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Vesicular trafficking has emerged as an important means by which eukaryotes modulate responses to microbial pathogens, likely by contributing to the correct localization and levels of host components necessary for effective immunity. However, considering the complexity of membrane trafficking in plants, relatively few vesicular trafficking components with functions in plant immunity are known. Here we demonstrate that  Arabidopsis thaliana  Dynamin-Related Protein 2B (DRP2B), which has been previously implicated in constitutive clathrin-mediated endocytosis (CME), functions in responses to flg22 (the active peptide derivative of bacterial flagellin) and immunity against flagellated bacteria  Pseudomonas syringae  pv.  tomato  ( Pto ) DC3000. Consistent with a role of DRP2B in Pattern-Triggered Immunity (PTI),  drp2b  null mutant plants also showed increased susceptibility to  Pto  DC3000  hrcC   − , which lacks a functional Type 3 Secretion System, thus is unable to deliver effectors into host cells to suppress PTI. Importantly, analysis of  drp2b  mutant plants revealed three distinct branches of the flg22-signaling network that differed in their requirement for  RESPIRATORY BURST OXIDASE HOMOLOGUE D (RBOHD) , the NADPH oxidase responsible for flg22-induced apoplastic reactive oxygen species production. Furthermore, in  drp2b , normal MAPK signaling and increased immune responses via the  RbohD /Ca 2+ -branch were not sufficient for promoting robust  PR1  mRNA expression nor immunity against  Pto  DC3000 and  Pto  DC3000  hrcC −  . Based on live-cell imaging studies, flg22-elicited internalization of the plant flagellin-receptor, FLAGELLIN SENSING 2 (FLS2), was found to be partially dependent on DRP2B, but not the closely related protein DRP2A, thus providing genetic evidence for a component, implicated in CME, in ligand-induced endocytosis of FLS2. Reduced trafficking of FLS2 in response to flg22 may contribute in part to the non-canonical combination of immune signaling defects observed in  drp2b . In conclusion, this study adds DRP2B to the relatively short list of known vesicular trafficking proteins with roles in flg22-signaling and PTI in plants.

Loss of Arabidopsis thaliana Dynamin-Related Protein 2B Reveals Separation of Innate Immune Signaling Pathways