Figure 1: Evolutionary history of the CYP98A8/9 clade in Brassicales/Brassicaceae.
From: Evolutionary interplay between sister cytochrome P450 genes shapes plasticity in plant metabolism
(a) Annotated protein maximum likelihood tree illustrating the phylogenetic relationships in the CYP98 family in Brassicales. The CYP98A3 clade is the parent to CYP98A8/9s. Evolution regimes (ω=dN/dS) computed with codeml Branch model and acting on different phylogenetic groups are indicated. Branch support in the tree was calculated from 1,000 bootstraps replicates. (b) In vitro assay of CYP98A8/A9 enzymes activities. Tri-coumaroyl-spermidine and tri-feruloyl-spermidine were used to assess the 3′- and 5′-hydroxylase activities of yeast-expressed proteins. CYP98A8/A9 from all species have 3′-hydroxylase activity. Only CYP98A8 from Brassicaceae have 5′-hydroxylase activity. (c) Search for sinapoylated (m/z 207) phenolamides by parent ion scan analysis of flower buds demonstrates the absence in vivo of the tri-hydroxylated phenolamide di-(hydroxyferuloyl)-sinapoyl-spermidine (m/z 736) in A. grandiflorum, while it is present in Brassicaceae from lineages I, II and III. Colour of each profile is according to the lineage colour code in a.
