Glucose-6-phosphate dehydrogenase (G6PDH or G6PD) is the key regulatory enzyme in the oxidative pentose phosphate pathway (OPPP). The cytosolic isoforms including G6PD5 and G6PD6 account for the major part of the G6PD total activity in plant cells. Here, we characterized the Arabidopsis single null mutant g6pd5 and g6pd6 and double mutant g6pd5/6. Compared to wild type, the mutant seeds showed a reduced germination rate and root elongation under salt stress. The seeds and seedlings lacking G6PD5 and G6PD6 accumulate more reactive oxygen species (ROS) than the wild type under salt stress. Cytosolic G6PD (cy-G6PD) affected the expression of NADPH oxidases and the G6PD enzymatic activities in the mutant atrbohD/F, in which the NADPH oxidases genes are disrupted by T-DNA insertion and generation of ROS is inhibited, were lower than that in the wild type. The NADPH level in mutants was decreased under salt stress. In addition, we found that G6PD5 and G6PD6 affected the activities and transcript levels of various antioxidant enzymes in response to salt stress, especially the ascorbate peroxidase and glutathione reductase. Exogenous application of ascorbate acid and glutathione rescued the seed and root phenotype of g6pd5/6 under salt stress. Interestingly, the cytosolic G6PD negatively modulated the NaCl-blocked primary root growth under salt stress in the root meristem and elongation zone.
Keywords: NADPH oxidases; NaCl; germination; glucose-6-phosphate dehydrogenase; reactive oxygen species; root system architecture.