NbCSPR underlies age-dependent immune responses to bacterial cold shock protein in Nicotiana benthamiana

Isabel M L Saur, Yasuhiro Kadota, Jan Sklenar, Nicholas J Holton, Elwira Smakowska, Youssef Belkhadir, Cyril Zipfel, John P Rathjen

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Abstract

Plants use receptor kinases (RKs) and receptor-like proteins (RLPs) as pattern recognition receptors (PRRs) to sense pathogen-associated molecular patterns (PAMPs) that are typical of whole classes of microbes. After ligand perception, many leucine-rich repeat (LRR)-containing PRRs interact with the LRR-RK BRI1-ASSOCIATED KINASE 1 (BAK1). BAK1 is thus expected to interact with unknown PRRs. Here, we used BAK1 as molecular bait to identify a previously unknown LRR-RLP required for the recognition of the csp22 peptide derived from bacterial cold shock protein. We established a method to identify proteins that interact with BAK1 only after csp22 treatment. BAK1 was expressed transiently in Nicotiana benthamiana and immunopurified after treatment with csp22. BAK1-associated proteins were identified by mass spectrometry. We identified several proteins including known BAK1 interactors and a previously uncharacterized LRR-RLP that we termed RECEPTOR-LIKE PROTEIN REQUIRED FOR CSP22 RESPONSIVENESS (NbCSPR). This RLP associates with BAK1 upon csp22 treatment, and NbCSPR-silenced plants are impaired in csp22-induced defense responses. NbCSPR confers resistance to bacteria in an age-dependent and flagellin-induced manner. As such, it limits bacterial growth and Agrobacterium-mediated transformation of flowering N. benthamiana plants. Transgenic expression of NbCSPR into Arabidopsis thaliana conferred responsiveness to csp22 and antibacterial resistance. Our method may be used to identify LRR-type RKs and RLPs required for PAMP perception/responsiveness, even when the active purified PAMP has not been defined.

Original languageEnglish
Pages (from-to)3389-3394
Number of pages6
JournalProceedings of the National Academy of Sciences of the United States of America (PNAS)
Volume113
Issue number12
Early online date4 Mar 2016
DOIs
Publication statusPublished - 22 Mar 2016

Keywords

  • plant immunity
  • PAMP
  • receptor
  • BAK1
  • Nicotiana benthamiana

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