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The N-terminal MADA motif of NRC3 is required for Cf-4/Avr4-triggered hypersensitive cell death.

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posted on 22.09.2022, 18:17 authored by Jiorgos Kourelis, Mauricio P. Contreras, Adeline Harant, Hsuan Pai, Daniel Lüdke, Hiroaki Adachi, Lida Derevnina, Chih-Hang Wu, Sophien Kamoun

A) NRC3 has an N-terminal MADA motif. Alignment of NRC2, NRC3, NRC4 and ZAR1 N-terminal MADA motif with the consensus sequence pattern of the MADA motif and the HMM score for each sequence. B) Schematic representation of NRC2, NRC3, and the respective NRC2L17E and NRC3L21E MADA mutants. C) NRC2 and NRC3 MADA mutants accumulate to similar levels as wild-type proteins. Immunoblot analysis of transiently expressed C-terminally 6xHA-tagged NRCs 5 days after agroinfiltration in wild-type N. benthamiana plants. D) NRC MADA mutants do not complement Cf-4/Avr4 or Pto/AvrPto-triggered hypersensitive cell death in the N. benthamiana nrc2/3/4 lines. Representative N. benthamiana leaves infiltrated with appropriate constructs were photographed 7–10 days after infiltration. The NRCs tested, NRC2 and NRC3, are labelled above the leaf of nrc2/3/4- The receptor/effector pair tested, Cf-4/Avr4 and Prf (Pto/AvrPto), are labelled on the leaf image. A representative leaf of the independent nrc2/3/4- line is shown in S7E Fig) Quantification of hypersensitive cell death. Cell death was scored based on a 0–7 scale between 7–10 days post infiltration. The results are presented as a dot plot, where the size of each dot is proportional to the count of the number of samples with the same score within each biological replicate. The experiment was independently repeated three times. The columns correspond to the different biological replicates. Significant differences between the conditions are indicated with an asterisk (*). Details of statistical analysis are presented in S7 Fig.