RESEARCH ARTICLE Diversification of (, ) in : Revised Phylogeny Reveals Species Accumulation through Time Rather than Rapid Radiation

Herinandrianina N. Andriananjamanantsoa1☯*, Shannon Engberg2☯, Edward E. Louis, Jr2☯, Luc Brouillet1☯

a11111 1 DeÂpartement de sciences biologiques, Universite de MontreÂal, MontreÂal, QueÂbec, Canada, 2 Omaha's Henry Doorly Zoo and Aquarium, Omaha, Nebraska, United States of America

☯ These authors contributed equally to this work. * [email protected]

OPEN ACCESS Abstract Citation: Andriananjamanantsoa HN, Engberg S, Angraecum is the largest of subtribe (Orchidaceae) with about 221 Louis EE, Jr, Brouillet L (2016) Diversification of species. Madagascar is the center of the diversity for the genus with ca. 142 species, of Angraecum (Orchidaceae, Vandeae) in which 90% are endemic. The great morphological diversity associated with species diversi- Madagascar: Revised Phylogeny Reveals Species Accumulation through Time Rather than Rapid fication in the genus on the island of Madagascar offers valuable insights for macroevolu- Radiation. PLoS ONE 11(9): e0163194. tionary studies. Phylogenies of the Angraecinae have been published but a lack of taxon doi:10.1371/journal.pone.0163194 and character sampling and their limited taxonomic resolution limit their uses for macroevo- Editor: William Oki Wong, Institute of Botany, lutionary studies. We present a new phylogeny of Angraecum based on chloroplast CHINA sequence data (matk, rps16, trnL), nuclear ribosomal (ITS2) and 39 morphological charac- Received: March 25, 2016 ters from 194 Angraecinae species of which 69 were newly sampled. Using this phylogeny, Accepted: September 6, 2016 we evaluated the monophyly of the sections of Angraecum as defined by Garay and investi- gated the patterns of species diversification within the genus. We used maximum parsi- Published: September 26, 2016 mony and bayesian analyses to generate phylogenetic trees and dated divergence times of Copyright: © 2016 Andriananjamanantsoa et al. the phylogeny. We analyzed diversification patterns within Angraecinae and Angraecum This is an open access article distributed under the terms of the Creative Commons Attribution with an emphasis on four floral characters ( color, flower size, labellum position, spur License, which permits unrestricted use, length) using macroevolutionary models to evaluate which characters or character states distribution, and reproduction in any medium, are associated with speciation rates, and inferred ancestral states of these characters. The provided the original author and source are credited. phylogenetic analysis showed the polyphyly of Angraecum sensu lato and of all Angraecum sections except sect. Hadrangis, and that morphology can be consistent with the phylog- Data Availability Statement: All sequences data are available from the GenBank database eny. It appeared that the characters (flower color, flower size, spur length) formerly used by (accession numbers KT826761, KT826966). many authors to delineate Angraecum groups were insufficient to do so. However, the Original matrices and trees are available from newly described character, position of the labellum (uppermost and lowermost), was the TreeBase (http://purl.org/phylo/treebase/phylows/ main character delimiting clades within a monophyletic Angraecum sensu stricto. This char- study/TB2:S19429). All pictures and voucher specimens are accessible from Canadensys (http:// acter also appeared to be associated with speciation rates in Angraecum. The macroevolu- data.canadensys.net/explorer/en/search). tionary model-based phylogeny failed to detect shifts in diversification that could be

Funding: This wo