StMPK7 phosphorylates and stabilizes a potato RNA-binding protein StUBA2a/b to enhance plant defence responses

Abstract

Mitogen-activated protein kinase (MAPK) cascades play pivotal roles in regulating plant immunity. MAPKs usually transduce signals and regulate plant immunity by phosphorylating the downstream defence-related components. Our previous study indicates that StMPK7 positively regulates plant defence to Phytophthora pathogens via SA signalling pathway. However, the downstream component of StMPK7 remains unknown. In this study, we employed GFP-StMPK7 transgenic potato and performed immunoprecipitation-mass spectrometry (IP-MS) to identify the downstream component of StMPK7. We found that an RNA binding protein StUBA2a/b interacted with StMPK7, as revealed by luciferase complementation imaging (LCI) and coimmunoprecipitation (co-IP) assays. Transient expression of StUBA2a/b in Nicociana benthamiana enhanced plant resistance to Phytophthora pathogens, while silencing of UBA2a/b decreased the resistance, suggesting a positive regulator role of UBA2a/b in plant immunity. Similar to StMPK7, StUBA2a/b was also involved in SA signalling pathway and induced SGT1-dependent cell death as constitutively activated (CA)-StMPK7 did. Immune blotting indicated that StMPK7 phosphorylates StUBA2a/b at thr248 and thr408 (T248/408) sites and stabilizes StUBA2a/b. Silencing of MPK7 in N. benthamiana suppressed StUBA2a/b-induced cell death, while co-expression with StMPK7 enhanced the cell death. Besides, StUBA2a/bT248/408A mutant showed decreased ability to trigger cell death and elevate the expression of PR genes, indicating the phosphorylation by StMPK7 enhances the functions of StUBA2a/b. Moreover, CA-StMPK7-induced cell death was largely suppressed by silencing of NbUBA2a/b, genetically implying UBA2a/b acts as the downstream component of StMPK7. Collectively, our results reveal that StMPK7 phosphorylates and stabilizes its downstream substrate StUBA2a/b to enhance plant immunity via the SA signalling pathway.

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