Accumulation of reactive oxygen species and oxidation of cytokinin in germinating soybean seeds

Abstract

Seed germination is an important developmental switch when quiescent seed cells initiate oxidative phosphorylation for further development and differentiation. During early imbibition of soybean seeds (Glycine max L. cv. Weber), a superoxide dismutase (SOD) activity peak was observed, in embryonic axes, after 6 h imbibition. Peroxidase activities, including catalase, were significantly increased after 12 h inhibition and during germination phase III. Catalase was the most efficient enzyme in catabolizing H2O2 in embryonic axes. When stored at 42 degrees C and 100% relative humidity, seeds were stressed and lost their viability in a time-dependent manner. A significant increase in the Cu, Zn-superoxide-dismutase activity, and to a lesser extent, Mn superoxide dismutase activity was observed during germination in low-viability (stressed) seeds as compared to high-viability (unstressed) seeds. Northern blot analysis confirmed that superoxide dismutase induction resulted from an accumulation of its transcripts in response to the production of O2-. The induction of catalase did not occur in low-viability seeds, resulting in dramatic accumulation of H2O2. Using capillary electrophoresis, HPLC and NMR we found that the endogenous cytokinin, zeatin riboside, was present in large quantities in the high-viability seeds, but it was oxidized into adenine in the low-viability seeds. In vitro superoxide anion could also oxidize the cytokinin. Zeatin riboside, but not adenine, was found to act as a scavenger of superoxide anions and may help to maintain seed viability by detoxifying reactive oxygen species. Germination of stressed seeds was partially restored by the addition of exogenous cytokinin (zeatin riboside). Protection against oxidative stress by cytokinin seemed to be a general phenomenon, as Escherichia coli cells were also protected against superoxide stress in the presence of cytokinin.