rRNA Gene ITS Region Research Articles

Copper Tolerance of Novel Rhodotorula sp. Yeast Isolated from Gold Mining Ore in Gia Lai, Vietnam

In this study, twenty-five yeast strains were isolated from soil samples collected in the gold mining ore in Gia Lai, Vietnam. Among them, one isolate named GL1T could highly tolerate Cu2+ up to 10 mM, and the isolates could also grow in a wide range of pH (3–7), and temperature (10–40 °C). Dried biomass of GL1 was able to remove Cu2+ effectively up to 90.49% with a maximal biosorption capacity of 18.1 mg/g at pH 6, temperature 30 °C, and incubation time 60 min. Sequence analysis of rDNA indicated this strain was closely related to Rhodotorula mucilaginosa but with 1.53 and 3.46% nucleotide differences in the D1/D2 domain of the 28S rRNA gene and the ITS1-5.8S rRNA gene-ITS2 region sequence, respectively. Based on phylogenetic tree analysis and the biochemical characteristics, the strain appears to be a novel Rhodotorula species, and the name Rhodotorula aurum sp. nov. is proposed. This study provides us with more information about heavy metal-tolerant yeasts and it may produce a new tool for environmental control and metal recovery operations.

Interannual dynamics of Tuber melanosporum and fungal communities in productive black truffle orchards amended with truffle nests.

Truffle growers devote great efforts to improve black truffle productivity, developing agronomic practices such as 'truffle nests' (peat amendments that are supplemented with truffle spore inoculum). It has been hypothesized that improved fruiting associated with nests is linked to stimulation of truffle mycelia previously established in soil or to changes generated in soil fungal community. To assess this, we used real-time PCR to quantify black truffle extraradical mycelium during 2 years after nests installation. We also characterized the fungal community via high-throughput amplicon sequencing of the ITS region of rRNA genes. We found that neither the abundance of truffle mycelium in nests nor in the soil-nest interphase was higher than in the bulk soil, which indicates that nests do not improve mycelial growth. The fungal community in nests showed lower richness and Shannon index and was compositionally different from that of soil, which suggests that nests may act as an open niche for fungal colonization that facilitates truffle fruiting. The ectomycorrhizal fungal community showed lower richness in nests. However, no negative relationships between amount of truffle mycelium and reads of other ectomycorrhizal fungi were found, thus countering the hypothesis that ectomycorrhizal competition plays a role in the nest effect.

Land utilization types impacted the species diversity and population distribution of nodulating and non-nodulating rhizobia in the Philippines

The influence of three land utilization types (LUT) on the diversity and population of soybean-nodulating rhizobia indicated how human activities affect the ecology of these microorganisms. This report revealed that based on LUT (LUT1-rice-based; LUT2-soybean-based; LUT3-legume-rice), different species of rhizobia can be more abundant than the other species. Targeting the 16S rRNA gene and the 16S-23S rRNA gene ITS region through Polymerase Chain Reaction - Restriction Fragment Length Polymorphisms (PCR-RFLP), it was found out that a majority of the nodulating rhizobia was classified under the genus Bradyrhizobium and the non-nodulating rhizobia was classified under the genus of Sinorhizobium. The species diversity of nodulating rhizobia on the LUT2 showed the lowest at 0.00 whereas the LUT1 showed the highest diversity at 0.87. Similarly, the lowest species diversity of the non-nodulating rhizobia was found on the LUT2 at 0.00 but the highest was obtained from the LUT3 at 0.69. These results suggest that the nodulating rhizobia in this study may have preference with rice-based cropping pattern than with the two cropping systems. On the other hand, the diversity of rhizobia observed on LUT2 indicated that soybean monocropping may have resulted to a lesser diversity of both nodulating and non-nodulating rhizobia.Thus, it can be stated that the change in land utilization even in an agriculture land-use, species diversification and population dominance of certain rhizobia may occur.

Microbiota characterization of Exaiptasia diaphana from the Great Barrier Reef

BackgroundCoral reefs have sustained damage of increasing scale and frequency due to climate change, thereby intensifying the need to elucidate corals’ biological characteristics, including their thermal tolerance and microbial symbioses. The sea anemone, Exaiptasia diaphana, has proven an ideal coral model for many studies due to its close phylogenetic relationship and shared traits, such as symbiosis with algae of the family Symbiodiniaceae. However, established E. diaphana clonal lines are not available in Australia thus limiting the ability of Australian scientists to conduct research with this model. To help address this, the bacterial and Symbiodiniaceae associates of four Great Barrier Reef (GBR)-sourced E. diaphana genotypes established in laboratory aquaria and designated AIMS1–4, and from proxies of wild GBR E. diaphana were identified by metabarcoding of the bacterial 16S rRNA gene and eukaryotic rRNA gene ITS2 region. The relationship between AIMS1–4 and their bacterial associates was investigated, as was bacterial community phenotypic potential. Existing data from two existing anemone clonal lines, CC7 and H2, were included for comparison.ResultsOverall, 2238 bacterial amplicon sequence variants (ASVs) were observed in the AIMS1–4 bacterial communities, which were dominated by Proteobacteria and Bacteroidetes, together comprising > 90% relative abundance. Although many low abundance bacterial taxa varied between the anemone genotypes, the AIMS1–4 communities did not differ significantly. A significant tank effect was identified, indicating an environmental effect on the microbial communities. Bacterial community richness was lower in all lab-maintained E. diaphana compared to the wild proxies, suggesting a reduction in bacterial diversity and community phenotypic potential due to culturing. Seventeen ASVs were common to every GBR lab-cultured anemone, however five were associated with the Artemia feedstock, making their specific association to E. diaphana uncertain. The dominant Symbiodiniaceae symbiont in all GBR anemones was Breviolum minutum.ConclusionDespite differences in the presence and abundance of low abundance taxa, the bacterial communities of GBR-sourced lab-cultured E. diaphana are generally uniform and comparable to communities reported for other lab-cultured E. diaphana. The data presented here add to the global E. diaphana knowledge base and make an important contribution to the establishment of a GBR-sourced coral model organism.

Morphological and molecular characterization of a species of Tetratrichomonas present in feces of Brazilian sheep (Ovis aries) and goats (Capra hircus).

The trichomonads form part of the phylum Parabasalia, a complex assemblage of diverse species of flagellated protists, with some members recognized as pathogens of men and/or animals. Associations, probably as commensals, between the species Tetratrichomonas ovis and sheep were reported in North America during the 1960s based on morphological and cultural characteristics. Intriguingly, no subsequent studies of this topic have been published. Feces, collected from sheep (n = 55) and goats (n = 14), reared on small-scale, production facilities in Southeastern Brazil, were examined for parabasalids. Protozoa, demonstrating morphologies and motility characteristic of trichomonads, were detected by direct microscopy in 64% of sheep and 43% of goat samples. In contrast to T. ovis, none of the samples could be cultured in Diamond's medium; however, cultures were obtained for three goat and seventeen sheep samples in peptonized broth. Based on morphological analyses, all isolates were classified as members of the genus Tetratrichomonas. Sequencing of the ITS1-5.8S rRNA gene-ITS2 region revealed three highly similar genotypes that were essentially identical to sequences reported for Tetratrichomonas spp. isolated from the preputial cavity of cattle in the USA and Southern Brazil. The findings of this study extend and enhance our knowledge of parasitism in small ruminants by parabasalids.

Clustering of <i>Vibrio vulnificus</i> Isolates Derived from the Coast of Nagasaki Prefecture and Clinical Samples Based on Restriction Fragment Length Polymorphism of 16S–23S rRNA Gene ITS Region (rITS-RFLP)

Clustering of <i>Vibrio vulnificus</i> Isolates Derived from the Coast of Nagasaki Prefecture and Clinical Samples Based on Restriction Fragment Length Polymorphism of 16S–23S rRNA Gene ITS Region (rITS-RFLP)

Constraints on Phylogenetic Interrelationships among Four Free-living Litostomatean Lineages Inferred from 18S rRNA gene-ITS Region sequences and Secondary Structure of the ITS2 molecule

We investigated interrelationships between four free-living litostomatean lineages, using 18S rRNA gene and ITS region sequences as well as the secondary structure of the ITS2 molecules. Our phylogenetic analyses confirmed the deep split of free-living litostomateans into Rhynchostomatia and Haptoria represented here by Haptorida, Pleurostomatida, and Spathidiida. This bifurcation is also corroborated by the signature of the rhynchostomatian and haptorian ITS2 molecules. Specifically, the consensus stems of helices II and III are longer by one base pair in Rhynchostomatia, while the terminal loops of both helices are longer by one or two nucleotide/-s in Haptoria. A close relationship of Pleurostomatida and Haptorida is favored by quartet likelihood-mapping and supported by a 5’-AG vs. CU-3’ motif in the variable part of helix II and by two morphological apomorphies, i.e., meridionally extending somatic kineties and a non-three-rowed dorsal brush. Although monophyletic origin of Spathidiida is poorly supported in phylogenetic trees, the unique motif 5’-GA vs. UC-3’ present in the consensus helix II stem could be an important molecular synapomorphy of spathidiids, apart from the ancestrally anteriorly curved somatic kineties and the three-rowed dorsal brush. The peculiar family Pseudoholophryidae has very likely found its phylogenetic home among spathidiids, as an early branching lineage.

Comparison of Cultivable Acetic Acid Bacterial Microbiota in Organic and Conventional Apple Cider Vinegar.

Organic apple cider vinegar is produced from apples that go through very restricted treatment in orchard. During the first stage of the process, the sugars from apples are fermented by yeasts to cider. The produced ethanol is used as a substrate by acetic acid bacteria in a second separated bioprocess. In both, the organic and conventional apple cider vinegars the ethanol oxidation to acetic acid is initiated by native microbiota that survived alcohol fermentation. We compared the cultivable acetic acid bacterial microbiota in the production of organic and conventional apple cider vinegars from a smoothly running oxidation cycle of a submerged industrial process. In this way we isolated and characterized 96 bacteria from organic and 72 bacteria from conventional apple cider vinegar. Using the restriction analysis of the PCR-amplified 16S-23S rRNA gene ITS regions, we identified four different HaeIII and five different HpaII restriction profiles for bacterial isolates from organic apple cider vinegar. Each type of restriction profile was further analyzed by sequence analysis of the 16S-23S rRNA gene ITS regions, resulting in identification of the following species: Acetobacter pasteurianus (71.90%), Acetobacter ghanensis (12.50%), Komagataeibacter oboediens (9.35%) and Komagataeibacter saccharivorans (6.25%). Using the same analytical approach in conventional apple cider vinegar, we identified only two different HaeIII and two different HpaII restriction profiles of the 16S‒23S rRNA gene ITS regions, which belong to the species Acetobacter pasteurianus (66.70%) and Komagataeibacter oboediens (33.30%). Yeasts that are able to resist 30 g/L of acetic acid were isolated from the acetic acid production phase and further identified by sequence analysis of the ITS1-5.8S rDNA‒ITS2 region as Candida ethanolica, Pichia membranifaciens and Saccharomycodes ludwigii. This study has shown for the first time that the bacterial microbiota for the industrial production of organic apple cider vinegar is clearly more heterogeneous than the bacterial microbiota for the industrial production of conventional apple cider vinegar. Further chemical analysis should reveal if a difference in microbiota composition influences the quality of different types of apple cider vinegar.

Jaminaea phylloscopi sp. nov. (Microstromatales), a basidiomycetous yeast isolated from migratory birds in the Mediterranean basin.

During a survey of yeasts vectored by migratory birds in the Mediterranean basin, isolations from the cloacae of members of the order Passeriformes collected in Ustica (Italy) were performed. Based on phylogenetic analysis of the D1/D2 domain of the 26S rRNA gene and the internal transcribed spacer ITS1-5.8S rRNA gene-ITS2 region, five yeast isolates clustered in a new lineage within the Microstromatales clade. The DNA sequences of these isolates differed from those of their closest relatives, Jaminaea angkorensis and Jaminaea lanaiensis, by 20 and 25 nt substitutions in the D1/D2 domain and 119 and 131 nt substitutions in the complete ITS region, respectively. In addition, the five isolates showed phenotypic characteristics not observed in their closest relatives, such as the ability to grow at 44 °C and at pH 2.5, which suggests a possible adaptation to the bird gastrointestinal tract. On the basis of the isolation source, phenotypic features and molecular strain typing carried out with randomly amplified polymorphic DNA (RAPD)-PCR and mini-satellite-primed (MSP)-PCR analysis, the five isolates were characterized as five distinct strains of a novel species formally described as Jaminaea phylloscopi sp. nov., with 551B6T ( = PYCC 6783T = CBS 14087T) as the type strain. The Mycobank accession number is MB811984.

The Diversity of Pea Microsymbionts in Various Types of Soils and Their Effects on Plant Host Productivity.

The growth and yield of peas cultivated on eight different soils, as well as the diversity of pea microsymbionts derived from these soils were investigated in the present study. The experimental plot was composed of soils that were transferred from different parts of Poland more than a century ago. The soils were located in direct vicinity of each other in the experimental plot. All soils examined contained pea microsymbionts, which were suggested to belong to Rhizobium leguminosarum sv. viciae based on the nucleotide sequence of the partial 16S rRNA gene. PCR-RFLP analyses of the 16S-23S rRNA gene ITS region and nodD alleles revealed the presence of numerous and diversified groups of pea microsymbionts and some similarities between the tested populations, which may have been the result of the spread or displacement of strains. However, most populations retained their own genetic distinction, which may have been related to the type of soil. Most of the tested populations comprised low-effective strains for the promotion of pea growth. No relationships were found between the characteristics of soil and symbiotic effectiveness of rhizobial populations; however, better seed yield was obtained for soil with medium biological productivity inhabited by high-effective rhizobial populations than for soil with high agricultural quality containing medium-quality pea microsymbionts, and these results showed the importance of symbiosis for plant hosts.

Comparative molecular genetic study of the Asian population of Drosophila mercatorum (Diptera; Drosophilidae) at the COI gene fragment and the ITS1–ITS2 region of the rRNA genes

Drosophila mercatorum is a species of neotropical origin. It is represented by two subspecies, marcatorum and pararepleta. Synanthropic subspecies D. mercatorum mercatorum spread into Eurasia in the mid-20th century, and to date, it is widespread over the territory of Europe and the former Soviet Union. The experiments were performed using the specimens of D. m. mercatorum from natural Asian populations, laboratory stock 2328 of D. mercatorum (displaying some morphological differences from Asian wild-type flies), the specimens of D. busckii and D. virilis, and the COI gene sequence of D. mercatorum from NCBI database, accession number, DQ471607. The specimens were investigated for the variability of the standard DNA sequences used for species identification (the cytochrome oxidase subunit I (COI) gene fragment and the ITS1–ITS2 region of rRNA genes). Sequences of the COI gene fragment from Asian D. m. mercatorum were identical between the specimens, but differed from the sequences of stock 2328, as well as from the NCBI sequence. Asian populations of D. mercatorum and stock 2328 were identical in the ITS1–ITS2 sequences. Low sequence divergence between the COI gene fragments, along with the absence of differences at the ITS1–ITS2 region of the rRNA genes indicate that the difference between the Asian populations of D. m. mercatorum and morphologically different stock 2328 are within the frames of intraspecies divergence.

Microbial community structure and disposal issues of table olive wastewaters generated from the fermentation of the olive cultivar ‘Kalamon’

Table olive wastewaters (TOWW) are produced worldwide in large quantities annually and are characterized by high COD concentration, salt and phenolic content. Although microbial communities in fermentation processes have been studied extensively, a limited number of studies have been performed on the disposed brine. In this study, molecular techniques were applied to examining the dominant bacterial and fungal communities in Kalamon TOWW and the findings are discussed in relation to disposal issues of these wastewaters. Based on fluorescence in situ hybridization (FISH), the bacterial population in TOWW was estimated to be 55 ± 8 % of the total microbial population, while total fungal counts were 11.7 ± 2.6 × 104 CFU mL−1. The main bacterial taxon identified in Kalamon TOWW was associated, but not closely, to Microvirga subterranea, while the dominant fungal isolates were related to the genus Pichia. However, these fungal isolates warrant further investigation since they were distantly related to their closest taxonomically described relative Pichia manshurica (96.0 % similarity in ITS1-5.8S rRNA gene-ITS2 region). Based on 16S rRNA gene sequencing, at least five novel phylogenetic linkages among Bacteria were identified. No lactic acid bacteria were detected in the brine, indicating that lactic acid fermentation was completed at an early stage. Moreover, no fecal contamination threat was found since coliforms and other enteric bacteria were not identified. However, physicochemical characterization and phytoxicity tests revealed that TOWW should be considered as a wastewater of high toxicity. It appears that oligotrophy, endospore-formation, osmotolerance and polyphenol-tolerance are adaptive strategies adopted by indigenous microbiota to resist adverse environmental conditions in TOWW.

Impacts of biochar on bioavailability of the fungicide azoxystrobin: A comparison of the effect on biodegradation rate and toxicity to the fungal community

There is great interest in using biochar (BC) as a soil amendment to provide a long-term repository of carbon to mitigate climate change. BC can have major impacts on soil biogeochemical cycling processes, largely by the sorption and protection of organic matter from microbial turnover. Application of BC to agricultural soil could also affect the efficacy, fate and environmental impact of pesticides. In the current study we investigated the effect of BC on bioavailability of the fungicide azoxystrobin in soil. We found that application of BC had no effect on sorption or degradation of azoxystrobin, even at a rate of 2% w/w. While azoxystrobin reduced dehydrogenase activity, BC addition greatly increased dehydrogenase, although the inhibitory effect of azoxystrobin was still evident in BC amended soil. Using Terminal Restriction Fragment Length Polymorphism of fungal SSU rRNA gene ITS regions it was found that azoxystrobin altered the structure of the soil fungal community, although this effect was dampened by BC addition. BC application had minor effects on fungal community structure. We conclude that measurement of the effect of BC on pesticide bioavailability by analysis of biodegradation rate and non-target effects on fungal community structure gave contrasting conclusions.

Identification and quantification of acetic acid bacteria in wine and vinegar by TaqMan–MGB probes

A Real-Time PCR (RT-PCR) assay was developed using TaqMan minor groove binder (MGB) probes for the specific detection and quantification of five acetic acid bacteria (AAB) species ( Acetobacter pasteurianus, Acetobacter aceti, Gluconacetobacter hansenii, Gluconacetobacter europaeus and Gluconobacter oxydans) in wine and vinegar. The primers and probes, designed from the 16S rRNA gene, showed good specificity with the target AAB species. The technique was tested on AAB grown in glucose medium (GY) and inoculated samples of red wine and wine vinegar. Standard curves were constructed with the five target species in all these matrices. Quantification was linear over at least 5 log units using both serial dilution of purified DNA and cells. When this technique was tested in GY medium and inoculated matrices, at least 10 2–10 3 cells/ml were detected. To quantify low populations of AAB in microbiologically complex samples, a PCR enrichment including part of the 16S–23S rRNA gene ITS region was needed to increase the amount of target DNA compared to non-target DNA. The RT-PCR assay used in this study is a reliable, specific and fast method for quantifying these five AAB species in wine and vinegar.

Cryptococcus shivajii sp. nov.: A Novel Basidiomycetous Yeast Isolated from Biogas Reactor

Five yeast morphotypes were isolated from biogas reactors at North Wyke Research, Okehampton, UK. Out of the five morphotypes, four were identified as known species. In contrast, the fifth morphotype strain, Bio10(T), was found to differ from Bullera dendrophila and Kwoniella mangroviensis, its closest phylogenetic neighbours, by 2.6-2.9% with respect to the nucleotide sequence of the D1/D2 domain of the 26S rRNA gene and by 5.6-6.2% with respect to the internal transcribed spacer 1 (ITS1)-5.8S rRNA gene-ITS2 region. Bio10(T) also differs from these two species by a number of phenotypic characteristics. Thus, based on the phenotypic differences and phylogenetic analysis, strain Bio10(T) is assigned the status of a new species of Cryptococcus, for which the name Cryptococcus shivajii sp. nov. is proposed. The type strain is Bio10(T) (NCYC 3541(T) = CBS 11374(T)).

Actinomucor elegans var. kuwaitiensis isolated from the wound of a diabetic patient

The new variety of Actinomucor elegans var. kuwaitiensis, isolated from an open necrotic wound of a diabetic patient is described. This strain differed from the two previously described varieties of A. elegans, A. elegans var. elegans and A. elegans var. meitauzae by nearly 1% or more sequence divergence within D1/D2 regions of 28S rRNA and the ITS region of rRNA genes. Like the other two varieties, no zygospores were observed; however, there was evidence suggesting intersexual diploidy, a feature not described previously in this species. Additionally, A. elegans var. kuwaitiensis was pathogenic to white mice causing 100% mortality within 5 days.

Rhodotorula himalayensis sp. nov., a novel psychrophilic yeast isolated from Roopkund Lake of the Himalayan mountain ranges, India

Twenty-five psychrophilic yeasts were isolated from the soil of Roopkund Lake, Himalayas, India. Two colony morphotypes were identified and representatives of 'morphotype 1' were identified as Cryptococcus gastricus. Representatives of 'morphotype 2', namely 3AT, 4A, 4B and Rup4B, showed similar phenotypic properties and are identical with respect to the nucleotide sequence of the ITS1-5.8S rRNA gene-ITS2 region and D1/D2 domain of the 26S rRNA gene. The sequence of D1/D2 domain of 3AT shows 97.6-98.8% similarity with Rhodotorula psychrophila CBS10440T, Rhodotorula glacialis CBS10437T and Rhodotorula psychrophenolica CBS10438T and in the neighbour-joining phylogenetic tree strains; 3AT, 4A, 4B and Rup4B form a cluster with Rhodotorula glacialis and Rhodotorula psychrophila. Strains 3AT, 4A, 4B and Rup4B also differ from their nearest phylogenetic relatives in several biochemical characteristics such as in assimilation of D-galactose, L-sorbose, maltose, citrate, D-glucuronate and creatinine. Thus, based on the phylogenetic analysis and the phenotypic differences 3AT, 4A, 4B and Rup 4B are assigned the status of a new species of Rhodotorula for which the name Rhodotorula himalayensis sp. nov. is proposed with 3AT as the type strain (=CBS10539T =MTCC8336T).

Blastobotrys serpentis sp. nov., isolated from the intestine of a Trinket snake (Elaphe sp., Colubridae)

Asporogenus yeast strains W113AT and W113B were isolated from the intestine of a dead Trinket snake. The two isolates showed 100% sequence similarity in the D1/D2 domain of the large-subunit (LSU) rRNA gene, internal transcribed spacer (ITS) 1-5.8S rRNA gene-ITS2 region and mitochondrial small-subunit rRNA gene and the cytochrome oxidase II gene sequence and also showed similar phenotypic characteristics. The nearest phylogenetic neighbors of W113AT and W113B based on the sequence of the D1/D2 domain of the LSU rRNA gene were Blastobotrys chiropterorum NRRL Y-17017T and Blastobotrys terrestris NRRL Y-17704T with about 98% similarity. The close affiliation of W113AT and W113B with B. chiropterorum NRRL Y-17017T and B. terrestris NRRL Y-17704T was also evident from the high similarity observed in the nucleotide sequences of the mitochondrial small subunit rRNA (96-97.8%) and the cytochrome oxidase II (95.5-95.6%) genes. In the neighbor-joining phylogenetic trees constructed based on the D1/D2 domain or cytochrome oxidase gene, the isolates clustered with the above-mentioned species. However, the isolates showed a number of differences in their phenotypic properties with B. chiropterorum NRRL Y-17017T and B. terrestris NRRL Y-17704T and hence are regarded as representing a novel member of the genus Blastobotrys, for which the name Blastobotrys serpentis sp. nov. is proposed.

Sequence polymorphism in the ribosomal DNA internal transcribed spacers differs among Theileria species

The genomic region spanning the two ribosomal RNA internal transcribed spacers (ITS1 and ITS2) and the 5.8S rRNA gene was cloned and sequenced from sixteen Theileria isolates. Each Theileria species possessed ITS1 and ITS2 of unique size(s) and species specific nucleotide sequences. Varying degrees of ITS1 and ITS2 intra- and inter-species sequence polymorphism were found among ruminant Theileria species. The spacers were most polymorphic in the agent of tropical theileriosis, Theileria annulata, and were more conserved in two benign species, Theileria buffeli and Theileria sergenti Chitose. Phylogenetic analysis of the rDNA ITS1-5.8S rRNA gene-ITS2 region clearly separated each taxon, placing them in three clusters. One held T. annulata, Theileria parva, and Theileria mutans, with the latter two most closely related. The second held T. sergenti Ikeda, T. sergenti Chitose, and T. buffeli, with the latter two most closely related. The third cluster held the Theileria ovis isolates.

Candida hyderabadensissp. nov., a novel ascomycetous yeast isolated from wine grapes

Three ascomycetous yeast strains were isolated from decaying green wine grapes, collected from Hyderabad city in India. Two strains, YS9 and YS21, were identified as Kodamaea ohmeri and Candida fermentati, respectively. The third strain, YS12(T), differs from Candida parapsilosis, Candida orthopsilosis and Candida metapsilosis, the nearest phylogenetic neighbours, by 1.6-1.9% with respect to the nucleotide sequence of the D1/D2 domain of the 26S rRNA gene and by 1.4-9.2% with respect to the nucleotide sequence of the internal transcribed spacer 1 (ITS1)-5.8S rRNA gene-ITS2 region. YS12(T) also differs from C. parapsilosis, C. metapsilosis and C. orthopsilosis by some phenotypic characteristics. Thus, based on the phenotypic differences and phylogenetic analysis, strain YS12(T) is assigned the status of a new species of Candida, for which the name C. hyderabadensis sp. nov. is proposed. The type strain is YS12(T) (NRRL Y-27953(T)=CBS10444(T)=IAM15334(T)).