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Lastly, we identified sites that experienced a selective shift along the three branches of interest.
These branches of interest are indicated in red in figure 1.
Only trees that show significant bootstraps for the branches of interest were considered.
To identify genes with higher dN/dS ratios in branches of interest, we used the branch model in Codeml [ 48].
Second, to investigate if the omega ratio varied among particular branches of interest in the phylogeny, we compared two different branch models.
Evolutionary rates of particular branches of interest (ω1) versus the background ratio (ω0) were computed using the branch model (model = 2).
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The haplotype relationships determined using a neighbor-joining algorithm are shown in Figure 2 with bootstrap values indicated above each branch of interest.
However, specifically allowing for a different rate in the branch of interest (branch number 5) and allowing ω to vary among sites within this branch suggested the existence of sites evolving under positive selection in this branch.
This was confirmed by a series of two-ratio branch models, in which each foreground branch of interest was in turn allowed to have a different ratio from the rest of the tree (background).
In this model the branch of interest (foreground) can have sites with an ω-value larger than one and all other branches (background) are restricted to ω-values below or equal to one [69].
In this model, one parameter represents the branch of interest and the other parameter represents the rest of the orthologous gene tree (i.e., Model = 2 and Nsites = 2).
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