Morphometrics and molecular phylogenetics of Emph

Authors: Murielle SimoDroissart Bonaventure Sonké Vincent Droissart Claire Micheneau Porter P Lowry Olivier J Hardy Gregory M Plunkett Tariq Stévart

Publish Date: 2016/06/15

Volume: 302, Issue: 8, Pages: 1027-1045

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Abstract

Species delimitation within Angraecum section Dolabrifolia is problematic due to morphological variability coupled with overlap in many of the characters traditionally used to distinguish species Recent molecular phylogenetic studies of the genus included three of the five currently described species of the Dolabrifolia group placing them as sister to continental African species of Angraecum sect Pectinaria In preparation for a taxonomic revision of section Dolabrifolia we analyzed morphological and molecular data to reassess the circumscription of each of the five currently described species examined the relationships among members of the section and assessed their position within the genus We used 172 alcoholpreserved specimens to perform multivariate analyses on 15 morphological characters We also collected molecular sequence data from 16 taxa including all members of the section using six DNA regions and analyzed these data with parsimony and Bayesian methods The morphometric study revealed five distinct groups of which four correspond to currently recognized species while the fifth represents a taxonomic novelty Angraecum podochiloides is the most distinctive morphologically recognizable by its narrow leaves bearing whiteyellowish flowers The often confused species Angraecum distichum and Angraecum bancoense are clearly distinguishable by flower size Molecular phylogenetic analyses indicated that section Dolabrifolia forms a wellsupported clade related to the continental African members of section Pectinaria Four species are well delimited while the accessions of Angraecum aporoides formed two wellsupported clades corresponding to two subclusters revealed by the morphometric study The recently published Angraecum poppendickianum is shown to be a synonym of A distichumThe authors are grateful to the curators of WAG and to Jean Philippe Biteau Libreville Gabon for making available their living and DNA collections and for allowing the authors access to their facilities We would like to thank Marie Noël Djuikouo Catherina Guiakam Gyslène Kamdem Narcisse Kamdem Sandrine Mayogo Charlemagne Nguembou Hermann Taedoumg and Lise Zemagho for the collection of specimens in the Yaoundé shadehouse and Eric Akouangou and Christelle Nyangala for the collection of specimens in the Sibang shadehouse Libreville We are grateful to the American Orchid Society for support of the first author’s work in Cameroon Fieldwork was also funded by the Central Africa Regional Program of the Environment CARPE “Sud Expert Plantes” project project 375 under French Ministry of Foreign Affairs DIVEAC and ECOFAC DNA sequencing was supported by the Belgian Fund for Scientific Research FRSFNRS through Grants FRFC 2457710 and MIS 451910 Additional fieldwork herbarium visits and part of laboratory studies were supported by the US National Science Foundation Grant 1051547 T Stévart as PI G M Plunkett as CoPIVoucher information and GenBank accession numbers for taxa used in the phylogenetic analysis of Angraecum sect Dolabrifolia including outgroups For each taxon voucher information between brackets is listed in the following order locality date collector name and collection number herbarium acronym Accession numbers are listed in the following order ITS1 matK rps16 trnCpetN trnLF ycf1 Hyphens indicate that no data are availableIngroup Angraecum aporoides Summerh South Cameroon Bifa 12 Jul 2007 Droissart et al Yaoundé shadehouse 592 BRLU KF672217 KF672266 KF672230 KF672308 KF662332 KF672336 A aporoides Summerh South Cameroon Bipindi 1 Oct 2009 Simo et al Yaoundé shadehouse 1839 BRLU KX060061 KX060081 KX060101 − KX060139 KX060159 A aporoides Summerh Gabon Andok Foula 2 Feb 2008 Jardin Botanique de Bambusa 144 BRLU KX060058 KX060078 KX060098 KX060117 KX060136 KX060156 A aporoides Summerh Gabon Rabi 8 Jul 2011 Jardin Botanique de Bambusa 193 BRLU KX060057 KX060077 KX060097 KX060116 KX060135 KX060155 A aporoides Summerh Gabon sloc 25 May 2007 Jardin Botanique de Bambusa 83 BRLU KX060055 KX060075 KX060095 KX060114 KX060133 KX060153 A aporoides Summerh Gabon sloc 6 Feb 2012 Jardin Botanique de Bambusa 259 BRLU KX060054 KX060074 KX060093 KX060113 KX060132 KX060152 A aporoides Summerh Gabon sloc 2 Mar 2010 Jardin Botanique de Bambusa 131 BRLU KX060046 KX060066 KX060086 KX060106 KX060124 KX060144 A aporoides Summerh Gabon Mont Songo 3 Mar 2008 Accession N° of Cult Tchimbélé shadehouse MBG 161 BRLU KX060059 KX060079 KX060099 KX060118 KX060137 KX060157 A aporoides Summerh São Tomé and Príncipe Praia da Lapa 1 Feb 2008 Accession N° of Cult Bom Successo shadehouse 946 BRLU KF672202 KF672276 KF672242 KF672292 KF662338 KF672340 A aporoides Summerh Gabon sloc 25 May 2007 Cultivated at JardiGab shadehouse N° BTO 30 BRLU KX060056 KX060076 KX060096 KX060115 KX060134 KX060154 Angraecum aff aporoides Gabon Rabi 3 Jun 2010 Accession N° of Cult BR 2009038738 BR KX060051 KX060071 KX060091 KX060110 KX060129 KX060149 A aff aporoides Gabon sloc sd Jardin Botanique de Bambusa 50 BRLU KF672200 KF672286 KF672232 KF672288 KF662340 KF672333 Angraecum atlanticum Stévart and Droissart Gabon Doudou Mountains 12 Sep 1986 van der Laan 1068 BRLU KF672213 KF672284 KF672243 KF672299 KF662343 KF672335 Angraecum bancoense Burg South Cameroon Nkolembonda 14 Oct 2010 Simo et al Yaoundé shadehouse 2462 BRLU KX060047 KX060067 KX060087 KX060107 KX060125 KX060145 A bancoense Burg Littoral Cameroon DoualaEdéa Reserve 6 Jun 2009 Simo et al Yaoundé shadehouse 1581 BRLU KX060053 KX060073 KX060093 KX060112 KX060131 KX060151 A bancoense Burg South Cameroon Monts des Eléphants 26 Jul 2007 Droissart et al Yaoundé shadehouse 620 BRLU KF672221 KF672280 KF67225 KF672311 KF662335 KF672320 A bancoense Burg South Cameroon Nkoltsia 28 Jun 2010 Simo et al Yaoundé shadehouse 2207 BRLU KX060060 KX060080 KX060100 KX060119 KX060138 KX060158 Angraecum distichum Lindl Centre Cameroon Mbam Minkom 2 Aug 2004 Droissart et al Yaoundé shadehouse 62 BRLU KX060062 KX060082 KX060102 KX060120 KX060140 KX060160 A distichum Lindl South Cameroon YokadoumaMoloundou road 23 Aug 2010 Simo et al Yaoundé shadehouse 2309 BRLU KX060049 KX060069 KX060089 KX060108 KX060127 KX060147 A distichum Lindl South Cameroon Ngoyla 17 Sep 2010 Simo et al Yaoundé shadehouse 2394 BRLU KX060048 KX060068 KX060088 − KX060126 KX060146 A distichum Lindl East Cameroon Dja Forest Reserve 24 Jan 2008 Droissart et al Yaoundé shadehouse 924 BRLU KF672227 KF672265 KF672231 KF672289 KF662348 KF672337 A distichum Lindl GuineaBissau Mounts Kalikouma 3 Jun 2010 Accession N° of Cult BR 19540481 BR KX060050 KX060070 KX060090 KX060109 KX060128 KX060148 Angraecum doratophyllum Summerh São Tomé and Príncipe sloc 3 Jun 2010 Accession N° of Cult BR 2009037526 BR KF672224 KF672261 KF672247 KF672296 KF662351 KF672328 Angraecum gabonense Summerh Gabon between Rabi 49 and 50 22 Jun 1992 Arends 957 BRLU KF672209 KF672279 KF672237 KF672295 KF662333 KF672344 Angraecum podochiloides Schltr sloc 3 Jun 2010 Accession N° of Cult BR 200938031 BR KX060052 KX060072 KX060092 KX060111 KX060130 KX060150 A podochiloides Schltr Southwest Cameroon Banyang Mbo wildlife sanctuary 8 Feb 2008 Droissart et al Yaoundé shadehouse 939 BRLU KF672225 KF672281 KF672238 KF672293 KF662330 KF672339 A podochiloides Schltr Gabon sloc sd Accession N° of Cult Tchimbélé shadehouse MBG 656 BRLU KX060045 KX060065 KX060085 KX060105 KX060123 KX060143 A podochiloides Schltr Gabon sloc 15 Feb 2012 Accession N° of Cult Tchimbélé shadehouse MBG 703 BRLU KX060044 KX060064 KX060084 KX060104 KX060122 KX060142 A podochiloides Schltr Gabon sloc 11 Jan 2011 Jardin Botanique de Bambusa 159 BRLU KX060043 KX060063 KX060083 KX060103 KX060121 KX060141 Angraecum pungens Schltr Southwest Cameroon Banyang Mbo wildlife sanctuary 31 May 2011 Simo et al Yaoundé shadehouse 2817 BRLU KF672216 KF672260 KF672249 KF672304 KF662328 KF672338 Angraecum subulatum Lindl Ivory Coast sloc 3 Jun 2010 Cultivated at BR Greenhouse Accession N° BR 2009038839 BR KF672206 KF672285 KF672251 KF672300 KF662355 KF672324 Diaphananthe odoratissima Rchbf PJCribb and Carlsward Rwanda Bugesera 3 Jun 2010 Cultivated at BR Greenhouse Accession N° BR 1991019269 BR KF672208 KF672282 KF672256 KF672315 KF662341 KF672345 Tridactyle aurantiopunctata PJCribb and Stévart São Tomé and Príncipe Pico Papagaio 1 Sep 1999 Stévart 656 BRLU KF672201 KF672287 KF672236 KF672290 KF662356 KF672319 Tridactyle bicaudata Lindl Schltr Democratic Republic of the Congo Kisantu sd Cultivated at Kisantu shadehouse Accession N° KIS 135 BRLU KF672210 KF672263 KF672234 KF672305 KF662346 − Outgroups Polystachya albescens Ridl subsp imbricata Rolfe Summerh South Cameroon Akom II 8 Nov 2010 Simo et al Yaoundé shadehouse 2553 BRLU KF672219 KF672259 KF672246 KF672310 KF662352 KF672317 Polystachya calluniflora Kraenzl Southwest Cameroon Rumpi Hills 2 Nov 2010 Simo et al Yaoundé shadehouse 2527 BRLU KF672214 KF672262 KF672229 − KF662331 KF672329 Polystachya pyramidalis Lindl South Cameroon Eboundja 25 Oct 2010 Simo et al Yaoundé shadehouse 2497 BRLU KF672212 KF672283 KF672245 KF672294 KF662339 KF672332

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