Extended Data Fig. 6: Molecular evolution of Fag e 2 genes and sequence of Fag e 2 knockout mutant.
a, Amino acid alignment of Fag e 2 genes in the three Fagopyrum species. The epitope sequence25 is indicated by a red square. Conserved cysteine residues of Fag e 2 homologues are indicated by red arrowheads. The background colour is proportional to the degree of similarity of each residue compared to its aligned column b, Maximum likelihood phylogenetic tree of Fag e 2 genes constructed with IQ-TREE based on the amino acid alignment of the gene family including Caryophyllales species identified by OrthoFinder. Tree is unrooted and bootstrap values of ≥80% are indicated by each node. Scale bar indicates branch length. Nodes corresponding to tandem duplications are indicated by blue diamonds. c, Amino acid alignment of the epitope sequence indicated by a red square in a. The position conserved across all sequences is indicated by asterisk. The three groups correspond to those in b. d, Upper panel shows the amino acid sequence encoded by the EMS-induced Fag e 2 gene. Red letters indicate epitope amino acids. Green letters indicate the eight Cys residues conserved within plant 2S albumins. Lower panel indicates the results of Sanger sequencing of the mutant/wild type heterozygote (left) and wild type homozygote (right).
