Aim Long-distance dispersal has contributed to the disjunct biogeographical distribution of rain forest plants – something that has fascinated biogeographers since Humboldt’s time. However, the dispersal ‘agent’ for these tropical plant lineages remains puzzling. Here, we investigate which frugivory-related traits may have facilitated past intercontinental long-distance dispersal in the custard apple family (Annonaceae), a major vertebrate-dispersed tropical plant family. We hypothesize that long-distance dispersal was associated with the evolution of traits related to dispersal by large-bodied mammals (e.g. large, dull-coloured, ‘megafaunal’ fruits) and strong-flying, ocean-crossing birds and bats (e.g. dehiscent, moniliform or cauliflorous fruits). Location Global Taxon Annonaceae Methods We used a fossil-calibrated phylogenetic framework to infer the biogeographic history of 234 Annonaceae species (10%, covering nearly all genera) in relation to the evolution of 15 frugivory-related traits, using maximum likelihood and Bayesian inferences. Furthermore, we used linear and generalized linear models and phylogenetic simulations to test whether ancestral fruit traits during intercontinental dispersal were different from those of other lineages not involved in long-distance dispersal. Results We inferred the ancestral Annonaceae fruits to be small with a single or few small seeds and a small number of fruitlets. These fruits were most probably apocarpous, indehiscent and/or moniliform (i.e. long beads of fruitlets). Furthermore, most of the long-distance dispersal events in Annonaceae occurred via the expanded tropical forests in the Early Cenozoic (‘geodispersal’), and were significantly associated with large (c. 3 cm long), dull-coloured fruits, and short stipes. Additionally, long-distance dispersal was also facilitated by dehiscent, moniliform and non-cauliflorous fruits. Main conclusions We suggest that the evolution of frugivory-related traits associated with dispersal by frugivores that frequently move across large distances and/or barriers, such as large-bodied mammals and strong-flying birds, has contributed to the disjunct tropical biogeographical distribution of Annonaceae, and probably of tropical rain forest plants more generally.

Functional trait data and GenBank sequences for AnnonaceaeThis file contains all of the data recorded in the PROTEUS database to produce the NEXUS trait matrix, including: PrimaryCharacters; PrimaryCharacterstates; References; PrimaryDataMatrix; Matrix for analysis; Updated taxonomy; GenBank sequences.Onstein_et_al_Appendix S1.xlsxAnnonaceae maximum clade credibility (MCC) phylogenetic treeWe reconstructed and dated the Annonaceae phylogenetic tree in BEAST 1.8.2. We used the uncorrelated lognormal clock model and a Yule birth prior. Substitution rates of markers were linked, and we used the GTR + gamma + invariant sites substitution rate model. To date the Annonaceae, we used the fossils Endressinia (most-recent common ancestor of Magnoliaceae and Annonaceae; 113 Ma) for the split between Magnoliaceae and Annonaceae, and Futabanthus (89 Ma) for the Annonaceae crown node (for more details, see Appendix S2). Please note that some of the taxonomic names have been changed (can be found in Appendix S1 [= Dryad file "Functional trait data and GenBank sequences in Annonaceae"]) and should be updated in future use of this datafile. In case you need 100 trees from the posterior distribution, please write to Renske Onstein (onsteinre@gmail.com).MCC_treescombineAnnonaceae Supermatrix AlignmentWe sampled 234 Annonaceae species (c. 10% of total) representing 105 (c. 97% of total) genera and all four subfamilies, and seven outgroup taxa from related families. For these species, we generated a molecular supermatrix of eight plastid markers as assembled by Chatrou et al. (2012) supplemented with 157 additional sequences for 37 species. For details on the methods, see Chatrou et al. (2012), GenBank numbers for the new sequences are provided in Appendix S1 (= Dryad file "Functional trait data and GenBank sequences in Annonaceae"). Please note that in the meantime some of the taxonomic names have changed (can also be found in Appendix S1) and should be updated in future use of this datafile.Annonaceae_Supermatrix_MP_3.02_forBEAST.nex