Cross-bracing uncalibrated nodes in molecular dating improves congruence of fossil and molecular age estimates

The practice of molecular dating is an essential tool for hypothesis testing in evolutionary biology. Vagaries of fossilization and taphonomic bias commonly engender high uncertainty in molecular dating in taxonomic groups wherein few fossils can be unambiguously assigned to phylogenetic nodes. A recent and novel implementation in molecular dating, “cross-bracing”, exploits gene duplications by formally linking calibrated node dates throughout the paralogous subtrees through hierarchical Bayesian models. An unexplored refinement of this method is cross-bracing nodes with unknown dates, in addition to calibrated nodes, such that all nodes representing the same cladogenetic events have linked priors. We applied such a refinement to molecular dating in chelicerates, one of the earliest groups of arthropods present in the fossil record, but whose molecular dating has been greatly inconsistent in the literature. We inferred divergence times using hemocyanin paralogs isolated from de novo assembled transcriptomic libraries, and multiple fossil calibrations. We show that extending cross-bracing to uncalibrated nodes greatly reduced variance in estimates of divergence times throughout the phylogeny, particularly for estimated diversification ages of spiders and scorpions, whereas cross-bracing calibrated nodes alone did not affect age estimation for uncalibrated, derived clades. Comparing ages inferred with extended cross-bracing to the fossil record, we observe smaller gaps between diversification and the first appearance of crown group fossils than have previously been inferred, particularly for spiders. Our dating indicates that scorpions have a Silurian origin, but diversification of extant lineages occurred near the Triassic-Jurassic boundary, falsifying previous inference of Permian diversification age based on extant distribution alone. The significant reduction of variance in divergence time estimates upon extending cross-bracing to uncalibrated nodes makes this approach greatly suited for evolutionary inference in groups with poor fossil records, with particular reference to terrestrial arthropods.