Cytosolic glyceraldehyde-3-phosphate dehydrogenase 2/5/6 increase drought tolerance via stomatal movement and reactive oxygen species scavenging in wheat

Drought is a major threat to wheat growth and crop productivity. However, there has been only limited success in developing drought-hardy cultivars. This lack of progress is due, at least in part, to a lack of understanding of the molecular mechanisms of drought tolerance in wheat. Here, we evaluated the potential role of three cytosolic glyceraldehyde-3-phosphate dehydrogenases (TaGAPC2/5/6) under drought stress in wheat and Arabidopsis. We found that TaGAPC2/5/6 all positively responded to drought stress via reactive oxygen species (ROS) scavenging and stomatal movement. The results of yeast co-transformation and electrophoretic mobility shift assay showed that TaWRKY33 acted as a direct regulator of TaGAPC2/5/6 genes. The dual luciferase reporter assay indicated that TaWRKY33 positively activated the expression of TaGAPC2/5/6. The results of bimolecular fluorescence complementation and yeast two-hybrid system demonstrated that TaGAPC2/5/6 interacted with phospholipase D delta (PLD delta). We then demonstrated that TaGAPC2/5/6 positively promoted the activity of TaPLD delta in vitro and in vivo. Furthermore, lower PLD delta activity in RNAi wheat could lead to less PA accumulation, causing higher stomatal aperture sizes under drought stress. In summary, our results establish a new positive regulatory mechanism of TaGAPCs which helps wheat fine-tune their drought responses.