The identification of TPS genes in wild Freesia species. A) Phylogenetic analysis of TPS proteins from Freesia and other plants. The percentage of replicate trees in which the associated taxa clustered together in the bootstrap test (1,000 replicates) are shown next to the branches. The tree is drawn to scale, with branch lengths in the same units as those of the evolutionary distances used to infer the phylogenetic tree. The evolutionary distances are computed in the units of the number of amino acid substitutions per site, as shown by the scale below the tree. Freesia TPS proteins are indicated by blue balls. GenBank accessions of TPS proteins are provided in Supplemental Table S3. B) Amino acid sequence alignment of Freesia TPS proteins. The conserved domains R(R, P, Q)(X)₈W, DDXXD, and NSE/DTE are highlighted. Numbers represent positions of the last residue in each line. *, identical amino acids; : or ·, similar amino acids. C) Subcellular localizations of the cloned TPS9 to TPS15 from representative wild Freesia species observed in Arabidopsis protoplasts. Bright light, bright field image; green, GFP fluorescence detected in the green channel; red, chlorophyll autofluorescence detected in the red channel; merged, merged green and red channel images. Bars = 25 µ M.