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Imbibition with H2O2 induced changes in the levels of ASC and GSH. Whereas H2O2 pre-treatments did not produce any change in reduced ASC, a significant decrease in reduced GSH was observed (Table?3). Regarding the oxidized forms of both anti-oxidants, a difference in behaviour was noticed: ASC experienced an oxidation (increase in DHA) that was progressive with the H2O2 concentration, whereas oxidized GSH did not accumulate in pea seedlings, and a decrease was even produced by the highest H2O2 concentration used in the experiment. The accumulation of DHA brought about a decrease in the redox state of ASC. A slight decrease in the redox state of GSH in http://www.selleckchem.com/products/INCB18424.html the pea seedlings was produced by the H2O2 pre-treatment (5?10?mm), the effect being due to the decrease in GSH (Table?3). Similar to the studies of the effect of H2O2 on the non-enzymatic anti-oxidants, we also used pea seedlings incubated in darkness for analysis of the endogenous contents of H2O2. The imbibition of pea seeds with H2O2?had increased its concentration in pea seedlings after 48?h of growth in darkness. http://www.selleck.cn/products/VX-770.html However, the increase was only statistically significant when 20?mm H2O2 was used. Under these conditions, a threefold increase in H2O2 was recorded both on a fresh weight and on a protein basis (Fig.?3a). When pea seedlings were stained with DAB reagent to locate H2O2 production, a red�Cbrown staining was observed. The red-brown intensity was progressive with the H2O2 concentration used in the seed imbibition (Fig.?3b, d�Cg). This staining seemed to be due to H2O2 as it was suppressed totally by 10?mm ASC (Fig.?3c,h). In order to study the effect of H2O2-imbibition of pea seeds on the differential protein expression from the pea seedlings, a proteomic approach was applied (Fig.?4). Because the more important changes in plant growth were produced when pea seeds were incubated in darkness, and particularly with the highest H2O2 concentration, this proteomic approach was carried out for crude extracts of pea seedlings developing from seeds imbibed in http://www.selleckchem.com/products/SB-431542.html the presence or absence of 20?mm H2O2 and grown in darkness for 48?h. After 2D electrophoresis and gel analysis, the spots whose intensity increased or decreased significantly, based on anova analysis (P?