Wickerhamomyces sinyiensis f.a., sp. nov., a new ascomycetous yeast species in the Wickerhamomyces clade isolated in Taiwan.

This study describes Wickerhamomyces sinyiensis, a new anamorphic ascomycetous yeast species, four strains of which were isolated from soil and the fruiting body of a mushroom in Taiwan between 2006 and 2007. Analysis of the sequences of the large-subunit rRNA, small-subunit rRNA and elongation factor-1α identified this species as a member of the Wickerhamomyces clade. The yeast strains of W. sinyiensis exhibited a 0-3 nucleotide difference in the sequences of the D1/D2 domain of the large subunit rRNA when compared to one another and a 10 and 11 nucleotide difference when compared to Candida sp. BG99-11-14-10-4-1 and NRRL Y-7574, the closest undescribed species, respectively. The yeast strains differed by 77 and 78 nucleotides from W. orientalis and W. bispora, the close Wickerhamomyces species, respectively. The internal transcribed spacer sequences of the four isolates exhibited a divergence of 106-108 substitutions from the recognized species W. xylosivorus. No sexual reproduction was observed. The strains differed from those of related species in terms of their carbon and nitrogen assimilation patterns. Therefore, this study proposes W. sinyiensis f.a., sp. nov. to accommodate these four strains, with W. sinyiensis BCRC 23185T (isotype CBS 11432T; MycoBank number MB563484) as the holotype.

Isolation of endophytic fungi with antimicrobial activity from medicinal plant Zanthoxylum simulans Hance.

Fungal endophytes have been found to exist in many plant species and appear to be important to their plant hosts. However, the diversity and biological activities of these fungi remain largely unknown. Zanthoxylum simulans Hance, a popular natural spice and medicinal plant, commonly known as Szechuan pepper or Chinese-pepper, grows on Kinmen Island, Taiwan. In this study, leaf and stem samples of Z. simulans, collected in summer and winter, were screened for antimicrobial and anti-inflammatory metabolite-producing endophytic fungi. A total of 113 endophytic strains were isolated and cultured from Z. simulans, among which 23 were found to possess antimicrobial activity, belonging to six fungal genera: Penicillium (26.09%, 6), Colletotrichum (21.74%, 5), Diaporthe (21.74%, 5), Daldinia (17.39%, 4), Alternaria (8.70%, 2), and Didymella (4.34%, 1). We also found that the number of species with antimicrobial activity and their compositions differed between summer and winter. Our study demonstrated that Z. simulans might contain large and diverse communities of endophytic fungi, and its community composition varies seasonally. In addition, fungal endophytes produce antimicrobial agents, which may protect their hosts against pathogens and could be a potential source of natural antibiotics.

Wickerhamiella osmotolerans sp. nov. and Wickerhamiella tropicalis sp. nov., novel ascomycetous yeast in the family Wickerhamiellaceae.

Seven yeast strains, DMKU VGT1-14T, DMKU VGT1-19T, DMKU-JMGT1-28, DMKU-JMGT1-32, DMKU VGT2-06, DMKU VGT2-19 and DMKU VGT6-14, were isolated from a grease trap in Thailand and two strains, SJ-1 and SN-102 were isolated from the sea surface microlayer in Taiwan. On the basis of phenotypic characteristics and sequence analysis of the D1/D2 region of the large subunit (LSU) rRNA gene and the internal transcribed spacer (ITS) region, these strains represented two novel yeast species of the genus Wickerhamiella. In terms of pairwise sequence similarity, four strains, DMKU VGT1-14, DMKU-JMGT1-32, DMKU VGT6-14 and SN-102, were closely related to Wickerhamiella infanticola NRRL Y-17858T but differed by 13 nucleotide substitutions with one gap (2.46 %) in the D1/D2 domain of the LSU rRNA gene and 15 nucleotide substitutions with 23 gaps (4.2 %) in the ITS region. The strains DMKU VGT1-19T, DMKU-JMGT1-28, DMKU VGT2-06, DMKU VGT2-19 and SJ-1, differed from the type strain of the most closely related species, Wickerhamiella sorbophila NRRL Y-7921T, by nine nucleotide substitutions with one gap (1.66 %) in the D1/D2 domain of the LSU rRNA gene and nine nucleotide substitutions with 17 gaps (2.52%) in the ITS region. Hence, the names Wickerhamiella osmotolerans sp. nov. and Wickerhamiella tropicalis sp. nov. are proposed to accommodate these species in the genus Wickerhamiella. The holotypes are W. osmotolerans DMKU VGT1-14T (ex-type strain TBRC 11425=PYCC 8359=CGMCC 2.6179; Mycobank number 833394) and W. tropicalis DMKU VGT1-19T (ex-type strain TBRC 11426=PYCC 8360=CGMCC 2.6180; Mycobank number 833393).

Teunomyces basidiocarpi sp. nov. and Teunomyces luguensis sp. nov., two ascomycetous yeast species related to the Candida kruisii clade isolated in Taiwan.

Four strains of anamorphic yeasts isolated from the fruiting bodies of mushrooms collected in Taiwan were found to represent two novel yeast species belonging to the genus Teunomyces, which was formally known as the Candida kruisii clade. Strains NY13M09T and NY14M14 were related to the type strains of Teunomyces panamensis, T. pallodes, T. tritomae and T. lycoperdinae, and strains GG4M07T and GG6M14 were related to T. kruisii NRRL Y-17087T and T. cretensis NRRL Y-27777T. However, strains NY13M09T and NY14M14 differed from their closest phylogenetic neighbours by 2.9-3.7 % in the D1/D2 domain sequence of the LSU rRNA gene and by 6.6-13.7 % in the internal transcribed spacer (ITS); GG4M07T and GG6M14 differed from their closest known species by 2.4 % in the D1/D2 domain sequence of the LSU rRNA gene and by 8.7-10.0 % in the ITS. Meanwhile, these strains were also clearly distinguished from their closest relatives based on the results of physiological tests. Based on the characteristics described above, the strains could be regarded as representing two novel species of the genus Teunomyces, for which the names Teunomyces basidiocarpi sp. nov. and Teunomyces luguensis sp. nov. are proposed. The holotypes are Teunomyces basidiocarpi BCRC 23475T and Teunomyces luguensis BCRC 23476T.

Taxonomy of the yeast genus Vanderwaltozyma and proposal of Vanderwaltozyma meishanica sp. nov., Vanderwaltozyma huisunica sp. nov., and Vanderwaltozyma molinica sp. nov.

Using electrophoretic karyotyping, RAPD fingerprinting and phylogenetic analysis of ribosomal RNA gene sequences, twenty-six Vanderwaltozyma strains were studied. Out of 19 strains isolated in mountainous areas of Taiwan, eighteen strains were isolated from soil and one strain was isolated from the fruiting body of mushroom, six were identified as V. polyspora and three as V. verrucispora. Based on the results of a multigene sequence analysis (D1/D2, ITS and mitochondrial COX II gene) and DNA-DNA reassociation, three new ascosporic members of the genus Vanderwaltozyma are formally described: V. huisunica sp. nov. (GA1S06T = CBS 12250T = BCRC 23260T), V. meishanica sp. nov. (EN4S02T = CBS 12249T = BCRC 23255T) and V. molinica sp. nov. (GJ8S05T = CBS 12251T = BCRC 23264T), and the holotypes of these novel species are assigned as BCRC 23260T, BCRC 23255T and BCRC 23264T, respectively.

Comparison of biofunctional activity of Asparagus cochinchinensis (Lour.) Merr. Extract before and after fermentation with Aspergillus oryzae.

Asparagus cochinchinensis root (ACR) is used in traditional Chinese medicine. In this study, ACR was first extracted with 25% ethyl acetate (EA) and then fermented by Aspergillus oryzae to enhance its antioxidant activity and evaluate its potential antityrosinase activity. The physiological activity and cytotoxicity of A. oryzae-fermented ACR extract, along with its antityrosinase activity and effects on melanogenic factor levels in human epidermal melanocytes (HEMs), were analyzed and compared with those of the unfermented extract. The results showed that the physiological activity of the fermented extract in vitro or in cells was significantly higher than that of the unfermented extract. The IC50 values for 2,2-diphenyl-1-picrylhydrazine radical scavenging activity, reducing power, and antityrosinase activity in vitro for the fermented extract were 250.6 ± 32.5, 25.7 ± 3.5, and 50.6 ± 3.1 mg/L, respectively. The fermented extract favored cellular antityrosinase activity with low melanin production in human melanoma cells compared with the unfermented extract. The inhibitory mechanism of melanin synthesis by unfermented extract was independent of the tested melanogenesis-related proteins. However, the inhibitory mechanism of the fermented extract was possibly caused by synergistic inhibition of these proteins. Thus, A. oryzae-fermented ACR extract may be used for developing new health food or cosmetic ingredients.

Cystobasidium keelungensis sp. nov., a novel mycosporine producing carotenogenic yeast isolated from the sea surface microlayer in Taiwan.

Cystobasidium keelungensis SN2T (CBS 6949 = BCRC 920080), a new anamorphic basidiomycetous yeast species, is described in this paper. The strains belonging to this species were isolated from the sea surface microlayer and underlying water in Taiwan. These strains were identified by examining nucleotide sequences in the species-specific D1/D2 domains of the large subunit (LSU) ribosomal RNA (rRNA) and by evaluating their physiological characteristics. Phylogenetic analyses of D1/D2 sequences revealed that C. keelungensis was most closely related to the species C. slooffiae (LSU rRNA gene sequence divergence of 1.5%), and it belonged to the Erythrobasidium clade. No sexual reproduction was observed. This species differed from related species in carbon and nitrogen assimilation patterns and growth at 35 °C. Screening C. keelungensis for the presence of UV-absorbing compounds showed that mycosporine-glutaminol-glucoside and mycosporine-glutamicol-glucoside (maximum absorption: 310 nm) were the major UV-absorbing compounds, which differ from the compounds present in some freshwater yeast strains reported in previous studies. After UV induction, SN2 had a higher level of mycosporine production than other carotenogenic yeasts in this study.

Vishniacozyma changhuana sp. nov., and Vishniacozyma taiwanica sp. nov., two novel yeast species isolated from mangrove forests in Taiwan.

Mangroves grow in the intertidal zone, which alternates between fresh water and sea water, with abundant bioresources. In mangrove habitats, yeasts play an important role in the decomposition of organic matter, and such nutrient cycling has high ecological importance. During a study of the diversity of yeast from Taiwan mangroves, seven strains of basidiomycetous yeasts were isolated and these yeasts represent two novel yeast species belonging to the genus Vishniacozyma. According to the sequences of the D1/D2 domain of large subunit (LSU) rRNA and the internal transcribed spacer (ITS) region, these seven strains could be clearly classified into two groups representing two individual, distinct species. Strains HM5L06, HM6L07, HM11L11 and BJ3S01, differed from their closest relative species Vishniacozyma phoenicis by 4-5 nt substitutions (no gaps) in the sequences of the D1/D2 domain of LSU rRNA and by 23 nt substitutions (10 gaps) in the ITS region. Strains HM6L11, HM7L02 and HM8L19, differed from their closest relative species Vishniacozyma penaeus by 7 nt substitutions (one gap) in the sequences of the D1/D2 domain of LSU rRNA and by 27 nt substitutions (12 gaps) in the ITS region. The scientific names of Vishniacozyma taiwanica sp. nov. and Vishniacozyma changhuana sp. nov. are proposed for these strains. The holotypes are Vishniacozyma taiwanica BCRC 23477T (ex-type HM5L06=CBS 16558; MycoBank number MB837428) and Vishniacozyma changhuana BCRC 23478T (ex-type HM6L11=CBS 16556; MycoBank number MB837429).

Diversity of yeasts associated with the sea surface microlayer and underlying water along the northern coast of Taiwan.

Yeast communities inhabiting the sea surface microlayer (SSML) on the northern coast of Taiwan were examined using a cultivation method and compared with those inhabiting the underlying water (UW) at a 50-cm depth. Culturable yeasts were recovered from the SSML and UW samples collected in the morning during 4 field campaigns, and 420 strains were isolated. The 420 isolates were grouped into 43 species using a polyphasic molecular approach, including sequence analysis of the 26S rDNA D1/D2 domain and 5.8S-ITS region. From the SSML samples, 12 genera and 39 species, including 7 new species of Cryptococcus sp. (1), Candida spp. (4), and Rhodotorula spp. (2), were isolated. From the UW samples, 10 genera and 21 species, including one new species of Rhodotorula sp. (1), were isolated. Rhodotorula mucilaginosa was the most abundant species present in the yeast community in SSML (37.6%) and UW (21.6%) samples. Basidiomycetous yeasts (63.6%) and pigmented yeasts (64.5%) comprised the major yeast population. The yeast community in the SSML had a higher species number and abundance than the UW. Moreover, although the majority of yeast community species were from the SSML, individual species distribution in the SSML was unequal.

Characterization of polyhydroxyalkanoate-producing bacteria isolated from sludge of commercial pig farms for producing methyl esters.

The objectives of this work were to isolate and characterize the polyhydroxyalkanoate (PHA) producing bacteria in enriched piggery sludge and make methyl esters from PHA for industrial applications. The strain ECAe24 isolated from piggery sludge with the highest PHA production was selected to produce PHA and then methyl ester by trans-esterification using glucose as substrate under mesophilic conditions. The final product after trans-esterification consisted of approximately 75.39% of fatty acid methyl ester and was identified as decanoic acid-3-hydroxy-methyl ester, octanoic acid-3-hydroxy-methyl ester, and some other contents. The novelty of this study is to use PHA-producing bacteria from piggery sludge to make fatty acid methyl esters which can be used as materials for producing biodiesel from piggery wastes.

Identifying and characterizing Yarrowia keelungensis sp. nov., an oil-degrading yeast isolated from the sea surface microlayer.

Ascomycetous yeast strain SM-22 was isolated from the sea-surface microlayer near the Keelung City off the northern coast of Taiwan. This strain showed a cell surface hydrophobicity higher than 90 %, moderate UV A/B resistance, and it degraded 68 % of the total petroleum hydrocarbon content of an artificial seawater medium containing 1 % (v v(-1)) diesel oil within 15 days at 25 °C. The closest phylogenetic relative of this strain is Candida oslonensis CBS 10146(T), but it differs from strain SM-22 by a 3.7 % divergence (including 18 nucleotide substitutions and 2 gaps) in the D1/D2 domain sequence of the large subunit rRNA gene. This difference clearly suggests that the strain SM-22 represents a distinct species. Strain SM-22 does not produce ascospores on common sporulation media and it can therefore be considered an anamorph of the genus Yarrowia. Thus, the name Yarrowia keelungensis sp. nov. (type strain SM-22(T) = BCRC 23110(T) = JCM 14894(T) = CBS 11062(T)) is proposed as a novel species of genus Yarrowia.

Tetrapisispora taiwanensis sp. nov. and Tetrapisispora pingtungensis sp. nov., two ascosporogenous yeast species isolated from soil.

Two novel yeast species, Tetrapisispora taiwanensis sp. nov. and Tetrapisispora pingtungensis sp. nov., belonging to the Saccharomycetaceae within the Saccharomycetales, are proposed to accommodate six strains isolated from samples of Taiwanese forest soil between 2005 and 2010. Sequence analysis of the internal transcribed spacer (ITS) and the D1/D2 domains of the large-subunit (LSU) rRNA gene revealed that the two species are phylogenetically closely related to species of the genus Tetrapisispora. Moreover, the assimilation spectrum of carbon and nitrogen sources and morphological characteristics are very similar to those of other Tetrapisispora species. The molecular, morphological and physiological characteristics described above indicate that these two species should be classified as members of the genus Tetrapisispora. The two species can be differentiated from each other and from other Tetrapisispora species based on their LSU D1/D2 rRNA gene and ITS sequences. Thus, the two species could be regarded as novel species of the genus Tetrapisispora, and the names Tetrapisispora taiwanensis sp. nov. (type strain SJ6S04(T) =CBS 10586(T) =NBRC 102652(T) =BCRC 23090(T)) and Tetrapisispora pingtungensis sp. nov. (type strain NC2S06(T) =CBS 12780(T) =BCRC 23409(T)) are proposed. The type strains of the two species were isolated from forest soil in Leinhuatsu Park, Nantou, in 2006 and from Hungchun, Pingtung, in 2009, respectively.

Five novel species of the anamorphic genus Candida in the Cyberlindnera clade isolated from natural substrates in Taiwan.

Twelve strains representing five novel yeast species were isolated from natural samples distributed in mountain areas in Taiwan during 2007 and 2009. Sequence analysis of the D1/D2 domain of the large subunit (LSU) rRNA gene revealed that these species are members of the Cyberlindnera clade. These five new species have a greater than 1% difference from their closest relatives in the sequences of the D1/D2 domain of the LSU rRNA gene and were well separated from their closest relatives in terms of physiological characteristics. Moreover, a sexual state could not be found in these five novel yeast species. Therefore, the scientific names of Candida maesa sp. nov. (type strain GJ8L01(T)), Candida takata sp. nov. (type strain EN25S01(T)), Candida taoyuanica sp. nov. (type strain GY15S07(T)), Candida hungchunana sp. nov. (type strain NC3W71(T)) and Candida stauntonica sp. nov. (type strain GY13L05(T)) were proposed to accommodate these yeasts.

Kloeckera taiwanica sp. nov., an ascomycetous apiculate yeast species isolated from mushroom fruiting bodies.

Three apiculate yeast strains, EJ7M09(T), GJ5M15 and GJ15M04, isolated from mushrooms in Taiwan were found to represent a novel species of the genus Kloeckera. The phylogenetically closest relative of this novel species is Hanseniaspora occidentalis, but the type strain of H. occidentalis differed by 4.6 % divergence (25 substitutions; 5 gaps) in the sequence of the D1/D2 domain of the large subunit rRNA gene. This difference clearly suggests that the three strains represent a distinct species. As none of the strains that were examined in this study produced ascospores or exhibited conjugation on common sporulation medium either alone or in a pairwise mixture, this species could be considered as an anamorphic member of the genus Hanseniaspora, and a novel species, Kloeckera taiwanica sp. nov., is proposed, with EJ7M09(T) ( = BCRC 23182(T) = CBS 11434(T)) as the type strain.

Candida theae sp. nov., a new anamorphic beverage-associated member of the Lodderomyces clade.

Four strains representing a novel yeast species belonging to the genus Candida were independently isolated in Taiwan and Ecuador. Two strains (G17(T) and G31) were isolated in Taiwan, by pellet precipitation from plastic-bottled tea drinks produced in Indonesia, while two additional strains (CLQCA 10-049 and CLQCA 10-062) were recovered from ancient chicha fermentation vessels found in tombs in Quito, Ecuador. These four strains were morphologically, and phylogenetically identical to each other. No sexual reproduction was observed on common sporulation media. Large-subunit (LSU) rRNA gene sequence analysis revealed the four strains to belong to the Lodderomyces clade, closely related to members of the Candida parapsilosis species complex. The four strains, which have identical LSU D1/D2 sequences, differ from their closest phylogenetic neighbors, Candida orthopsilosis and Candida parapsilosis, by 6-9 nt substitutions, respectively. Physiologically, the four strains are similar to Candida parapsilosis, although they can be distinguished from their closest relative by the assimilation of arbutin, nitrite, and creatine. The Indonesian and Ecuadorian strain sets can also be distinguished from one another based on ITS sequencing, differing by 4 substitutions in ITS1 and 1 single nucleotide indel in ITS2. Collectively, the results indicate that the four strains represent a previously unrecognized species of Candida. The name Candida theae sp. nov. is proposed to accommodate these strains, with G-17(T) (BCRC 23242(T)=CBS 12239(T)=ATCC MYA-4746(T)) designated as the type strain.

The relationship of cytomorphology of medullary thyroid carcinomas between family members with the same RET proto-oncogene mutation.

OBJECTIVE: To evaluate the relationship of the cytomorphology of medullary thyroid carcinomas (MTC) between family members with the same RET proto-oncogene mutation. STUDY DESIGN: Review of the fine-needle aspiration slides of 13 cases with MTC proven by surgery and pathology from 5 unrelated families with either multiple endocrine neoplasia (MEN) type 2A or familial MTC (FMTC). RESULTS: Small, round, and abundant large oval-to-polygonal cells were major cytomorphologic findings in 66.7% of family members with exon 11, codon 634 TGC → CGC germline mutation. Small, round cells and only a few or no polygonal cells were found in 66.7% of family members with exon 11, codon 634 TGC → TTC germline mutation and in 100% of family members with codon 634 TGC → TGG germline mutations, as well as in 100% of family members with exon 10, codon 620 TGC → GGC germline mutation. CONCLUSIONS: The high rate of similarity of cytomorphology (66.7-100%) in the family members with MEN type 2A or FMTC might be related to the same etiology in the production of MTC in the same family. The relationship of the respective cytomorphology with the long-term prognosis is worth elucidating further.

Candida gosingica sp. nov., an anamorphic ascomycetous yeast closely related to Scheffersomyces spartinae.

During surveys on yeast diversity in forest soils from Taiwan and Thailand, ten yeast strains isolated from different samples were found to have similar molecular and physiological characteristics. Sequence analysis of small subunit (SSU) rDNA, the D1/D2 domain of large subunit (LSU) rDNA and internal transcribed spacer (ITS)-5.8S rDNA demonstrated that these strains were closely related to Scheffersomyces spartinae. The novel strains could be differentiated from S. spartinae by a 0.9  % sequence divergence (5 substitutions, 0 gaps) in the D1/D2 domain of LSU rDNA, a 1.5  % divergence (8 substitutions, 0 gaps) in the ITS-5.8S rDNA and a 0.7  % divergence (12 substitutions, 2 gaps) in the SSU rDNA. The novel strains also showed specific patterns of electrophoretic karyotypes that differed from that of S. spartinae. Therefore, a novel yeast species, Candida gosingica sp. nov., is proposed to accommodate these strains. The type strain SJ7S11(T) (=BCRC 23194(T)=CBS 11433(T)) was assigned and deposited in the Bioresource Collection and Research Center (BCRC), Food Industry Development and Research Institute, Hsinchu, Taiwan, and Centraalbureau voor Schimmelcultures (CBS), Utrecht, The Netherlands.

Asterotremella meifongana sp. nov. and Asterotremella nantouana sp. nov., two anamorphic basidiomycetous yeasts isolated from soil and mushrooms.

Two novel yeast species, Asterotremella meifongana sp. nov. and Asterotremella nantouana sp. nov., belonging to Asterotremellaceae within the Trichosporonales, are proposed to accommodate five strains isolated from Taiwanese samples of soil and basidiocarps during 2006-2007. Sequence analysis of the D1/D2 domains of the large subunit rRNA gene revealed that the two species are phylogenetically closely related to species of the genus Asterotremella, which indicates that they are members of the genus. The two species can be further differentiated from other Asterotremella species based on their internal transcribed spacer sequences, as well as on a few physiological test responses. The type strains are A. meifongana SN4S04(T) (=CBS 11424(T) = BCRC 23196(T)) and A. nantouana SN5S06(T) (=CBS 10890(T) = BCRC 23135(T)).