Materials and Methods Sarah K . Morris1†*; Oscar A. Pérez-Escobar1,2†*; Bente B. Klitgård1; Alexandre Antonelli1,2; Alejandro Zuluaga3; Monica Carlsen4; Eve J. Lucas1; Simon Mayo1; Anna Haigh1; Steven Dodsworth5; Olivier Maurin1; Grace Brewer1; Robyn Cowan1; Ilia J. Leitch1; Felix Forest1; William J. Baker1 The origin and diversification of the hyper-diverse flora in the Chocó biogeographic region Objectives Anthurium obtusilobum Silica-gel dried leaf fragments 13 outgroup 27 Anthurium DNA extraction, shearing, library preparation, pooling, hybridization and sequencing as described in Johnson et al. (2018) with Angiosperm353 bait kit HybPiper v 1.3.1 (with Spathiphyllum kochii reference) skimmed plastid genome data (39 Anthurium) multiple sequence alignment use full genomic alignment to constrain combined alignment for Maximum Likelihood tree Secondary calibration age of Pothoideae13 Petrocardium cerrejonense (55.8 Myr)8 divergence time estimation with BEAST v2.0 geographic occurrence data from GBIF ancestral range estimation comparing biogeographical models with R-package BioGeoBEARS v0.2.1 1Royal Botanic Gardens, Kew. TW9 3AE, London, UK 2Gothenburg Global Biodiversity Centre, Sweden 3Universidad del Valle, Colombia 4Missouri Botanical Garden, USA 5University of Bedfordshire, Luton, UK †these authors contributed equally to this study *corresponding authors: o.perez-escobar@kew.org, s.morris@kew.org Alwyn H. Gentry (Annals of the Missouri Botanical Garden, 1996) Geographic extension of the Chocó biogeographical region in Central and South America12 and its compound geological origin14 Extract loci from genomic alignments that match Sanger sequences from Carlsen & Croat (2013) Angiosperm 353 kit successfully enriched 50-333 nuclear genes (& recovered 93 plastid genes) species-tree coalescence analyses revealed extensive incongruence near the MRCA of Anthurium the timing and origin of the MRCA of Anthurium remains unclear multiple migrations into Chocó: 3 at ~9 Mya from Central America 1 at ~5 Mya from Amazonia multiple recent migrations within 1 My from Central America and northern Andes Were Gentry’s ideas about the mode and tempo of plant evolution into the Chocó correct? Chocoan flora is likely older than Gentry assumed (perhaps late Miocene) migrations occurred after rapid mountain building had begun in the northern Andes allopatric speciation due to Andean uplift unlikely origin of Chocoan flora from Central America and northern Andes is supported migrations split between 9 Mya and < 1 Mya, suggesting a burst of migration after the closure of the CAS and again more recently Gentry believed the habit type, dispersal system and pollination syndrome of the Chocoan flora allowed for rapid speciation of endemics7: shorter generation time of shrubby and herbaceous groups bird and mammal (more localized) dispersal specific pollinator relationships To investigate these biotic factors, we are conducting similar genomic studies of other representative groups, including Blakea (Melastomataceae) and Neotropical orchids14 key geologic events in mid-late Miocene: 1. accretion of the Chocó Block4 2. Northern Andean orogeny9 3. closure of the Central American Seaway (CAS)10 The Chocó Introduction Gentry from available floristic, geological and palynological data in the 80s: Chocoan flora originated in the mid- Pleistocene from Amazonian lineages which migrated around the northern Andes, through southern Central America 6 “the historical accident of the Andean uplift, with the concomitant opportunity for explosive speciation among certain taxa … may largely explain the “excess” plant species diversity of the Neotropics” 5 Test Gentry’s hypothesis with genomic data of Anthurium using phylogeny reconstruction, molecular dating and ancestral area estimation We expect to see: a MRCA for the Anthurium lineage in Amazonia a MRCA of Chocoan clades in the northern Andes and Central America migration into the Chocó in the Pleis