proteins involved in plant defense mechanisms endochitinase I (CHIT 1) and phenylalanine ammonia-lyase (PAL) were down-regulated indicating a cross-talk between defense and heavy metal signaling pathways. Taken together, these results showed that the grape cultivar Limberger is highly Zn 2+ -responsive and could be used as a model plant for studying physiological and molecular responses to heavy metal excess.

Effects of ZnCl 2 on ROS generation, plasma membrane properties, and changes in protein expression in grapevine root explants