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In each analysis, 4 categories of rate variation were used.
ML and BI analyses of the amino acid alignment (i.e., alignment 4) was analyzed under a WAG model of substitution considering corrections for site-to-site rate variation (gamma) with eight categories of rate variation and proportion of invariable sites.
The best likelihood trees were searched under the GTR-GAMMA model with six categories of rate variation (500 bootstrap replicates were undertaken for estimation of node support).
We utilised four categories of rate variation across the amino acid sites and 500 bootstrap replicates were generated for the estimation of node support.
Nucleotide substitution models were obtained from jModeltest (version 2.1, [ 38, 39], testing 11 substitution schemes, each with 4 categories of rate variation among sites) and using the corrected AIC criterion (AICc) to find the best fitting model.
We performed this inference on both untrimmed and BMGE-trimmed versions of the concatenated matrix, using a combination of NNI and SPR heuristic searches, and inferring under the LG+F model with four 'free' (--free-rates) categories of rate variation.
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To test for the presence of LBA we calculated 8 categories of rates of evolution for all sites, from the most rapidly evolving to the most slowly evolving.
Both experiments, under GTR and CAT, used four discrete categories of rates across sites and were run twice in order to check chains' convergence.
Phylogenetic trees were inferred using maximum likelihood available in Physher [ 26] employing the generalised time-reversible (GTR) substitution model and a discretised gamma distribution (4 categories) of rates across sites (Γ4).
MrAIC [ 39] estimated that, among 56 models of nucleotide substitution, the best AIC score [ 40] was achieved by the most general homogeneous and reversible model (GTR, "general time reversible" [ 41]), combined with four discrete gamma categories of "rates across sites" [ 42], plus an additional rate category for invariant sites (model GTR nt + Γ4 + I, where subscript nt stands for nucleotides).
Another major category of rate separation methods is based on the diffusion of molecules through semipermeable barriers.
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