Arabidopsis pavement cell shape formation involves spatially confined ROPGAP regulators

In many plant species, pavement cell development relies on the coordinated formation of interdigitating lobes and indentations. Polarity signaling via the activity of antagonistic Rho-related GTPases from plants (ROPs) was implicated in pavement cell development, but the spatiotemporal regulation remained unclear. Here, we report on the role of the PLECKSTRIN HOMOLOGY GTPase ACTIVATING PROTEINS (PHGAPS) during multipolar growth in pavement cell shape establishment. Loss of function in phgap1phgap2 double mutants severely affected the shape of Arabidopsis leaf epidermal pavement cells. Predominantly, PHGAPs interacted with ROP2 and displayed a distinct and microtubule-dependent enrichment along the anticlinal cell face and transfacial boundary of pavement cell indentation regions. This localization was established upon undulation initiation and was maintained throughout the expansion of the cell. Our data suggest that PHGAP1/REN2 and PHGAP2/REN3 are key players in the establishment of ROP2 activity gradients and underscore the importance of locally controlled ROP activity for the orchestrated establishment of multipolarity in epidermal cells.