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<p>In the ongoing plant–pathogen arms race, plants use pattern recognition receptors (PRRs) to recognize pathogen-associated molecular patterns (PAMPs), while in successful pathogens, PAMPs can evolve to evade detection. Engineering PRRs to recognize evading PAMPs could potentially generate broad-spectrum and durable disease resistance. Here we reverse-engineered two natural variants of the PRR FLAGELLIN SENSING 2 (FLS2), VrFLS2XL and GmFLS2b, with extended recognition specificities towards evading flg22 variants. We identified minimal gain-of-function residues enabling blind FLS2s to recognize otherwise evading flg22 variants. We uncovered two strategies: (1) optimizing FLS2–flg22 interaction around flg22’s key evasion sites and (2) strengthening direct FLS2–BAK1 interaction to overcome weak agonistic and antagonistic flg22s, respectively. In addition, we leveraged polymorphisms that enhance recognition through unknown mechanisms to engineer a superior recognition capability. These findings offer basic design principles to engineer PRRs with broader recognition spectra, paving the way for PRR engineering to generate precisely gene-edited disease-resistant crops.</p>

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Reverse engineering of the pattern recognition receptor FLS2 reveals key design principles of broader recognition spectra against evading flg22 epitopes

  • 期刊论文

  • 出版模式：

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  • 发表日期：

    2025年7月28日

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  • Songyuan Zhang,
  • Songyuan Liu,
  • Hung-Fei Lai,
  • Kyle W. Bender,
  • Gijeong Kim,
  • Amedeo Caflisch,
  • Cyril Zipfel

摘要

In the ongoing plant–pathogen arms race, plants use pattern recognition receptors (PRRs) to recognize pathogen-associated molecular patterns (PAMPs), while in successful pathogens, PAMPs can evolve to evade detection. Engineering PRRs to recognize evading PAMPs could potentially generate broad-spectrum and durable disease resistance. Here we reverse-engineered two natural variants of the PRR FLAGELLIN SENSING 2 (FLS2), VrFLS2XL and GmFLS2b, with extended recognition specificities towards evading flg22 variants. We identified minimal gain-of-function residues enabling blind FLS2s to recognize otherwise evading flg22 variants. We uncovered two strategies: (1) optimizing FLS2–flg22 interaction around flg22’s key evasion sites and (2) strengthening direct FLS2–BAK1 interaction to overcome weak agonistic and antagonistic flg22s, respectively. In addition, we leveraged polymorphisms that enhance recognition through unknown mechanisms to engineer a superior recognition capability. These findings offer basic design principles to engineer PRRs with broader recognition spectra, paving the way for PRR engineering to generate precisely gene-edited disease-resistant crops.