Phylogenetic investigation and divergence dating of Poa (Poaceae,tribe Poeae) in the Australasian region

The Australasian region contains a significant proportion of worldwide Poa diversity, but the evolutionary relationships of taxa from this region are incompletely understood. Most Australasian species have been placed in a monophyletic Poa subgenus, Poa supersection Homalopoa section Brizoides clade, but with limited resolution of relationships. In this study, phylogenetic relationships were reconstructed for Australasian Poa, using three plastid (rbcL and matK genes and the rpl32‐trnL intergenic spacer) and two nuclear [internal/external transcribed spacer (ITS/ETS)] markers. Seventy‐five Poa spp. were represented (including 42 Australian, nine New Guinean, nine New Zealand and three Australian/New Zealand species). Maximum parsimony, maximum likelihood and Bayesian inference criteria were applied for phylogenetic reconstruction. Divergence dates were estimated using Bayesian inference, with a relaxed clock applied and rates sampled from an uncorrelated log‐normal distribution. Australasian Poa spp. are placed in three lineages (section Brizoides, section Parodiochloa and the ‘X clade’), each of which is closely related to non‐Australasian taxa or clades. Section Brizoides subsection Australopoa is polyphyletic as currently circumscribed. In Australasia, Poa has diversified within the last 4.3 Mya, with divergence dating results broadly congruent with fossil data that record the appearance of vegetation with a prominent grassland understorey or shrubland/grassland mosaic vegetation dating from the mid‐Pliocene. © 2014 The Linnean Society of London, Botanical Journal of the Linnean Society, 2014, 175 , 523–552.     

Genetic characterization of Mortierella alpina by sequencing the 18S‐28S ribosomal gene internal transcribed spacer region

Aims: The aim of this study was to establish a genetic background of Mortierella alpina, which is a cosmopolitan, soil‐inhabiting Zygomycete that also has important biotechnology potential. Methods and Results: A total of 44 18S‐28S ribosomal gene internal transcribed spacer (ITS1‐5·8S rDNA‐ITS2 DNA) regions of M. alpina from three diverse locations (Far East Asia, North America and West‐Central Europe) were sequenced and investigated. The sequences between M. alpina and the three closely related species (Mortierella macrocystis, Mortierella gamsii and Mortierella humilis) showed 74–84% sequence identity. When a phylogenetic tree was constructed with a neighbour‐joining algorithm, four clades of M. alpina isolates were clearly distinct with high bootstrap values. In addition, isolates from the West‐Central Europe were found to have the highest gene and nucleotide diversities. Conclusions: The ITS region was a suitable tool for distinguishing M. alpina from other closely related species. The region also provided information of the diversity of M. alpina. Significance and Impact of the Study: The study established a genetic background of M. alpina for identification and the diversity of M. alpina provided information for further isolation and screening of the fungus.     

Molecular phylogeny of Osmanthus (Oleaceae) based on non‐coding chloroplast and nuclear ribosomal internal transcribed spacer regions

Abstract The phylogenetic relationships of Osmanthus Lour. were investigated using the nuclear ribosomal internal transcribed spacer (ITS) regions and non‐coding chloroplast regions (psbA‐trnH, trnL‐F). The two datasets support the conclusion that Osmanthus is polyphyletic, with some species of the subtribe Oleinae nested within Osmanthus. Osmanthus didymopetalus P. S. Green is nested within the clade formed by species of section Osmanthus in two trees. Osmanthus attenuatus P. S. Green, O. yunnanensis P. S. Green, and O. gracilinervis R. L. Lu of traditional section Osmanthus are clearly divergent from other accessions, and do not form a monophyletic group with other Osmanthus accessions. Osmanthus marginatus Hemsl. is embedded in the clade formed by species of section Osmanthus in the ITS tree. In cpDNA trees all species of section Osmanthus are placed in the large clade and all species of section Leiolea formed a group. The taxonomic incongruence among trees for ITS and cpDNA indicate hybridization, as introgression may have occurred among some species of sections Osmanthus and Leiolea. Phylogeny of Osmanthus is discussed in light of molecular and morphological data, and a revised infrageneric classification with three sections (Leiolea, Siphosmanthu, and Osmanthus) is presented. The section Linocieroides is abandoned and united with section Osmanthus.     

Flower morphology and relationships of Schefflera subintegra (Araliaceae,Apiales): an evolutionary step towards extreme floral polymery

Gross morphology and the development of flowers in Schefflera subintegra (Araliaceae) are examined. The floral groundplan of this species is found to be very similar to that of Tupidanthus calyptratus representing a case of most extreme floral polymery within Araliaceae. Schefflera subintegra differs from T. calyptratus with respect to a lower floral merism (19–43 versus 60–172 stamens and 15–33 versus 60–138 carpels respectively) and by transformation from polysymmetry to disymmetry of flower in the course of its development. Close relationships between S. subintegra, T. calyptratus, and Schefflera hemiepiphytica have been confirmed by phylogenetic analysis based on nuclear ribosomal internal transcribed spacer sequences. These species form a subclade within the Asian Schefflera clade, with T. calyptratus as a sister taxon to two other species. Apart from more or less pronounced floral polymery, the species of this subclade share calyx and corolla without any traits of individual sepals and petals, and also a massive calyptra. As these data suggest, the extremely polymerous flowers of Tupidanthus apparently evolved in two steps: (1) the saltational multiplication of floral elements together with a loss of individuality of sepals in the calyx and petals in the corolla and (2) further polymerization of androecium and gynoecium. Mutation(s) in CLAVATA‐like gene(s) are suggested as a possible mechanism of the saltation event. © 2014 The Linnean Society of London, Botanical Journal of the Linnean Society, 2014, 175 , 553–597.     

Molecular phylogenetics and taxonomic revision of Habenaria section Pentadactylae (Orchidaceae,Orchidinae)

As a step towards a revision of the sectional classification of Neotropical species of Habenaria, we focus here on section Pentadactylae. In its current delimitation, this is the largest of the 14 New World sections and embraces a group of 34 morphologically heterogeneous species. We expanded the sampling of Neotropical species currently placed in this section and performed Bayesian, maximum likelihood and parsimony analyses using nucleotide sequences from one nuclear (internal transcribed spacer, ITS) and three plastid (matK, trnK intron, rps16–trnK) DNA regions. In addition, morphological features of these species were reassessed. Based on our analyses, we propose that Habenaria section Pentadactylae should be recircumscribed to include only seven species: H. pentadactyla (the type species of the section), H. dutrae, H. ekmaniana, H. exaltata, H. henscheniana, H. megapotamensis and H. montevidensis. Thirty‐two species previously assigned to the section grouped within unrelated clades and are therefore excluded from the section. There are no unambiguous morphological synapomorphies for the section, but the group can be confidently recircumscribed and identified on the basis of a combination of diagnostic morphological vegetative and floral characters. Morphological floral features in Habenaria montevidensis are distinct from those of other species in the section, probably as a result of a shift to diurnal pollinators. Following a taxonomic revision of the group, H. crassipes is placed under the synonymy of H. exaltata and neotypes are designated for H. crassipes, H. montevidensis and H. recta (= H. ekmaniana). All species in the section live in marshes or wet grasslands from northern Argentina to central Brazil; most species are concentrated in southern Brazil. Most species are probably rare, and five may be threatened according to the World Conservation Union (IUCN) criteria. © 2014 The Linnean Society of London, Botanical Journal of the Linnean Society, 2014, 175 , 47–73.     

Molecular phylogenetics of Paphiopedilum (Cypripedioideae; Orchidaceae) based on nuclear ribosomal ITS and plastid sequences

Phylogenetic relationships in the genus Paphiopedilum were studied using nuclear ribosomal internal transcribed spacer (ITS) and plastid sequence data. The results confirm that the genus Paphiopedilum is monophyletic, and the division of the genus into three subgenera Parvisepalum, Brachypetalum and Paphiopedilum is well supported. Four sections of subgenus Paphiopedilum (Pardalopetalum, Cochlopetalum, Paphiopedilum and Barbata) are recovered as in a recent infrageneric treatment, with strong support. Section Coryopedilum is also recovered, with low bootstrap but high posterior probability values for support of monophyly. Relationships in section Barbata remain unresolved, and short branch lengths and the narrow geographical distribution of many species in the section suggest that it possibly underwent rapid radiation. Mapping chromosome and genome size data (including some new genome size measurements) onto the phylogenetic framework shows that there is no clear trend in increase in chromosome number in the genus. However, the diploid chromosome number of 2n = 26 in subgenera Parvisepalum and Brachypetalum suggests that this is the ancestral condition, and higher chromosome numbers in sections Cochlopetalum and Barbata suggest that centric fission has possibly occurred in parallel in these sections. The trend for genome size evolution is also unclear, although species in section Barbata have larger genome sizes than those in other sections. © 2012 The Linnean Society of London, Botanical Journal of the Linnean Society, 2012, 170 , 176–196.     

Morphological phylogenetic analysis of Ophrys (Orchidaceae): insights from morpho‐anatomical floral traits into the interspecific relationships in an unresolved clade

Reconstructing the phylogeny of the sexually deceptive orchid genus Ophrys is crucial to our understanding of the evolution of its complex floral morphology. Molecular phylogenetic analyses showed that section Pseudophrys forms a well supported clade with Ophrys bombyliflora, O. tenthredinifera and O. speculum, but were unable to elucidate the relationships between these four groups of taxa. Here we conduct a morphological phylogenetic analysis of this unresolved clade of Ophrys based on a data matrix of 45 macro‐ and micromorphological and anatomical floral characters, using maximum parsimony and Bayesian inference. Our cladistic analysis yielded a single most parsimonious tree and a Bayesian 50% majority‐rule consensus tree which differed in their overall topology but agreed that O. tenthredinifera and O. bombyliflora are not sister groups. The phylogenetic placement of O. tenthredinifera was ambiguous since it shares six valid synapomorphies each with the cluster of O. speculum–O. bombyliflora and with section Pseudophrys. In contrast, O. bombyliflora is most likely the sister group to O. speculum, a finding that rejects an earlier morphological phylogenetic hypothesis and favours the existing molecular trees based on nuclear ITS rather than plastid data. © 2015 The Linnean Society of London, Botanical Journal of the Linnean Society, 2015, 179 , 454–476.     

A Comparison of ITS Nuclear rDNA Sequence Data and AFLP Markers for Phylogenetic Studies in Phyllostachys (Bambusoideae, Poaceae)

Phyllostachys , a large, economically important genus of woody bamboos. DNA sequences of the internal transcribed spacer (ITS) region of nuclear ribosomal DNA (nrDNA) were used in a parsimony analysis. Phyllostachys was well supported as monophyletic with Chimonobambusa as its closest allied genus. The 5S spacer region of nrDNA was investigated but found unsuitable for this purpose. The AFLP analysis showed much higher discriminating power between species and was more useful for phylogenetic reconstruction at this taxonomic level. The combined data were used to review the previous infra-generic classifications. Section Heteroclada Wang & Ye is strongly supported and can be further divided into sub-groups. A group within section Phyllostachys is strongly supported, but a further group of taxa previously included in this section is difficult to place. The ability of the methods to help separate species such as P. sulphurea and investigate genetic diversity at the infra-specific level was also assessed. It is argued that AFLPs could often be the method of choice for phylogenetic studies of closely related taxa for which DNA sequence data provide insufficient resolution. Received 18 April 2000/ Accepted in revised form 10 June 2000     

Phylogeny and evolution of the South African genus Metalasia (Asteraceae–Gnaphalieae) inferred from molecular and morphological data

Metalasia is a genus in tribe Gnaphalieae (Asteraceae), endemic to South Africa and with its main distribution in the Cape Floristic Region. The genus comprises 57 species and, with a number of closely related genera, it constitutes the ‘Metalasia clade’. A species‐level phylogenetic analysis is presented, based on DNA sequences from two nuclear (internal and external transcribed spacer: ITS, ETS) and two plastid (psbA‐trnH, trnL‐trnF) regions together with morphological data. Analyses combining molecular and morphological data attempt not only to resolve species interrelationships, but also to detect patterns in character evolution. Phylogenetic analyses corroborate our earlier study and demonstrate that Metalasia is formed of two equally sized, well‐supported sister groups, one of which is characterized by papillose cypselas. The results differ greatly from earlier hypotheses based on morphology alone, as few morphological characters support the phylogenetic patterns obtained. The two clades of Metalasia do, however, appear to differ in distribution, corresponding to the different rainfall regimes of South Africa. Analyses show a few taxa to be problematic; one example is the widely distributed M. densa which appears to be an intricate species complex. © 2013 The Linnean Society of London, Botanical Journal of the Linnean Society, 2014, 174 , 173–198.     

An initial investigation of the taxonomic status of Saussurea esthonica Baer ex Rupr. utilising DNA markers and sequencing

Saussurea esthonica Baer ex Rupr. is a species of the genus Saussurea that is found in Estonia, Latvia and the Leningrad region in Russia. Only two populations have been identified in Latvia, and it is protected at the national level. The taxonomic relationships within this genus are unclear, and the majority of phylogenetic studies of this genus have been undertaken on species found in Asia. The relationship of S. esthonica to the other Saussurea species is equally unclear, with some suggestions that this species should be regarded as a subspecies of S. alpina. Two different molecular marker techniques, iPBS and ITS sequencing, were utilised to investigate the phylogeny of S. esthonica and its relationship with other Saussurea species, including three European species: S. alpina, S. pygmaea and S. discolor. The phylogeny obtained with the iPBS markers was similar to that obtained with the ITS sequences. The phylogenies obtained were broadly consistent with previous morphological classification, with S. alpina, S. esthonica and S. discolor clustering together. S. pulchella, S. albescens and S. candicans also formed one discrete cluster. The taxonomic status of S. esthonica was not unambiguously determined, with Norwegian and Italian S. alpina populations clustering with Latvian and Estonian S. esthonica populations, and the Austrian S. alpina populations clustering with Austrian S. discolor populations.     

Assessing what is needed to resolve a molecular phylogeny: simulations and empirical data from emydid turtles

Background  

Section Calochroi is one of the most species-rich lineages in the genus Cortinarius (Agaricales, Basidiomycota) and is widely distributed across boreo-nemoral areas, with some extensions into meridional zones. Previous phylogenetic studies of Calochroi (incl. section Fulvi) have been geographically restricted; therefore, phylogenetic and biogeographic relationships within this lineage at a global scale have been largely unknown. In this study, we obtained DNA sequences from a nearly complete taxon sampling of known species from Europe, Central America and North America. We inferred intra- and interspecific phylogenetic relationships as well as major morphological evolutionary trends within section Calochroi based on 576 ITS sequences, 230 ITS + 5.8S + D1/D2 sequences, and a combined dataset of ITS + 5.8S + D1/D2 and RPB1 sequences of a representative subsampling of 58 species.     

Floral scent evolution in the section Pseudophrys: pollinator‐mediated selection or phylogenetic constraints?

• Sexually deceptive orchid species from the Mediterranean genus Ophrys usually interact with one or a few pollinator species by means of specific floral scents. In this study, we investigated the respective role of pollinator‐mediated selection and phylogenetic constraints in the evolution of floral scents in the section Pseudophrys.
• We built a phylogenetic tree of 19 Pseudophrys species based on three nuclear loci; we gathered a dataset on their pollination interactions from the literature and from our own field data; and we extracted and analysed their floral scents using solid phase microextraction and gas chromatography‐mass spectrometry. We then quantified the phylogenetic signal carried by floral scents and investigated the link between plant–pollinator interactions and floral scent composition using phylogenetic comparative methods.
• We confirmed the monophyly of the section Pseudophrys and demonstrated the existence of three main clades within this section. We found that floral scent composition is affected by both phylogenetic relationships among Ophrys species and pollination interactions, with some compounds (especially fatty acid esters) carrying a significant phylogenetic signal and some (especially alkenes and alkadienes) generating dissimilarities between closely related Pseudophrys pollinated by different insects.
• Our results show that in the section Pseudophrys, floral scents are shaped both by pollinator‐mediated selection and by phylogenetic constraints, but that the relative importance of these two evolutionary forces differ among compound classes, probably reflecting distinct selective pressures imposed upon behaviourally active and non‐active compounds.

     

Unravelling the stylar polymorphism in Melochia (Malvaceae): reciprocity and ancillary characters

Heterostyly is a genetic polymorphism in which plant populations possess two (distyly) or three (tristyly) morphs with flowers differing reciprocally in stigma and anther height. Sex organ deployment has been described as being highly variable among and within species of several distylous taxa belonging to different taxonomic groups. However, the number of studies considering within‐species disparities is still limited. For a better overview of the existing amount of variation that can occur within and between heterostylous species, we sampled 46 populations of six Melochia spp., a style‐polymorphic genus in Cuba. We characterized the floral morphology in all populations and described a set of ancillary characters per species. All of these Melochia spp. are distylous, except for the monomorphic M. nodiflora. The S‐morph produces fewer, larger pollen than L‐morphs, and has verrucose ornamentation. The L‐morph produces reticulate pollen and has larger stigmatic papillae than the S‐morph. The monomorphic M. nodiflora shows ancillary characteristics that are similar to the L‐morph individuals in the related species. As expected, there are differences in ancillary characters among species and also dissimilarities in reciprocity among and within species of Melochia. Our results highlight the importance of considering intraspecific variation in the morphometric characterization of heterostylous taxa. © 2014 The Linnean Society of London, Botanical Journal of the Linnean Society, 2014, 176 , 147–158.     

mtDNA sequences of Sphyraena viridensis (Perciformes: Sphyraenidae) from Italy: insights into historical events and the phylogeny of the genus

The yellowmouth barracuda, Sphyraena viridensis, is a Mediterranean native species whose exact distribution is uncertain due to a long‐term taxonomic confusion with Sphyraena sphyraena. Records of this species in the Mediterranean Sea have recently increased, and a northwards expansion of its distribution has been suggested. Three mtDNA regions, namely cytochrome oxidase I, cytochrome b and the control region, were analysed in S. viridensis samples from Italian coastal regions to provide molecular markers useful in species identification, in phylogenetic analysis and in detecting the distribution of genetic variability of the yellowmouth barracuda in this area. The data clearly distinguish S. viridensis from S. sphyraena and the other four (one native and three Lessepsian) Mediterranean Sphyraena species and identify two clearly distinct lineages that diverged during the Pleistocene but are currently panmictic in the investigated area. Both lineages retain signatures of historical population expansion. © 2014 The Linnean Society of London, Biological Journal of the Linnean Society, 2014, 113 , 635–641.     

Phylogenetic analysis of the genus Hexachlamys (Myrtaceae) based on plastid and nuclear DNA sequences and their taxonomic implications

Myrtaceae are one of the most species‐rich families of flowering plants in the Neotropics. They include several complex genera and species; Hexachlamys is one of the complex genera. It has not been recognized as a distinct genus and has been included in Eugenia, based on morphological grounds. Therefore, molecular systematic studies may be useful to understand and to help to solve these relationships. Here, we performed a molecular phylogenetic analysis using plastid and nuclear data in order to check the inclusion of Hexachlamys in Eugenia. Plastid (accD, rpoB, rpoC1, trnH‐psbA) and nuclear (ITS2) sequence data were analysed using Bayesian and maximum parsimony methods. The trees constructed using ITS2 and trnH‐psbA were the best able to resolve the relationships between species and genera, revealing the non‐monophyly of Hexachlamys. The molecular phylogenetic analyses were in agreement with previous morphological revisions that have included Hexachlamys in Eugenia. These results reinforce the importance of uniting knowledge and strategies to understand better issues of delimitation of genera and species in groups of plants with taxonomic problems. © 2013 The Linnean Society of London, Botanical Journal of the Linnean Society, 2013, 172 , 532–543.     

Plastid DNA sequence data help to clarify phylogenetic relationships and reticulate evolution in Lycoris (Amaryllidaceae)

The temperate East Asian genus Lycoris is a well known lineage of ornamental geophytes consisting of at least 20 species, some of which are thought to be of natural hybrid origin. Previous genetic studies have supported this hypothesis, but these have relied solely on the use of karyology and/or nuclear ribosomal ITS sequences. No plastid DNA data have been available to address interspecific relationships within Lycoris until now. In this study, 500 individuals from 29 populations representing 16 of the 20 published Lycoris spp. were sampled, and DNA sequences were generated for two plastid markers (trnS‐trnfM and trnC‐ycf6). From these data we inferred phylogenetic relationships among the sampled taxa at the species and population levels using concatenated phylogenetic methods. A well resolved and strongly supported phylogenetic reconstruction for Lycoris was obtained. Although the plastid DNA topology differs from that derived previously using ITS, both genomes produce trees that cluster Lycoris spp. into three clades. One of these, containing polyploid taxa such as L. albiflora, L. caldwellii, L. straminea and L. houdyshelii, shows strong evidence of reticulation, and we discuss the identity of potential parents of these allopolyploids. In contrast, we offer evidence that challenges the hypothesis that triploid individuals of L. radiata are the result of hybridization. Instead, they appear to be autotriploids that have arisen in more than one location. By comparing the phylogenetic results obtained using nuclear genomic data to those from the plastid genome, a much clearer picture of the role that hybridization and reticulation have played in the evolution of Lycoris is emerging. © 2014 The Linnean Society of London, Botanical Journal of the Linnean Society, 2014, 176 , 115–126.     

Molecular phylogenetic analysis of Tulipa (Liliaceae) based on noncoding plastid and nuclear DNA sequences with an emphasis on Turkey

We investigated the phylogenetic relationships in Tulipa in Turkey using DNA sequences from the plastid trnL‐trnF region and the internal transcribed spacer (ITS) of nuclear ribosomal DNA. We generated trnL‐trnF and nuclear ITS sequences for 11 Tulipa spp. from Turkey and compared the utility of trnL‐trnF and ITS sequences for phylogenetic analysis. Neighbor‐joining, Bayesian and maximum parsimony methods were implemented using the same matrices. Our study of Tulipa based on molecular data revealed congruent results with previous studies. Despite the relatively lower resolution of trnL‐trnF than that of ITS, both sequence matrices generated similar results. Three clades were clearly distinguished, corresponding to subgenera Tulipa, Eriostemones and Orithyia. It is not fully resolved whether Clusianae should be recognized as a separate section of subgenus Tulipa or a distinct subgenus. © 2013 The Linnean Society of London, Botanical Journal of the Linnean Society, 2013, 172 , 270–279.     

Complex evolutionary relationships in Origanum section Majorana (Lamiaceae)

Origanum (Lamiaceae) comprises a number of essential oil‐rich species that have been used by humans for centuries. Today, the four species of section Majorana (O. onites, O. dubium, O. majorana and O. syriacum) are amongst the most widely used. Despite the importance of this section, phylogenetic relationships and species boundaries amongst its four taxa are unclear. In the present investigation, we used DNA sequence data from two nuclear regions [internal transcribed spacer (ITS) and 1‐deoxy‐d ‐xylulose 5‐phosphate synthase (DXS)] as well as five microsatellite loci to test the taxonomic status of the four species of section Majorana. The combined DNA data revealed O. onites and O. syriacum as the older species in the section. Origanum majorana descends directly from O. syriacum. Origanum dubium was found to be of hybridogenous origin showing attributes of O. onites, O. syriacum and a third, unknown, Origanum species. Both sequence and microsatellite analyses provided evidence for recent hybridization between O. onites and O. dubium in Turkey. © 2013 The Linnean Society of London, Botanical Journal of the Linnean Society, 2013 , 171 , 667–686.     

Phylogenetics and evolution of phyllotaxy in the Solomon's seal genus Polygonatum (Asparagaceae: Polygonateae)

Polygonatum is the largest and most complex genus in tribe Polygonateae, comprising approximately 57 species widely distributed in the warm temperate, subtropical and boreal zones of the Northern Hemisphere. However, phylogenetic relationships in the genus remain poorly understood. The objectives of this study were to reconstruct the phylogenetic relationships of the genus using four plastid markers, and to examine the evolution of leaf arrangement in Polygonatum in the phylogenetic context of its closely related taxa. Thirty Polygonatum species were sampled to infer phylogenetic relationships using maximum‐likelihood and Bayesian analyses. The evolution of leaf arrangements was reconstructed using Bayesian, parsimony and likelihood methods. The phylogenetic analyses supported the current generic delimitation of Polygonatum, with Heteropolygonatum recognized as a distinct genus. Three major lineages in Polygonatum were well supported, largely correlated with geographical distribution and the most recent classification at the sectional level. However, our results did not support the currently recognized series, especially the two large series Verticillata and Alternifolia. Bayesian analyses support the alternate‐leaf arrangement as the ancestral state for Polygonatum, but parsimony and maximum‐likelihood analyses suggest an equivocal state for crown Polygonatum. Leaf arrangement was found to be evolutionarily labile. A new nomenclatural combination was made: P olygonatum section S ibirica (L.I.Abramova) Y.Meng, comb. nov. © 2014 The Linnean Society of London, Botanical Journal of the Linnean Society, 2014, 176 , 435–451.