Nitric oxide and hydrogen cyanide as regulating factors of enzymatic antioxidant system in germinating apple embryos

Short-term (3 or 6 h) pre-treatment of apple (Malus domestica Borkh.) embryos with nitric oxide (NO) or hydrogen cyanide (HCN) induces transient accumulation of reactive oxygen species (ROS) leading to dormancy removal and germination. We demonstrated that enhanced NO emission by apple embryos during early phase of germination “sensu stricto” is required for seed transition from dormant into non-dormant state, and may be described by the model of “nitrosative door”, analogous to “oxidative window”. Cellular ROS concentration, resulting from NO or HCN embryo pre-treatment, seems to be under severe control of antioxidant system. Activity of superoxide dismutase (SOD), catalase (CAT), glutathione reductase (GR), glutathione peroxidase (GPX) and total peroxidases (Prxs) was determined during NO and HCN-mediated germination “sensu stricto” of embryos. CAT and SOD activity increased transiently 24 h after embryos pre-treatment, while GR and Prx activity was stimulated mainly after 96 h. The most evident alterations were detected in GPX activity, being more than threefold stimulated by NO or HCN. Based on this results, we conclude that these reactive molecules act simultaneously crossing their signaling pathways and we propose that ROS, reactive nitrogen species, HCN at accurate level are essential during seed germination as signaling factors.