Phylogenetic assessment and taxonomic revision of <Emphasis Type="Italic">Mariannaea</Emphasis>

Four new species of Mariannaea were described in this paper, namely M. chlamydospora, M. cinerea, M. fusiformis, and M. lignicola. Mariannaea chlamydospora is characterized by its cream-colored, zonate colonies on PDA, smooth conidiophores, fusiform conidia, and abundant chlamydospores. Mariannaea cinerea forms grey colonies and ellipsoidal to subglobose conidia. Mariannaea fusiformis forms purple colonies and fusiform to subglobose conidia. Mariannaea lignicola has brown conidiophores and broad hyphae. The molecular phylogeny was inferred using ITS, LSU, and TUB-2 loci. The type species of Mariannaea (M. elegans) is epitypified. The variety M. elegans var. punicea is raised to species rank. Mariannaea clavispora is excluded from Mariannaea because of its cylindrical phialides, straight conidial chains and deviating phylogenetic affinity. Mariannaea nipponica did not fit well the generic concept of Mariannaea based on their morphological characters, and its generic placement remains uncertain. A key to the currently accepted 15 species of Mariannaea is provided.     

Molecular phylogeny and morphology of <Emphasis Type="Italic">Ophiocordyceps unituberculata</Emphasis> sp. nov. (Ophiocordycipitaceae), a pathogen of caterpillars (Noctuidae,Lepidoptera) from Yunnan,China

Ophiocordyceps unituberculata, a new insect pathogenic fungus from southwestern China, is described using molecular phylogenetic and morphological data. This fungus differs from other Ophiocordyceps species by its enormously long monophialidic conidiogenous cells and a large periclinal protuberance often growing near the apex of conidiogenous cells, single conidia (lanceolate to fusiform) embedded in mucous sheath, and much larger conidial dimensions. Four-locus (nrSSU, nrLSU, tef-1α and rpb1) and ITS data phylogenetic analyses show that O. unituberculata belongs to the Hirsutella nodulosa clade within the genus Ophiocordyceps of Ophiocordycipitaceae and is a separate clade from other allied species. Molecular phylogeny and morphology both strongly support the distinctiveness of this taxon. The interspecific relationships in the H. nodulosa clade are discussed.     

New and interesting chaetothyrialean fungi from Spain

In the course of taxonomic studies on saprobic microfungi from Spain, several slow-growing, dematiaceous hyphomycetes were isolated from soil, submerged plant material and river sediments. Sixteen of these strains were identified as members of the ascomycete order Chaetothyriales on the basis of morphology and DNA sequence analyses of the internal transcribed spacer region and partial large subunit ribosomal RNA gene. These included three novel species (Cladophialophora pseudocarrionii, Cyphellophora chlamydospora, and Rhinocladiella amoena) and five interesting, little-known or clinically-relevant species (Cyphellophora suttonii, Exophiala aquamarina, E. lacus, E. radicis, and Rhinocladiella similis). In addition, Exophiala oligosperma, an emerging opportunistic fungus, was found for the first time in an aquatic freshwater environment (river sediments). Cladophialophora pseudocarrionii resembles C. carrionii in the branching pattern of its conidial chains, but differs from the latter species in its inability to grow at 30 °C. Cyphellophora chlamydospora differs from other species of the genus in the absence of conidiation, producing only chlamydospores in vitro. Rhinocladiella amoena shows branched conidiophores similar to those of R. anceps, R. atrovirens, R. basitona and R. similis, but differs from them in conidial shape and size. The ex-type strain of Phialophora livistonae, included in the phylogenetic study, clustered with high statistical support with members of the genus Cyphellophora and is transferred to this genus.     

Multilocus phylogeny of <Emphasis Type="Italic">Clonostachys</Emphasis> subgenus <Emphasis Type="Italic">Bionectria</Emphasis> from Brazil and description of <Emphasis Type="Italic">Clonostachys chloroleuca</Emphasis> sp. nov.

Phylogenetic analyses based on protein-encoding gene exons and introns of ATP citrate lyase (ACL1), beta tubulin (TUB), the largest subunit of RNA polymerase II (RPB1), and translation elongation factor 1-α (TEF1) are used for inferring the existence of a new Clonostachys species from the Cerrado biome in Brazil, described here as C. chloroleuca. The species produces dimorphic, primary, and secondary conidiophores that form consistently greenish conidial masses on artificial media. It resembles therefore C. rosea f. catenulata although it differs from this species by less adpressed branches in the secondary conidiophores. The new species is also phylogenetically related to C. byssicola and C. rhizophaga. Our inventory suggests that C. byssicola, C. chloroleuca, C. pseudochroleuca, C. rhizophaga, C. rogersoniana, and C. rosea commonly occur in native and agriculturally used soils of the Cerrado and Amazon Forest. Using sequences available from two genome-sequenced strains employed as biological control agents, we confirm the identity of the European strain IK726 as C. rosea and identify strain 67-1 from China as C. chloroleuca.     

Molecular taxonomy and morphological characterization reveal new species and new host records of <Emphasis Type="Italic">Torula</Emphasis> species (Torulaceae,Pleosporales)

Four new species and two new host records of Torula (Torulaceae, Pleosporales) are described and illustrated from herbaceous litter collected in Italy and Thailand. The new species possess colony, conidiophore and conidial characteristics that fit within the generic concept of Torula. Detailed morphological observations clearly demarcate four of these from extant species and are hence described as new (Torula chiangmaiensis sp. nov., Torula pluriseptata sp. nov., Torula chromolaenae sp. nov., Torula mackenziei sp. nov.). Details of asexual morphs are described, and justifications for establishing these new species are provided. The nuclear are sequenced ribosomal RNA genes as well as protein coding genes to infer phylogenetic relationships and discuss phylogenetic affinities with morphologically similar species. Our morphological distinction is further supported by phylogenetic discrimination. In particular, phylogenies depict a close relationship of Torula chiangmaiensis and T. pluriseptata to T. hollandica, while T. chromolaenae and T. mackenziei constitute an independent phylogenetic lineage basal to T. herbarum and T. ficus. Torula ficus and T. masonii are also described and their phylogeny investigated as new host records from Bidens pilosa and Iris germanica, respectively.     

A taxonomic and phylogenetic study of the <Emphasis Type="Italic">Golovinomyces biocellatus</Emphasis> complex (Erysiphales,Ascomycota) using asexual state morphology and rDNA sequence data

The Golovinomyces biocellatus complex consists of powdery mildew (Erysiphales) species restricted to hosts of the family Lamiaceae. Previous authors used minor morphological features of the sexual state and host range data to split the complex. The data, however, were not sufficient to define a convincing species concept. Our taxonomic study is based on molecular phylogenetic and asexual state morphology data. For morphological studies, mainly features of the asexual morph (conidiophores, conidia, germination patterns) were studied using light and scanning electron microscopy. Detailed line drawings of asexual state features are provided. For phylogenetic analyses, two markers (rDNA: ITS, LSU) of 64 specimens were applied. The phylogeny resulted in two major clades. Clade I consists of specimens with Lamiaceae hosts and three specimens of Verbena. Clade II consists of two sister groups, the first (IIa) with Salvia spp. and the second (IIb) with Lycopus europaeus (the type host G. biocellatus) and Glechoma. Clades I and IIb and two subclades of IIa with Salvia hosts are characterized by specific morphological traits (differences in conidiophore length, conidial shape, width, and germination patterns). Based on these data, we suggest to consider specimens of clades I (including specimens on Verbena) and IIb and the two subclades of IIa as distinct species, namely G. monardae, G. biocellatus, G. salviae, and G. neosalviae sp. nov. A key for the identification of species based on asexual state features is provided. The results are discussed with respect to host range, jumps, co-evolutionary aspects, and distribution patterns.     

Further progress towards the delimitation of <Emphasis Type="Italic">Cheilanthes</Emphasis> (Cheilanthoideae,Pteridaceae), with emphasis on South American species

Cheilanthoid ferns (Cheilanthoideae sensu PPG 1 2016) constitute an important group within the Pteridaceae and are cosmopolitan in distribution. In South America, there are 155 species distributed in 13 genera, among which the largest are Adiantopsis (35), Cheilanthes (27), and Doryopteris (22). Most of the cheilanthoid species are morphologically adapted to grow in arid to semi-arid conditions and show convergent evolution, which has implied difficulties in defining the genera throughout their taxonomic history (Copeland 1947, Tryon & Tryon 1973, Gastony & Rollo 1995, 1998, Kirkpatrick Systematic Botany, 32: 504–518, 2007, Rothfels et al. Taxon, 57: 712–724, 2008). Here, we sequenced two plastid markers (rbcL?+?trnL-F) of 33 South American cheilanthoid species, most of which have not been included in phylogenetic analyses previously. The South American species were analyzed together with South African and Australasian Cheilanthes and representatives of related cheilanthoid genera. The phylogenetic analysis showed that most Cheilanthes species are related to the genus Hemionitis, constituting different groups according to their distribution; moreover, three species—C. hassleri, C. pantanalensis, and C. obducta—appear as the sister clade of Hemionitis. Cheilanthes micropteris, the type species, is strongly supported in a clade with Australasian Cheilanthes plus five South American Cheilanthes species, all of which show a reduction in the number of spores per sporangium; this feature would be a synapomorphy for core Cheilanthes s.s. We found no support uniting other South American Cheilanthes to either the group of South African Cheilanthes or to core Cheilanthes s.s. On the other hand, C. geraniifolia, C. goyazensis, and C. bradei formed a clade related to Doryopteris that, with further study, could be considered as a new genus. The phylogenetic hypotheses presented here contribute substantially to the delimitation of Cheilanthes s.s. and related groups and provide the basis for re-examining the generic taxonomy.     

Two new <Emphasis Type="Italic">Cordyceps</Emphasis> species from a community forest in Thailand

Two new species found in northern Thailand, Cordyceps chiangdaoensis and Cordyceps morakotii, pathogenic to Coleoptera larvae and Odontomachus ant pupae, respectively, are described using morphological and molecular phylogenetic data. Both species produce narrowly ovoid superficial perithecia at the end of a cylindrical stroma, bola ascospores, Evlachovaea-like conidial morph, and cylindrical conidia with rounded ends. These two species differ from other bola ascospore-producing species in the genus Cordyceps in the sizes of the ascospores and perithecia, as well as the host. Phylogenetic analyses based on internal transcribed spacer (ITS) regions of the rDNA and partial sequences of translation elongation factor 1-alpha (TEF1) data strongly support these two fungi as two distinct new species in the Cordycipitaceae.     

Myrothecium-like (Ascomycota,Hypocreales) species from tropical areas: <Emphasis Type="Italic">Digitiseta</Emphasis> gen. nov. and additions to <Emphasis Type="Italic">Inaequalispora</Emphasis> and <Emphasis Type="Italic">Parvothecium</Emphasis>

Inaequalispora and Parvothecium are two myrothecium-like, closely related genera of Hypocreales. They are also morphologically similar, sharing sporodochial conidiomata, penicillate conidiophores, fusiform to ellipsoidal conidia accumulating in a green slimy drop, and hypha-like setoid extensions emerging through the conidial mass. During a revision of myrothecium-like isolates originating from rainforest areas of South America (Ecuador, Brazil) and Southeast Asia (Singapore), multilocus phylogenetic inferences (based on DNA sequence data of ITS, partial nuc 28S, and partial tef1a, rpb2 and tub2) and morphological studies concordantly revealed the occurrence of two undescribed species of Inaequalispora (I. longiseta sp. nov. and I. cylindrospora sp. nov.) and one undescribed species of Parvothecium (P. amazonensesp. nov.). Myrothecium setiramosum, M. dimorphum, and two undescribed taxa form the base of a new lineage, sister to the current Parvothecium lineage. This lineage is recognized as Digitiseta gen. nov., typified by D. setiramosa comb. nov. Digitiseta dimorpha comb. nov. is also proposed, and the new species D. parvodigitata sp. nov. and D. multidigitata sp. nov. are described.     

Characterization of the velvet regulators in <Emphasis Type="Italic">Aspergillus flavus</Emphasis>

Fungal development and secondary metabolism are closely associated via the activities of the fungal NK-kB-type velvet regulators that are highly conserved in filamentous fungi. Here, we investigated the roles of the velvet genes in the aflatoxigenic fungus Aspergillus flavus. Distinct from other Aspergillus species, the A. flavus genome contains five velvet genes, veA, velB, velC, velD, and vosA. The deletion of velD blocks the production of aflatoxin B1, but does not affect the formation of sclerotia. Expression analyses revealed that vosA and velB mRNAs accumulated at high levels during the late phase of asexual development and in conidia. The absence of vosA or velB decreased the content of conidial trehalose and the tolerance of conidia to the thermal and UV stresses. In addition, double mutant analyses demonstrated that VosA and VelB play an inter-dependent role in trehalose biosynthesis and conidial stress tolerance. Together with the findings of previous studies, the results of the present study suggest that the velvet regulators play the conserved and vital role in sporogenesis, conidial trehalose biogenesis, stress tolerance, and aflatoxin biosynthesis in A. flavus.     

Diversity of <Emphasis Type="Italic">Colletotrichum</Emphasis> spp. isolated from chili pepper fruit exhibiting symptoms of anthracnose in Thailand

Colletotrichum spp. are causal agents of anthracnose disease in chili fruits and other tropical crops. The disease is increasing in chili fruits in Thailand and significantly reduces fruit quality and fruit production. Forty-eight isolates of Colletotrichum spp. associated with chili anthracnose were collected from different areas of Thailand during 2010–2015. Based on morphological characteristic identification, 10 isolates were shown to belong to the C. gloeosporioides species complex, 24 isolates belong to the C. acutatum species complex and 14 isolates to C. capsici. For molecular identification, two primer sets, ITS1/ITS4 and ACT528/ACT738, were used for amplification of the internal transcribed spacer of rRNA gene (ITS1–5.8S–ITS2) and partial region actin gene (ACT), respectively. The phylogenetic analysis of individual and combined ITS region and actin nucleotide sequences identified the collected isolates into 4 species: C. gloeosporioides, C. siamense, C. acutatum and C. capsici. The pathogenicity test demonstrated that all four species were pathogenic on intact unwounded and healthy fruits. These results indicated that C. capsici, C. acutatum, C. gloeosporioides and C. siamense were the causal agents of chili anthracnose disease.     

<Emphasis Type="Italic">Cantharellus</Emphasis> sect. <Emphasis Type="Italic">Amethystini</Emphasis> in Asia

In this contribution on the genus Cantharellus in Asia, C. subvaginatus is described from the Republic of Korea as a close relative to the Chinese C. vaginatus, which is here reported for the first time from India. Both species are here placed in Cantharellus subg. Cantharellus sect. Amethystini, together with the Indian C. pseudoformosus (syn.: C. umbonatus) and the Malayan C. subamethysteus. As such, Asia has suddenly become the continent with the highest diversity for Amethystini. Species delimitation in sect. Amethystini is molecularly supported by a combined phylogenetic analysis of rDNA sequences obtained for LSU and ITS and additionally suggests the existence of a still undescribed species in North America. Character variability is discussed for all known members of Amethystini, including atypical specimens of the North American C. lewisii that are morphologically more reminiscent of the South Korean C. subvaginatus.     

Relationships within <Emphasis Type="Italic">Capitotricha bicolor</Emphasis> (Lachnaceae,Ascomycota) as inferred from ITS rDNA sequences,including some notes on the <Emphasis Type="Italic">Brunnipila</Emphasis> and <Emphasis Type="Italic">Erioscyphella</Emphasis> clades

DNA sequences of Capitotricha bicolor from Quercus, Fagus sylvatica, Alnus alnobetula, and Nothofagus, and C. rubi from Rubus idaeus were obtained from apothecia to establish whether specimens from different hosts belong to separate species. The obtained ITS1–5.8S–ITS2 rDNA sequences were examined with Bayesian and parsimony phylogenetic analyses. Intra- and interspecific variation was also investigated based on molecular distances in the ITS region. The phylogenetic analyses supported the specific distinctness of Capitotricha rubi and the Capitotricha from Nothofagus, but also suggest specific distinctness between samples from Quercus, Fagus, and Alnus. The interspecific distances were larger than intraspecific distances for all examined units. The smallest distance was found between the “Alnus alnobetula” and “Fagus sylvatica” units. Two new sequences of Brunnipila are published. Capitotricha, Lachnum, and Erioscyphella are compared to each other based on hair and excipulum characteristics.     

Two novel <Emphasis Type="Italic">Aspergillus</Emphasis> species from hypersaline soils of The National Park of Lake Urmia,Iran

Two novel Aspergillus species, one belonging to the section Terrei and the other to section Flavipedes, were isolated from hypersaline soils of The National Park of Lake Urmia (Iran) and are here described as Aspergillus iranicus and Aspergillus urmiensis. A polyphasic taxonomic approach comprising extrolite profiles, phenotypic characters and molecular data (beta-tubulin, calmodulin and ribosomal polymerase II second largest subunit gene sequences) was applied to determine their novel taxonomic status. Aspergillus iranicus (CBS 139561^T) is phylogenetically related to A. carneus, A. niveus, A. allahabadii and A. neoindicus, and it can be differentiated from those species by a unique extrolite pattern (citrinin, gregatins, and a terrequinone) and its conidial colour. Aspergillus urmiensis (CBS 139558^T) shares a most recent common ancestor with A. templicola. The former species produces globose vesicles, and those of A. templicola are predominantly elongate. The Aspergillus urmiensis isolates produce several uncharacterized extrolites. Two other strains obtained during this study reside in a clade, together with the type strain of A. movilensis (CCF 4410^T), and are identified accordingly. Based on the phylogenetic data presented in this study, A. frequens is reduced to synonymy with A. micronesiensis and A. mangaliensis is considered to be a synonym of A. templicola.     

Integrative analysis of the West African <Emphasis Type="Italic">Ceraceosorus africanus</Emphasis> sp. nov. provides insights into the diversity,biogeography, and evolution of the enigmatic Ceraceosorales (Fungi: Ustilaginomycotina)

The order Ceraceosorales (Ustilaginomycotina) currently includes the single genus Ceraceosorus, with one species, Ceraceosorus bombacis, parasitic on Bombax ceiba in India. The diversity, biogeography, evolution, and phylogenetic relationships of this order are still relatively unknown. Here, a second species of Ceraceosorus is described from West Africa as a novel species, Ceraceosorus africanus, infecting Bombax costatum in Benin, Ghana, and Togo. This species produces conspicuous fructifications, similar to corticioid basidiomata when mature, but sorus-like in early stages of ontogenetic development. The fructifications cover much of the leaf surface and resemble leaf blight. This contrasts with the inconspicuous fructifications of C. bombacis comprising small spots scattered over the lower leaf surface that resemble leaf spot. Both species of Ceraceosorus differ in several micromorphological traits, infect different host plant species in widely separated geographical areas, and are separated by a considerable genetic distance in 28S rDNA and RPB2 genes. The distinct corticioid fructification of C. africanus is a unique morphological trait within the Ustilaginomycotina. Molecular phylogenetic analyses of a single gene dataset (D1/D2 28S rDNA) supported the monophyly of the two Ceraceosorus species and the Ceraceosorales and their placement within the Ustilaginomycotina. Molecular phylogenetic analyses of a multigene dataset (18S/5.8S/28S rDNA/RPB2/TEF1) revealed Exobasidium rhododendri (Exobasidiales) as the closest relative of Ceraceosorus, both clustering together with Entyloma calendulae (Entylomatales), indicating affinities to the Exobasidiomycetes. This phylogenetic placement is in agreement with ultrastructural characteristics (presence of local interaction zone and interaction apparatus) reported for the Ceraceosorales, Entylomatales, and Exobasidiales.     

Phylogenetic analysis of <Emphasis Type="Italic">Plectosphaerella</Emphasis> species based on multi-locus DNA sequences and description of <Emphasis Type="Italic">P. sinensis</Emphasis> sp. nov.

Species in Plectosphaerella are well known as pathogens of several plant species causing fruit, root and collar rot and collapse. In an investigation of endophytic fungi associated with cucurbit plants in China, we isolated 77 strains belonging to the genus Plectosphaerella. To identify the isolated strains, we collected the type or reference strains of all currently accepted species in Plectosphaerella except P. oratosquillae and conducted a phylogenetic analysis. Phylogenetic analysis of the partial 28S rDNA sequences showed that all species in Plectosphaerella were located in one clade of Plectosphaerellaceae. Based on multi-locus phylogenetic analysis of the ITS, CaM, EF1, TUB and morphological characteristics, all species in Plectosphaerella were well separated. Three endophytic strains from stems of Cucurbita moschata, Citrullus lanatus and Cucumis melo from North China were assigned to a new species described as P. sinensis in this paper. The new species differs morphologically from other Plectosphaerella species by irregular chlamydospores, and the dimensions of phialides and conidia. The other endophytic strains from several cucurbit plants were identified as P. cucumerina.     

Genetic polymorphism,haplotype distribution,and phylogeny of <Emphasis Type="Italic">Daphnia</Emphasis> (Cladocera: Anomopoda) species from the water bodies of russia as inferred from the 16S mtDNA gene sequencing

The data on the genetic polymorphism of the most widespread Daphnia species occupying different water bodies of Russia are presented. The phylogenetic relationships between the examined species were established, and the haplotype networks were constructed. A fragment of the 16S mitochondrial DNA gene was used as a genetic marker. The results of molecular phylogenetic analysis generally coincided with modern concepts in the systematics of the genus Daphnia. The representatives of the divergent mitochondrial lineages within the D. longispina, D. pulex, and D. magna complex remain poorly investigated morphologically. For D. dentifera, a new habitat on the territory of Russia, namely, the water bodies of the Lake Baikal basin, was identified. A conclusion was made that the 16S mtDNA gene could be successfully used in phylogeographic analysis of the genus Daphnia.