Nucleotide-binding leucine-rich repeat(NLR)proteins assemble into genetically linked pairs to mediate effectortriggered immunity(ETI)in plants.Here,we characterize the paired NLRs NRCX and NARY(NRCX adjacent resistanc...Nucleotide-binding leucine-rich repeat(NLR)proteins assemble into genetically linked pairs to mediate effectortriggered 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.展开更多
基金supported by the National Natural Science Foundation of China(32230089,32402315 and 32270208)the China Agriculture Research System(CARS-21).
文摘Nucleotide-binding leucine-rich repeat(NLR)proteins assemble into genetically linked pairs to mediate effectortriggered 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.
