Pathogen effector recognition-dependent association of NRG1 with EDS1 and SAG101 in TNL receptor immunity

Xinhua Sun,Paul Derbyshire,Hirofumi Nakagami,Jaqueline Bautor,Servane Blanvillain-Baufumé,Dmitry Lapin,Sara C Stolze,Jane E Parker,Joanna M Feehan,Joram A Dongus,Anne Harzen,Jonathan D G Jones,Frank L H Menke,Katharina Kramer,Iris Finkemeier,Jakub Rzemieniewski

doi:10.1038/s41467-021-23614-x

Abstract

Plants utilise intracellular nucleotide-binding, leucine-rich repeat (NLR) immune receptors to detect pathogen effectors and activate local and systemic defence. NRG1 and ADR1 “helper” NLRs (RNLs) cooperate with enhanced disease susceptibility 1 (EDS1), senescence-associated gene 101 (SAG101) and phytoalexin-deficient 4 (PAD4) lipase-like proteins to mediate signalling from TIR domain NLR receptors (TNLs). The mechanism of RNL/EDS1 family protein cooperation is not understood. Here, we present genetic and molecular evidence for exclusive EDS1/SAG101/NRG1 and EDS1/PAD4/ADR1 co-functions in TNL immunity. Using immunoprecipitation and mass spectrometry, we show effector recognition-dependent interaction of NRG1 with EDS1 and SAG101, but not PAD4. An EDS1-SAG101 complex interacts with NRG1, and EDS1-PAD4 with ADR1, in an immune-activated state. NRG1 requires an intact nucleotide-binding P-loop motif, and EDS1 a functional EP domain and its partner SAG101, for induced association and immunity. Thus, two distinct modules (NRG1/EDS1/SAG101 and ADR1/EDS1/PAD4) mediate TNL receptor defence signalling.

Highlights

Plants utilise intracellular nucleotide-binding, leucine-rich repeat (NLR) immune receptors to detect pathogen effectors and activate local and systemic defence
We show in Arabidopsis that enhanced disease susceptibility 1 (EDS1)–senescenceassociated gene 101 (SAG101)–N requirement gene 1 (NRG1) and EDS1–phytoalexin-deficient 4 (PAD4)–activated disease resistance 1 (ADR1) operate genetically as non-interchangeable signalling nodes in effector-triggered immunity (ETI)
We find in Arabidopsis stable transgenic lines and in N. benthamiana reconstitution assays that pattern-triggered immunity (PTI) activation is insufficient for NRG1induced association with EDS1 and SAG101

Summary

Introduction

Plants utilise intracellular nucleotide-binding, leucine-rich repeat (NLR) immune receptors to detect pathogen effectors and activate local and systemic defence. NRG1 and ADR1 “helper” NLRs (RNLs) cooperate with enhanced disease susceptibility 1 (EDS1), senescenceassociated gene 101 (SAG101) and phytoalexin-deficient 4 (PAD4) lipase-like proteins to mediate signalling from TIR domain NLR receptors (TNLs). Plants and animals have evolved structurally and functionally related cell surface and intracellular receptors that detect pathogen-derived molecules and activate innate immune responses In both kingdoms, pathogen recognition by intracellular nucleotide-binding/leucine-rich repeat (NLR) receptors restricts disease[1]. Whereas mammals tend to have few functional NLR receptors, many plants have expanded and diversified NLR gene repertoires, likely in response to evolutionary pressure from host-adapted pathogens and pests[1,2] Despite these different trajectories, plant and mammalian NLRs behave as conformational switches for triggering defence and immune-related death pathways[3]. CNLs and TNLs rely on a network of signalling NLRs (generically referred to as helper NLRs) to promote immunity and host cell death[17,18,19]

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Results

Conclusion