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).     

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.     

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).     

Sporobolomyces diospyroris sp. nov., Sporobolomyces lophatheri sp. nov. and Sporobolomyces pyrrosiae sp. nov., three new species of ballistoconidium-forming yeasts in the Agaricostilbum lineage isolated from plants in Taiwan

Three strains of xylose-lacking and ubiquinone-10-having ballistoconidium-forming yeasts isolated from plant leaves collected in Taiwan were found to represent respective new species. In phylogenetic trees constructed based on the nucleotide sequences of 18S rDNA and D1/D2 domain of 26S rDNA, they were located in the Agaricostilbum lineage (Agaricostilbum/Bensingtonia cluster). Since the taxonomic properties of these species coincide with those of the genus Sporobolomyces, they are described as Sporobolomyces diospyroris sp. nov., Sporobolomyces lophatheri sp. nov. and Sporobolomyces pyrrosiae sp. nov., respectively.     

Candida kazuoi sp. nov. and Candida hasegawae sp. nov., two new species of ascomycetous anamorphic yeasts isolated from insect frass in Thailand

Two strains of anamorphic yeasts isolated from insect frass collected in southern Thailand were assigned to the genus Candida based on the conventional taxonomic criteria used for yeast classification. In the phylogenetic tree based on the D1/D2 domain of the 26S rDNA, these strains are distant from the known species of yeasts and considered to represent two different new species. They are named Candida kazuoi sp. nov. and Candida hasegawae sp. nov.     

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.     

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.     

Three new thermotolerant methylotrophic yeasts, Candida krabiensis sp. nov., Candida sithepensis sp. nov., and Pichia siamensis sp. nov., isolated in Thailand

Five strains of thermotolerant methylotrophic yeasts isolated in Thailand were found to represent three new species in the genera Pichia and Candida, based on phylogenetic analysis of D1/D2 domain of 26S rDNA, in addition to the morphological, physiological, biochemical and chemotaxonomic characterization. Three strains, FS96 and FS101 from flowers and M02 from tree flux, were characterized by ubiquinone Q7, multilateral budding, and the formation of hat-shaped ascospores that are liberated at maturation. These strains showed identical nucleotide sequences in the D1/D2 domain and formed a cluster with Candida thermophila, "Pichia salicis" and Pichia angusta. They differed by 1.9% of nucleotide substitutions from Candida thermophila, the nearest species. They were considered to represent a single new species and are described as Pichia siamensis sp. nov. Two strains, N051 and S023, isolated from soil did not produce ascospores, proliferated by multilateral budding, did not demonstrate urease or DBB color reaction, and lacked sexual stages. These characteristics correspond to the genus Candida. Strains N051 and S023 differed by 2.8% and 1.9% of nucleotide substitutions in the D1/D2 domain from the nearest species, Candida nemodendra and Candida ovalis, respectively, and are considered to represent respective new species. N051 and S023 are described as Candida krabiensis sp. nov. and Candida sithepensis sp. nov., respectively.     

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.     

Sporobolomyces magnisporus sp. nov., a new yeast species in the Erythrobasidium cluster isolated from plants in Taiwan

Two strains of yeasts that contain Q-10 as the major ubiquinone, lack cellular xylose and produce large bilaterally symmetrical ballistoconidia were isolated from plants collected in a protected subtropical rain forest in Taiwan and were found to represent a new species. The taxonomic properties of this species coincide with the genus Sporobolomyces, so it is described as Sporobolomyces magnisporus sp. nov. In phylogenetic trees based on the nucleotide sequences of 18S ribosomal DNA and D1/D2 domain of 26S ribosomal DNA, this species is located in the Erythrobasidium cluster.     

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)).     

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.     

Candida easanensis sp. nov., Candida pattaniensis sp. nov. and Candida nakhonratchasimensis sp. nov., three new species of yeasts isolated from insect frass in Thailand

Six strains of anamorphic yeasts isolated from insect frass collected in several regions of Thailand were assigned to the genus Candida based on the conventional taxonomic criteria used for yeast classification. These strains have Q-7 as the major ubiquinone and are suggested to have close relationships to the genus Pichia. Three strains, ST-225, ST-228 and ST-229, have identical nucleotide sequences in the D1/D2 domain of 26S rDNA and are closely related to Pichia japonica, but differ by six nucleotides (1.1% ) from this species. These three strains are considered to represent a single new species, which is described as Candida easanensis sp. nov. Two strains, ST-311 and ST-320, have identical sequences in the D1/D2 domain and resemble Pichia veronae and Pichia fabianii but differ from them by nine nucleotides (1.6%) in D1/D2 sequences. The two strains are described as Candida pattaniensis sp. nov. The remaining strain, ST-37, is related to Pichia americana and Pichia bimundalis but differs by six(1.1%) and seven (1.2%) nucleotides from these species, respectively. This strain is described as Candida nakhonratchasimensis sp. nov.     

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.     

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).     

Pichia koratensis sp. nov., a new ascomycetous yeast related to Pichia acaciae isolated from insect frass in Thailand

Two yeast strains (ST-235 and ST-237) isolated from insect frass collected in northeastern Thailand, were identified as Pichia acaciae and a hitherto undescribed species based on the sequence analysis of the D1/D2 domain of 26S rDNA, ITS regions, and DNA-DNA reassociation experiments. The latter strain, ST-237 (=BCC 11769=NBRC 103638=JCM 12576), was described as Pichia koratensis sp. nov. Pichia koratensis is closely related to Pichia acaciae in the D1/D2 domain sequence of 26S rDNA and phenotypical characteristics but differs by 5 nucleotides (0.9%) from the type strain of P. acaciae and is clearly distinguished by the assimilation and fermentation of sucrose.     

Identification and phylogeny of ascomycetous yeasts from analysis of nuclear large subunit (26S) ribosomal DNA partial sequences

Approximately 500 species of ascomycetous yeasts, including members of Candida and other anamorphic genera, were analyzed for extent of divergence in the variable D1/D2 domain of large subunit (26S) ribosomal DNA. Divergence in this domain is generally sufficient to resolve individual species, resulting in the prediction that 55 currently recognized taxa are synonyms of earlier described species. Phylogenetic relationships among the ascomycetous yeasts were analyzed from D1/D2 sequence divergence. For comparison, the phylogeny of selected members of the Saccharomyces clade was determined from 18S rDNA sequences. Species relationships were highly concordant between the D1/D2 and 18S trees when branches were statistically well supported.     

Zygosaccharomyces kombuchaensis,a new ascosporogenous yeast from 'Kombucha tea'

A new ascosporogenous yeast, Zygosaccharomyces kombuchaensis sp. n. (type strain NRRL YB-4811, CBS 8849), is described; it was isolated from Kombucha tea, a popular fermented tea-based beverage. The four known strains of the new species have identical nucleotide sequences in domain D1/D2 of 26S rDNA. Phylogenetic analysis of D1/D2 and 18S rDNA sequences places Z. kombuchaensis near Zygosaccharomyces lentus. The two species are indistinguishable on standard physiological tests used for yeast identification, but can be recognized from differences in restriction fragment length polymorphism patterns obtained by digestion of 18S-ITS1 amplicons with the restriction enzymes DdeI and MboI.     

Two new species of the genus Candida in the Zygoascus clade, Candida lundiana sp. nov. and Candida suthepensis sp. nov., isolated from raw honey in Thailand

During a survey of yeasts associated with raw honey collected in Thailand, two strains of the Zygoascus clade were isolated from the Asian cavity-nesting honeybee Apis cerana and the stingless bee Homotrigona fimbriata. Phylogeny based on 26S rDNA D1/D2 sequences placed these yeasts as members of a clade including Candida bituminiphila, Candida patagonica and Candida polysorbophila. The strains of the two novel species, CBS 12271^T and CBS 12270^T, respectively, could be unquestionably distinguished from their relatives by rDNA sequences and other taxonomic characteristics. Therefore, the novel anamorphic species, Candida lundiana sp. nov. (type strain CBS 12271^T = JCM 16823^T) and Candida suthepensis sp. nov. (type strain CBS 12270^T = JCM 16822^T) are described.     

Pichia nongkratonensis sp. nov., a new species of ascomycetous yeast isolated from insect frass collected in Thailand

A yeast strain isolated from insect frass collected in Thailand was found to represent a new species of the genus Pichia. It is described as Pichia nongkratonensis sp. nov. In the phylogenetic tree based on the D1/D2 domain sequences of 26S rDNA, this yeast constitutes a cluster with Pichia dryadoides with high bootstrap confidence level; however, it differs from the latter species by 5.6% base substitutions. Pichia nongkaratonensis resembles P. dryadoides also in the phenotypic characteristics but is distinguished from this species by the assimilation of several carbon and nitrogen compounds.