Phylloplane yeasts from Portugal: seven novel anamorphic species in the Tremellales lineage of the Hymenomycetes (Basidiomycota) producing orange-coloured colonies

A survey of epiphytic yeasts on leaves of selected Mediterranean plant species collected at the 'Arrábida Natural Park' (Portugal) yielded about 850 isolates, mostly of basidiomycetous affinity. Amongst the basidiomycetes, 35 strains showed the following characteristics: production of orange-coloured colonies, ability to produce starch-like compounds, assimilation of D-glucuronic acid and/or inositol, inability to utilize nitrate, and formation of ballistoconidia by many of the isolates. This group of yeasts was assigned to the Tremellales lineage of the Hymenomycetes and was further characterised using a combination of conventional phenotypic identification tests with molecular methods, namely PCR fingerprinting and rDNA sequencing. Eight additional strains presumptively identified as Bullera armeniaca, B. crocea or Cryptococcus hungaricus were also studied. Twenty-eight strains could be assigned to or were phylogenetically related to recognised species of Dioszegia in the 'Luteolus clade', but the 15 remaining strains belonged to other clades within the Tremellales. Ten phylloplane isolates were identified as Dioszegia hungarica, one as D. aurantiaca, another as D. crocea and three others were ascribed to the recently described species D. zsoltii. Seven novel species, viz. Cryptococcus amylolyticus, C. armeniacus, C. cistialbidi, Dioszegia buhagiarii, D. catarinonii, D. fristingensis and D. takashimae, are proposed for the remaining strains that did not correspond to any of the hitherto recognised species.     

Two new ballistoconidium-forming yeast species, Bullera melastomae and Bullera formosana, found in Taiwan

Two yeast strains, the cells of which contained xylose and Q-10 as the major ubiquinone, were isolated from a plant leaf collected in Taiwan. These yeasts were found to represent two new species of the genus Bullera in the Hymenomycetes. Identification was based on the sequence analysis of the 18S rDNA, the internal transcribed spacer (ITS) regions and the D1/D2 domain of 26S rDNA. The yeasts are named Bullera melastomae sp. nov. and Bullera formosana sp. nov. In the phylogenetic trees based on 18S rDNA and D1/D2 domain of 26S rDNA sequences, these two species constitute a cluster connected with Dioszegia cluster in the Cryptococcus luteolus lineage.     

Dioszegia zsoltii sp. nov., a new ballistoconidium-forming yeast species with two varieties

As a result of conventional characterization of yeasts isolated from various plant leaves collected in Yunnan, China, six ballistoconidium-forming strains with orange-colored colonies were grouped together. Molecular phylogenetic analysis based on 18S rDNA sequencing showed that two representative strains of this group of yeasts, CH 2.068 and CH 2.497, were closely related to the species in the genus Dioszegia and had signature sequences typical of this genus. However, the six strains from Yunnan differed from the described Dioszegia species remarkably (14.5-17.7% nucleotide divergences) in the ITS (internal transcribed spacer) region sequences, which indicated that they represent a distinct species. Furthermore, among the six strains studied, the ITS region sequence comparison allowed the recognition of two subgroups represented by CH 2.068 and CH 2.497, which differ from each other in three bases in the ITS 2 region. DNA-DNA relatedness revealed that the two subgroups represent two varieties of a new species in the genus Dioszegia, for which Dioszegia zsoltii sp. nov. var. zsoltii and Dioszegia zsoltii var. yunnanensis var. nov. are proposed.     

Genetic diversity in the red yeast Cryptococcus hungaricus and its phylogenetic relationship to some related basidiomycetous yeasts

Cryptococcus hungaricus is a basidiomycetous yeast with the abilities to synthesize carotenoid pigments and to grow under psychrophile conditions. Six C. hungaricus strains have been isolated so far from different habitats. In this study we wished to clarify the relationships amongst them. Morphological and physiological characters, mitochondrial DNA restriction profiles, and the presence of mycoviruses were examined. Internal transcribed spacers together with the 5.8S rDNA, the D1/D2 region of 26S rDNA, and partial sequences of the 18S rRNA gene were also analysed. On the basis of the phylogenetic analyses the type strain CBS 4214(T) together with four other C. hungaricus isolates were closely related to Bullera armeniaca and Bullera crocea, while strain CBS 6569 was much more similar to Cystofilobasidium than to the other C. hungaricus isolates.     

Molecular phylogeny of basidiomycetous yeasts in the Cryptococcus luteolus lineage (Tremellales) based on nuclear rRNA and mitochondrial cytochrome b gene sequence analyses: proposal of Derxomyces gen. nov. and Hannaella gen. nov., and description of eight novel Derxomyces species

Phylogenetic relationships among the hymenomycetous yeasts in the Cryptococcus luteolus lineage of the Tremellales were examined based on sequence analyses of the 18S rRNA gene, 26S rRNA gene D1/D2 domain, internal transcribed spacer (ITS) region including 5.8S rRNA gene and mitochondrial cytochrome b gene. In addition to the Dioszegia clade, two clades represented by Bullera mrakii and Bullera sinensis, respectively, were revealed to be well-separated monophyletic groups in the lineage. These clades also exhibited distinguishable colony characters. Two new genera, Derxomyces gen. nov. (type species: Derxomyces mrakii comb. nov.) and Hannaella gen. nov. (type species: Hannaella sinensis comb. nov.), are proposed to accommodate the species in the B. mrakii and B. sinensis clades, respectively. Mainly based on D1/D2 and ITS sequence comparison, eight novel Derxomyces species were recognized from ballistoconidium-forming strains isolated from plant leaves. The new species and their type strains are as follows: Derxomyces boekhoutii (AS 2.3758(T)=CBS 10824(T)), Derxomyces hainanensis (AS 2.3467(T)=CBS 10820(T)), Derxomyces linzhiensis (AS 2.2668(T)=CBS 10827(T)), Derxomyces pseudocylindrica (AS 2.3778(T)=CBS 10826(T)), Derxomyces qinlingensis (AS 2.2446(T)=CBS 10818(T)), Derxomyces simaoensis (AS 2.3571(T)=CBS 10822(T)), Derxomyces wuzhishanensis (AS 2.3760(T)=CBS 10825(T)) and Derxomyces yunnanensis (AS 2.3562(T)=CBS 10821(T)).     

Diversity of basidiomycetous phylloplane yeasts belonging to the genus Dioszegia (Tremellales) and description of Dioszegia athyri sp. nov., Dioszegia butyracea sp. nov. and Dioszegia xingshanensis sp. nov

From approximately 200 basidiomycetous yeast isolates forming orange or orange-red colonies isolated from senescent leaves collected in different regions of China, 29 representative strains varying in their geographic distribution and ballistoconidium forming ability were selected for further phenotypic and molecular taxonomic studies. Sequence analysis of the large subunit (26S) rDNA D1/D2 domain and the internal transcribed spacer (ITS) region including 5.8S rRNA from the strains resulted in the recognition of seven Dioszegia species, including four described species, namely D. aurantiaca, D. fristingensis, D. hungarica and D. zsoltii var. zsoltii and D. zsoltii var. yunnanensis, and three undescribed species. The three new species are described as Dioszegia athyri sp. nov. (type strain: CB 159(T) = AS 2.2559(T) = CBS 10119(T)), Dioszegia butyracea sp. nov. (type strain: CB 261(T) = AS 2.2600(T) = CBS 10122(T)) and Dioszegia xingshanensis sp. nov. (type strain: HB 1.4(T) = AS 2.2481(T) = CBS 10120(T)) in the present study.     

Diversity of Cryptococcus and Dioszegia yeasts (Basidiomycota) inhabiting arbuscular mycorrhizal roots or spores

The genera Cryptococcus and Dioszegia contain basidiomycetous yeasts found in a wide range of habitats. Primers to amplify the internal transcribed spacer (ITS) region of the nuclear ribosomal DNA (nrDNA) of arbuscular mycorrhizal fungi (AMF) also allow detecting members of this yeast group. Here we report the results of a sequence analysis using maximum parsimony on a set of 50 ITS sequences of yeasts associated with AMF structures (roots of 26 plant species, AM spores) from six field sites in Central Germany. Among 10 separated taxa, respectively five in the Tremellales and two in the Filobasidiales had unknown sequences. Therefore it was not possible to assign these sequences to any known species. The study indicates that exploring the diversity of Cryptococcus and Dioszegia in soil habitats with molecular methods might enlarge the actually estimated biodiversity of the group.     

Bullera koratensis sp. nov. and Bullera lagerstroemiae sp. nov., two new ballistoconidium-forming yeast species in the Trichosporonales clade isolated from plant leaves in Thailand

Seven strains of ballistoconidiogenous yeasts that contain xylose, form Q-10 ubiquinone, propagate by budding and don't produce stalk conidia were isolated from plant leaves collected in Thailand and were found to represent two new species. The taxonomic properties of the two species coincided with the genus Bullera so they are described as Bullera koratensis sp. nov. and Bullera lagerstroemiae sp. nov. In phylogenetic trees based on the nucleotide sequences of 18S ribosomal DNA and the D1/D2 domain of 26S rDNA, these two species are located in the Trichosporonales clade (Cryptococcus humicola-Trichosporon lineage).     

Description of Bullera kunmingensis sp. nov., and clarification of the taxonomic status of Bullera sinensis and its synonyms based on molecular phylogenetic analysis

A ballistoconidium-forming yeast strain, CH 2.506, isolated from a semi-dried leaf of Parthenocissus sp. collected near Kunming City in Yunnan, China, was shown to be closely related to the non-ballistoconidium-forming species Cryptococcus luteolus (Saito) C.E. Skinner and the ballistoconidium-forming species Bullera sinensis Li by molecular phylogenetic analysis based on 18S rDNA sequencing. This strain was demonstrated to represent a distinct undescribed yeast species by internal transcribed spacer (ITS) region sequence and G+C content comparison and DNA-DNA relatedness, for which the name Bullera kunmingensis sp. nov. is proposed. Meanwhile, the taxonomic relationships among Bullera sinensis and its synonyms B. derxii Nakase & Suzuki and B. alba (Hanna) Derx var. lactis Li, were clarified on the basis of molecular phylogenetic analysis and DNA-DNA reassociation. B. derxii was confirmed to be conspecific with B. sinensis, while B. alba var. lactis was shown to represent a variety of B. sinensis. A new combination, Bullera sinensis Li var. lactis (Li) Bai, Takashima et Nakase, is therefore proposed. Comparative analysis of different types of molecular criteria employed in the present study suggested that when inferring phylogenetic relationships among sibling taxa, sequence data from ITS regions should be interpreted with caution.     

Bullera taiwanensis sp nov and Bullera formosensis sp nov,two new ballistoconidium-forming yeast species isolated from plant leaves in Taiwan

Two strains of ballistoconidiogenous yeasts that contain xylose and form Q-10 ubiquinone were isolated from plant leaves collected in Taiwan and were found to represent two new species. The taxonomic properties of both coincide with the genus Bullera so they are described as Bullera taiwanensis sp. nov. and Bullera formosensis sp. nov. In a phylogenetic tree based on the nucleotide sequences of 18S ribosomal DNAs, these two species are distant from the clusters where the remaining members of the genus Bullera are located, i.e., Bullera taiwanensis is located in the Filobasidium lineage (Filobasidiales clade) and Bullera formosensis is located in the Cryptococcus humicola-Trichosporon lineage (Trichosporonales clade).     

Bullera hoabinhensis sp. nov., a new ballistoconidiogenous yeast isolated from a plant leaf collected in Vietnam

VY-68, a ballistoconidiogenous yeast strain, isolated from a plant leaf at Cuc Phuong National Park of Ninh Binh Province, Vietnam, was assigned to the genus Bullera based on morphological and chemotaxonomical characteristics. Based on the sequence analyses of 18S rDNA, D1/D2 region of 26S rDNA, and internal transcribed spacer regions (ITS), VY-68 was phylogenetically closely related to Bullera pseudoalba and Cryptococcus cellulolyticus. DNA-DNA reassociation experiments among VY-68, B. pseudoalba and C. cellulolyticus revealed that strain VY-68 is a distinct species, and the latter two are conspecific. Bullera hoabinhensis is proposed for VY-68.     

Cryptococcus bestiolae and Cryptococcus dejecticola, two new yeast species isolated from frass of the litchi fruit borer Conopomorpha sinensis Bradley

Two new yeast species, Cryptococcus bestiolae and Cryptococcus dejecticola, were discovered in the frass of the litchi fruit borer Conopomorpha sinensis Bradley. The yeasts utilize inositol, hydrolyze urea, produce starch-like substance, and contain CoQ10. Phylogenetic analyses of D1/D2 26S rDNA and internal transcribed spacer (ITS) sequences indicate that the yeasts are closely related to Bullera dendrophila and an undescribed species of Cryptococcus (strain CBS 8507). The two new species differed from each other by 17 nucleotides in the D1/D2 region and by 68 nucleotides in the ITS region. Cryptococcus bestiolae is a sister species to Cryptococcus sp. CBS 8507, from which it differs by eight nucleotides in the D1/D2 region and 59 nucleotides in the ITS region. Cryptococcus dejecticola and B. dendrophila differed by 13 nucleotides in the D1/D2 and 57 nucleotides in the ITS region. Cryptococcus bestiolae and Cr. dejecticola formed with B. dendrophila a well defined clade consisting of insect associated species. The type strain of Cr. bestiolae is TH3.2.59 (=CBS 10118=NRRL Y-27894), and the type strain of Cr. dejecticola is Litch 17 (=CBS 10117=NRRL Y-27898).     

Phylogenetic and taxonomic heterogeneity of Cryptococcus humicolus by analysis of the sequences of the internal transcribed spacer regions and 18S rDNA, and the phylogenetic relationships of C. humicolus, C. curvatus, and the genus Trichosporon

The phylogenetic and taxonomic heterogeneity of a rare opportunistic yeast pathogen, Cryptococcus humicolus, was revealed by analysis of the sequence of the internal transcribed spacer (ITS) region. Sixteen strains of C. humicolus showed a wide diversity in their ITS sequences. In addition, their 18S rDNA sequences were determined and used to analyze the phylogenetic relationships among C. humicolus and related yeasts. On trees constructed by the Neighbor-Joining and Maximum Parsimony methods, C. humicolus strains were phylogenetically closely related to each other with the exception of one strain, and they clustered with C. curvatus and Trichosporon species with high bootstrap values. Three C. humicolus strains obtained from humans belonged to the group of Trichosporon serotype I species. The results suggest that C. humicolus is a genetically heterogeneous species which should be reclassified on the basis of DNA sequence data.     

Reinstatement of Rhodotorula colostri (Castelli) Lodder and Rhodotorula crocea Shifrine & Phaff, former synonyms of Rhodotorula aurantiaca (Saito) Lodder

Rhodotorula aurantiaca (Saito) Lodder is an anamorphic basidiomycetous yeast species that belongs to the so-called "Erythrobasidium lineage" of the Urediniomycetes, according to molecular phylogenetic studies based on nucleotide sequence analyses of different ribosomal DNA regions. In the most recent editions of the yeast taxonomy treatises the species Rhodotorula colostri (Castelli) Lodder and Rhodotorula crocea Shifrine & Phaff were listed as synonyms of R. aurantiaca. Taxonomic heterogeneity within R. aurantiaca was demonstrated in a study based on whole-cell protein profiles and is also hinted at by the observed differences in physiological and biochemical characteristics among the different strains under that species name. We determined partial nucleotide sequences of the 26S rRNA gene (D1/D2 domains) of strains maintained in the CBS culture collection under R. aurantiaca, including the type strains of its synonyms. The results showed that R. colostri and R. crocea are clearly distinct from R. aurantiaca and from any other currently recognised basidiomycetous yeast species. Furthermore, phylogenetic analysis of the sequence data placed the former two species in separate lineages of the Microbotryomycetidae: R. colostri in the "ruineniae clade" (Sporidiobolus lineage or Sporidiobolales) and R. crocea loosely linked to Rhodotorula javanica (Microbotryum lineage).     

Two new species of Botryosphaeria with brown, 1-septate ascospores and Dothiorella anamorphs

Botryosphaeria sarmentorum sp. nov. and B. iberica sp. nov. are described and illustrated. These two species are unusual in this genus because of their brown, 1-septate ascospores. Phylogenetic analysis based on ITS and EF1-alpha sequences place them within the clade containing species with Fusicoccum anamorphs. The brown, 1-septate conidia, however, do not conform to Fusicoccum. Therefore phylogenetically and morphologically the anamorphs of these two species belong in a genus distinct from any of the currently accepted anamorph genera assigned to Botryosphaeria. Through a study of the type species of Dothiorella this genus is resurrected to accommodate anamorphs of Botryosphaeria with brown, 1-septate conidia. Botryosphaeria sarmentorum is shown to be the teleomorph of Diplodia sarmentorum, which in turn is transferred to Dothiorella. Otthia quercus is transferred to Botryosphaeria as B. quercicola nom. nov.     

Bullera begoniae sp. nov. and Bullera setariae sp. nov., two new species of ballistoconidium-forming yeasts in the Bullera variabilis (Bulleribasidium) cluster isolated from plants in Taiwan

Two strains of xylose-containing and Q-10-having ballistoconidiogenous yeasts isolated from plant leaves collected in Taiwan were found to represent two new species of the genus Bullera. In the phylogenetic trees based on the sequence analysis of 18S rDNA and D1/D2 domain of 26S rDNA, these species are located in the Bullera variabilis (Bulleribasidum) cluster in Hymenomycetes. They are described as Bullera begoniae sp. nov. and Bullera setariae sp. nov., respectively.     

Cryptococcus foliicola sp. nov. and Cryptococcus taibaiensis sp. nov., novel basidiomycetous yeast species from plant leaves

A molecular taxonomic investigation performed on basidiomycetous yeast strains isolated from plant leaves collected in two areas of China revealed two novel species, Cryptococcus foliicola sp. nov. (type strain HS 23.3(T) = AS 2.2471(T) = CBS 9920(T)) and Cryptococcus taibaiensis sp. nov. (type strain ST 7.9(T) = AS 2.2444(T) = CBS 9919(T)), among the non ballistoconidium-forming strains producing cream-colored colonies. These new species differed markedly from closely related species in the internal transcribed spacer (ITS) and 26S rRNA D1/D2 region sequences. They clustered in a strongly supported clade represented by Cryptococcus victoriae in the Tremellales group in the phylogenetic trees drawn from ITS and D1/D2 sequences.     

丝虫草及其无性型关系的分子确证

目的对丝虫草（Ophiocordyceps filiform is）及从子囊孢子分离所得的无性型进行测序和比较分析,确证二者之间的对应关系。方法采用PCR技术,以rDNA-ITS区为分子指标。结果系统进化树显示丝虫草的无性型归为被毛孢属较为合理,同时通过形态特征比较的结果表明,其无性型为荔波被毛孢（Hirsutella liboensis）。结论本研究首次确立了丝虫草的有性型与无性型的对应关系,并且成功培育出了与天然丝虫草相同的虫草子实体。     

Bullera anomala sp. nov. and Bullera pseudovariabilis sp. nov., two new ballistoconidium-forming yeast species from Yunnan,China

Two ballistoconidium-forming yeast strains, CH 2.504 and CH 2.509, were isolated from the wilting leaves of Parthenocissus sp. collected in Yunnan, China in 1996. These two strains were assigned to the genus Bullera Derx by the characteristics of forming rotationally symmetric ballistoconidia, containing xylose in the cell hydrolysates and having Q-10 as the major ubiquinone. Molecular phylogenetic analysis based on small subunit ribosomal RNA gene (18S rDNA) and internal transcribed spacer (ITS) region sequences revealed that CH 2.504 is closely related to B. variabilis Nakase & Suzuki, while CH 2.509 is located in a clearly separate clade among the described Bullera species and related taxa. The results indicated that CH 2.509 represents a new species, for which Bullera anomala sp. nov. is the proposed name (type strain AS 2.2094^T). Further DNA-DNA reassociation data showed that CH 2.504 also represents a distinct species, for which the name Bullera pseudovariabilis sp. nov. is given (type strain AS 2.2092^T). This revised version was published online in June 2006 with corrections to the Cover Date.     

Xylogone ganodermophthora sp. nov., an ascomycetous pathogen causing yellow rot on cultivated mushroom Ganoderma lucidum in Korea

Yellow rot, caused by an ascomycetous fungus having a distinctive arthroconidial anamorph, is the most destructive disease of cultivated Ganoderma lucidum in Korea, but the identity of the yellow rot pathogen (YRP) remains uncertain. Isolates have been identified as Xylogone sphaerospora (with putative anamorph Sporendonema purpurascens) or as Arthrographis cuboidea. Therefore we used morphological features, pathogenicity tests and phylogenetic analyses of DNA sequences from the nuclear ribosomal genes, including partial small subunit and internal transcribed spacer regions, and from the gene encoding RNA polymerase second largest subunit to evaluate the relationship between YRP isolates and these species. YRP isolates formed a distinct subgroup within a clade that included X. sphaerospora, A. cuboidea and Scytalidium lignicola, the type species of Scytalidium, but the disposition of the clade within the Leotiomycetes was uncertain. We describe Xylogone ganodermophthora sp. nov. and Scytalidium ganodermophthorum sp. nov. for the teleomorph and anamorph of YRP respectively. Arthrographis cuboidea is reclassified as Scytalidium cuboideum comb. nov., and the anamorph of X. sphaerospora is named Scytalidium sphaerosporum sp. nov. In pathogenicity tests only X. ganodermophthora caused disease in Ganoderma lucidum. Amplified fragment length polymorphism analyses showed that X. ganodermophthora populations from diseased fruiting bodies or from oak wood in Korea consisted of two clonal groups.