FIGURE 2.

A time-calibrated phylogeny of Neotropical Freshwater Fishes (NFF) illustrating evolutionary relationships among genera from all recognized families, except within Characiformes (shown in detail in Figure 3), Siluriformes (shown in detail in Figure 4), Cichliformes (shown in detail in Figure 5), and Cyprinodontiformes (shown in detail in Figure 6). Divergence time estimates indicate that the evolutionary history of the NFF has its roots in the Ordovician, around 480 million years ago (mya), a period of high diversification in the oceans, particularly among agnathans. Pie charts on nodes indicate bootstrap support, with the proportion of black space corresponding to the percentage of bootstrap replicates in support. Full black circles represent 100% support. All GenBank accession numbers, alignments, species-level relationships, divergence clade estimates and species representativeness percentages are provided in the Supplementary Material. This phylogeny, as well as one with nodes bearing bootstrap values below 70% collapsed, is available as a Newick string in the Supplementary Material and can be found in the “phy” folder.