Wheat DOF transcription factors TaSAD and WPBF regulate glutenin gene expression in cooperation with SPA

Grain storage proteins (GSPs) quantity and composition determine the end-use value of wheat flour. GSPs consists of low-molecular-weight glutenins (LMW-GS), high-molecular-weight glutenins (HMW-GS) and gliadins. GSP gene expression is controlled by a complex network of DNA-protein and protein-protein interactions, which coordinate the tissue-specific protein expression during grain development. The regulatory network has been most extensively studied in barley, particularly the two transcription factors (TFs) of the DNA binding with One Finger (DOF) family, barley Prolamin-box Binding Factor (BPBF) and Scutellum and Aleurone-expressed DOF (SAD). They activate hordein synthesis by binding to the Prolamin box, a motif in the hordein promoter. The BPBF ortholog previously identified in wheat, WPBF, has a transcriptional activity in expression of some GSP genes. Here, the wheat ortholog of SAD, named TaSAD, was identified. The binding of TaSAD to GSP gene promoter sequences in vitro and its transcriptional activity in vivo were investigated. In electrophoretic mobility shift assays, recombinant TaSAD and WPBF proteins bound to cis-motifs like those located on HMW-GS and LMW-GS gene promoters known to bind DOF TFs. We showed by transient expression assays in wheat endosperms that TaSAD and WPBF activate GSP gene expression. Moreover, co-bombardment of Storage Protein Activator (SPA) with WPBF or TaSAD had an additive effect on the expression of GSP genes, possibly through conserved cooperative protein-protein interactions.

Automated summary

The quantity and composition of grain storage proteins (GSPs) play a crucial role in determining the value of wheat flour. The expression of GSP genes is regulated by a complex network of DNA-protein and protein-protein interactions. In this study, the wheat ortholog of the transcription factor SAD, named TaSAD, was identified and its binding to GSP gene promoters was investigated. The results showed that TaSAD and another transcription factor WPBF activate GSP gene expression, and their co-bombardment with Storage Protein Activator (SPA) had an additive effect. These findings provide important insights into the regulation of GSP gene expression in wheat.