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<p>Nucleotide-binding leucine-rich repeat (NLR) proteins assemble into genetically linked pairs to mediate effector-triggered immunity (ETI) in plants. Here, we characterize the paired NLRs <i>NRCX</i> and <i>NARY</i> (NRCX adjacent resistance gene Y) in <i>Nicotiana benthamiana</i>. CRISPR/Cas9 knockout of <i>NRCX</i> caused severe dwarfism and constitutively activated immunity, marked by <i>PR1</i> upregulation and enhanced resistance to <i>Phytophthora capsici</i>. Co-silencing or double knockout of the adjacent NLR <i>NARY</i> partially rescued the <i>nrcx</i> phenotype, revealing NARY as a compensatory regulator that modulates growth and immunity. Structural analysis revealed that NARY harbors non-canonical Walker B and MHD motifs, which lack autoactivation capacity despite their divergence from canonical NLR executors. Split-luciferase and co-immunoprecipitation assays showed that NRCX and NARY interact exclusively through their CC domains, forming a non-canonical regulatory complex. Notably, simultaneous silencing of <i>NRC2/3</i> and <i>NARY</i> incompletely restored growth in <i>nrcx</i> mutants, implicating additional factors in immune modulation. Our findings establish NARY as a compensatory NLR partner of NRCX that fine-tunes immunity without triggering cell death, revealing a novel mechanism for balancing growth and defense in Solanaceae.</p>

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A CNL protein forms an NLR pair with NRCX to modulate plant immunity

  • 期刊论文

  • 出版模式：

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  • 发表日期：

    2025年9月3日

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  • Xiaohua Dong,
  • Xiaoyan Zhang,
  • Xu Lu,
  • Yufeng Yang,
  • Chuyan Xia,
  • Weiye Pan,
  • Zhiyuan Yin,
  • Yaning Zhao,
  • Gan Ai,
  • Daolong Dou

摘要

Nucleotide-binding leucine-rich repeat (NLR) proteins assemble into genetically linked pairs to mediate effector-triggered immunity (ETI) in plants. Here, we characterize the paired NLRs NRCX and NARY (NRCX adjacent resistance gene Y) in Nicotiana benthamiana. CRISPR/Cas9 knockout of NRCX caused severe dwarfism and constitutively activated immunity, marked by PR1 upregulation and enhanced resistance to Phytophthora capsici. Co-silencing or double knockout of the adjacent NLR NARY partially rescued the nrcx phenotype, revealing NARY as a compensatory regulator that modulates growth and immunity. Structural analysis revealed that NARY harbors non-canonical Walker B and MHD motifs, which lack autoactivation capacity despite their divergence from canonical NLR executors. Split-luciferase and co-immunoprecipitation assays showed that NRCX and NARY interact exclusively through their CC domains, forming a non-canonical regulatory complex. Notably, simultaneous silencing of NRC2/3 and NARY incompletely restored growth in nrcx mutants, implicating additional factors in immune modulation. Our findings establish NARY as a compensatory NLR partner of NRCX that fine-tunes immunity without triggering cell death, revealing a novel mechanism for balancing growth and defense in Solanaceae.