Fluctuation of oxidative stress indicators in Salix nigra seeds during priming

Salix nigra seeds subjected to increased humidification show a decrease in normal germination (NG) during early imbibition followed by a recovery in that parameter at increasing imbibition times. Since photo-oxidized seeds contain high levels of reactive oxygen species (ROS), it is possible to infer that the atypical decrease in NG is a consequence of a higher ROS mobilization at early imbibition and the subsequent recovery from an increase in antioxidant activity. In this study, several oxidative stress indicators were evaluated in photo-oxidized seeds subjected to priming. ROS production was studied using electronic spin resonance spectroscopy, spontaneous chemiluminescence (SCL), spectrophotometry (with XTT), and histochemical (with DAB and NBT) and cytochemical (with CeCl(3)) techniques. Four indicators of molecular damage were monitored: lipid peroxidation, pigment destruction, protein oxidation, and membrane integrity. Antioxidant activity was evaluated by changes in the enzymes SOD, CAT, APX, and POX. The results revealed that the decrease in NG at the beginning of priming occurs by an oxidative burst, as determined by increases in both SCL and superoxide anion radical (O2(·-)) Such oxidative burst generates lipid peroxidation, protein oxidation, and a decrease in both pigment content and enzyme activities. With increasing hydration, damages are progressively reversed and NG restored, which coincides with the increased activity of antioxidant defences. It is proposed that these novel observations regarding the occurrence of an oxidative burst are related to the high basal ROS levels and the high membrane content retained in the mature embryo tissues.