Paleontological and neontological systematics seek to answer evolutionary questions with different data sets. Phylogenies inferred for combined extant and extinct taxa provide novel insights into the evolutionary history of life. Primates have an extensive, diverse fossil record and molecular data for living and extinct taxa are rapidly becoming available. We used two models to infer the phylogeny and divergence times for living and fossil primates, the tip-dating (TD) and fossilized birth–death process (FBD). We collected new morphological data, especially on the living and extinct endemic lemurs of Madagascar. We combined the morphological data with published DNA sequences to infer near-complete (88% of lemurs) time-calibrated phylogenies. The results suggest that primates originated around the Cretaceous–Tertiary boundary, slightly earlier than indicated by the fossil record and later than previously inferred from molecular data alone. We infer novel relationships among extinct lemurs, and strong support for relationships that were previously unresolved. Dates inferred with TD were significantly older than those inferred with FBD, most likely related to an assumption of a uniform branching process in the TD compared with a birth–death process assumed in the FBD. This is the first study to combine morphological and DNA sequence data from extinct and extant primates to infer evolutionary relationships and divergence times, and our results shed new light on the tempo of lemur evolution and the efficacy of combined phylogenetic analyses.
Online Supplemental Materials v2
Description of online supplementary materials files, figures, tables, methods, results and morphological character descriptions.
Online Supplemental Materials.docx
OSM File S1. Full 421 character morphological data matrix
Morphological data matrix in nexus format with all 421 characters coded for 81 taxa. The file can be opened in the software Mesquite for easy editing, or read directly with any text editor. Data are also available on MorphoBank project # 2167 (www.morphobank.org).
OSM File S2. Character dissimilarity analysis code
R code for running the character dissimilarity analysis described in the text. Comments throughout the code should allow the user to perform the analysis "out-of-the-box". Analysis used to identify characters with 0 dissimilarity, indicating they were possibly not independent.
OSM File S3. Reduced 369 character morphological data matrix
Reduced morphological data matrix of 369 characters after excluding those characters found to have 0 dissimilarity from the Gower analysis.
OSM File 3. Reduced 369 character morphological data matrix.nex
OSM File S4. Morphological data sources GenBank accession numbers and dataset tabulations
Sources of morphological and molecular data. Morphological data were either taken from the publication of Seiffert et al. 2015 or scored in this study. GenBank accession numbers are given for each gene used.
OSM File S5. Concatenated molecular dataset w gene partitions
Nexus file of DNA sequence alignments concatenated with the gene partitions indicated
OSM File S6. Total evidence dataset with reduced morphological data matrix including MrBayes blocks for FBD and TD analyses
Nexus file with total evidence dataset (369 morphological characters, 5767 molecular characters) and the MrBayes blocks of code to run the tip-dating and fossilized birth-death process analyses described in the text. The optimal data partitions, including the two molecular partitions identified by PartitionFinder analysis, are specified. The codes for the divergence time analyses are included as comments (enclosed by square brackets []). The first block runs the fossilized birth-death process analysis, remove the [] to run. The second block includes the tip-dating analysis. The individual matrices and codes are also available in the MorphoBank project # 2167.
OSM Fig S1 MCC FBD full morph
PDF version of Figure 3 of the main text, enlarged. Mean clade credibility tree from the fossilized birth death process, total evidence dataset with full morphological dataset
OSM Fig S2 MCC FBD reduced morph
Maximum clade credibility tree from fossilized birth-death process with 369 morphological characters and 5767 molecular characters. Posterior probabilities of nodes are labelled.
OSM Fig S3 MCC TD full morph
Online Supplemental Material Figure S3. Maximum clade credibility tree from tip-dating method with 421 morphological characters and 5767 molecular characters posterior probabilities of nodes are labelled.
OSM Fig 3 MCC TD full morph.pdf
OSM Fig S4 MCC TD reduced morph
Maximum clade credibility tree from tip-dating method with 369 morphological characters and 5767 molecular characters posterior probabilities of nodes are labelled.
OSM Fig S5 Node age estimates with fixed and wide priors
Comparison of node age estimates for 21 select nodes, using two divergence time estimation techniques, two datasets (full and reduced morphological datasets), and wide age range or fixed age priors
Online Supplemental Material File S7.
1000 trees from the posterior distribution of trees inferred from the total evidence dataset (full morphological dataset) under the fossilized birth-death model in MrBayes.
1000fbd421trees.tre
Online Supplemental Material File S8.
1000 trees from the posterior distribution of trees inferred from the total evidence dataset (reduced morphological dataset) under the fossilized birth-death model in MrBayes.
1000fbd369trees.tre
Online Supplemental Material File S9.
1000 trees from the posterior distribution of trees inferred from the total evidence dataset (full morphological dataset) using tip-dating under the uniform clock model in MrBayes.
1000td421trees.tre
Online Supplemental Material File S10.
1000 trees from the posterior distribution of trees inferred from the mtDNA dataset in MrBayes.
1000mtdna_trees_combined.tre
Online Supplemental Material File S11.
1000 trees from the posterior distribution of trees inferred from the nDNA dataset in MrBayes.
1000ndna_trees_combined.tre
Online Supplemental Material File S12.
1000 trees from the posterior distribution of trees inferred from the morphological dataset in MrBayes.
1000morph_trees_combined.tre
Online Supplemental Material File S13.
The primary concordance tree inferred using Bayesian concordance analysis (BUCKy) of the posterior distributions of trees inferred from individual analysis of the mtDNA, nDNA and morphology.
PCT_rooted.tre
Online Supplemental Material File S14.
Tree topology inferred from total evidence dataset and reduced morphological dataset using maximum likelihood (RaxML).
RAxML_bestTree_total_evidence.tree
Online Supplemental Material File S15.
55 most parsimonious tree topologies inferred from total evidence dataset and reduced morphological dataset using the parsimony ratchet (TNT).
pars_trees2.tre
Online Supplemental Material Figure S6.
Maximum clade credibility tree topology for unconstrained (non-clock) Bayesian analysis of mtDNA in MrBayes. Circles on nodes represent the posterior probability of that node. The tree was rooted for drawing purposes only.
mtDNA tree w node probs.pdf
Online Supplemental Material Figure S7.
Maximum clade credibility tree topology for unconstrained (non-clock) Bayesian analysis of nDNA in MrBayes. Circles on nodes represent the posterior probability of that node. The tree was rooted for drawing purposes only.
nDNA tree w node probs.pdf
Online Supplemental Material Figure S8.
Maximum clade credibility tree topology for unconstrained (non-clock) Bayesian analysis of the reduced morphological dataset (369 characters) in MrBayes. Circles on nodes represent the posterior probability of that node. The tree was rooted for drawing purposes only.
morph tree w node probs.pdf
Online Supplemental Material Figure S9.
The primary concordance tree topology inferred using Bayesian concordance analysis (BUCKy) of the posterior distributions of trees inferred from individual analysis of the mtDNA, nDNA and morphology. Numbers on branches indicate the concordance factor, a measure of the average number of data types that support the given branch. The tree was rooted for drawing purposes only.
PCT w CFs.pdf
Online Supplemental Material Figure S10.
Box plots of Robinson-Foulds distances (RF) from the primary concordance tree (PCT) for tree sets inferred from different data types. mtDNA = mitochondrial DNA, nDNA = nuclear DNA, TE = total evidence, Null1 and Null2 = randomized trees to generate null distributions. See main text for further details.
box plot RF distances PCT vs other trees.pdf
Online Supplemental Material Figure S11.
The tree topology with the highest likelihood found by RaxML with the bootstrap support values from 1000 pseudoreplicates indicated on branches.
RaxML tree w bootstraps.pdf
Online Supplemental Material Figure S12.
50% majority rule consensus of 55 most parsimonious tree topologies inferred from the total evidence dataset (reduced morphological dataset) using the parsimony ratchet (TNT).
Parsimony 50 percent majority rule.pdf
Online Supplemental Material File S16
Mean clade credibility tree from FBD analysis of full morphological and full molecular dataset.
fbd421agerange.tre
Online Supplemental Material File S17.
Mean clade credibility tree from FBD analysis of reduced morphological and full molecular dataset.
fbd369agerange_gooddates.tre
Online Supplemental Material File S18.
Mean clade credibility tree from TD analysis of full morphological and full molecular dataset.
td421fixed.tre
Online Supplemental Material File S19.
Mean clade credibility tree from TD analysis of reduced morphological and full molecular dataset.
td369fixed.tre
Online Supplemental Material Figure S13.
Box plots of Robinson-Foulds distances (RF) among trees inferred from the total evidence dataset using different tree inference techniques. ML = maximum likelihood (using RaxML), parsimony = maximum parsimony (using TNT, 55 most parsimonious trees), Bayes = Bayesian inference (using MrBayes v3.2.6).
box plot RF distances ml pars bayes.pdf