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1 Department of Microbiology, Faculty of Science, Kasetsart University, 50 Paholyothin Rd, Bangkok 10900, Thailand
2 Department of Botany, University of East Yangon, Thanyin Township, Myanmar
3 The International Center for Biotechnology, Osaka University, 2-1 Yamada-oka, Suita-City, Osaka 565-0871, Japan
Correspondence
Savitree Limtong
fscistl{at}ku.ac.th
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ABSTRACT |
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The GenBank/EMBL/DDBJ accession numbers for the D1/D2 domains of the LSU rDNA sequences of strains S-34T and S-35 are AB258462 and AB258463, respectively.
A neighbour-joining phylogenetic tree based on SSU rDNA sequences is available as a supplementary figure in IJSEM Online.
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MAIN TEXT |
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TOPABSTRACTMAIN TEXTREFERENCES |
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). In 2003, several species previously assigned to Saccharomyces and Kluyveromyces were transferred to Kazachstania on the basis of a multigene sequence analysis that included genes of the rDNA repeat (18S, 26S, internal transcribed spacer), single-copy nuclear genes (translation elongation factor 1
, actin-1, RNA polymerase II) and mitochondrially encoded genes [small-subunit (SSU) rDNA, cytochrome oxidase II] (Kurtzman, 2003; Kurtzman & Robnett, 2003). Recently, three non-pathogenic species (Lu et al., 2004; Wu & Bai, 2005) and four pathogenic species (Kurtzman et al., 2005) of the genus Kazachstania were described. During an investigation of yeasts in soil, two novel strains, S-34T and S-35, were isolated and found to represent a novel species of the genus Kazachstania. The novel species is closely related to Kazachstania aquatica, Kazachstania aerobia, Kazachstania solicola, Kazachstania servazzii and Kazachstania unispora; the first three of these were described only recently (Lu et al., 2004; Wu & Bai, 2005) and the last two were transferred from the genus Saccharomyces (Kurtzman, 2003; Kurtzman & Robnett, 2003). K. aquatica and K. solicola were identified as novel species by sequence analysis of the SSU rDNA, the internal transcribed spacer region (including 5.8S rDNA) and the large-subunit (LSU) D1/D2 domain (Wu & Bai, 2005), whereas K. aerobia was circumscribed on the basis of the internal transcribed spacer and D1/D2 sequences and electrophoretic karyotypes (Lu et al., 2004). K. servazzii and K. unispora were transferred from the genus Saccharomyces on the basis of a multigene sequence analysis (Kurtzman, 2003; Kurtzman & Robnett, 2003). Strains S-34T and S-35 were isolated from two soil samples collected from a mixed deciduous forest in Amphoe Wang Nam Khiao, Nakhon Ratchasima Province, Thailand, by using an enrichment technique. Each soil sample (5 g) was added to 50 ml acidified yeast extract/malt extract (YM) broth (0.3 % yeast extract, 0.3 % malt extract, 0.5 % peptone, 1 % glucose; adjusted to pH 3.7–3.8 with 1 M HCl) supplemented with 0.025 % sodium propionate and 200 mg chloramphenicol l–1 in a 250 ml Erlenmeyer flask and incubated at room temperature for 3–4 days on a rotary shaker. The enrichment culture was then spread out on YM agar supplemented with 0.025 % sodium propionate and 200 mg chloramphenicol l–1. When necessary, colonies were restreaked onto YM agar.
The sequences of the D1/D2 domain of the LSU rDNA and the SSU rDNA were determined from PCR products from genomic DNA extracted from yeast cells by using a slightly modified version of the method described by Lachance et al. (1999). The D1/D2 domain of the LSU rDNA was amplified by a PCR with the forward primer NL-1 and the reverse primer NL-4 (O'Donnell, 1993); amplification of the SSU rDNA was done with the forward primer P1 and the reverse primer P2 (Sjamsuridzal et al., 1997). The PCR product was checked by agarose gel electrophoresis, purified using the QIAquick purification kit (Qiagen) and cycle-sequenced using the ABI BigDye terminator cycle sequencing kit, version 3.1 (Applied Biosystems), with the external primers NL-1 and NL-4 for the D1/D2 domain (Kurtzman & Robnett, 1998) and eight primers, P1–P8, for the SSU rDNA (Yamada et al., 1999). The sequences were determined with an ABI PRISM 3100 automated DNA sequencer (Applied Biosystems) according to the instructions of the manufacturer. The sequences were compared pairwise by the BLASTN homology search program (Altschul et al., 1990) and were aligned with the sequences of related species retrieved from GenBank using the multiple alignment program CLUSTAL X, version 1.81 (Thompson et al., 1997). A phylogenetic tree was constructed from the evolutionary distance data according to the two-parameter method of Kimura (1980) and the neighbour-joining method (Saitou & Nei, 1987). The robustness of the phylogenetic trees was estimated using bootstrap analysis (1000 replicates) (Felsenstein, 1985).
The strains were characterized morphologically, biochemically and physiologically according to the standard methods described by Yarrow (1998). Assimilation of nitrogen compounds was examined on solid media with starved inocula, using the method of Nakase & Suzuki (1986). Growth at various temperatures was determined by cultivation on yeast extract/peptone/dextrose (YPD) agar (1 % yeast extract, 2 % peptone, 2 % glucose and 2 % agar). Ubiquinones were extracted from intact packed cells cultivated in YPD broth on a rotary shaker at 28 °C for 24–48 h and then purified according to the method described by Yamada & Kondo (1973). The isoprenologues were identified by HPLC using a Cosmosil (5C18; Waters) 4.6x250 mm column and methanol/2-propanol (2 : 1) at 1 ml min–1 as the elution system, with spectrophotometric detection (275 nm wavelength), according to the method of Kuraishi et al. (1985).
The sequences of the D1/D2 domains of the LSU rDNAs of strain S-34T and S-35 were identical. The two strains clustered with K. aquatica, K. unispora, K. aerobia, K. servazzii and K. solicola with high bootstrap values and were separate from Saccharomyces species (Fig. 1) and as well as from the pathogenic species of the Kazachstania telluris complex (Kazachstania bovina, Kazachstania heterogenica, Kazachstania pintolopesii and Kazachstania slooffiae) recently described by Kurtzman et al. (2005) (data not shown). Strain S-34T and S-35 showed 3 % divergence (14 nucleotide substitutions and three gaps out of 566 nt) from K. aquatica, the closest species in terms of pairwise sequence similarity, indicating that the two strains could represent a novel species. According to Kurtzman & Robnett (1998), yeast strains that show nucleotide substitution greater than 1 % in the D1/D2 domain of the LSU rDNA usually represent different species. However, to confirm the novelty of the two strains, their SSU rDNA sequences were determined: they were found to be identical. The closest relative of the two strains in terms of pairwise sequence similarity was K. aquatica, which differed by nine nucleotide substitutions and three gaps out of 1781 nt. In the phylogenetic analysis based on the SSU rDNA sequences, the two novel strains clustered with K. aquatica, K. unispora, K. aerobia, K. servazzii and K. solicola (see Supplementary Fig. S1 available in IJSEM Online), which is similar to what is seen in the tree based on the D1/D2 LSU rDNA sequences. These results lend further support to the conclusion that the two strains represent a novel species closely related to K. aquatica.
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), proliferated by multilateral budding (Fig. 2), lacked arthrospores and ballistospores, fermented glucose but did not assimilate nitrate, gave negative reactions for the diazonium blue B and urease tests and contained Q-6 as the major ubiquinone. These characteristics fit well with those of species of the genus Kazachstania. The two strains also shared the same conventional taxonomic characteristics, as shown in Table 1. We conclude, therefore, that the two strains represent a single novel species of the genus Kazachstania, for which the name Kazachstania siamensis is proposed.
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Latin diagnosis of Kazachstania siamensis Limtong, Yongmanitchai, Tun, Kawasaki et Seki sp. nov.
In agaro YM post dies 3 ad 28 °C cellulae ovoideae aut ellipsoideae, (2.4–4.8x2.9–8.1 µm), singulae, aut binae, per germinationem multipolarem reproducentes. Cultura cremea, butyrosa, initida, glabra, nitida et margine undulato. In agaro farinae Zea mays post dies 21 ad 28 °C et agaro YM post dies 21 ad 25 °C pseudomycelium nec mycelium non formantur. In agaro aceti et agaro Gorodkowa post dies 7 ad 28 °C asci formantur. Asci inconjugatio fiunt. Ascosporae globosae, 1 in quoque asco. In medio liquido YM post unum mensem ad 28 °C, sedimentum formantur et pellicula non formantur. Glucosum et galactosum (infirme) fermentatur at non maltosum, sucrosum, trehalosum, lactosum nec raffinosum. Glucosum, galactosum et trehalosum assimilantur at non L-sorbosum, D-ribosum, D-xylosum, L-arabinosum, D-arabinosum, L-rhamnosum, sucrosum, maltosum, cellobiosum, salicinum, melezitosum, inulinum, amylum solubile, glycerolum, erythritolum, ribitolum, glucitolum, D-mannitolum, galactitolum, inositolum, acidum 2-ketogluconicum, acidum D-gluconicum, acidum D-glucuronicum, acidum D-galacturonicum, acidum DL-lacticum, acidum succinicum, acidum citricum, methanolum nec ethanolum. Ammonium sulfatum assimilatur at non natrum nitrosum, kalium nitricum, ethylaminum, L-lysinum nec cadaverinum. Crescere potest in temperatura 40 °C at non in 42 °C. Crescit in 0.01 % cycloheximido, 0.1 % cycloheximido et 50 % glucosum (infirme). Non crescit in 60 % glucosum nec 10 % NaCl/5 % glucosum (10 : 5). Ureum non hydrolysatur. Diazonium caeruleum B non respondens. Ubiquinonum majum: Q-6.
Typus stirpis S-34T (=CBS 10361T=NBRC 101968T=BCC 21230T) isolatus e terea, Nakhon Ratchasima Provincia, Thailandia, conservatur in collectionibus culturarum quas Centraalbureau voor Schimmelcultures (Utrecht, The Netherlands), NITE Biological Resource Center, National Institute of Technology and Evaluation (Chiba, Japan) et BIOTEC Culture Collection, National Center for Genetic Engineering and Biotechnology, Thailand (Pathumthani, Thailand) deposita est.
Description of Kazachstania siamensis Limtong, Yongmanitchai, Tun, Kawasaki & Seki sp. nov.
Kazachstania siamensis (si.am.en'sis. N.L. fem. adj. siamensis referring to Siam, the old name of Thailand, where the two strains were isolated).
After 3 days growth on YM agar at 28 °C, cells are ovoid to ellipsoid (2.4–4.8x2.9–8.1 µm) and occur singly or in pairs (Fig. 2). Budding is multilateral. Streak culture on YM agar after 3 days at 28 °C is butyrous, cream-coloured, glossy, smooth and raised with undulate margins. In Dalmau plate culture on cornmeal agar at 28 °C and YM agar at 25 °C after 3 weeks, pseudohyphae and true hyphae are not formed. Neither arthrospores nor ballistospores are produced. Ascospores form on Forwell's acetate agar and Gorodkowa agar after 7 days at 28 °C; asci are unconjugated, persistent and each contain a single globose ascospore (Fig. 2). After 1 month in YM broth at 28 °C, sediment is present. A pellicle is not present during growth on the surface of assimilation medium. The major ubiquinone is Q-6. Phenotypic characteristics of the species are shown in Table 1.
The type strain, S-34T (CBS 10361T=NBRC 101968T=BCC 21230T), was isolated from soil collected in a mixed deciduous forest in Amphoe Wang Nam Khiao, Nakhon Ratchasima Province, Thailand.
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ACKNOWLEDGEMENTS |
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REFERENCES |
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TOPABSTRACTMAIN TEXTREFERENCES |
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Felsenstein, J. (1985). Confidence limits on phylogenies: an approach using the bootstrap. Evolution 39, 783–791.[CrossRef]
Kimura, M. (1980). A simple method for estimating evolutionary rates of base substitutions through comparative studies of nucleotide sequences. J Mol Evol 16, 111–120.[CrossRef][Medline]
Kuraishi, H., Katayama-Fujimura, Y., Sugiyama, J. & Yokoyama, T. (1985). Ubiquinone systems in fungi I. Distribution of ubiquinones in the major families of ascomycetes, basidiomycetes, and deuteromycetes, and their taxonomic implications. Trans Mycol Soc Jpn 26, 383–395.
Kurtzman, C. P. (2003). Phylogenetic circumscription of Saccharomyces, Kluyveromyces and other members of the Saccharomycetaceae, and the proposal of the new genera Lachancea, Nakaseomyces, Naumovia, Vanderwaltozyma and Zygotorulaspora. FEMS Yeast Res 4, 233–245.[CrossRef][Medline]
Kurtzman, C. P. & Robnett, C. J. (1998). Identification and phylogeny of ascomycetous yeasts from analysis of nuclear large-subunit (26S) ribosomal DNA partial sequences. Antonie van Leeuwenhoek 73, 331–371.[CrossRef][Medline]
Kurtzman, C. P. & Robnett, C. J. (2003). Phylogenetic relationships among yeasts of the ‘Saccharomyces complex’ determined from multigene sequence analysis. FEMS Yeast Res 3, 417–432.[CrossRef][Medline]
Kurtzman, C. P., Robnett, C. J., Ward, J. M., Brayton, C., Gorelick, P. & Walsh, T. J. (2005). Multigene phylogenetic analysis of pathogenic Candida species in the Kazachstania (Arxiozyma) telluris complex and description of their ascosporic states as Kazachstania bovina sp. nov., K. heterogenica sp. nov., K. pintolopsii sp. nov., and K. slooffiae sp. nov. J Clin Microbiol 43, 101–111.
Lachance, M. A., Bowles, J. M., Starmer, W. T. & Baker, S. F. (1999). Kodamaea kadaduensis and Candida tolerans, two new ascomycetous yeast species from Australian Hibiscus flowers. Can J Microbiol 45, 172–177.[CrossRef][Medline]
Lu, H.-Z., Cai, Y., Wu, Z.-W., Jia, J.-H. & Bai, F.-Y. (2004). Kazachstania aerobia sp. nov., an ascomycetous yeast species from aerobically deteriorating corn silage. Int J Syst Evol Microbiol 54, 2431–2435.
Nakase, T. & Suzuki, M. (1986). Bullera megalospora, a new species of yeast forming larger ballistospores isolated from dead leaves of Oryza sativa, Miscanthus sinensis and Sasa sp. in Japan. J Gen Appl Microbiol 32, 225–240.
O'Donnell, K. (1993). Fusarium and its near relatives. In The Fungal Holomorph: Mitotic, Meiotic and Pleomorphic Speciation in Fungal Systematics, pp. 225–233. Edited by D. R. Reynolds & J. W. Taylor. Wallingford, UK: CAB International.
Saitou, N. & Nei, M. (1987). The neighbor-joining method: a new method for reconstructing phylogenetic trees. Mol Biol Evol 4, 406–425.[Abstract]
Sjamsuridzal, W., Tajiri, Y., Nishida, H., Thuan, T. B., Kawasaki, H., Hirata, A., Yokota, A. & Sugiyama, J. (1997). Evolutionary relationships of members of the genera Taphrina, Protomyces, Schizosaccharomyces and related taxa within the Archiascomycetes: integrated analysis of genotypic characteristics. Mycoscience 38, 267–280.[CrossRef]
Thompson, J. D., Higgins, D. G., Gibson, T. J., Plewniak, F., Jeanmougin, F. & Higgins, D. J. (1997). The CLUSTAL_X windows interface: flexible strategies for multiple sequence alignment aided by quality analysis tools. Nucleic Acids Res 25, 4876–4882.
Wu, Z.-W. & Bai, F.-Y. (2005). Kazachstania aquatica sp. nov. and Kazachstania solicola sp. nov., novel ascomycetous yeast species. Int J Syst Evol Microbiol 55, 2219–2224.
Yamada, Y. & Kondo, K. (1973). Coenzyme Q system in the classification of the yeast genera Rhodotorula and Cryptococcus and the yeast-like genera Sporobolomyces and Rhodosporidium. J Gen Appl Microbiol 19, 59–77.
Yamada, Y., Kawasaki, H., Nagatsuka, Y., Mikata, K. & Seki, T. (1999). The phylogeny of the catophilic yeasts based on the 18S ribosomal RNA gene sequences: the proposals of Phaffomyces antillensis and Starmera caribaea, new combinations. Biosci Biotechnol Biochem 63, 827–832.[CrossRef][Medline]
Yarrow, D. (1998). Methods for the isolation, maintenance and identification of yeasts. In The Yeasts, a Taxonomic Study, 4th edn, pp. 77–100. Edited by C. P. Kurtzman & J. W. Fell. Amsterdam: Elsevier.
Zubkova, R. D. (1971). Genus novum Saccharomycetacearum e Kazachstania. Bot Mater Gerb Inst Bot Akad Nauk Kazakh SSR 7, 53–56 (in Russian).
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