Integrative taxonomy and phylogeny of leafless Vanilla orchids from the South‐West Indian Ocean region reveal two new Malagasy species

The leafless Vanilla species complex from the South‐West Indian Ocean (SWIO) region has long been a taxonomic challenge, due to limited patterns of morphological differentiation and an absence of variation within chloroplast sequences. This complex includes seven known morphospecies: V. madagascariensis, V. bosseri, V. decaryana, and V. perrieri endemic to Madagascar, V. humblotii presumed as endemic to the Comoros Archipelago, but also present in Madagascar, V. roscheri from the East African coast, and V. phalaenopsis endemic to Seychelles. A previous population genetic study using microsatellite markers allowed us to distinguish, in addition to the five recognized Malagasy taxa, two other genetic clusters present in the East of the island. An integrative taxonomy approach was therefore conducted by combining microsatellite and morphological data used in the previous study with new data sets, and by adding ITS sequencing data, to validate the taxonomic level of these Malagasy genetic clusters and unravel phylogenetic relationships between SWIO species. As a result, based on phylogenetic, genotypic and morphological evidence, nine species were discriminated in the SWIO region, including seven in Madagascar, with two new eastern species. The leafless Vanilla group originated and diversified in Madagascar, from an ancestor of African descent, with three subsequent independent colonization events from Madagascar to the other territories of SWIO within the two main lineages (white versus yellow flower species). The new Malagasy species, V. allorgeae Andriamihaja & Pailler sp. nov., and V. atsinananensis Andriamihaja & Pailler sp. nov., are described and a new identification key is proposed.

[1]  D. Monos,et al.  Next-generation sequencing technologies: An overview. , 2021, Human immunology.

[2]  P. Besse,et al.  Drivers of population divergence and species differentiation in a recent group of indigenous orchids (Vanilla spp.) in Madagascar , 2021, Ecology and evolution.

[3]  P. Besse,et al.  Plant DNA Barcoding Principles and Limits: A Case Study in the Genus Vanilla. , 2021, Methods in molecular biology.

[4]  P. Besse,et al.  Genotyping-by-Sequencing Technology in Plant Taxonomy and Phylogeny. , 2021, Methods in molecular biology.

[5]  A. Damián,et al.  A reappraisal of Neotropical Vanilla. With a note on taxonomic inflation and the importance of alpha taxonomy in biological studies , 2020, Lankesteriana.

[6]  Haibao Tang,et al.  A phased Vanilla planifolia genome enables genetic improvement of flavour and production , 2020, Nature Food.

[7]  P. Besse,et al.  The Leafless Vanilla Species-Complex from the South-West Indian Ocean Region: A Taxonomic Puzzle and a Model for Orchid Evolution and Conservation Research , 2020, Diversity.

[8]  L. Gigord,et al.  Bridging the Mozambique Channel: first record of Dombeya rosacea (Dombeyoideae, Malvaceae) for Mayotte (Comoros Archipelago, Indian Ocean) , 2020 .

[9]  F. Hilgen,et al.  The Paleogene Period , 2012 .

[10]  A. W. Ferreira,et al.  Elucidating cryptic sympatric speciation in terrestrial orchids , 2019, bioRxiv.

[11]  S. Reddy,et al.  Phylogeography of the Rufous Vanga and the role of bioclimatic transition zones in promoting speciation within Madagascar. , 2019, Molecular phylogenetics and evolution.

[12]  Alexander Damián Parizaca Vanilla yanesha (Orchidaceae), a new species of the membranaceous-leaved group from the central rainforest of Peru , 2019, Willdenowia.

[13]  Jianquan Liu,et al.  Nuclear simple sequence repeat markers are superior to DNA barcodes for identification of closely related Rhododendron species on the same mountain , 2018, Journal of Systematics and Evolution.

[14]  Deren A. R. Eaton,et al.  Reconciling species diversity in a tropical plant clade (Canarium, Burseraceae) , 2018, PloS one.

[15]  Sudhir Kumar,et al.  MEGA X: Molecular Evolutionary Genetics Analysis across Computing Platforms. , 2018, Molecular biology and evolution.

[16]  M. Suchard,et al.  Posterior summarisation in Bayesian phylogenetics using Tracer , 2022 .

[17]  F. Pinheiro,et al.  Plant Species Complexes as Models to Understand Speciation and Evolution: A Review of South American Studies , 2018 .

[18]  Amal Y. Aldhebiani,et al.  Species concept and speciation , 2017, Saudi journal of biological sciences.

[19]  B. Bremer,et al.  Island hopping, long‐distance dispersal and species radiation in the Western Indian Ocean: historical biogeography of the Coffeeae alliance (Rubiaceae) , 2017 .

[20]  G. C. Fernández-Concha,et al.  Wild Vanilla planifolia and its relatives in the Mexican Yucatan Peninsula: Systematic analyses with ISSR and ITS , 2017 .

[21]  J. A. García-García,et al.  A new vanilla species from Costa Rica closely related to V. planifolia (Orchidaceae) , 2017 .

[22]  E. Pansarin,et al.  Two new species of Vanilla (Orchidaceae) in the Brazilian Atlantic Forest , 2017 .

[23]  R. Francis,et al.  pophelper: an R package and web app to analyse and visualize population structure , 2017, Molecular ecology resources.

[24]  C. Cruaud,et al.  Molecular phylogenetics of Sarcolaenaceae (Malvales), Madagascar's largest endemic plant family , 2016 .

[25]  N. S. Flanagan,et al.  An integrated strategy for the conservation and sustainable use of native Vanilla species in Colombia , 2016 .

[26]  P. Besse,et al.  Combining pollination ecology and fine-scale spatial genetic structure analysis to unravel the reproductive strategy of an insular threatened orchid , 2016 .

[27]  E. Pansarin,et al.  A new species of Vanilla (Orchidaceae: Vanilloideae) from Brazil , 2016 .

[28]  S. Buerki,et al.  Generic delimitations, biogeography and evolution in the tribe Coleeae (Bignoniaceae), endemic to Madagascar and the smaller islands of the western Indian Ocean. , 2016, Molecular phylogenetics and evolution.

[29]  H. Comes,et al.  Spatial patterns of AFLP diversity in Bulbophyllum occultum (Orchidaceae) indicate long-term refugial isolation in Madagascar and long-distance colonization effects in La Réunion , 2016, Heredity.

[30]  Alexander Schwartz,et al.  Systematics And The Origin Of Species , 2016 .

[31]  D. T. Blackburn,et al.  Fire and Late Oligocene to Mid-Miocene peat mega-swamps of south-eastern Australia: a floristic and palaeoclimatic interpretation , 2016 .

[32]  P. Besse,et al.  Active sexual reproduction but no sign of genetic diversity in range-edge populations of Vanilla roscheri Rchb. f. (Orchidaceae) in South Africa , 2014, Conservation Genetics.

[33]  G. Bringmann,et al.  Microsatellites facilitate species delimitation in Congolese Ancistrocladus (Ancistrocladaceae), a genus with pharmacologically potent naphthylisoquinoline alkaloids , 2014 .

[34]  Christian Olsen,et al.  Geneious R8: A Bioinformatics Platform for Biologists , 2014 .

[35]  M. Gaudeul,et al.  Plant taxonomy: a historical perspective, current challenges, and perspectives. , 2014, Methods in molecular biology.

[36]  P. Besse Guidelines for the choice of sequences for molecular plant taxonomy. , 2014, Methods in molecular biology.

[37]  R Core Team,et al.  R: A language and environment for statistical computing. , 2014 .

[38]  Pascale Besse,et al.  Nuclear ribosomal RNA genes: ITS region. , 2014, Methods in molecular biology.

[39]  Rafe M. Brown,et al.  THE CHALLENGE OF SPECIES DELIMITATION AT THE EXTREMES: DIVERSIFICATION WITHOUT MORPHOLOGICAL CHANGE IN PHILIPPINE SUN SKINKS , 2013, Evolution; international journal of organic evolution.

[40]  M. Arenas,et al.  A new infrageneric classification and synopsis of the genus Vanilla Plum. ex Mill. (Orchidaceae: Vanillinae) , 2013 .

[41]  M. Arenas,et al.  A revision of the Mexican and Central American species of Vanilla Plumier ex Miller with a characterization of their ITS region of the nuclear ribosomal DNA , 2013 .

[42]  Cindy E. Hauser,et al.  Quantifying Plant Colour and Colour Difference as Perceived by Humans Using Digital Images , 2013, PloS one.

[43]  Christopher Phillips,et al.  An overview of STRUCTURE: applications, parameter settings, and supporting software , 2013, Front. Genet..

[44]  D. N. Stamos Darwin’s Species Concept Revisited , 2013 .

[45]  M. Chase,et al.  In and out of Madagascar: Dispersal to Peripheral Islands, Insular Speciation and Diversification of Indian Ocean Daisy Trees (Psiadia, Asteraceae) , 2012, PloS one.

[46]  M. Alves,et al.  Integrating different tools to disentangle species complexes: A case study in Epidendrum (Orchidaceae) , 2012 .

[47]  Rod Peakall,et al.  GenAlEx 6.5: genetic analysis in Excel. Population genetic software for teaching and research—an update , 2012, Bioinform..

[48]  S. Kumaria,et al.  Assessment of phylogenetic inter-relationships in the genus Cymbidium (Orchidaceae) based on internal transcribed spacer region of rDNA. , 2012, Gene.

[49]  M. Suchard,et al.  Bayesian Phylogenetics with BEAUti and the BEAST 1.7 , 2012, Molecular biology and evolution.

[50]  Alexei J. Drummond,et al.  Calibrated Tree Priors for Relaxed Phylogenetics and Divergence Time Estimation , 2011, Systematic biology.

[51]  D. Kenfack,et al.  Testing species delimitation in sympatric species complexes: the case of an African tropical tree, Carapa spp. (Meliaceae). , 2012, Molecular phylogenetics and evolution.

[52]  A. Risterucci,et al.  Nineteen polymorphic microsatellite markers from two African Vanilla species: across-species transferability and diversity in a wild population of V. humblotii from Mayotte , 2012, Conservation Genetics Resources.

[53]  G. Rodolphe,et al.  Biodiversity and Evolution in the Vanilla Genus , 2011 .

[54]  O. Grillo,et al.  The Dynamical Processes of Biodiversity - Case Studies of Evolution and Spatial Distribution , 2011 .

[55]  De‐Zhu Li,et al.  Comparative analysis of a large dataset indicates that internal transcribed spacer (ITS) should be incorporated into the core barcode for seed plants , 2011, Proceedings of the National Academy of Sciences.

[56]  B. Leikind,et al.  An Assessment of , 2011 .

[57]  T. Chauhan,et al.  Molecular markers and their applications in fisheries and aquaculture , 2010 .

[58]  C. Thébaud,et al.  Why does the biota of the Madagascar region have such a strong Asiatic flavour? , 2010, Cladistics : the international journal of the Willi Hennig Society.

[59]  B. Bremer,et al.  Historical biogeography of the coffee family (Rubiaceae, Gentianales) in Madagascar: case studies from the tribes Knoxieae, Naucleeae, Paederieae and Vanguerieae , 2010 .

[60]  P. Besse,et al.  Evidence of transoceanic dispersion of the genus Vanilla based on plastid DNA phylogenetic analysis. , 2010, Molecular phylogenetics and evolution.

[61]  P. Cribb,et al.  Field Guide to the Orchids of Madagascar , 2010 .

[62]  T. Sieber,et al.  Microsatellite size homoplasies and null alleles do not affect species diagnosis and population genetic analysis in a fungal species complex , 2010, Molecular ecology resources.

[63]  M. Huber,et al.  Mammalian biodiversity on Madagascar controlled by ocean currents , 2010, Nature.

[64]  J. Tregear,et al.  Species Delimitation in the Genus Phoenix ( Arecaceae ) Based on SSR Markers , with Emphasis on the Identity of the Date Palm ( Phoenix dactylifera ) , 2010 .

[65]  J. Wells,et al.  DIVERSITY, NATURAL HISTORY, AND CONSERVATION OF VANILLA (ORCHIDACEAE) IN AMAZONIAN WETLANDS OF MADRE DE DIOS, PERU , 2010 .

[66]  P. Besse,et al.  Evolutionary processes and diversification in the genus #Vanilla# , 2010 .

[67]  Bernhard Seifert,et al.  Integrative taxonomy: a multisource approach to exploring biodiversity. , 2010, Annual review of entomology.

[68]  M. Combes,et al.  Adaptive radiation in Coffea subgenus Coffea L. (Rubiaceae) in Africa and Madagascar , 2010, Plant Systematics and Evolution.

[69]  M. Di Michele,et al.  Plant species delimitation: A comparison of morphological and molecular markers , 2009 .

[70]  M. Vences,et al.  Madagascar as a model region of species diversification. , 2009, Trends in ecology & evolution.

[71]  T. Saxena The Species Concept , 2009 .

[72]  M. Vences,et al.  Phylogenetic relationships of a new species of pseudoxyrhophiine snake (Reptilia: Lamprophiidae: Thamnosophis) suggest a biogeographical link between western and northern Madagascar , 2009 .

[73]  G. Duncan Atlas of the Vegetation of Madagascar - by Justin Moat and Paul Smith , 2008 .

[74]  K. Cameron,et al.  Neotropical roots of a Polynesian spice: the hybrid origin of Tahitian vanilla, Vanilla tahitensis (Orchidaceae). , 2008, American journal of botany.

[75]  D. Posada jModelTest: phylogenetic model averaging. , 2008, Molecular biology and evolution.

[76]  Sébastien Lê,et al.  FactoMineR: An R Package for Multivariate Analysis , 2008 .

[77]  M. Chase,et al.  Phylogenetics and biogeography of Mascarene angraecoid orchids (Vandeae, Orchidaceae). , 2008, Molecular phylogenetics and evolution.

[78]  P. Besse,et al.  Biodiversity and preservation of vanilla: present state of knowledge , 2008, Genetic Resources and Crop Evolution.

[79]  S. S. Veena,et al.  Genetic variations and interrelationships in Vanilla planifolia and few related species as expressed by RAPD polymorphism , 2008, Genetic Resources and Crop Evolution.

[80]  Mark P. Simmons,et al.  A phylogeny of Anisophylleaceae based on six nuclear and plastid loci: ancient disjunctions and recent dispersal between South America, Africa, and Asia. , 2007, Molecular phylogenetics and evolution.

[81]  David J. Lohman,et al.  Cryptic species as a window on diversity and conservation. , 2007, Trends in ecology & evolution.

[82]  K. de Queiroz,et al.  Species concepts and species delimitation. , 2007, Systematic biology.

[83]  R. Petit,et al.  Blind population genetics survey of tropical rainforest trees , 2006, Molecular ecology.

[84]  K. N. Babu,et al.  Interspecific hybridization in vanilla and molecular characterization of hybrids and selfed progenies using RAPD and AFLP markers , 2006 .

[85]  A. Yoder,et al.  Mouse Lemur Phylogeography Revises a Model of Ecogeographic Constraint in Madagascar , 2006 .

[86]  C. Chou,et al.  Molecular phylogeny of Phalaenopsis Blume (Orchidaceae) based on the internal transcribed spacer of the nuclear ribosomal DNA , 2005, Plant Systematics and Evolution.

[87]  G. Evanno,et al.  Detecting the number of clusters of individuals using the software structure: a simulation study , 2005, Molecular ecology.

[88]  B. Dayrat,et al.  Towards integrative taxonomy , 2005 .

[89]  M. Möller,et al.  Phylogeny and biogeography of exacum (gentianaceae): a disjunctive distribution in the Indian ocean basin resulted from long distance dispersal and extensive radiation. , 2005, Systematic biology.

[90]  K. Queiroz**,et al.  A unified concept of species and its consequences for the future of taxonomy , 2005 .

[91]  T. Ricketts,et al.  Terrestrial Ecoregions of Africa and Madagascar: A Conservation Assessment , 2004 .

[92]  S. Renner Plant Dispersal across the Tropical Atlantic by Wind and Sea Currents , 2004, International Journal of Plant Sciences.

[93]  Michael D. Abràmoff,et al.  Image processing with ImageJ , 2004 .

[94]  L. Forrest,et al.  A recircumscription of Begonia based on nuclear ribosomal sequences , 2003, Plant Systematics and Evolution.

[95]  Miguel V Ences,et al.  Phylogeography, systematics and conservation status of boid snakes from Madagascar (Sanzinia and Acrantophis) , 2003 .

[96]  L. Gallo,et al.  Genetic diversity and differentiation in a southern beech subjected to introgressive hybridization , 2001, Heredity.

[97]  L. Nielsen Natural hybridization between Vanilla claviculata (W.Wright) Sw. and V. barbellata Rchb.f. (Orchidaceae): genetic, morphological, and pollination experimental data , 2000 .

[98]  P. Donnelly,et al.  Inference of population structure using multilocus genotype data. , 2000, Genetics.

[99]  H. Siegismund,et al.  Interspecific differentiation and hybridization in Vanilla species (Orchidaceae) , 1999, Heredity.

[100]  J. Miller,et al.  Taxonomic Revision of the Genus Sarcolaena (Sarcolaenaceae) , 1999 .

[101]  L. Rieseberg,et al.  Plant hybridization. , 1998, The New phytologist.

[102]  J. Goudet FSTAT (Version 1.2): A Computer Program to Calculate F-Statistics , 1995 .

[103]  J Mallet,et al.  A species definition for the modern synthesis. , 1995, Trends in ecology & evolution.

[104]  Robert R. Sokal,et al.  PHENETIC TAXONOMY: Theory and Methods , 1986 .

[105]  E. Wiley,et al.  The Evolutionary Species Concept Reconsidered , 1978 .

[106]  L. V. Valen,et al.  Ecological Species, Multispecies, and Oaks , 1976 .

[107]  J. L. Larson,et al.  The Species Concept of Linnaeus , 1968, Isis.

[108]  R. A. Rolfe A Revision of the Genus Vanilla , 1896 .

[109]  A. Bennett The Origin of Species by means of Natural Selection; or the Preservation of Favoured Races in the Struggle for Life , 1872, Nature.

[110]  Augustin Pyramus de Candolle,et al.  Théorie élémentaire de la botanique, ou exposition des principes de la classification naturelle et de l'art de décrire et d'étudier les végétaux , 1813 .