Prolonged dark period modulates the oxidative burst and enzymatic antioxidant systems in the leaves of salicylic acid-treated tomato

Salicylic acid (SA) is an important plant growth regulator playing a role in the hypersensitive reaction (HR) and the induction of systemic acquired resistance. Since the SA-mediated signalling pathways and the formation of reactive oxygen species (ROS) are light-dependent, the time- and concentration-specific induction of oxidative stress was investigated in leaves of tomato plants kept under light and dark conditions after treatments with 0.1 mM and 1 mM SA. The application of exogenous SA induced early superoxide- and H2O2 production in the leaves, which was different in the absence or presence of light and showed time- and concentration-dependent changes. 1 mM SA, which induced HR-like cell death resulted in two peaks in the H2O2 production in the light but the first, priming peak was not detected in the dark. Unlike 0.1 mM SA, 1 mM SA application induced NADPH oxidase activity leading to increased superoxide production in the first hours of SA treatments in the light. Moreover, SA treatments inhibited catalase (CAT) activity and caused a transient decline in ascorbate peroxidase (APX), the two main enzymes responsible for H2O2 degradation, which led to a fast H2O2 burst in the light. Their activity as well as the expression of some isoenzymes of SOD and APX increased only from the 12th h in the illuminated samples. The activity of NADPH oxidase and expression SlRBOH1 gene encoding a NADPH oxidase subunit was much lower in the dark. In spite of low CAT and APX activity after SA treatments in the dark, the activation of guaiacol-dependent peroxidase (POD) could partially substitute H2O2 scavenging activity of these enzymes in the dark, which reduced the ROS burst and development of lesion formation in the leaves.