Emendation of Dioszegia with redescription of Dioszegia hungarica and two new combinations,Dioszegia aurantiaca and Dioszegia crocea

During phylogenetic analyses of hymenomycetous yeasts based on 18S rDNA sequences, we found that Bullera armeniaca showed an extremely close phylogenetic relationship to Cryptococcus hungaricus. The analyses of internal transcribed spacer (ITS) regions of the two yeasts and the phylogenetically related species, Bullera aurantiaca and Bullera crocea, showed that B. armeniaca and C. hungaricus had identical sequences, indicating that these were conspecific. B. aurantiaca and B. crocea also showed high sequence similarity, 97.1% for ITS1, 100% for ITS2, and 98.7% for overall ITS regions. A DNA-DNA reassociation experiment revealed that B. armeniaca and C. hungaricus were conspecific and B. aurantiaca and B. crocea were two distinct species. These species occurred at a phylogenetically different lineage from that of Bulleromyces albus (anamorph: Bullera alba, type species of Bullera) and Filobasidiella neoformans (anamorph: Cryptococcus neoformans, neotype species of Cryptococcus). Based on these results, we emend the genus Dioszegia to include both ballistoconidium-forming and non-ballistoconidium-forming yeasts and redescribe the species Dioszegia hungarica. B. aurantiaca and B. crocea are also transferred to Dioszegia as Dioszegia aurantiaca comb. nov. and Dioszegia crocea comb. nov.     

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).     

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.     

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.     

Candida khmerensis sp. nov., a novel cation-tolerant yeast isolated from dry salted shrimp and sewage in Cambodia

Two cation-tolerant yeasts with powdered colonies, K28-3-2T and K26-1-4, were isolated from dry salted shrimp and sewage, respectively, in Siem Reap province, Cambodia. The D1/D2 sequences of the 26S rDNA data showed that the two isolates were conspecific and related to the Pichia burtonii and Candida fennica. Two isolates were examined by a polyphasic taxonomic approach, including molecular phylogenetic analysis, morphological, physiological and biochemical tests, DNA hybridization and MSP-PCR fingerprinting, in comparison with P. burtonii and C. fennica. The two isolates were found to grow by multilateral budding with true and pseudo-mycelium, to not produce ascospores, and to contain ubiquinone Q-8 similar to that of P. burtonii and C. fennica. The two isolates were not differentiated from the two closest species, P. burtonii and C. fennica, by the phenotypic character examined, except for the cation (Li+)-tolerance. From DNA-DNA reassociation studies, however, the two isolates showed low similarities to the closest two species. Based on D1/D2 sequences of 26S rDNA and DNA-DNA reassociation data, they were shown to be a new distinct species from P. burtonii and C. fennica. Therefore, a novel species is proposed, Candida khmerensis sp. nov., represented by strain K28-3-2T (=JCM 13262(T)=CBS 9784T). The novel species, Candida khmerensis sp. nov. can be clearly distinguished from P. burtonii and C. fennica by either the 26S rDNA D1/D2 or ITS region with 5.8S rDNA sequencing, or by the MSP-PCR fingerprinting pattern.     

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.     

The taxonomic position of Saccharomyces boulardii as evaluated by sequence analysis of the D1/D2 domain of 26S rDNA, the ITS1-5.8S rDNA-ITS2 region and the mitochondrial cytochrome-c oxidase II gene

A taxonomic study was carried out on eight strains of Saccharomyces boulardii. Morphological and physiological characteristics were consistent with those of Saccharomyces cerevisiae. Sequences of the D1/D2 domain of the 26S rDNA were identical for all strains examined and had a similarity value of 100% compared to sequences of the type strain of S. cerevisiae (CBS 1171T) and strain S288c. For all S. boulardii isolates was found the exact same ITS1-5.8S rDNA-ITS2 sequence, which displayed a close resemblance with the sequences published for S288c (99.9%), CBS 1171(T) (99.3%) and other S. cerevisiae strains. Sequence analysis of the mitochondrial cytochrome-c oxidase II gene (COX2) also resulted in identical sequences for the S. boulardii isolates and comparisons with available nucleotide sequences revealed close relatedness to strains of S. cerevisiae including S288c (99.5%) and CBS 1171(T) (96.6%). The electrophoretic karyotypes of the S. boulardii strains appeared quite uniform and although very typical of S. cerevisiae, they formed a cluster separate from strains of this species. The results of the present study strongly indicate a close relatedness of S. boulardii to S. cerevisiae and thereby support the recognition of S. boulardii as a member of S. cerevisiae and not as a separate species.     

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).     

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.     

斑马鱼肠道中一株酵母菌菌株的鉴定与系统发育分析

从斑马鱼肠道中分离到一株酵母菌,编号为ZF-5,进行了形态学观察、生理特征测定和26S rDNA D1/D2序列分析,并构建系统发育树。结果表明ZF-5菌株细胞呈卵圆形或杆状,为芽殖,有假菌丝;除乳糖外,能够发酵葡萄糖、蔗糖、麦芽糖等多种碳源;26S rDNA D1/D2区序列分析表明与季也蒙毕赤酵母Pichia guilliermondii的序列相似性最高,构建的系统发育进化树显示菌株ZF-5与Pichia guilliermondii模式菌株CBS 2030（= NRRL Y-2075）亲缘关系最近,     

Genetic diversity of the Pichia membranifaciens strains revealed from rRNA gene sequencing and electrophoretic karyotyping, and the proposal of Candida californica comb. nov

The genetic diversity of the types or authentic strains of 20 facultative synonyms of Pichia membranifaciens (E.C. Hansen) E.C. Hansen was revealed on the basis of large-subunit (26S) rDNA D1/D2 domain and internal transcribed spacer region sequencing and electrophoretic karyotyping. At least five separate species were recognized among the strains studied. Fourteen strains were confirmed to belong to P. membranifaciens. Strain CBS 241, an authentic strain of Zygosaccharomyces chevalieri Guilliermond var. fermentati Saito, should be assigned to Pichia manshurica Santa María. Strain CBS 243, an authentic strain of Zygopichia chiantigiana Castelli, is conspecific with CBS 2287, the type strain of Pichia fluxuum (Phaff & Knapp) Kreger-van Rij. Strain CBS 1367, the type of Zygosaccharomyces bisporus Anderson, belongs to Pichia kluyveri Bedford var. kluyveri. Strain CBS 989, the type of Cryptococcus californicus Anderson & Skinner, represents a distinct species, for which a new combination, Candida californica comb. nov., is proposed. The taxonomic status of strains CBS 189, the type of Pichia calliphorae Kl?cker, and CBS 214, the type of Pichia derossii Castelli, remain to be studied further. Their D1/D2 sequences and chromosomal DNA banding patterns were similar to those of P. membranifaciens, but their internal transcribed spacer sequences differed significantly.     

Phylogenetic identification of n-alkane assimilating Candida yeasts based on nucleotide divergence in the 59 end of LSU rDNA gene

Phylogenetic relationships of several species within the n-alkane assimilating Candida yeasts were investigated by using characters from the nucleotide sequence of the variable D1/D2 region at the 5' end of a large-subunit (26S) ribosomal DNA (rDNA) gene. First the nucleotide sequences of D1/D2 domain of Candida sp. 1098 (formerly identified as C. tropicalis 1098) and its dicarboxylic acid-producing-mutant strain M1210 were investigated. These two nucleotide sequences were identical and lacked only one base pair compared with that of C. maltosa CBS 5611 (type strain), and they were identified as C. maltosa. We then showed that C. maltosa IFO 1978 (formerly identified as C. cloacae) and C. maltosa IFO 1975 (formerly identified as C. subtropicalis) had the same nucleotide sequence and had only one base pair substitution compared with C. maltosa CBS 5611 (type strain), which is consistent with conventional classification. We also found that another widely studied n-alkane assimilating Candida yeast, C. tropicalis pk233, to be C. viswanathii.     

Fellomyces mexicanus sp. nov., a new member of the yeast genus Fellomyces isolated from lichen Cryptothecia rubrocincta collected in Mexico

Two strains isolated from the lichen Cryptothecia rubrocincta collected in Mexico are described as new members of the sterigmatoconidia-producing genus Fellomyces. Based on the identical sequences of the D1/D2 regions of 26S rDNA, as well as the results of the cluster analysis of the AFLP fingerprints, the strains have been shown to be conspecific. The 26S rDNA-based phylogeny has indicated that Fellomyces mexicanus clusters within a clade including species such as four other Fellomyces isolates from lichens: F. chinensis, F. lichenicola, F. sichuanensis and F. thailandicus. F. mexicanus is characterised by the presence of xylose in the cell walls and CoQ10 in the mitochondrial membrane. The type culture is strain HB25=CBS8279.     

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.     

Saccharomyces bacillaris is not a synonym of Candida stellata: reinstatement as Starmerella bacillaris comb. nov.

Torulopsis bacillaris (Kroemer and Krumbholz) Lodder (basionym Saccharomyces bacillaris Kroemer and Krumbholz) was frequently detected in oenological works on yeast ecology conducted in the mid-1950s in different wine regions of the world, before its unification with Torulopsis stellata (Kroemer and Krumbholz) Lodder. Most of the phenotypic characteristics pointed out for T. bacillaris are currently attributed to Candida zemplinina Sipiczki. In the present work isoenzyme profiles and rDNA restriction profiles of the neotype of S. bacillaris from two yeast culture collections (CBS 843 and PYCC 3044) and of the type strain of C. zemplinina (CBS 9494) were determined and similar profiles were detected. Moreover, the sequences of the D1/D2 region of the 26S rRNA gene of the three strains were 100 % identical. Different profiles were observed for the type strain of C. stellata (CBS 157) both for isoenzyme and rDNA restriction analysis and only 91 % similarity was found between the D1/D2 sequence of this strain and that of the neotype of S. bacillaris. In view of the newly obtained data and the fact that all above-mentioned species belong to the Starmerella clade, only distantly related to Candida tropicalis (the type species of the genus), S. bacillaris is hereby reinstated as Starmerella bacillaris comb. nov., with C. zemplinina as an obligate synonym.     

Two new species in the Pichia guilliermondii clade: Pichia caribbica sp. nov., the ascosporic state of Candida fermentati, and Candida carpophila comb. nov

Pichia caribbica sp. nov. (type strain DBVPG 4519, NRRL Y-27274, CBS 9966) is described as the ascosporic state of Candida fermentati, and Candida guilliermondii var. carpophila (type strain DBVPG 7739, NRRL Y-17905, CBS 5256) is elevated to species status as Candida carpophila comb. nov. These new taxa, which are indistinguishable on the basis of conventional taxonomic criteria, differ from one another and from Pichia guilliermondii by low DNA base sequence relatedness, different electrophoretic karyotypes, and nucleotide divergence in domains D1/D2 of 26S rDNA. Pichia caribbica produces one, rarely two, saturn-shaped ascospores in persistent asci. On the basis of molecular criteria, C. carpophila comb. nov., C. fukuyamaensis, and C. xestobii are conspecific, with the name C. carpophila having taxonomic priority.     

Six new anamorphic ascomycetous yeasts near Candida tanzawaensis

Six new species of the yeast genus Candida are described from their unique nucleotide sequences in the D1/D2 domain of 26S rDNA. Five of these species form a clade with Candida tanzawaensis, and the sixth is basal to this group. The new species and their sources of isolation are the following: Candida ambrosiae (type strain NRRL YB-1316, CBS 8844), from insect frass, rotted wood and mushroom fruiting bodies; Candida canberraensis (type strain NRRL YB-2417, CBS 8846), from soil; Candida caryicola (type strain NRRL YB-1499, CBS 8847), from a pignut hickory tree; Candida prunicola (type strain NRRL YB-869, CBS 8848), from exuded gum of a black cherry tree; Candida pyralidae (type strain NRRL Y-27085, CBS 5035), from insect frass; Candida xylopsoci (type strain NRRL Y-27066, CBS 6037), from insect frass.