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Rhizobia induce SYMRK endocytosis in Phaseolus vulgaris root hair cells

Raúl Dávila-Delgado,Karen Flores-Canúl,M. A. Juárez-Verdayes,R. Sánchez-López

Published 2023 in Planta

ABSTRACT

Main conclusion Pv SYMRK-EGFP undergoes constitutive and rhizobia-induced endocytosis, which rely on the phosphorylation status of T589, the endocytic YXXØ motif and the kinase activity of the receptor. Abstract Legume-rhizobia nodulation is a complex developmental process. It initiates when the rhizobia-produced Nod factors are perceived by specific LysM receptors present in the root hair apical membrane. Consequently, SYMRK (Symbiosis Receptor-like Kinase) becomes active in the root hair and triggers an extensive signaling network essential for the infection process and nodule organogenesis. Despite its relevant functions, the underlying cellular mechanisms involved in SYMRK signaling activity remain poorly characterized. In this study, we demonstrated that Pv SYMRK-EGFP undergoes constitutive and rhizobia-induced endocytosis. We found that in uninoculated roots, Pv SYMRK-EGFP is mainly associated with the plasma membrane, although intracellular puncta labelled with Pv SymRK-EGFP were also observed in root hair and nonhair-epidermal cells. Inoculation with Rhizobium etli producing Nod factors induces in the root hair a redistribution of Pv SYMRK-EGFP from the plasma membrane to intracellular puncta. In accordance, deletion of the endocytic motif YXXØ (YKTL) and treatment with the endocytosis inhibitors ikarugamycin (IKA) and tyrphostin A23 (TyrA23), as well as brefeldin A (BFA), drastically reduced the density of intracellular Pv SYMRK-EGFP puncta. A similar effect was observed in the phosphorylation-deficient (T589A) and kinase-dead (K618E) mutants of Pv SYMRK-EGFP, implying these structural features are positive regulators of Pv SYMRK-EGFP endocytosis. Our findings lead us to postulate that rhizobia-induced endocytosis of SYMRK modulates the duration and amplitude of the SYMRK-dependent signaling pathway.

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