Abstract
Cellular redox status plays critical roles in cell division and differentiation, but the underlying mechanism is unclear. Here we explored the effect of redox status on stem cell identity in distal stem cells (DSCs) of Arabidopsis (Arabidopsis thaliana) roots. Treatment with the reductive reagent glutathione and the oxidative reagent H2O2 inhibited DSC differentiation, as did endogenously altering reactive oxygen species production via various mutations. This suggests that both highly reductive and oxidative environments inhibit specification of stem cell identity. In our observations of mutant components of the CLAVATA3/ENDOSPERM SURROUNDING REGION 40 (CLE40)-ARABIDOPSIS CRINKLY4 (ACR4)/CLAVATA1 (CLV1)-WUSCHEL RELATED HOMEOBOX5 (WOX5) module, both reductive and oxidative reagents influenced DSC differentiation in wox5-1 and clv1-1, but not in acr4-2 or cle40 mutant plants. The stability of the receptor-like kinase ACR4 is modulated by redox status through endocytosis in root tips. ACR4 with multiple Cys mutations in the tumor necrosis factor receptor (TNFR) extracellular domain failed to undergo endocytosis. ACR4 with a complete deletion of the TNFR domain was localized directly to endosomes, bypassing the plasma membrane. Both mutations affected DSC differentiation, but not seed filling. Conversely, the intracellular domain of the ACR4 protein is partially required for seed filling, but not for DSC differentiation. Our study uncovers an important biological role of the TNFR domain in redox-mediated endocytosis of ACR4 in root DSC differentiation.