Ancestral sequence reconstruction of NRC3.

(A) Phylogenetic tree of the solanaceous NRC3 clade. The NLR sequences were extracted from 124 genomes in the Solanaceae family using the NLRtracker software. The phylogenetic tree was reconstructed based on the codon-based nucleotide sequence alignment of NRCX, NRC1, NRC2, and NRC3 clades using IQ-TREE. Ancestral sequence reconstruction was performed using FastML. The nodes of ancestral variants tested in this study were indicated with star shapes. N3: the ancestral variant before the divergence of the three allelic groups; N4: the ancestral variant of NRC3a; N88: the ancestral variant before the divergence of NRC3b/c; N95: the ancestral variant of NRC3b; N89: the ancestral variant of NRC3c. (B) Polymorphisms at the six positions identified across NRC3 natural variants. (JPF)