TaERFL1a enhances drought resilience through DHAR-mediated ASA-GSH biosynthesis in wheat

Wheat is one of the important cereal crops around the world, but it often suffers from abiotic stresses, which threaten food security. Thus, it is critical to identify the genes that determine drought tolerance in wheat. AP2/ERFs are known to regulate drought stress in various crops. In this study, TaERFL1a-overexpressing wheat transgenic lines (TaERFL1a-OEs) were used to determine drought resilience mechanism. After 12 d without watering, the growth phenotype of TaERFL1a-OEs was better than that of the wild type (WT), whose activities of superoxide dismutase and catalase, and contents of ascorbate acid (ASA) and glutathione (GSH) were significantly increased, while malondialdehyde content was significantly decreased. Transcriptome analysis revealed that 28,520 genes were differentially expressed between TaERFL1a-OEs and WT under drought condition. Further analysis found that these DEGs were involved in multiple stress-response processes, especially in the ASA-GSH pathway. qPCR revealed that the expression levels of GPX, DHAR, and MDHAR, which are suggested to be participated in ASA-GSH biosynthesis, were significantly up-regulated in TaERFL1a-OEs under drought stress, especially the DHAR gene. Moreover, dual-luciferase and luciferase complementation imaging revealed that TaERFL1a was more promoted DHAR transcription to a greater extent than other genes. Furthermore, yeast one-hybrid, electrophoretic mobility shift assay, and chromatin immunoprecipitation combined with qPCR revealed that TaERFL1a regulates DHAR expression by binding to the cis-element ERF in DHAR promoter and promotes the transcription of later in vivo and in vitro. Overall, our results provided molecular regulatory evidence for TaERFL1a in wheat drought stress and suggested candidate genes for improving drought-tolerant wheat breeding.