Extended Data Fig. 2: Functional testing and structural analysis of FoTO1 homologues.

a, FoTO1 homologues from T. media and A. thaliana have no effect on taxane yield increase. TDS2 and the first three pathway oxidases were co-expressed in N. benthamiana with either GFP, FoTO1, At5g04830 (A. thaliana homologue), or ctg8326.3 (T. media homologue). Masses corresponding to the expected products, 5α,10β,13α-triol and 5α,10β-diol 13-one, are displayed as EICs and bar graph quantified across three independent replicate leaves. Unlike FoTO1, neither FoTO1 homologue alters the product profile of this early subpathway. Data are shown as the mean ± standard deviation, n = 3. b, FoTO1 structural homologues identified in other land plants. Using FoldSeek v4⁷¹ we searched all pre-folded protein databases in FoldSeek for full-length structural homologues of FoTO1. The top five hits were proteins from the genomes of model plants Oryza sativa Japonica (rice), Zea mays (corn), A. thaliana and Glycine max (soy). After the NTF2 ___domain, each of these homologues contain the α-helical C terminus (highlighted in transparent turquoise) that we found to be crucial for FoTO1’s phenotype in vivo and for binding to TDS and T5αΗ. The identification of these structural homologues indicates that FoTO1 is not just restricted to gymnosperms, but has structural analogues across both angiosperms.