Drought is one of the most common abiotic stresses, and can limit wheat yield, crops and productivity. GAPCs play vital roles under drought stress conditions in multiple species. The aim of this experiment was to determine the regulatory mechanism of TaGAPC5 under drought stress. In this study, the genes and promoters of TaGAPC5 in diverse drought-tolerant cultivars were cloned. The amino acid sequences were conserved, while the promoter fragments were not identical. Under abiotic stress, the expression level of TaGAPC5 was substantially different among the diverse drought-tolerant cultivars and the promoter activities were significantly improved. The yeast one-hybrid system and Electrophoretic mobility shift assay (EMSA) demonstrated that TaWRKYs bound to specific W-boxes: TaWRKY28, TaWRKY33, TaWRKY40 and TaWRKY47 bind to G/ATGACG/C/A, C/G/ATGACG, C/ATGACC and C/ATGACC/G, respectively. By analyzing different 5' deletion mutants of these promoters, it was determined that these W-boxes in CW-TaGAPC5 promoter (-1262, -1202, -904, -880 and -207) and ZY-TaGAPC5 promoter (-697 and -220) bound by these four TaWRKYs and were functional under drought stress. The deletion or addition of specific W-boxes in the promoter fragments significantly restrained or advanced the promoter activity under drought stress, and these results further confirmed that these W-boxes play vital roles in improving transcription levels under drought stress. The W-boxes in CW-TaGAPC5P (-1262, -1202, -904, -880 and -207) and ZY-TaGAPC5P (-697 and -220) were identified as the key cis-elements for responding to drought stress and were bound by the transcription factor TaWRKY.

Drought stress is one of the major factors that affects wheat yield. Glyceraldehyde-3-Phosphate dehydrogenase (GAPDH) is a multifunctional enzyme that plays the important role in abiotic stress and plant development. However, in wheat, limited information about drought-responsive GAPC genes has been reported, and the mechanism underlying the regulation of the GAPC protein is unknown. In this study, we evaluated the potential role of GAPC1 in drought stress in wheat and Arabidopsis. We found that the overexpression of TaGAPC1 could enhance the tolerance to drought stress in transgenic Arabidopsis. Yeast one-hybrid library screening and EMSA showed that TaWRKY40 acts as a direct regulator of the TaGAPC1 gene. A dual luciferase reporter assay indicated that TaWRKY40 improved the TaGAPC1 promoter activity. The results of qRT-PCR in wheat protoplast cells with instantaneous overexpression of TaWRKY40 indicated that the expression level of TaGAPC1 induced by abiotic stress was upregulated by TaWRKY40. Moreover, TaGAPC1 promoted H2O2 detoxification in response to drought. These results demonstrate that the inducible transcription factor TaWRKY40 could activate the transcription of the TaGAPC1 gene, thereby increasing the tolerance of plants to drought stress.