THE MITOCHONDRIAL AND NUCLEAR GENETIC HOMOGENEITY OF THE PHENOTYPICALLY DIVERSE DARWIN'S GROUND FINCHES

The most extensively studied group of Darwin's finches is the genus Geospiza, the ground finches, and yet little is known about the evolutionary history and genetic relationships of these birds. Studies using either allozyme or morphological data have been unable to resolve relationships between the six species and numerous populations of ground finches. In this paper we report the results of a study using mitochondrial control region and nuclear internal transcribed spacer (ITS) 1 sequence data. The differentiation of the ground finch species based on morphological data is not reflected in either mitochondrial or nuclear DNA sequence phylogenies. Furthermore, there is little concordance between the mitochondrial haplotypes and ITS alleles found within individuals. We suggest that the absence of species-specific lineages can be attributed to ongoing hybridization involving all six species of Geospiza. There are no long term selective pressures against hybridization within this genus, and therefore a genetically homogenous genus may be maintained indefinitely. Hybridization has apparently played a role in the adaptive radiation of Darwin's finches.     

Population structure, history and gene flow in a group of closely related land snails: genetic variation in Partula from the Society Islands of the Pacific

Previous studies of Partula land snails from the Society Islands, French Polynesia, have shown that populations within species are highly differentiated in terms of their morphology, behaviour, ecology and molecular genetic variation. Despite this level of variability, differences between species are sometimes small, possibly reflecting the fact that reproductive isolation is not always complete and there exists the opportunity for genetic exchange between taxa through hybridization. The present study uses sequence data from a mitochondrial gene to further investigate genetic variation in Society Island Partula. Most populations are found in this study to be highly differentiated, but within individual species there seems to be no simple relationship either between genetic distance and geographical proximity, or between variation in mitochondria and that in allozymes or morphological characteristics. Among species there appears to be no simple correlation between degrees of reproductive isolation and genetic relatedness according to mitochondrial DNA. The results suggest that past events as well as ongoing drift and selection may have been important in affecting patterns of variation. Similarities among species at specific localities suggest that there must have been some genetic exchange in the past, although this may not necessarily reflect ongoing rates of hybridization. The discrepancy between results for different markers probably reflects the differential effects of drift and selection on mitochondrial and nuclear genes.     

Its evolution in Platanus (Platanaceae): homoeologues, pseudogenes and ancient hybridization

BACKGROUND AND AIMS: Platanaceae is an old family of angiosperms extending back to the Early Cretaceous but consisting of a single extant genus, Platanus. Species of Platanus have long been known to hybridize, and the London plane, Platanus x hispanica, is a well-known example of a hybrid species that formed in historical times. In addition, morphological studies have suggested past interspecific or interlineage hybridization and reticulation as possibly important factors in the evolution of the genus. This study aims at unravelling the complex evolutionary information contained in internal transcribed spacer (ITS) sequences. METHODS: The ITS of the 35S nuclear ribosomal DNA are biparentally inherited, multi-copy markers with a high potential for resolving intrageneric relationships especially when ancient hybridization (reticulation) is involved. Phylogenetic trees, splits graphs and motif analysis are used to extract phylogenetic information from 223 cloned ITS sequences, representing ten species and varieties of Platanus. Non-pseudogenous and pseudogenous sequence motives are assessed to explain how different evolutionary modes contribute to possibly conflicting character state patterns in the ITS. KEY RESULTS: It was found that putative non-functional ITS copies ('pseudogenes') form distinct groups in phylograms and splits graphs, and that pseudogenous lineages reflect ancient hybridization events conserved in the ITS. Specifically, pseudogenous clones of an 'Atlantic' North American clade share sequence motives with non-pseudogenous clones of the western ('Pacific') North American P. racemosa species aggregate. In addition, evidence was found for recent lateral gene flow as a possible factor in the evolution of the central Mexican P. rzedowskii. CONCLUSIONS: Broad ITS data sets that cover intra- and interindividual variability reveal past and ongoing speciation processes in Platanus. Evolutionary pathways can be visualized with splits graphs, but not with bifurcating trees.     

Mechanical barriers to introgressive hybridization revealed by mitochondrial introgression patterns in Ohomopterus ground beetle assemblages

To reveal the role of diverged body size and genital morphology in reproductive isolation among closely related species, we examined patterns of, and factors limiting, introgressive hybridization between sympatric Ohomopterus ground beetles in central Japan using mitochondrial NADH dehydrogenase subunit 5 (ND5) gene sequences. We sampled 17 local assemblages that consisted of two to five species and estimated levels of interspecific gene flow using the genetic distance, D(A), and maximum-likelihood estimates of gene flow. Sharing of haplotypes or haplotype lineages was detected between six of seven species that occurred in the study areas, indicating mitochondrial introgression. The intensity and direction of mitochondrial gene flow were variable among species pairs. To determine the factors affecting introgression patterns, we tested the relationships between interspecific D(A) and five independent variables: difference in body size, difference in genital size, phylogenetic relatedness (nuclear gene sequence divergence), habitat difference, and species richness of the assemblage. Body and genital size differences contributed significantly to preventing gene flow. Thus, mechanical isolation mechanisms reduce the chance of introgressive hybridization between closely related species. Our results highlight the role of morphological divergence in speciation and assemblage formation processes through mechanical isolation.     

Extreme gender flexibility: using a phylogenetic framework to infer the evolution of variation in sex allocation, phylogeography, and speciation in a genus of bidirectional sex changing fishes(Lythrypnus, Gobiidae)

The genus Lythrypnus is a group of marine gobies that exhibit extreme gender flexibility as bidirectional sex changers. The genus consists of 20 described species and several undescribed species that are distributed in the Americas. Five species have been characterized with respect to sex allocation and gonad morphology. The hormonal, morphological, and behavioral aspects of sex change have been studied extensively for one species, L. dalli. These data, however, have not been interpreted in an evolutionary context because a phylogenetic hypothesis has not previously been proposed for the genus Lythrypnus. We propose the first phylogenetic hypothesis for the genus based on molecular data from three mitochondrial genes (12s, ND2, and Cytb), one nuclear gene (Rag1) and one nuclear intron (S7). We also include three previously undescribed Lythrypnus species. Our results support the monophyly of the genus with L. heterochroma, an Atlantic species, as the basal taxon. After the divergence of L. heterochroma, there are two main clades, one comprised of species distributed in the Atlantic, the other comprised of species distributed in the Pacific. These data indicate an Atlantic origin for the genus, followed by divergence after the closure of the Isthmus of Panama. Our data also support the monophyly of three previously described species complexes, the L. rhizophora complex and L. dalli complex in the Pacific, and the L. mowbrayi complex in the Atlantic. We mapped patterns of sex allocation within this genus onto the fully resolved and supported topology, and found that sexual plasticity and gender flexibility is likely a synapomorphy for the genus. Overall our results create a well-supported framework to understand the phylogeography of the genus, and to interpret the evolution of sex allocation in Lythrypnus gobies.     

A combination of molecular and morphological approaches resolves species in the taxonomically difficult genus Procladius Skuse (Diptera: Chironomidae) despite high intra-specific morphological variation

Molecular approaches for identifying aquatic macroinvertebrate species are increasingly being used but there is ongoing debate about the number of DNA markers needed to differentiate species accurately. Here, we use two mitochondrial genes (cytochrome oxidase I, cytochrome b) and a nuclear gene (carbamoylphosphate synthetase) to differentiate species variation within the taxonomically challenging chironomid genus Procladius from southern Australia, a genus which is important for pollution monitoring. The mitochondrial genes indicated cryptic species that were subsequently linked to morphological variation at the larval and pupal stage. Two species previously described based on morphological criteria were linked to molecular markers, and there was evidence for additional cryptic species. Each genetic marker provided different information, highlighting the importance of considering multiple genes when dissecting taxonomically difficult groups, particularly those used in pollution monitoring.     

Falling apart and merging: diversification of slender salamanders (Plethodontidae: Batrachoseps) in the American West

The plethodontid genus Batrachoseps , the slender salamanders, is the most diverse clade of salamanders in western North America, but it has posed taxonomic difficulties because it contains many morphologically cryptic species. A segment of the mitochondrial DNA gene cytochrome b was studied for 278 individuals densely sampled from throughout the range of all 18 described species and several undescribed species. Phylogenetic analyses of the mtDNA data identify six major clades, one corresponding to the subgenus Plethopsis and five within a monophyletic subgenus Batrachoseps. All major clades and most species within these clades display strong phylogeographic structuring. Comparisons of mtDNA and allozyme data show that several allozymically cohesive groups are not monophyletic with respect to mtDNA. We suggest that this phenomenon results from fragmentation of populations, divergence in allopatry, and then recontact and gradual merging of units caused predominantly by male-mediated gene flow. The mtDNA offers evidence that populations were once more isolated than they are now, while the patterns of allozyme variation reflect recent and current interactions among populations. The complex patterns of morphological, allozymic and mtDNA variation associated with the constantly changing geological landscape give insight into the nature of processes responsible for species formation in Batrachoseps .  © 2002 The Linnean Society of London. Biological Journal of the Linnean Society , 2002, 76 , 361–391.     

Sexual hybridization experiments and phylogenetic relationships as inferred from rubisco spacer sequences in the genus alaria (phaeophyceae)

Alaria (Alariaceae, Phaeophyceae) is a common genus of kelps generally found in the lower intertidal and shallow subtidal regions of rocky shores subject to strong wave exposure. Fourteen species are currently recognized, of which 11 are found in the cold–temperate North Pacific Ocean. Alaria esculenta (L.) Greville, the type species described from the North Atlantic, exhibits a range of biogeographically correlated morphotypes indicating the possibility of multiple species, subspecies, and/or hybrids. This has led to an unstable taxonomy. We compared five species from the Atlantic and Pacific, including six North Atlantic isolates of A. esculenta. Phylogenetic analyses based on Rubisco spacer sequences resulted in a well-resolved topology of these five species, but did not distinguish between the six biogeographic isolates of A. esculenta. Laboratory hybridization experiments among four A. esculenta isolates showed partial intrafertility. Among five tested Alaria species, interfertility as well as fertility barriers were encountered, inconsistent with reproductive isolation. The data reject both a biological and morphological species concept and support only a phylogenetic species concept for Alaria , demonstrating that morphological variation has evolved independently of molecular variation in the genes under investigation in the species of the genus Alaria.     

Determination of evolutionary units in European representatives of the crab genus Pilumnus

Bristle crabs of the genus Pilumnus (Brachyura: Heterotremata: Pilumnidae) are common inhabitants of European waters. They are easily identifiable as a genus, but with the exception of P. inermis, intrageneric classification turns out to be quite complex. There is no general agreement on the number and distinction of species. Therefore, this genus is well-suited for comparative molecular studies. Specimens of the Pilumnus hirtellus complex, here defined as including Pilumnus hirtellus, P. villosissimus, P. spinifer, P. aestuarii, and an undescribed species, were gathered from throughout the Mediterranean Sea and the eastern Atlantic Ocean. DNA sequence data were obtained from the barcoding region of the cytochrome oxidase 1 mitochondrial gene and used for reconstruction of a phylogenetic tree and a haplotype network. The morphology of the gastric ossicles was compared in the search of separating characters. Our results give evidence for five genetic clusters within the P. hirtellus complex. There is negligible geographic variation within these clusters. Unambiguous mtDNA sequences within morphologically variable local populations argue against possible hybridization. The here encountered evolutionary units are relatively young and possibly allow to study ongoing processes of morphological, genetic, and ecological differentiation, leading to speciation and radiations in the coastal marine environment.     

Population genetic divergence corresponds with species-level biodiversity patterns in the large genus Begonia

Begonia is one of the largest angiosperm genera, containing over 1500 species. Some aspects of the distribution of biodiversity in the genus, such as the geographical restrictions of monophyletic groups, the rarity and morphological variability of widespread species, and a preponderance of narrow endemics, suggest that restricted gene flow may have been a factor in the formation of so many species. In order to investigate whether this inference based on large-scale patterns is supported by data at the population level, we examined the distribution of genetic variation within Begonia sutherlandii in the indigenous forests of Kwazulu-Natal, South Africa, using microsatellite markers. Despite the species being predominantly outbreeding, we found high and significant levels of population structure (standardized      = 0.896). Even within individual populations, there was evidence for clear differentiation of subpopulations. There is thus congruence in evolutionary patterns ranging from interspecific phylogeny, the distribution of individual species, to the levels of population differentiation. Despite this species-rich genus showing a pan-tropical distribution, these combined observations suggest that differentiation occurs over very local scales. Although strongly selected allelic variants can maintain species cohesion with only low levels of gene flow, we hypothesize that in Begonia , gene flow levels are often so low, that divergence in allopatry is likely to be a frequent occurrence, and the lack of widespread species may in part be attributable to a lack of a mechanism for holding them together.     

Nuclear-mitochondrial discordance and gene flow in a recent radiation of toads

Natural hybridization among recently diverged species has traditionally been viewed as a homogenizing force, but recent research has revealed a possible role for interspecific gene flow in facilitating species radiations. Natural hybridization can actually contribute to radiations by introducing novel genes or reshuffling existing genetic variation among diverging species. Species that have been affected by natural hybridization often demonstrate patterns of discordance between phylogenies generated using nuclear and mitochondrial markers. We used Amplified Fragment Length Polymorphism (AFLP) data in conjunction with mitochondrial DNA in order to examine patterns of gene flow and nuclear-mitochondrial discordance in the Anaxyrus americanus group, a recent radiation of North American toads. We found high levels of gene flow between putative species, particularly in species pairs sharing similar male advertisement calls that occur in close geographic proximity, suggesting that prezygotic reproductive isolating mechanisms and isolation by distance are the primary determinants of gene flow and genetic differentiation among these species. Additionally, phylogenies generated using AFLP and mitochondrial data were markedly discordant, likely due to recent and/or ongoing natural hybridization events between sympatric populations. Our results indicate that the putative species in the A. americanus group have experienced high levels of gene flow, and suggest that their North American radiation could have been facilitated by the introduction of beneficial genetic variation from admixture between divergent populations coming into secondary contact after glacial retreats.     

What is in a name? Terfezia classification revisited

Desert truffles (mycorrhizal hypogeous Ascomycota) are found in arid and semi-arid areas of the globe and have great ecological and economic importance. Terfezia is undoubtedly the most diversified of all desert truffle genera, but its taxonomy is far from resolved. Specifically, the large number of newly described species plus the high intraspecific morphological variability observed within some Terfezia lineages as rendered the use of molecular techniques mandatory for specimen's discrimination. But the subsequent increasing amount of sequence data produced also a huge number of undescribed taxa that required determination. We compiled and used the public available ITS data on Terfezia spp. on the custom-curated UNITE database to reconstruct the genus phylogeny. We found at least 17 distinct lineages within the genus and successfully resolved some of the more pressing taxonomic issues, namely the T. leptoderma/olbiensis complex and some misapplied synonymy. Based on this resolved phylogeny, and motivated by the recent new described species, we proposed an identification key to Terfezia genus highlighting the importance of morphological and ecological characterization.     

Genetic Variability of the Grey Wolf Canis lupus in the Caucasus in Comparison with Europe and the Middle East: Distinct or Intermediary Population?

Despite continuous historical distribution of the grey wolf (Canis lupus) throughout Eurasia, the species displays considerable morphological differentiation that resulted in delimitation of a number of subspecies. However, these morphological discontinuities are not always consistent with patterns of genetic differentiation. Here we assess genetic distinctiveness of grey wolves from the Caucasus (a region at the border between Europe and West Asia) that have been classified as a distinct subspecies C. l. cubanensis. We analysed their genetic variability based on mtDNA control region, microsatellite loci and genome-wide SNP genotypes (obtained for a subset of the samples), and found similar or higher levels of genetic diversity at all these types of loci as compared with other Eurasian populations. Although we found no evidence for a recent genetic bottleneck, genome-wide linkage disequilibrium patterns suggest a long-term demographic decline in the Caucasian population – a trend consistent with other Eurasian populations. Caucasian wolves share mtDNA haplotypes with both Eastern European and West Asian wolves, suggesting past or ongoing gene flow. Microsatellite data also suggest gene flow between the Caucasus and Eastern Europe. We found evidence for moderate admixture between the Caucasian wolves and domestic dogs, at a level comparable with other Eurasian populations. Taken together, our results show that Caucasian wolves are not genetically isolated from other Eurasian populations, share with them the same demographic trends, and are affected by similar conservation problems.     

Natural hybridization in the genus Colletia (Rhamnaceae)

Natural hybridization in the genus Colletia (Rhamnaceae). Natural hybridization between two Chilean species of Colletia is assumed on the basis of morphological evidence. The characters of both species, C. hystrix Clos and C. ulicina Gill. & Hook., and those of their putative hybrids are described. The morphological intermediacy of the assumed hybrids is tested through Wells' distance diagram.     

地卷属地衣1中国新记录种——栗色地卷(新拟)

本研究以采集于河北省小五台山自然保护区地卷属地衣为试验材料,通过形态解剖学、地衣次生代谢产物和nrDNA的ITS分子系统学分析,对地卷属地衣进行鉴定,发现小五台山分布地卷属1中国地衣新记录种——栗色地卷（新拟）(Peltigera castanea Goward, Goffinet & Mi?dl.)。该种的识别特征为：地衣体上表面灰褐色至暗褐色,中部稍具光泽;裂片顶端边缘稍具绒毛;粉芽颗粒状,褐色,并聚集成不规则圆形;下表面脉纹扁平,淡白色、淡褐色至淡黑色;假根密集,束状、丛生,上部呈柔毛状。     

Molecular systematics in the genus Mucor with special regards to species encountered in cheese

The genus Mucor, a member of the order Mucorales, comprises different species encountered in cheeses. Although fungi play a fundamental role in cheese manufacturing and ripening, the taxonomy of many fungal species found in cheese is poorly defined; indeed, this is the case for Mucor spp. In the present study, we assessed the phylogenetic relationships among 70 Mucor strains, including 36 cheese isolates, by using a five gene phylogenetic approach combined with morphological analyses. Overall, at least six species of Mucor were identified among the cheese isolates including a possible new taxon. The present study also suggests that the genus Mucor comprises undescribed taxa and needs to be properly defined.     

ddRAD‐seq data reveal significant genome‐wide population structure and divergent genomic regions that distinguish the mallard and close relatives in North America

Recently evolved species typically share genetic variation across their genomes due to incomplete lineage sorting and/or ongoing gene flow. Given only subtle allele frequency differences at most loci and the expectation that divergent selection may affect only a tiny fraction of the genome, distinguishing closely related species based on multi‐locus data requires substantial genomic coverage. In this study, we used ddRAD‐seq to sample the genomes of five recently diverged, New World “mallards” (Anas spp.), a group of dabbling duck species characterized by diagnosable phenotypic differences but minimal genetic differentiation. With increased genomic sampling, we aimed to characterize population structure within this group and identify genomic regions that may have experienced divergent selection during speciation. We analyzed 3,017 autosomal ddRAD‐seq loci and 177 loci from the Z‐chromosome. In contrast to previous studies, the ddRAD‐seq data were sufficient to assign individuals to their respective species or subspecies and to generate estimates of gene flow in a phylogenetic framework. We find limited evidence of contemporary gene flow between the dichromatic mallard and several monochromatic taxa, but find evidence for historical gene flow between some monochromatic species pairs. We conclude that the overall genetic similarity of these taxa likely reflects retained ancestral polymorphism rather than recent and extensive gene flow. Thus, despite recurring cases of hybridization in this group, our results challenge the current dogma predicting the genetic extinction of the New World monochromatic dabbling ducks via introgressive hybridization with mallards. Moreover, ddRAD‐seq data were sufficient to identify previously unknown outlier regions across the Z‐chromosome and several autosomal chromosomes that may have been involved in the diversification of species in this recent radiation.     

The modality of nine Triturus newt hybrid zones assessed with nuclear,mitochondrial and morphological data

The shape of hybrid zones provides insight into genetic isolation between the evolutionary lineages involved: the greater the limitation to gene flow, the farther along the unimodal to bimodal continuum. We study hybrid zone modality in a group of closely related species at a variety of levels of genetic divergence. We explore the degree and the geography of hybridization in 12 transects of Triturus newts, for nine of the ten secondary contact zones in Europe, using nuclear and mitochondrial encoded genetic data and morphological characteristics. Species status is verified by examining the correlation between the three marker systems in a geographical context. At 40 allozyme loci, two transects classify as unimodal, three as bimodal and two as intermediate, while the signal for hybridization is weak or absent in five transects. One zone studied in duplicate was classified as intermediate in one region and as bimodal in another region. mtDNA introgression is frequent and extends beyond nuclear introgression in two transects. Morphology provides additional evidence for hybridity, including one transect for which the signal of nuclear gene flow is weak. Compared to simulations allowing panmixia, the observed allozyme transitions at contact zones show a deficit of backcrossing to various degrees. Over all transects, there is a weak negative relationship between the level of hybridization and allozyme genetic distance for species pairs, consistent with Bateson–Dobzhansky–Muller effects. This observation, based upon highly comparable data for a single genus, supports conclusions derived from analyses over a wide variety of other taxa. © 2014 The Authors. Biological Journal of the Linnean Society published by John Wiley & Sons Ltd on behalf of The Linnean Society of London, Biological Journal of the Linnean Society, 2014, 113 , 604–622.