NADPH oxidase-dependent hydrogen peroxide production, induced by salinity stress, may be involved in the regulation of total calcium in roots of wheat

SummaryHydrogen peroxide (H2O2) is often generated by cells and tissues under environmental stress. In this work, we provide evidence that plasma membrane (PM) NADPH oxidase-dependent H2O2 production might act as an intermediate step in the NaCl-induced elevation of calcium (Ca) in roots of wheat. Remarkable increases in the content of total Ca were observed not only in roots exposed to NaCl but also in roots of seedlings exposed to exogenous H2O2. In roots, H2O2 production increased upon exposure to salt stress. PM vesicles were isolated from roots, and NADPH oxidase activity was determined by measuring superoxide anion (O2−) production. NADPH oxidase-dependent O2− production was 11.6 nmol mg−1 protein min−1 in control vesicles, but 19.6 nmol after NaCl treatment (24 h), indicating that salt stress resulted in the activation of the PM NADPH oxidase. Furthermore, the NaCl-induced increase in total Ca was partially abolished by the addition of 150 U/mL catalase (CAT), a H2O2 scavenger, and also by 10 μM diphenylane iodonium (DPI), a NADPH oxidase inhibitor. This data suggest that NADPH oxidase-dependent H2O2 production might be involved in the modulation of the Ca content in wheat roots. In conclusion, our results show that salinity stress increases the total Ca content of wheat roots, which is partly due to PM NADPH oxidase-dependent H2O2 generation.