Species delimitation in the green algal genus Codium (Bryopsidales) from Korea using DNA barcoding

Codium, one of the largest marine green algal genera, is difficult to delimit species boundary accurately based on morphological identification only. DNA barcoding is a powerful tool for discriminating species of seaweeds. The plastid elongation factor TU （tufA） is considered as maker to perform DNA barcoding of green algal species than rbcL gene due to universality and rapid evolution rate. We conducted DNA barcoding application to Codium specimens from the Jeju Island, Korea to overcome the limit of morphological identification and to confirm the species diversity. As a result of applying tufA marker, we newly generated fifty-five tufA barcodes to resolve eight species. TufA marker exhibited 6.1%–21.8% interspecific divergences, wider than the gap of rbcL exon 1, 3.5%–11.5%. Molecular analysis of rbcL exon 1 sequences of Codium revealed eight distinct species like tufA analysis separated in five phylogenetic groups. DNA barcoding of the genus Codium using tufA marker is more helpful to overcome the limit of morphological identification, and this is more potential to reveal cryptic species and to resolve the relationships among subspecies than rbcL analysis alone. The complement of tufA barcoding and rbcL analyses including morphology for the genus Codium in the northwestern Pacific will give much more reliable achievement for discovering species diversity and resolving the phylogenetic relationships.

Historical mitochondrial genome introgression confounds species delimitation: evidence from phylogenetic inference in the Odorrana grahami species complex

Species delimitation is essential to informing conservation policy and understanding ecological and evolutionary processes.Most of our recent gains in knowledge on animal diversity rely on morphological characteristics and mitochondrial(mt)DNA variation.Concordant results based on both have led to an unprecedented acceleration in the identification of new species and enriched the field of taxonomy.However,discordances are also found commonly between morphological and mtDNA evidence.This confounds species delimitation,especially when gene flow or mt genome introgression has occurred.Here,we illustrate how mt genome introgression among species of the Odorrana grahami complex confounds species delimitation using the combined evidence of morphological characters,mt variation,and thousands of nuclear single-nucleotide polymorphisms(SNPs)from genotyping-by-sequencing(GBS).Fifty-eight samples across the distribution of the O.grahami complex were included.The mtDNA matrilineal genealogy indicated 2 clades,with O.grahami and Odorrana junlianensis clustered together.In contrast,all nuclear evidence including gene trees,species trees,and genetic structure analyses based on GBS data support 3 species with distinct genetic clusters.These 3 distinct genetic clusters also correspond to distinct morphological characters.They affirm the distinct taxonomic entities of both O.grahami and O.junlianensis,as well as a third clade distinct from either.Which species the third clade belongs to remains unclear and will require further testing.The nuclear genomic loci contradict the COI evidence,with indications of rampant historical mt genome introgression among the species of the O.grahami complex.These discordant signals previously confused species delimitation efforts in this group.Based on these findings,we recommend the integration of independent data,especially nuclear genomic evidence,in species delimitation so as to be robust against the pitfalls of mt introgression.

Molecular phylogeographic analyses and species delimitations reveal that Leopoldamys edwardsi(Rodentia:Muridae)is a species complex

Leopoldamys edwardsi is a species with wide distribution ranges in southern China but is not discussed in stud­ies on geographic variation and species differentiation.We used 2 mitochondrial(Cytb,CO1)and 3 nuclear(GHR,IRBP and RAG1)genes to clarify species phylogeography and geographical differentiation.Maximum likelihood(ML)and Bayesian phylogenetic inference(BI)trees consistently indicated that L.edwardsi is a spe­cies complex containing 3 main lineages with high Kimura-2-parameter(K2P)divergences(i.e.lineages LN,LS and LHN)found in the northern and southern China and Hainan Island,respectively.The 3 species delimitation methods,automated barcoding gap discovery,Bayesian poisson tree process analysis and Bayesian phylogenet­ics and phylogeography,consistently supported the existence of cryptic species.Divergence times among the main lineages were inferred to be during the Pleistocene,with LHN/LS split at 1.33 Ma and LN/(LHN+LS)at 2.61 Ma;the diversifications of L.edwardsi complex might be caused by the rapid uplifts of Tibetan Plateau,paleo­climate change and complex topography.The divergence between LHN and LS was probably related to the sepa­ration of Hainan Island from the mainland via the formation of the Qiongzhou Strait.Lineages LN and(LS+LHN)likely diverged due to the Wuyi-Nanling mountain range forming a dispersal barrier.Our results suggested that L.edwardsi complex contains at least 3 distinct species:LHN represents L.hainanensis,endemic to Hainan Island and previously considered as a subspecies L.e.hainanensis;LS represents a cryptic species distributed through­out the southern Chinese continent;and LN represents the nominotypical species L.edwardsi.

Continent-wide vocal leapfrog pattern in Collared Scops Owls obfuscates species boundaries

Much confusion has surrounded the taxonomy of the Collared Scops Owl(Otus bakkamoena)complex,distributed widely across tropical and subtropical Asia.Often divided into three species,modern taxonomies disagree on its treatment,as a lack of deep mitochondrial DNA divergence is seemingly in contrast with patterns of vocal differentiation.Analysing a dataset of territorial calls of 122 individuals across Asia,we identified a vocal leapfrog pattern in which owl populations at the eastern and western ends of the distribution sound more similar to each other but are separated by different-sounding populations in between.The boundaries between vocal types are not congruent with traditional species boundaries.Our data allow for a re-drawing of species boundaries and reveal that Collared Scops Owls follow a biogeographic pattern rare in Southeast Asian birds.However,this pattern is shared with several other owl complexes in that peninsular Malaysian populations form part of a continental Asian species centred further north in Southeast Asia rather than forming part of a Sundaic species centred in the adjacent Indonesian Archipelago.

Phylogeography and Cryptic Species Diversity of Paramesotriton caudopunctatus Species Group(Salamandridae: Paramesotriton) in Guizhou, China

The Paramesotriton caudopunctatus species group is mainly distributed in the karst mountain ecosystems of Guizhou, China. Although some species have been included in previous phylogenetic studies, the evolutionary relationships and divergence-time of members of this species group as a whole remain unexplored. In this study, we report the sequencing of one protein coding mitochondrial gene fragment(ND2) and one nuclear gene(POMC), and use a combination of phylogenetic analyses and coalescent simulations to explore the cryptic diversity and evolutionary history of the P. caudopunctatus species group. Phylogenetic relationships revealed that the P. caudopunctatus species group is composed of two major groups, i. e., East Clade and Western-South Clade. The divergence-time and ancestral area estimation suggested that the P. caudopunctatus species group likely originated in the Doupeng Mountains in Guizhou, China at 12.34 Ma(95% HPD: 8.30–14.73), and intraspecific divergence began at about 2.17 Ma(95% HPD: 1.39–2.97). This timing coincides with the orogenesis of the Miaoling Mountains during the Late Miocene to early Pleistocene. The delimitation of species in the P. caudopunctatus species group supports the existence of the currently identified species, and consensus was confirmed across methods for the existence of least to two cryptic species within what has been traditionally considered to be P. caudopunctatus species group. This study is of significance for understanding the species formation, dispersal, and diversity of the tailed amphibians in the karst mountains ecosystem of Guizhou and the role of the Miaoling Mountains as a geographical barrier to species dispersal.

Multiple lines of evidence supports the two varieties of Halenia elliptica(Gentianaceae)as two species

Delimiting species requires multiple sources of evidence.Here,we delimited two varieties of Halenia elliptica(Gentianaceae)using several lines of evidence,including morphological traits and mating system in a sympatric population,phylogenetic relationships based on nrITS and cpDNA(rpl16)data,and complete chloroplast genome sequences.Comparative analysis of 21 morphological traits clearly separates the two varieties of H.elliptica.Examination of the flowering process and pollination treatments indicate that H.elliptica var.grandiflora produces seeds via outcrossing,whereas H.elliptica var.elliptica produces seeds via mixed mating.Furthermore,hand-pollinated hybridization of the two varieties produced no seeds.Observations of pollinators showed that when bees began a pollination bout on H.elliptica var.grandiflora they preferred to continue pollinating this variety;however,when they began a pollination bout on H.elliptica var.elliptica,they showed no preference for either variety.Phylogenetic analysis confirmed the monophyly of H.elliptica,which was further divided into two monophyletic clades corresponding to the two varieties.A large number of variants from the chloroplast genomes reflected remarkable genetic dissimilarities between the two varieties of H.elliptica.We recommend that the two varieties of H.elliptica should be revised as two species(H.elliptica and H.grandiflora).Our findings indicate that H.elliptica varieties may have split into two separate species due to a shift in mating system,changes in flowering phenology and/or post-pollination reproductive isolation.

DNA barcoding of fishes from Zhoushan coastal waters using mitochondrial COI and 12S rRNA genes

Accurate species identification is a key component of biodiversity research.DNA barcoding is an effective molecular method used for fish species identification.We aimed to study the DNA barcoding of fish in Zhoushan coastal waters,explore the differences and applicability of two gene fragments(12S rRNA and COI)of DNA barcoding in fish species identification,and established a comprehensive fish barcoding reference database.Two hundred and eighty-seven captured fish samples from Zhoushan coastal waters were identified using morphological characteristics and DNA barcoding.A total of 26412S rRNA sequences(belonging to eight orders,31 families,55 genera,and 66 species)and 188 COI sequences(belonging to seven orders,30 families,48 genera,and 58 species)were obtained.The lengths of the 12S rRNA sequences ranged from 165 to 178 bp,and the guanine-cytosine(GC)content was 45.37%.The average 12S rRNA interspecific and intraspecific genetic distances(K2P)were 0.10%and 26.66%,respectively.The length of the COI sequence ranged 574–655 bp,and the content of GC was 45.97%.The average 12S rRNA interspecific and intraspecific genetic distances(K2P)were 0.16%and 27.45%,respectively.The minimum interspecific genetic distances of 12S rRNA and COI(1.23%and 1.86%)were both greater than their maximum intraspecific genetic distances(2.42%and 8.66%).Three molecular analyses(NJ tree,ABGD,and GMYC)were performed to accurately identify and delineate species.Clustering errors occurred when the 12S rRNA sequences were delimited using the NJ tree method,and the delimitation results of ABGD and GMYC are consistent with the final species identification results.Our results demonstrate that DNA barcoding based on 12S rRNA and COI can be used as an effective tool for fish species identification,and 12S rRNA has good application prospects in the environmental DNA(eDNA)metabarcoding of marine fish.

Comprehensive DNA barcode reference library and optimization of genetic divergence threshold facilitate the exploration of species diversity of green lacewings (Neuroptera: Chrysopidae)

Chrysopidae are a family of Neuroptera of significant importance in biocontrol against agricultural pests because of their predatory larvae.Currently,the taxonomy of Chrysopidae lacks a comprehensive revision,which impedes the exploration of species diversity as well as the selection and the conservation of green lacewings as biocontrol agents.We have established a DNA barcode reference library of the Chinese green lacewings based on an approximately complete sampling(95.63%)in 25 of the 34 provincial regions in China,comprising 1119 barcodes of 25 genera and 197 species(representing 85%genera and 43.62%species from China).Combining other 1049 high quality green lacewing DNA barcodes,we first inferred the optimal threshold of interspecific genetic divergence(1.87%)for successful species identification in multiple simulated scenarios based on present data.We further inferred the threshold of genetic divergence(7.77%)among genera with biocontrol significance.The inference and performance of the threshold appears to be mainly associated with the completeness of sampling,the proportion of closely related species,and the analytical approaches.Six new combinations,Apertochrysa platypa(Yang&Yang,1991)comb.nov.,Apertochrysa shennongana(Yang&Wang,1990)comb.nov.,Apertochrysa pictifacialis(Yang,1988)comb.nov.,Apertochrysa helana(Yang,1993)comb.nov.,Plesiochrysa rosulata(Yang&Yang,2002)comb.nov.,and Signochrysa hainana(Yang&Yang,1991),are proposed according to integrative species delimitation.Our library and optimal threshold will effectively facilitate the exploration of species diversity of green lacewings.Our study also provides a methodological reference in molecular delimitation of other insects.

Morphological study and molecular epidemiology of Anisakis larvae in mackerel fish

Objective:To examine and study the morphology,epidemiology,and molecular phylogeny of Anisakis larvae in blue mackerel[Scomber australasicus(Cuvier,1832)]and Indian mackerel[Rastrelliger kanagurta(Cuvier,1816)]using light microscope,scanning electron microscope,molecular phylogeny,and species delimitation methods for confirmation and investigation of Anisakis species and their evolutionary relationship.Methods:A total of 90 fish(45 per species)were purchased from a department store in Chiang Mai,Thailand.Anisakis samples were investigated for morphological characteristics using light and scanning electron microscopes.Molecular phylogeny and species delimitation methods based on the cox2 gene were performed.Results:The prevalence,mean intensity(Mean±SEM),and mean abundance of Anisakis larvae(Mean±SEM)in blue mackerel were 77.78%,6.74±1.320,and 5.24±1.107,respectively,and in Indian mackerel,these values were 13.33%,2.50±0.764,and 0.33±0.159,respectively.Scanning electron microscopy showed the detail of morphological characteristics and provided the different shapes of mucron and excretory pores in Anisakis larvae congruent with the phylogenetic tree.The species tree was congruent with the phylogenetic tree.Conclusions:The prevalence,mean intensity,and mean abundance of Anisakis larvae were higher in blue mackerel.To the best of our knowledge,this is the first time that Anisakis pegreffii was found in blue mackerel in Thailand.The phylogenetic tree also supported the morphological data of Anisakis larvae.However,species delimitation based on cox2 revealed 1-3 possible cryptic species in this genus.Anisakis spp.contamination of fish products is unpleasant and a health concern considering human infection with larvae(anisakiasis)arises.

Gracilariopsis lemaneiformis(Gracilariaceae,Rhodophyta)in the Mexican Coasts:A Case of Disjunct Distribution?

Gracilariopsis lemaneiformis is a widely reported species of marine red algae, with records from around the world. Particularly in Mexico, it has been recorded along the coasts of the Gulf of Mexico and the Mexican Caribbean as well as the Tropical Mexican Pacific;so, its current distribution on the Mexican coasts is disjunct. From its molecular characterization with COI-5P and rbcL sequences of specimens collected at sites in Mexico where this species has been most frequently recorded, our aim was to re-evaluate the current distribution of Gp. lemaneiformis on the Mexican coasts and discuss the taxonomic implications. Phylogenetic analysis, supported by DNA species delimitation methods, genetic distances and morphological comparisons, showed that the current disjunct distribution of Gp. lemaniformis in Mexico is a consequence of taxonomic misidentifications. From our results, Gp. lemaneiformis is the only species of the genus with a distribution in the Tropical Mexican Pacific, whereas Gp. tenuifrons is the only species with a distribution in the Gulf of Mexico and the Mexican Caribbean. Also, we propose that Gp. cata-luziana is to be merged with Gp. tenuifrons, while Gp. costarisensis is to be merged with Gp. lemaneiformis.

The genus Thelonectria(Nectriaceae,Hypocreales,Ascomycota)and closely related species with cylindrocarpon-like asexual states

The genus Thelonectria and closely related species with cylindrocarpon-like asexual states are a group of perithecial ascomycetes in the family Nectriaceae that occur as saprobes and in few cases as pathogens of hardwood trees,shrubs or other plants.Although a key component of forest ecosystems around the world,species relationships and distributions of these fungi are largely unknown.The objectives of this study were to:1)infer species rank phylogenetic relationships of the genus Thelonectria and closely related species with cylindrocarpon-like asexual states and test the monophyly of each of the groups studied;2)delimit taxa establishing taxon circumscriptions;3)resolve nomenclatural issues by identifying redundantly used names and synonyms;and 4)provide an updated outline to the genus,geographical distributions data and identification tools,specifically diagnostic keys and molecular data that can be used as molecular barcodes.The recovered consensus phylogeny resulted in a narrow circumscription of the genus Thelonectria,based on the type T.discophora,excluding one of the common species T.jungneri.According to the phylogenetic analyses,T.jungneri belongs in a segregate clade that should be recognized as a different genus.In the genus Thelonectria,a total of four new species and three new combinations are recognized.Additionally,three new genera,closely related to Thelonectria,are described to accommodate species displaying a morphological resemblance to those of Thelonectria:Cinnamomeonectria gen.nov.with C.cinnamomea as type species,Macronectria gen.nov.with M.jungneri as type species and including four additional newly described species,and Tumenectria gen.nov.with T.laetidisca as type species.

What is a species in fungal plant pathogens?

Scientific names are crucial for communicating knowledge concerning fungi and fungus-like organisms.In plant pathology,they link information regarding biology,host range,distribution and potential risk to agriculture and food security.In the past,delimitation among pathogenic taxa was primarily based on morphological characteristics.Due to distinct species sharing overlapping characteristics,the morphological identification of species is often neither straightforward nor reliable.Hence,the phylogenetic species concept based on molecular phylogenetic reconstructions gained importance.The present opinion discusses what a fungal species is and how identification of species in plant pathology has changed over the past decades.In this context,host-specialization and species complexes are discussed.Furthermore,species concepts in plant pathology are examined using case studies from Bipolaris,Colletotrichum,Curvularia,Diaporthe,Diplodia,Meliola,Plasmopara,rust fungi and Trichoderma.Each entry contains a brief introduction to the genus,concepts used in species identification so far and the problems in describing a species followed by recommendations.The importance of correctly naming and identifying a species is addressed in the context of recent introductions,and we also discuss whether the introduction of new species in pathogenic genera has been overestimated.We also provide guidelines to be considered when introducing a new species in a plant pathogenic genus.

Application of DNA barcoding to the identification of Hymenoptera parasitoids from the soybean aphid （Aphis glycines） in China

Aphis glycines Matsumura is an important pest of soybean in Asia and North America. Hymenoptera parasitoids play a key role in the control of the soybean aphid. Thecorrect identification of parasitoids is a critical step that precedes the assessment of their potential biological control agents. Accurate identification of the majority of the speciesattacking the soybean aphid often requires elaborate specimen preparation and expert taxonomic knowledge. In this study, we facilitated the identification of soybean aphidparasitoids by applying a DNA barcoding approach following a preliminary morphological identification. We generated DNA sequence data from the mitochondrial COI gene andthe D2 region of 28S rDNA to assess the genetic variation within and between parasitoid species emerging from the soybean aphid in China. Fifteen Hymenoptera parasitoid speciesbelonging to 10 genera of five families were identified with little intra-specific variation （0.09% ± 0.06% for 28S and 0.36% ± 0. 18% for COI） and large inter-specific divergence（30.46% ± 3.42% for 28S and 20.4% ± 1.20% for COI）.

The quest for a globally comprehensible Russula language

Since 2007,the quality of Russula descriptions has improved and the use of molecular support for species delimitation and the number of published new species has increased.However,the description style is not consistent and has regional or author-specific patterns.Most recent publications still favour descriptions of spores compared to hymenium and pileipellis elements,and usually only the spore size is provided with statistical support.This study proposes standards for descriptions of the microscopic structure of Russula species(Russulaceae,Agaricomycetes).We present the description template,the template measurements table,the specific terminology and the essential chemical reagents.The proposed standards were tested by mycologists from 11 countries who met at the Russula Microscopy Workshop in Slovakia.Descriptions of 26 species from 9 countries and four continents were prepared,among them R.amarissima,R.castanopsidis,R.seperina and R.subtilis are re-described and 15 species are introduced as new:R.abietiphila,R.amerorecondita,R.aurantioflava,R.echidna,R.flavobrunnescens,R.fluvialis,R.fortunae,R.garyensis,R.gemmata,R.laevis,R.madrensis,R.olivaceohimalayensis,R.purpureogracilis,R.sancti-pauli and R.wielangtae.Seven descriptions for candidate new species are provided without a formal name assignment.Pairwise comparison of species described in this study with available similar descriptions of related species suggests that microscopic characters from all parts of the basidiomata can be equally important for species recognition and they deserve the same treatment including number of measurements and statistics.The majority of recent studies does not recognise differences between the pileus margin and centre,but more than one-third of the species described in this study show distinct differences between the pileus areas,emphasizing the importance to specify the origin of pileipellis observations.This study proved that there is frequently insufficient difference in the ITS barcode between closely related species and that it is necessary to use more genetic markers combined with ecological and geographical data.

Taxonomic revisionofthegenus Lactarius(Russulales,Basidiomycota)in Korea

The genus Lactarius Pers.(Russulales)is a cosmopolitan group of Basidiomycota that forms ectomycorrhizal relationships primarily with both deciduous and coniferous trees.Although the genus has been well-studied in Europe and North America,only fragmentary researches have been carried out on Asian species.In particular,the distribution of Lactarius species in South Korea is poorly understood due to insufficient morphological descriptions and a lack of DNA sequence data.In addition,the misuse of European and North American names has added to confusion regarding the taxonomy of Asian Lactarius species.In this study,the diversity of Lactarius in South Korea was evaluated by employing both morphological and phylogenetic approaches.A multi-locus phylogenetic analysis of 729 Lactarius specimens collected between 1960 and 2017 was performed using the internal transcribed spacer(ITS)region,partial nuclear ribosomal large subunit(nrLSU),partial second largest subunit of RNA polymerase II(rpb2),and minichromosome maintenance complex component 7(mcm7).49 Lactarius species were identified in three Lactarius subgenera:L.subg.Russularia(17 spp.),L.subg.Lactarius(22 spp.),and L.subg.Plinthogalus(10 spp.).Among them,28 Lactarius species were identified as new to science,while just 17 were previously described Lactarius species.Four of the taxa remain un-named due to paucity of materials.A key to Korean Lactarius species,molecular phylogenies,a summary of diversity,and detailed description are provided.

Diversity of nivicolous myxomycetes of the Teberda State Biosphere Reserve (Northwestern Caucasus,Russia)

Nivicolous myxomycete assemblages were surveyed on the northwest of the Greater Caucasian ridge in May-June 2010 and 2011 at a north facing transect between 1,700 and 2,920 m elevation of the summit Malaya Khatipara situated within the Teberda State Biosphere Reserve.Morphological characters of 396 collections representing 45 taxa(39 species,3 varieties,and 3 forms)of myxomycetes in 8 genera and 5 families were recorded.Many(13)taxa are classified as rare(a species represents<0.5%of all records).Only seven species were found to be widely distributed(present in 50%or more of the 17 studied localities).To confirm the assignment of specimens to morphospecies,we obtained independently from determination 145 partial sequences of the 18S SSU rRNA gene from 35 taxa of Lamproderma,Meriderma,Physarum and Diderma,which turned out to represent 58 genotypes.Most of the taxa represented by more than one sequence had several genotypes,with an average of 1.7 genotypes per taxon.Except for three taxonomically difficult groups of species,partial SSU sequences did well correspond with the respective morphospecies and where similar or identical to sequences of specimens from the European Alps,making this marker a good candidate for barcoding in myxomycetes.Species richness and diversity increased from subalpine crooked-stem birch forests(23 species,2 varieties,H′02.8,E00.88,D00.08)to alpine dwarf shrub communities(34 species and 2 varieties,2 forms,H′03.2,E00.89,D00.05)but decreased again for alpine meadows(27 species and 2 varieties,2 forms,H′03.1,E00.91,D00.06).Species richness and alpha-diversity reached maximum values for ground litter,whereas leaves and stems of living shrubs above ground harboured a more depauperate myxomycete assemblage.

Trapped on the Roof of the World:taxonomic diversity and evolutionary patterns of Tibetan Plateau endemic freshwater snails(Gastropoda:Lymnaeidae:Tibetoradix)

The high-elevation Tibetan Plateau(western China)is inhabited by a unique,though not particularly species-rich,community of organisms.We explored the species content and evolutionary history of the Tibetan Plateau endemic freshwater snail genus Tibetoradix.Phylogenetic relationships within the genus were reconstructed based on available sequence data.We used a single-rate Poisson Tree Processes approach for species delimitation and compared putative species-level clades with already described taxa.We found that the genus consists of at least 6 species,of which we described 4 as new to science.Shell and soft body morphology was examined and the radula in Tibetoradix was described for thefirst time.Based on ourfindings,the diversification of the genus did not result in a prominent morphological differentiation and a number of species can be regarded as morphologically cryptic.Single species found in different drainage areas indicate relatively good passive dispersal abilities of the snails.The allopatric distribution of the species could result from competitive exclusion between them.The absence of Tibetoradix spp.outside the Tibetan Plateau could be explained by a scenario of an“evolutionary trap”,where adaptations to high elevation conditions prevented the taxa from a successful colonization of lower elevations.

Cantharellus(Cantharellales,Basidiomycota)revisited in Europe through a multigene phylogeny

Resolving species delimitation issues of European Cantharellus is crucial to correctly name chanterelles around the globe.Thirty names referring to Cantharellus s.str.have been described in Europe,some of which are used in other continents.Based on combined analyses of ITS2,LSU,RPB2 and TEF-1,merely eight species are here recognized in Europe applying the genealogical concordance phylogenetic species recognition criteria,one of which,C.roseofagetorum,is described as new.Morphological characters used in species delimitation are mapped and their variability evaluated.The colour of the hymenophore in young specimens is found to be a rather constant morphological character of taxonomic use.European species of Cantharellus are morphologically distinguished by unique combinations of characters,such as the presence of a pink pileal coating,pileus and hymenophore colour when young,and in some cases,the mean spore length and ecology.Eighteen type specimens from Europe are sequenced.Based on revised species concepts sixteen novel taxonomic synonyms are here proposed for European chanterelles:C.alborufescens(=C.henrici,C.ilicis,C.lilacinopruinatus),C.amethysteus(=C.cibarius subsp.squamulosus,C.cibarius var.umbrinus,C.rufipes),C.cibarius(=C.cibarius var.atlanticus,C.parviluteus),C.ferruginascens(=C.cibarius var.flavipes),C.friesii(=C.ignescens),C.pallens(=C.cibarius var.albidus,C.cibarius var.bicolor,C.subpruinosus),and C.romagnesianus(=C.pseudominimus,C.lourizanianus,C.romagnesianus var.parvisporus).The type of Cantharellus,C.cibarius,is epitypified.Descriptions,colour illustrations and a key to all European species are provided.

Hidden diversity revealed in the freshwater snails,Bythinella and Pseudamnicola,in the Island of Crete

Crete with its complex geomorphological history is the island with the highest number of endemism observed in animal and plant taxa throughout the Aegean archipelago.While other groups of organisms within Crete are well-studied,the freshwater gastropod fauna still remains poorly investigated.Bythinella and Pseudamnicola,are 2 genera of freshwater springsnails,both present on the island,inhabiting springs and other freshwater habitats.Here,we conduct a comprehensive study on the distribution of the different genetic lineages of the 2 gastropod genera in order to assess the mode of their differentiation on the island and infer the actual number of species present in the island.Towards these aims,sequence data from the mitochondrial gene were used and analyzed within a phylogenetic framework.For Bythinella,our results strongly support at least 5 delineated Bythinella spp.inhabiting Crete,which correspond to the already described species from previous studies with the addition of a new one.Bythinella analyses reveal an old time-frame of differentiation with vicariant phenomena being more likely the main drivers shaping the present-day distribution of the genus’genetic lineages.For Pseudamnicola,our data indicate the presence of at least 2 delineated Pseudamnicola spp.with a differentiation more consistent to an isolation-by-distance pattern of a relatively recent origin.Dispersion processes followed by isolation of the populations and/or recent speciation,seem to be the underlying process for the current distribution of Pseudamnicola lineages.