Directed Evolution of a Plant Immune Receptor for Broad Spectrum Effector Recognition

Overview

Journal bioRxiv

Date 2025 Feb 20

PMID 39975188

Authors

Ellen Y Rim

Oscar D Garrett

Alexander J Howard

Yejin Shim

Yuanyuan Li

Jonathan E Van Dyke

Ryan C Packer

Nguyen Ho

Rashmi Jain

Valley Stewart

Savithramma P Dinesh-Kumar

James H Notwell

Pamela C Ronald

Affiliations

Soon will be listed here.

Abstract

Rapid development of immune receptors that protect crops from emerging pathogens is a critical challenge . While novel immune receptors that recognize previously undetected pathogen effectors could provide protection against a wider range of pathogens, engineering such receptors has been constrained by the low throughput and speed of testing. We established yeast surface display as a high throughput platform to recapitulate plant immune receptor-ligand interactions and evolve new binding capabilities. Using this directed evolution platform, we engineered the ligand binding domain of the rice immune receptor Pik-1 to recognize diverse effectors from the fast-evolving fungal pathogen Our approach yielded Pik-1 ligand binding domains with affinity for variants of the effector Avr-Pik that previously escaped detection by known rice alleles of Pik-1, with assays confirming functional recognition of these effectors. Additional rounds of mutagenesis and selection led to a Pik-1 domain that binds all tested Avr-Pik variants as well as the evolutionarily divergent effector AvrPiz-t. These results demonstrate the potential of directed evolution to engineer immune receptors with new-to-nature recognition of a wide range of pathogen-derived ligands and accelerate development of broad spectrum resistance in crops.

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