Species delimitation of three species within the Russula subgenus Compacta in Korea: R. eccentrica,R. nigricans,and R. subnigricans

Distinguishing individual Russula species can be very difficult due to extensive phenotypic plasticity and obscure morphological and anatomical discontinuities. In this study, we use the internal transcribed spacer (ITS) and 28S nuclear ribosomal large subunit (LSU) markers to identify and study the genetic diversity of species in the Russula subgenus Compacta in Korea. We focus on two morphologically similar species that are often misidentified for each other: R. nigricans and R. subnigricans. Based on molecular phylogenetic analyses, we identify three subgroups of R. nigricans, with two from Asia and one from Europe/North America. Surprisingly, we find Korean R. subnigricans are more closely related to R. eccentrica from North America than the type specimen of R. subnigricans from Japan. These molecular data, along with habitat data, reveal that Korean R. subnigricans had previously been misclassified and should now be recognized as R. eccentrica. Both ITS and LSU exhibit high interspecific and low intraspecific variation for R. eccentrica, R. nigricans, and R. subnigricans. These markers provide enough resolutional power to differentiate these species and uncover phylogeographic structure, and will be powerful tools for future ecological studies of Russula.     

Two new species of Russula subgenus Compactae from Indian Himalaya based on morphology and molecular phylogenetic inferences

The identity of blackening Russula (R. subg. Compactae) from Indian Himalaya has since long remained a mystery and they are often called after their European look-alikes. Here, a combined approach including morphology and ITS phylogenetic inference resolved the identification of some of these taxa and revealed the discovery of two novel species. Thus, Russula ashihoi sp. nov., found under Abies in subalpine Himalaya, and R. indonigra sp. nov. occurring under Quercus in subtropical to temperate Himalaya are proposed herein with their morphological details, illustrations and ITS-based phylogeny. Similarities with allied taxa are also discussed.     

Phylogenetic diversity of true morels (Morchella), the main edible non-timber product from native Patagonian forests of Argentina

Morchella species are edible fungi in high demand and therefore command high prices in world markets. Phenotypic-based identification at the species-level remains inadequate because of their complex life cycles, minor differences and plasticity of morphological characteristics between species, and the lack of agreement between scientific and common names. In Patagonia–Argentina, morels are associated with native forests of Austrocedrus chilensis (Cordilleran or Chilean cypress) and Nothofagus antarctica (ñire) and several exotic conifers that were introduced from western North America. Little is known about their taxonomy and phylogenetic relationships with other species in the genus. This work focused on the identification of collections of Morchella from Patagonia and their phylogenetic relationships with other species from the Northern Hemisphere. The comparison was made by analysis of DNA sequences obtained from four loci: the nuclear ribosomal internal transcribed spacer region (ITS) and the partial RNA polymerase I gene (RPB1) for the complete collection; and ITS, RPB1, RNA polymerase II gene (RPB2), and translation elongation factor (EF1-α) for the species-rich Elata Subclade. Analyses of individual and combined data sets revealed that Patagonian morels belong to the Elata Clade and comprised three strongly supported species-level lineages from both Patagonian native forest, and exotic trees introduced from western North America. One lineage was identified as Morchella frustrata phylogenetic species Mel-2, which is known from the USA and Canada. The second lineage, which appeared to be ‘fire-adapted’, was identified as Morchella septimelata phylogenetic species (Mel-7), which is also known from the USA. This species was collected from burned native forests mainly composed of A. chilensis and N. antarctica but also Pseudotsuga menziesii (Mirb.) Blanco, which is native to western North America. The phylogenetic analyses suggested that the third species from Patagonia was nested within the species-rich Elata Subclade and represents a new species-level lineage (informally designated Mel-37) within Elata Clade. The present collections from Patagonia constitute the southernmost latitude from which Morchella has been reported to date. The identification of two Argentine morels as North American taxa is therefore a remarkable biogeographic pattern. In view of the hypothesis that the Elata Clade originated in western North America, we speculate that at least two of the lineages colonized South America from North America via long distance dispersal, migration or, more likely, they were introduced with the exotic tree species that they were collected near.     

Polyphyly of the Padus group of Prunus (Rosaceae) and the evolution of biogeographic disjunctions between eastern Asia and eastern North America

Prunus subgenus Padus is a group with a wide distribution in temperate eastern Asia and eastern North America with one species extending to Europe and one to Central America. Phylogenetic relationships of subgenus Padus were reconstructed using sequences of nuclear ribosomal ITS, and plastid ndhF gene, and rps16 intron and rpl16 intron. Prunus subgenus Padus is shown to be polyphyletic. Taxa of subgenus Padus and subgenus Laurocerasus are highly intermixed in both the ITS and the plastid trees. The results support two disjunctions between eastern North America and Eurasia within the Padus group. One disjunction is between Prunus virginiana of eastern North America and P. padus of Eurasia, estimated to have diverged at 2.99 (95 % HPD 0.59–6.15)–4.1 (95 % HPD 0.63–8.59) mya. The other disjunction is between P. serotina and its Asian relatives. The second disjunction may have occurred earlier than the former one, but the age estimate is difficult due to the unresolved phylogenetic position of the P. serotina complex.     

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.     

Coltriciella minuscula sp. nov., a new species of poroid fungus on Pinus merkusii from an Indonesian tropical forest

Coltriciella minuscula sp. nov. is described and illustrated from a specimen collected on Pinus merkusii from Bogor, Indonesia. This species is characterized by the size of its basidiocarp up to 4 mm in diam, with long hair on the stipe and with ornamented spores. Both morphological distinctiveness and phylogenetic separation based upon analyses of nrDNA ITS sequences support the establishment of this new species. Morphological dissimilarities between C. minuscula and closely related species are discussed.     

Hidden diversity uncovered in Hygrophorus sect. Aurei (Hygrophoraceae), including the Mediterranean H. meridionalis and the North American H. boyeri, spp. nov.

For many years, the binomial Hygrophorus hypothejus was widely applied to collections from various geographical regions in different continents, assuming a circum-boreal and circum-mediterranean distribution for this species. This hypothesis, however, had never been put to the test. To assess the diversity and species-limits within this complex of yellow-coloured waxcaps, a phylogenetic, morphological and taxonomical investigation into Hygrophorus sect. Aurei and similar species in sect. Olivaceoumbrini was carried out, including material of pan-European origin, as well as the east and west coasts of North America. Following sequencing of the ITS rDNA locus, nine lineages are confirmed in sect. Aurei, most of them highly continentalised. Of these, two are new to science, introduced here as Hygrophorus boyeri sp. nov., from Pinus banksiana and P. rigida forests in eastern North America and from P. muricata and P. contorta forests in western North America, and Hygrophorus meridionalis sp. nov., from Pinus brutia and Pinus halepensis forests in the island of Cyprus and mainland Greece. H. hypothejus is lectotypified and epitypified, and here resolved as a strictly European species, with the old forgotten taxon Hygrophorus siccipes revived as its North American vicariant. The placement of Hygrophorus fuligineus in sect. Aurei is phylogenetically confirmed and detailed comparisons between morphologically similar and phylogenetically affiliated taxa in sect. Aurei and sect. Olivaceoumbrini are provided. The chronic confusion associated with Hygrophorus fuscoalbus, a highly controversial taxon described from Germany nearly two centuries ago and variously interpreted since, is discussed, concluding that this name is too ambiguous to be applied to any currently recognized species.     

Phylogenetic relationships of Korean Sparassis latifolia based on morphological and ITS rDNA characteristics

Recent studies based on morphological characteristics and molecular analyses have revealed that the characteristics of Sparassis crispa from Asia are not concordant with those of collections from Europe and North America. Consequently, the Asian isolate was redefined as Sparassis latifolia. This study is the first report of Sparassis latifolia collected in Korea. The taxonomic relationships and replacement of Sparassis species were inferred from a comparison of the morphological characteristics and by molecular sequence analysis of the internal transcribed spacer (ITS) rDNA regions. In particular, this study focused on the phylogenetic relationships inferred from the biogeographical distribution of isolates within the genus Sparassis.     

An introduced population of Chorda asiatica (Chordaceae,Laminariales) in Puget Sound,Pacific coast of North America

Northeastern Pacific Ocean and northwestern Atlantic Ocean populations of Chorda species, which have not been examined in previous phylogenetic studies, were investigated. All specimens that were collected in Hood Canal, Puget Sound, WA, USA, Pacific coast of North America, showed identical ITS‐5.8S rDNA sequences, and they were included in the clade of Japanese Chorda asiatica. With morphological data added to the molecular data, they were identified as C. asiatica and were concluded to be non‐indigenous populations, most likely introduced with oyster spat together with Sargassum muticum. Specimens collected in New York, NY, USA, Atlantic coast of North America, were genetically closest to C. filum from Newfoundland and were identified as C. filum. The genetic divergence of the North Atlantic populations of C. filum was relatively small compared to that of Japanese C. asiatica considering their broader distributional ranges on both sides of the Atlantic.     

Molecular kinship analyses of the agaricoid Russulaceae: Correspondence with mycorrhizal anatomy and sporocarp features in the genus Russula

A data set of LSU DNA sequences of mainly European Russula and Lactarius species was subjected to molecular phylogenetic analysis. Species could be allocated to six clades, with an unresolved phylogeny. One of these clades represents the genus Lactarius. The only analysed species of the section Archaeinae (Russula) was placed basal to both genera. Thus Lactarius appears to be derived from Russula. Russula was divided into four clusters, corresponding to the sections Plorantinae and Nigricantinae, subgenus Heterophyllidia including the section Foetentinae, and a cluster representing the remaining subgenera of the “Genuinae”. Even though the resulting groups can be considered as valid classificatory groups, species associations resulting from molecular analyses neither support the division of Russula into the subgenus Compacta (including the sections Nigricantinae, Plorantinae, and Archaeinae) and the “Genuinae” (including all remaining taxa), nor do they support previously proposed evolutionary lineages within the “Genuinae”. Ribosomal ITS DNA sequences of Russula species were analysed to achieve better infrageneric resolution. The results are discussed in relation to current classification systems and to what is known about the mycorrhizae formed by Russula species. While the systematic value attached to many macroscopic and microscopic sporocarp features was not supported by sequence data, mycorrhizal anatomy is in good correspondence with many of the results from the phylogenetic analysis.     

Ectomycorrhizal iconoclasts: the ITS rDNA diversity and nitrophilic tendencies of fetid Russula

Fetid Russula are frequently dominant ectomycorrhizal fungi, and some appear to be especially nitrophilic. However, little is known about their phylogenetic relationships or how common nitrophilic traits are in this group. This study addresses this gap and presents a phylogenetic analysis of ITS rDNA sequences and a meta-analysis of studies that examine ectomycorrhizal fungi response to nitrogen increase. The phylogenetic analysis indicates that (i) this lineage contains numerous unidentified taxa; (ii) the taxa have distinct geographic distributions; and (iii) the misuse of names such as R amoenolens, R. foetens or R. pectinatoides is common. Twenty-three well supported phylotypes were identified and include clades specific to western North America, eastern North America, Europe and Asia, while morphologically similar collections from tropical-equatorial regions are distinct. The metaanalysis shows that nitrophilic tendencies appear throughout fetid Russulas suggesting that this character is not an isolated trait within this subgenus but instead is a more general feature of the group overall. Mapping these tendencies across a broader portion of the Russulaceae shows that this trait is more regularly found in the basal Russula lineages and Lactarius spp., suggesting that this ability evolved early in these fungi.     

Three new records of powdery mildews found in Mexico with one genus and one new species proposed

Cystotheca lanestris on the new host Quercus canbyi, Microidium bauhiniicola on the new host Bauhinia macranthera and an undescribed species of Phyllactinia on Mimosa aculeaticarpa have recently been collected in Mexico. Analyses of morphological traits and molecular sequence data led to identifications of the causal agents of the powdery mildew diseases involved. Microidium bauhiniicola, hitherto only known from Argentina and Brazil, is new to Mexico. The phylogenetic analysis revealed an isolated position distant from Microidium indicating an undescribed genus, which is introduced as Bulbomicroidium gen. nov. Sequences derived from a Mexican specimen of C. lanestris on Quercus canbyi agree completely with other North American sequences, but differ from sequences retrieved from Asian collections, suggesting that C. lanestris in Asia is not conspecific with C. lanestris s. str. in North America. A new species of Phyllactinia on Mimosa malacophylla is morphologically similar to P. dalbergiae but readily distinguishable by differences in the asexual morph. Based on morphological peculiarities and results of molecular sequence analyses, this species is described as Phyllactinia mimosae sp. nov.     

An ITS-based phylogenetic framework for the genus Vorticella: finding the molecular and morphological gaps in a taxonomically difficult group

Vorticella includes more than 100 currently recognized species and represents one of the most taxonomically challenging genera of ciliates. Molecular phylogenetic analysis of Vorticella has been performed so far with only sequences coding for small subunit ribosomal RNA (SSU rRNA); only a few of its species have been investigated using other genetic markers owing to a lack of similar sequences for comparison. Consequently, phylogenetic relationships within the genus remain unclear, and molecular discrimination between morphospecies is often difficult because most regions of the SSU rRNA gene are too highly conserved to be helpful. In this paper, we move molecular systematics for this group of ciliates to the infrageneric level by sequencing additional molecular markers—fast-evolving internal transcribed spacer (ITS) regions—in a broad sample of 66 individual samples of 28 morphospecies of Vorticella collected from Asia, North America and Europe. Our phylogenies all featured two strongly supported, highly divergent, paraphyletic clades (I, II) comprising the morphologically defined genus Vorticella. Three major lineages made up clade I, with a relatively well-resolved branching order in each one. The marked divergence of clade II from clade I confirms that the former should be recognized as a separate taxonomic unit as indicated by SSU rRNA phylogenies. We made the first attempt to elucidate relationships between species in clade II using both morphological and multi-gene approaches, and our data supported a close relationship between some morphospecies of Vorticella and Opisthonecta, indicating that relationships between species in the clade are far more complex than would be expected from their morphology. Different patterns of helix III of ITS2 secondary structure were clearly specific to clades and subclades of Vorticella and, therefore, may prove useful for resolving phylogenetic relationships in other groups of ciliates.     

Pseudozyma vetiver sp. nov., a novel anamorphic ustilaginomycetous yeast species isolated from the phylloplane in Thailand

Three strains representing one novel yeast species were isolated from the phylloplanes of the vetiver grasses (DMKU-LV90 and DMKU-LV99^T) and sugarcane (DMKU-SP260) collected in Thailand by leaf washing followed by a plating technique. On the basis of morphological, biochemical, physiological and chemotaxonomic characteristics and the sequence analysis of the D1/D2 region of the large subunit (LSU) rRNA gene and the internal transcribed spacer region (ITS), the three strains were found to represent a single novel anamorphic ustilaginomycetous yeast species in the genus Pseudozyma. The name Pseudozyma vetiver sp. nov. is proposed for this novel species. The type strain is DMKU-LV99^T (BCC 61021 = CBS 12824). The novel species showed phylogenetic relationships to the other members of the genus Pseudozyma and to teleomorphic fungal genera, namely Ustilago, Sporisorium and Anomalomyces in Ustilaginaceae, Ustilaginales. The three strains showed identical sequences both in the D1/D2 and ITS regions. The Pseudozyma species closest to the novel species in terms of pairwise sequence similarity in the D1/D2 region was Pseudozyma pruni but with 2.3 % nucleotide substitutions (14 nucleotide substitutions and no gaps out of 606 nt). The novel species and P. pruni differed by 10.9 % nucleotide substitutions (75 nucleotide substitutions and 31 gaps out of 691 nt) in the ITS region. The phylogenetic analysis based on the combined sequences of the ITS region and the D1/D2 region of the LSU rRNA gene showed that the novel species was found to be most closely related to Pseudozyma fusiformata but with 2.9 % nucleotide substitutions in the D1/D2 region and 7.4 % nucleotide substitutions in the ITS region.     

Intercontinental dispersal of Typha angustifolia and T. latifolia between Europe and North America has implications for Typha invasions

The full effects of biological invasions may be underestimated in many areas because of cryptogenic species, which are those that can be identified as neither native nor introduced. In North America, the cattails Typha latifolia, T. angustifolia, and their hybrid T. × glauca are increasingly aggressive invaders of wetlands. There is a widespread belief that T. latifolia is native to North America and T. angustifolia was introduced from Europe, although there has so far been little empirical support for the latter claim. We used microsatellite data and chloroplast DNA sequences to compare T. latifolia and T. angustifolia genotypes from eastern North America and Europe. In both species, our data revealed a high level of genetic similarity between North American and European populations that is indicative of relatively recent intercontinental dispersal. More specifically, the most likely scenario suggested by Approximate Bayesian Computation was an introduction of T. angustifolia from Europe to North America. We discuss the potential importance of our findings in the context of hybridization, novel genomes, and increasingly invasive behaviour in North American Typha spp.     

Cuticular terminology in Russula (agaricales)

The pileus cuticle is one-layered in a few species ofRussula, but in the others it can be interpreted as composed of two layers—the epicutis uppermost and below this, the subcutis. The basic difference between the two layers is in hyphal orientation. Cell inflation may obscure the orientation in the cuticles of some species. This has given rise to differing interpretations of cuticular structure, especially in regard to the number of nameable layers present, and thus to differing terms for the layers. No Russulas illustrate this better than those of subsectionVirescentinae, of which the two most common North American species,R. crustosa Peck andR. virescens (Schaeffer ex Secretan) Fries, are discussed and described.     

Thysanophora penicillioides includes multiple genetically diverged groups that coexist respectively in Abies mariesii forests in Japan

We investigated intraspecific diversity and genetic structures of a saprotrophic fungus--Thysanophora penicillioides--based on sequences of nuclear ribosomal internal transcribed spacer (ITS) in 15 discontinuous Abies mariesii forests of Japan. In such a well-defined morphological species, numerous unexpected ITS variations were revealed: 12 ITS sequence types detected in 254 isolates collected from 15 local populations were classified into five ITS sequence groups. Maximally, four ITS groups consisted of seven ITS types coexisting in one population. However, group 1 was dominant with approximately 65%; in particular, one haplotype, 1a, was most dominant with approximately 60% in respective populations. Therefore, few differences were recognized in genetic structure among local populations, implying that the gene flow of each lineage of the fungus occurs among local populations without geographic limitations. However, minor haplotypes in some ITS groups were found only in restricted areas, suggesting that they might expand steadily from their places of origin to neighboring A. mariesii forests. Aggregating sequence data of seven European strains and four North American strains from various substrates to those of Japanese strains, 18 ITS sequence types and 28 variable sites were recognized. They were clustered into nine lineages by phylogenetic analyses of the beta-tubulin and combined ITS and beta-tubulin datasets. According to phylogenetic species recognition by the concordance of genealogies, respective lineages correspond to phylogenetic species. Plural phylogenetic species coexist in a local population in an A. mariesii forest in Japan.     

Re-evaluation of Japanese Phytophthora isolates based on molecular phylogenetic analyses

Over the past 40 years in Japan, Phytophthora isolates have been collected from various diseased host tissues and infested soils and identified using morphological characters. In order to develop a molecular method for the characterization of Japanese Phytophthora species, we obtained nuclear ribosomal ITS and LSU and mitochondrial coxI DNA sequences from 151 isolates representing 21 known species and 10 unidentified isolates. These were compared with similar sequences from representative isolates of known species. Of these, 124 isolates were found to have been correctly identified. Among the remaining 37 isolates, 19 showed high homology with other described species. The remaining 18 isolates showed only low levels of homology with any known species, and generated monophyletic sub-clades in a phylogenetic tree based on the ITS and nLSU regions and the coxI gene. Therefore, these isolates are candidates for new species, falling into six groups. Together, the Japanese isolates were found to represent phylogenetically diverse groups of species. In a sequence variation analysis, the ITS regions and the coxI genes were found to be more variable than the nLSU sequences, suggesting that they will be more useful for Phytophthora identification.     

<Emphasis Type="Italic">Cortinarius</Emphasis> sect. <Emphasis Type="Italic">Riederi</Emphasis>: taxonomy and phylogeny of the new section with European and North American distribution

Cortinarius is one of the most species-rich genera of mushroom-forming fungi. Based on phylogenetic and morphological evidence, Cortinarius, sect. Riederi, is introduced at sectional level (= subsect. Riederi sensu Brandrud & Melot). The taxonomy, phylogeny, ecology and distribution of not only mainly European but also including some North American taxa of this section are treated, which includes nine species and two varieties. Of these, three taxa are described as new (C. burlinghamiae, C. pallidoriederi and C. argenteolilacinus var. dovrensis). The sect. Riederi species possess morphological features similar to Phlegmacium group(s) and forms a phylogenetically isolated lineage, with no supported affinity to other phlegmacioid groups. Three taxa are known from both Europe and North America, two species are known only from North America and five only from Europe. Altogether, eight of the ten taxa are associated with conifers or northern (boreal-subalpine) deciduous trees (Betula spp.). Only two species occur in more temperate forests (Fagus forests), and no species have so far been found in thermophilous Quercus forests     

Inocybe praetervisa group – A clade of four closely related species with partly different geographical distribution ranges in Europe

Sequence data from a broad geographical region and different habitats show that the Inocybe praetervisa clade is comprised of four closely related species. These species of section Marginatae are characterized by having nodulose spores and a stipe that is abundantly pruinose only in the upper half. Inocybe praetervisa occurs in Southern Europe in mountainous mixed coniferous forests, and is not confirmed from Northern Europe. Inocybe rivularis occurs in northern boreal forests up to the lower alpine zone, associated with Betula in moist habitats, and is not confirmed from Southern Europe. Inocybe taxocystis is confirmed as a later synonym of I. favrei. The species has a wide geographical distribution range in Europe, mainly restricted to the alpine zone and moist soils, associated with Salix herbacea. Inocybe arctica is here described as a new species. It occurs in the arctic and higher alpine zones, associated with Dryas octopetala, Salix polaris, S. reticulata and S. herbacea. All species except I. arctica are shown to have an intercontinental distribution range and are confirmed from North America. Sequence data suggest the occurrence of one additional species in the alpine zone of China. A key to the species in the I. praetervisa group is provided.