Dataset Information

Engineering the plant intracellular immune receptor Sr50 to restore recognition of the AvrSr50 escape mutant.

ABSTRACT: Sr50, an intracellular nucleotide-binding leucine-rich repeat receptor (NLR), confers resistance of wheat against stem rust caused by the fungal pathogen Puccinia graminis f. sp. tritici. The receptor recognizes the pathogen effector AvrSr50 through its C-terminal leucine-rich repeat domain, initiating a localized cell death immune response. However, this immunity is compromised by mutations in the effector, as in the escape mutant AvrSr50^QCMJC, which evades Sr50 detection. In this study, we employed iterative computational structural analyses and site-directed mutagenesis for rational engineering of Sr50 to gain recognition of AvrSr50^QCMJC. Following an initial structural hypothesis driven by molecular docking, we identified the Sr50^K711D single mutant, which induces an intermediate immune response against AvrSr50^QCMJC without losing recognition against AvrSr50. Increasing gene expression with a stronger promoter enabled the mutant to elicit a robust response, indicating weak effector recognition can be complemented by enhanced receptor expression. Further structural refinements led to the creation of five double mutants and two triple mutants with dual recognition of AvrSr50 and AvrSr50^QCMJC with greater immune response intensities than Sr50^K711D against the escape mutant. All effective mutations against AvrSr50^QCMJC required the K711D substitution, indicating that multiple solutions exist for gain of recognition, but the path to reach these mutations may be confined. Furthermore, this single substitution alters the prediction of AlphaFold 2, allowing it to model the complex structure of Sr50^K711D and AvrSr50 that match our final structural hypothesis. Collectively, our study outlines a framework for rational engineering of NLR systems to overcome pathogen escape mutations and provides datasets for future computational models for NLR resurrection.

SUBMITTER: Seong K

PROVIDER: S-EPMC11326300 | biostudies-literature | 2024 Aug

REPOSITORIES: biostudies-literature

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Publications

Resurrection of the Plant Immune Receptor Sr50 to Overcome Pathogen Immune Evasion.

Seong Kyungyong K Wei Wei W Sent Sophie C SC Vega Brandon B Dee Amanda A Ramirez-Bernardino Griselda G Kumar Rakesh R Parra Lorena L Saur Isabel Ml IM Krasileva Ksenia K

bioRxiv : the preprint server for biology 20250211

Pathogen-driven plant diseases cause significant crop losses worldwide. The introgression of intracellular nucleotide-binding leucine-rich repeat receptor (NLR) genes into elite crop cultivars is a common strategy for disease control, yet pathogens rapidly evolve to evade NLR-mediated immunity. The NLR gene Sr50 protects wheat against stem rust, a devastating disease caused by the fungal pathogen Puccinia graminis f. sp. tritici (Pgt). However, mutations in AvrSr50 al ...[more]

PMID: 39149390

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Dataset Information

Engineering the plant intracellular immune receptor Sr50 to restore recognition of the AvrSr50 escape mutant.

Publications

Resurrection of the Plant Immune Receptor Sr50 to Overcome Pathogen Immune Evasion.

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