Abstract

Relaxed molecular clock methods allow the use of genomic data to estimate divergence times across the tree of life. This is most commonly achieved in Bayesian analyses where the molecular clock is calibrated a priori through the integration of fossil information. Alternatively, fossil calibrations can be used a posteriori, to transform previously estimated relative divergence times that were inferred without considering fossil information, into absolute divergence times. However, as branch length is the product of the rate of evolution and the duration in time of the considered branch, the extent to which a posteriori calibrated, relative divergence time methods can disambiguate time and rate, is unclear. Here, we use forward evolutionary simulations and compare a priori and a posteriori calibration strategies using different molecular clock methods and models. Specifically, we compare three Bayesian methods, the strict clock, uncorrelated clock and autocorrelated clock, and the non-Bayesian algorithm implemented in RelTime. We simulate phylogenies with multiple, independent substitution rate changes and show that correct timescales cannot be inferred without the use of calibrations. Under our simulation conditions, a posteriori calibration strategies almost invariably inferred incorrect rate changes and divergence times. The a priori integration of fossil calibrations is fundamental in these cases to improve the accuracy of the estimated divergence times. Relative divergence times and absolute timescales derived by calibrating relative timescales to geological time a posteriori appear to be less reliable than a priori calibrated, timescales.

Highlights

  • Evolutionary biologists have long sought the timescale of life (Simpson 1944; Betts et al 2018)

  • We explore the performance of a priori and a posteriori approaches to calibrating molecular clock methods under the challenging conditions of multiple independent substitution rate changes correlated in time

  • We found that the use of relative timescales and the a posteriori calibration of relative timescales systematically fails to correctly estimate rate variation and divergence times. These results suggest that, irrespective of the implementation, relaxed relative divergence time approaches are not reliable for dating the tree of life— unless we can be confident that the data evolved under a clock, or we can rule out the possibility that multiple, independent substitution rate changes affect the data

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Summary

Introduction

Evolutionary biologists have long sought the timescale of life (Simpson 1944; Betts et al 2018). Traditionally this was done using fossils, the molecular clock (Zuckerkandl and Pauling 1965a, 1965b) has emerged as a powerful tool for extending the value of the fossil record to calibrating molecular evolution to geologic time and, thereby approaching an accurate timescale for the history of life (Avise et al 1992; Erwin et al 2011; Claramunt and Cracraft 2015; Dos Reis et al 2015; Betts et al 2018; Morris et al 2018). The new millennium ushered in a diversity of “relaxed” molecular clock models (Sanderson 1997; Thorne et al 1998; Drummond et al 2006), motivated by these findings, promising better estimation of divergence times when evolutionary rates are variable

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