Biogeography and genetic consequences of anagenetic speciation of Rhaphithamnus venustus (Verbenaceae) in the Juan Fernández archipelago,Chile: insights from AFLP and SSR markers

The genus R haphithamnus (Verbenaceae) consists of two species, one in South America and another endemic to the Juan Fernández archipelago, Chile. The genus represents an example of anagenetic speciation in which the island populations have diverged from their colonizing ancestors to the point where they are recognized as a distinct species. The island species R haphithamnus venustus differs from the continental R . spinosus primarily by floral traits associated with adaptation to hummingbird pollination. Two molecular markers, amplified fragment length polymorphisms (AFLPs) and microsatellites, were used to estimate divergence between the continental and insular species, and to compare diversity in the two species. The comparable or greater diversity in the insular species observed in some diversity indices of AFLPs would support the hypothesis that during the course of anagenetic speciation it has recovered from any reduction of genetic diversity associated with colonization of the archipelago. This pattern of comparable or higher diversity in insular species is seen with other instances of anagenetic speciation in the Juan Fernández archipelago. By contrast, the lower genetic diversity in the insular R . venustus found in microsatellites is likely to be the result of a founder effect from the original colonization of the archipelago; prior molecular studies suggest recent colonization of the Juan Fernández archipelago by R haphithamnus . The seeming non‐concordance between the present results and the widely accepted biogeography of R haphithamnus inferred from other data is discussed and an explanation is presented.     

Phylogeography and molecular phylogeny of Macaronesian island Tarphius (Coleoptera: Zopheridae): why are there so few species in the Azores?

Aim We used a phylogenetic framework to examine island colonization and predictions pertaining to differentiation within Macaronesian Tarphius (Insecta, Coleoptera, Zopheridae), and explain the paucity of endemics in the Azores compared with other Macaronesian archipelagos. Specifically, we test whether low diversity in the Azores could be due to recent colonization (phylogenetic lineage youth), cryptic speciation (distinct phylogenetic entities within species) or the young geological age of the archipelago. Location Macaronesian archipelagos (Azores, Madeira and the Canary Islands), northern Portugal and Morocco. Methods Phylogenetic analyses of mitochondrial and nuclear genes of Tarphius beetles of the Azores, other Macaronesian islands and neighbouring continental areas were used to investigate the origin of island biodiversity and to compare patterns of colonization and differentiation. A comparative nucleotide substitution rate test was used to select the appropriate substitution rate to infer clade divergence times. Results Madeiran and Canarian Tarphius species were found to be more closely related to each other, while Azorean taxa grouped separately. Azorean taxa showed concordance between species and phylogenetic clades, except for species that occur on multiple islands, which segregated by island of origin. Divergence time estimates revealed that Azorean Tarphius are an old group and that the most recent intra‐island speciation event on Santa Maria, the oldest island, occurred between 3.7 and 6.1 Ma. Main conclusions Our phylogenetic approach provides new evidence to understand the impoverishment of Azorean endemics: (1) Tarphius have had a long evolutionary history within the Azores, which does not support the hypothesis of fewer radiation events due to recent colonization; (2) the current taxonomy of Azorean Tarphius does not reflect common ancestry and cryptic speciation is responsible for the underestimation of endemics; (3) intra‐island differentiation in the Azores was found only in the oldest island, supporting the idea that young geological age of the archipelago limits the number of endemics; and (4) the lack of evidence for recent intra‐island diversification in Santa Maria could also explain the paucity of Azorean endemics. Phylogenetic reconstructions of other species‐rich taxa that occur on multiple Macaronesian archipelagos will reveal whether our conclusions are taxon specific, or of a more general nature.     

Higher efficiency of ISSR markers over plastid psbA‐trnH region in resolving taxonomical status of genus Ocimum L.

High level of morphological as well as chemical variability exists within the genus Ocimum, and its taxonomy and phylogenetic relationships are still doubtful. For evaluating interspecific genetic relationships among the Ocimum species, genotyping with intersimple sequence repeat (ISSR) markers and sequence analyses of noncoding psbA‐trnH intergenic region belonging to chloroplast DNA were carried out. Although ISSR markers are highly efficient and reproducible, they have not been used extensively in phylogenetic studies. The use of the plastidial barcode candidate was expected to provide more variable and informative insight into evolutionary rates, and was thus employed as a phylogenetic marker to assess interspecific relationships. This study revealed that the ISSR markers were more efficient than psbA‐trnH sequences in resolving the current status of Ocimum L. genus. Distance‐ and character‐based methodological approaches applied on the molecular data with biparental and maternal inheritance were used for deducing the phylogenetic relationships among Ocimum species. Average polymorphic information content (0.344) and resolving power (6.285) depicted through ISSR markers proved to be efficient in discriminating the studied species of Ocimum. The primers used in this study revealed 99.585% polymorphism across the species demonstrating the polymorphic nature of ISSR markers.     

DNA-DNA and rRNA-DNA hybridization studies in the genus Ectothiorhodospira and other purple sulfur bacteria

DNA-DNA hybridization reveals low DNA homologies (about 14%) between the species of Ectothiorhodospira genus and indicate clearly that the degree of divergence within this genus exceeds the interspecific level. The degree of genome similarities of E. mobilis and E. vacuolata (more than 80% homology) is high and characteristic for the strains of one and the same species.The results of rRNA-DNA and secondary DNA-DNA hybridization indicate the following: Ectothiorhodospira and Thiocapsa are far less related than the genera of one and the same family; the genus Ectothiorhodospira is equidistant from both families of purple sulfur and nonsulfur bacteria. Thus Ectothiorhodospira is a taxon of a higher rank than a genus; we agree with Imhoff's proposal of a new family Ectothiorhodospiraceae.     

Increased molecular divergence of two endemic Trachelipus (Isopoda,Oniscidea) species from Greece reveals patterns not congruent with current taxonomy

In the present study, we employed three mitochondrial DNA genetic markers in a phylogenetic analysis aiming at the delineation of the relationships amongst nominal Trachelipus kytherensis populations, as well as between populations of this species and of Trachelipus aegaeus and a new form, occurring syntopically with the latter. Both the phylogenetic analysis and the genetic distances separating populations, show the presence of several distinct and well differentiated clades that undermine the monophyly of T. kytherensis. On the other hand, despite the insular distribution of T. aegaeus populations, their divergence is low and the monophyly of this taxon can be rescued by the inclusion of two more insular populations previously assigned to T. kytherensis. The patterns of genetic divergence among clades are only partially congruent with the geographic distribution of populations. The validity of taxonomic characters used so far in the genus appears to be questionable; therefore, a more comprehensive phylogenetic study at a population level is deemed necessary for understanding the divergence of Trachelipus lineages. © 2008 The Linnean Society of London, Biological Journal of the Linnean Society, 2008, 95 , 361–370.     

Genetic structure of Micromeria (Lamiaceae) in Tenerife,the imprint of geological history and hybridization on within‐island diversification

Geological history of oceanic islands can have a profound effect on the evolutionary history of insular flora, especially in complex islands such as Tenerife in the Canary Islands. Tenerife results from the secondary connection of three paleo‐islands by a central volcano, and other geological events that further shaped it. This geological history has been shown to influence the phylogenetic history of several taxa, including genus Micromeria (Lamiaceae). Screening 15 microsatellite markers in 289 individuals representing the eight species of Micromeria present in Tenerife, this study aims to assess the genetic diversity and structure of these species and its relation with the geological events on the island. In addition, we evaluate the extent of hybridization among species and discuss its influence on the speciation process. We found that the species restricted to the paleo‐islands present lower levels of genetic diversity but the highest levels of genetic differentiation suggesting that their ranges might have contracted over time. The two most widespread species in the island, M. hyssopifolia and M. varia, present the highest genetic diversity levels and a genetic structure that seems correlated with the geological composition of the island. Samples from M. hyssopifolia from the oldest paleo‐island, Adeje, appear as distinct while samples from M. varia segregate into two main clusters corresponding to the paleo‐islands of Anaga and Teno. Evidence of hybridization and intraspecific migration between species was found. We argue that species boundaries would be retained despite hybridization in response to the habitat's specific conditions causing postzygotic isolation and preserving morphological differentiation.     

Exon primed intron-crossing (EPIC) markers reveal natural hybridization and introgression in Actinidia (Actinidiaceae) with sympatric distribution

Interspecific hybridization is widespread among plants. Understanding the phylogenetic relationships among species is necessary for revealing the potential hybridization events. Actinidia, best known as kiwifruit genus found throughout a wide range in eastern Asian from Indonesia to Siberia. In this study, phylogenetic relationships of Actinidia species with sympatric distributions were investigated using three chloroplast introns (trnL-F, atpB-rbcL and rpl32-trnL) and three Exon primed intron-crossing (EPIC) markers. Chloroplast phylogeny supports non-monophyly of the five species studied excluding Actinidia fulvicoma var. lanata. The non-monophyly was also revealed by EPIC markers. Our results showed EPIC markers are more variable and informative for phylogenetic inference than that of chloroplast markers. The incongruences between loci from the plastid and nuclear DNA phylogenic trees may stem from incomplete lineage sorting or historical introgression hybridization. Incomplete lineage sorting may explain the non-monophyly between Actinidia chrysantha (section Maculatae) and other four species (section Stellatae), and introgression hybridization and high level of interspecific gene flow may explain the non-monophyly among the species of sect. Stellatae. Thus, natural hybridization and introgression may be common in Actinidia with sympatric distribution.     

Molecular diversity of Amansieae (Ceramiales,Rhodophyta) from the Hawaiian Islands: a multi‐marker assessment reveals high diversity within Amansia glomerata

Sixty‐one Hawaiian algal specimens corresponding to members of the tribe Amansieae (Amansia and Osmundaria) were compared through DNA sequence analysis. Short DNA barcode‐like sequences of mitochondrial cytochrome c oxidase subunit I (COI) and universal plastid amplicon (UPA) markers were obtained for as many of the specimens as possible, and a subset of specimens was also used for amplification and sequencing of the nuclear small‐subunit rRNA (SSU) gene for phylogenetic inference in a broader taxonomic context. Statistical parsimony analysis of the COI and UPA markers for A. glomerata produced relationships among the samples that were largely congruent with each other, although the UPA marker was more conserved. The COI marker yielded three lineages, and nucleotide divergences for these three lineages were intermediate to those typically reported for intraspecific and interspecific comparisons, suggesting that they represent either incipient species or a complex of closely related species. The COI and UPA sequences demonstrated little to no divergence for Osmundaria obtusiloba and the taxon referred to as Amansia fimbrifolia. In contrast, specimens identified as A. daemelii were identical in sequence to lineage 3 sequences of A. glomerata, and it is recommended that this taxon no longer be included in species lists for the Hawaiian flora. Phylogenetic reconstruction based on the SSU gene was largely unresolved, indicating that this marker may be of limited utility for this purpose in this group of algae, but a small amount of nucleotide variation was found for samples of A. glomerata.     

Colonization and speciation on volcanic islands: phylogeography of the flightless grasshopper genus Arminda (Orthoptera,Acrididae) on the Canary Islands

Abstract Volcanic archipelagos represent excellent areas to study colonization and speciation processes. The grasshopper genus Arminda is one of many endemic taxa of the Canary Islands. It consists of seven wingless species, most of which are single‐island endemics. We sequenced two mitochondrial (12s rRNA, ND5) and two nuclear gene fragments (28s rRNA, ITS2) to reconstruct the colonization pattern of the genus. Our results are in accordance with a stepping‐stone colonization model from east to west, corresponding to the prevailing ocean currents, but alternative hypotheses cannot be fully rejected. The populations of A. brunneri from Tenerife belong to two different lineages (east and west) consistent with the geological history of the island. It remains to be tested whether these lineages represent different species and whether further lineages exist on this island. The five clades of the four western islands (A. brunneri group) have approximately similar branch lengths. The short internodes between these lineages resulted in a poorer phylogenetic resolution. Specimens from La Palma were genetically distinct and are subsequently described as a new species, Arminda palmae sp.n. Our results suggest in situ speciation on Gran Canaria, which was accompanied by a stronger degree of morphological diversification than the inter‐island speciation processes. The aberrant species A. canariensis has formerly been assigned to a monotypic subgenus Chopardminda, which is now synonymized with Arminda syn.n. based on its phylogenetic position. Gran Canaria seems to be the only island where Arminda species occur sympatrically, although allopatric speciation seems likely due to the long history of volcanism and erosion on the island.     

Hybridization relics complicate barcode‐based identification of species in earthworms

Introgressive hybridization results in mito‐nuclear discordance which could obscure the delimitation of closely related taxa. Although such events are increasingly reported, they have been poorly studied in earthworms. Here, we propose a method for investigating the degree of introgressive hybridization between three taxa of the Allolobophora chlorotica aggregate within two field populations (N = 67 and N = 105) using a reference data set including published DNA barcoding and microsatellite data of all known A. chlorotica lineages (N = 85). For this, we used both molecular phylogenetic and population genetic approaches. The test of correspondence between mitochondrial cytochrome c oxidase I (COI) lineages and clusters of nuclear microsatellite genotypes allowed individuals to be sorted in three categories (matching, admixed and nonmatching) and additional markers (mitochondrial NADH dehydrogenase subunit 1, nuclear Histone 3 and Internal transcribed Spacer Region 2) were used for phylogenetic reconstructions in order to check assignments. Although 15 admixed individuals were observed, no early‐generation hybrids were detected within the two populations. Interestingly, 14 nonmatching individuals (i.e. with a mtDNA haplotype that did not correspond to their nuclear cluster) were detected, a pattern that would result after multiple generations of unidirectional hybridization of female from one taxon to male of the other taxon. Because earthworms are simultaneous hermaphrodites, these events of unidirectional hybridization suggest sterility of the male function in several crosses and highlight that some individuals can be misidentified if reliance is placed on COI barcodes alone. These findings could improve the use of these barcodes in earthworms for species delineation.     

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.     

Dynamic colonization exchanges between continents and islands drive diversification in paradise‐flycatchers (Terpsiphone,Monarchidae)

Aim We use parametric biogeographical reconstruction based on an extensive DNA sequence dataset to characterize the spatio‐temporal pattern of colonization of the Old World monarch flycatchers (Monarchidae). We then use this framework to examine the role of dispersal and colonization in their evolutionary diversification and to compare plumages between island and continental Terpsiphone species. Location Africa, Asia and the Indian Ocean. Methods We generate a DNA sequence dataset of 2300 bp comprising one nuclear and three mitochondrial markers for 89% (17/19) of the Old World Monarchidae species and 70% of the Terpsiphone subspecies. By applying maximum likelihood and Bayesian phylogenetic methods and implementing a Bayesian molecular clock to provide a temporal framework, we reveal the evolutionary history of the group. Furthermore, we employ both Lagrange and Bayes‐ Lagrange analyses to assess ancestral areas at each node of the phylogeny. By combining the ancestral area reconstruction with information on plumage traits we are able to compare patterns of plumage evolution on islands and continents. Results We provide the first comprehensive molecular phylogenetic reconstruction for the Old World Monarchidae. Our phylogenetic results reveal a relatively recent diversification associated with several dispersal events within this group. Moreover, ancestral area analyses reveal an Asian origin of the Indian Ocean and African clades. Ancestral state reconstruction analyses of plumage characters provide an interpretation of the plumage differentiation on islands and continents. Ancestral plumage traits are inferred to be close to those of the Asian paradise‐flycatcher (Terpsiphone paradisi), and island species display a high degree of plumage autapomorphy compared with continental species. Main conclusions Terpsiphone paradisi is polyphyletic and comprises populations that have retained the ancestral plumage of the widespread Terpsiphone genus. The genus appears to have colonized south‐west Asia, the Indian Ocean and Africa from eastern Asia. The phylogeny and divergence time estimates indicate multiple simultaneous colonizations of the western Old World by Terpsiphone. These results reinforce a hypothesis of range expansions of a Terpsiphone paradisi‐like ancestor into eastern Asia and the western Old World.     

Mitonuclear discordance as a confounding factor in the DNA taxonomy of monogonont rotifers

Discordance between mitochondrial and nuclear phylogenies is being increasingly recognized in animals and may confound DNA‐based taxonomy. This is especially relevant for taxa whose microscopic size often challenges any effort to distinguish between cryptic species without the assistance of molecular data. Regarding mitonuclear discordance, two strikingly contrasting scenarios have been recently demonstrated in the monogonont rotifers of the genus Brachionus. While strict mitonuclear concordance was observed in the marine B. plicatilis species complex, widespread hybridization‐driven mitonuclear discordance was revealed in the freshwater B. calyciflorus species complex. Here, we investigated the frequency of occurrence and the potential drivers of mitonuclear discordance in three additional freshwater monogonont rotifer taxa, and assessed its potential impact on the reliability of DNA taxonomy results based on commonly used single markers. We studied the cryptic species complexes of Keratella cochlearis, Polyarthra dolichoptera and Synchaeta pectinata. Phylogenetic reconstructions were based on the mitochondrial barcoding marker cytochrome c oxidase subunit I gene and the nuclear internal transcribed spacer 1 locus, which currently represent the two most typical genetic markers used in rotifer DNA taxonomy. Species were delimited according to each marker separately using a combination of tree‐based coalescent, distance‐based and allele‐sharing‐based approaches. Mitonuclear discordance was observed in all species complexes with incomplete lineage sorting and unresolved phylogenetic reconstructions recognized as the likely drivers. Evidence from additional sources, such as morphology and ecology, is thus advisable for deciding between often contrasting mitochondrial and nuclear species scenarios in these organisms.     

Molecular phylogeny of the genus Asparagus (Asparagaceae) explains interspecific crossability between the garden asparagus (A. officinalis) and other Asparagus species

The genus Asparagus comprises approximately 200 species, some of which are commercially cultivated, such as the garden asparagus (A. officinalis). Many Asparagus species, including A. officinalis, are dioecious and have been grouped into a subgenus distinct from that of hermaphroditic species. Although many interspecific crossings have been attempted to introduce useful traits into A. officinalis, only some of the dioecious species were found to be cross-compatible with A. officinalis. Here, molecular phylogenetic analyses were conducted to determine whether interspecific crossability is proportional to the genetic distance between the crossing pairs and to further clarify the evolutionary history of the Asparagus genus. A clade with all cross-compatible species and no cross-incompatible species was recovered in the phylogenetic tree based on analyses of non-coding cpDNA regions. In addition, a sex-linked marker developed for A. officinalis amplified a male-specific region in all cross-compatible species. The phylogenetic analyses also provided some insights about the evolutionary history of Asparagus; for example, by indicating that the genus had its origin in southern Africa, subsequently spreading throughout the old world through intensive speciation and dispersal. The results also suggest that dioecious species were derived from a single evolutionary transition from hermaphroditism in Asparagus. These findings not only contribute towards the understanding of the evolutionary history of the genus but may also facilitate future interspecific hybridization programs involving Asparagus species.     

PHYLOGENOMICS REVEALS EXTENSIVE RETICULATE EVOLUTION IN XIPHOPHORUS FISHES

Hybridization is increasingly being recognized as a widespread process, even between ecologically and behaviorally divergent animal species. Determining phylogenetic relationships in the presence of hybridization remains a major challenge for evolutionary biologists, but advances in sequencing technology and phylogenetic techniques are beginning to address these challenges. Here we reconstruct evolutionary relationships among swordtails and platyfishes (Xiphophorus: Poeciliidae), a group of species characterized by remarkable morphological diversity and behavioral barriers to interspecific mating. Past attempts to reconstruct phylogenetic relationships within Xiphophorus have produced conflicting results. Because many of the 26 species in the genus are interfertile, these conflicts are likely due to hybridization. Using genomic data, we resolve a high‐confidence species tree of Xiphophorus that accounts for both incomplete lineage sorting and hybridization. Our results allow us to reexamine a long‐standing controversy about the evolution of the sexually selected sword in Xiphophorus, and demonstrate that hybridization has been strikingly widespread in the evolutionary history of this genus.     

Phylogenetics of the common raven complex (Corvus: Corvidae) and the utility of ND4, COI and intron 7 of the β-fibrinogen gene in avian molecular systematics

The common raven (Corvus corax) is one of the most widely distributed and recognizable avian species in the world. Recent molecular work, however, described two mitochondrial lineages of the common raven, termed the Holarctic clade and the California clade, and questioned the monophyly of this taxon by placing the Chihuahuan raven (C. cryptoleucus) sister to the California clade. We evaluated this phylogenetic hypothesis with additional sequence data and increased taxon sampling. We used ～3.7 kb of DNA sequence data from sections of the mitochondrial coding genes COI, cyt b and ND4, a fragment of the non‐coding mitochondrial DNA control region, and the entire intron 7 of the nuclear β‐fibrinogen gene (β‐fibint 7). We combined these DNA sequence data to erect hypotheses of relationships for lineages of the common raven and related taxa. Maximum parsimony, maximum likelihood, and Bayesian methods yield a paraphyletic common raven. These analyses nest the Chihuahuan raven within the common raven, with strong support for a sister relationship between the Chihuahuan raven and the California clade. In addition, the pied crow (C. albus) is also nested within the common raven, and is sister to the Holarctic clade. Our analyses reveal the challenge of determining phylogenetic relationships and species boundaries in this morphologically conservative genus, and suggest that future molecular work with increased taxon sampling will uncover cryptic species and novel evolutionary relationships. Lastly, this survey is one of a growing number of avian phylogenetic studies to employ either β‐fibint 7 or COI, and the first to use ND4. We developed a simple procedure for comparing rates of evolution in molecular markers, and show that in Corvus the nuclear intron β‐fibint 7 is evolving at a considerably slower pace than the mitochondrial markers, while COI is evolving at a slower rate than cyt b, and ND4 approximately the same rate as cyt b. Hence, β‐fibint 7 and other individual nuclear introns may have limited utility in resolving relationships among recently evolved taxa, whereas both COI and ND4 should be useful in a wide range of avian molecular genetic investigations.     

Evidence of taxon cycles in an Indo-Pacific passerine bird radiation (Aves: Pachycephala)

Many insular taxa possess extraordinary abilities to disperse but may differ in their abilities to diversify and compete. While some taxa are widespread across archipelagos, others have disjunct (relictual) populations. These types of taxa, exemplified in the literature by selections of unrelated taxa, have been interpreted as representing a continuum of expansions and contractions (i.e. taxon cycles). Here, we use molecular data of 35 out of 40 species of the avian genus Pachycephala (including 54 out of 66 taxa in Pachycephala pectoralis (sensu lato), to assess the spatio-temporal evolution of the group. We also include data on species distributions, morphology, habitat and elevational ranges to test a number of predictions associated with the taxon-cycle hypothesis. We demonstrate that relictual species persist on the largest and highest islands across the Indo-Pacific, whereas recent archipelago expansions resulted in colonization of all islands in a region. For co-occurring island taxa, the earliest colonists generally inhabit the interior and highest parts of an island, with little spatial overlap with later colonists. Collectively, our data support the idea that taxa continuously pass through phases of expansions and contractions (i.e. taxon cycles).     

Phylogenetic relationships in <Emphasis Type="Italic">Betula</Emphasis> (Betulaceae) based on AFLP markers

The genus Betula comprises various species in boreal and temperate climate zones of the Northern Hemisphere. The taxonomy of Betula is controversial and complicated by parallel evolution of morphological traits, polyploidization events, and extensive hybridization and introgression among species. Multilocus molecular data from AFLPs were used to provide phylogenetic information. A large number of polymorphic markers (321 variable bands) were produced in 107 Betula accessions from 23 species and 11 hybrids. The AFLP results were largely congruent with the results from previously examined nuclear DNA markers. Four distinct subgenera were identified within the genus Betula. These subgenera were partly in disagreement with the traditional (but disputed) division of the genus. In addition, the results indicated several groups of conspecific taxa. The majority of the species fell within subgenus Betula and shared a high degree of similarity with B. pendula. All hybrids were associated with this group, and the AFLP data contained signals on putative parents for some of the interspecific hybrids. Subgenus Chamaebetula and part of the Neurobetula species should be merged with Betula. The subgenera Betulenta, Betulaster, and the remaining part of Neurobetula are distinct and well supported. Although our results indicate that four major taxonomic groups can be recognized within the genus Betula, the relationship between them remains unclear. This may be due to the occurrence of hybridization and introgression, which would have a homogenizing effect on the relationships between species. Naturally occurring Betula species of hybrid origin may explain the low bootstrap values within the Betula clade. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users.