A species tree and macrosynteny relationships between P. trichocarpa, P. deltoides, and S. purpurea and alignments of de novo genes Potri.001G124250 and Potri.001G400201 against their respective syntenic intergenic regions in P. deltoides and S. purpurea. Highlighted bases and residues indicate disagreements to the de novo gene in P. trichocarpa. Red boxes indicate disabling mutations in the intergenic syntenic sequence. Both alignments follow the sequential structure of P. trichocarpa (de novo gene), P. deltoides (primary syntenic region), S. purpurea (primary syntenic region), P. trichocarpa (secondary syntenic region), P. deltoides (secondary syntenic region), and S. purpurea (secondary syntenic region). (A) A species tree of the phylogeny of P. trichocarpa, P. deltoides, and S. purpurea, the red point on the root of the tree is indicative of the WGD which occurred in the ancestor of Populus and Salix. (B) Macrosynteny relationships between P. trichocarpa and S. purpurea, and P. trichocarpa and P. deltoides. (C) Alignment of Potri.001G124250 against intergenic syntenic regions. (D) Alignment of Potri.001G400201 against intergenic syntenic regions.