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Systematic Mycology and Lichenology Laboratory, Institute of Microbiology, Chinese Academy of Sciences, Beijing 100080, China
Correspondence
Feng-Yan Bai
baify{at}sun.im.ac.cn
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ABSTRACT |
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Published online ahead of print on 23 May 2003 as DOI 10.1099/ijs.0.02807-0.
The GenBank accession numbers for the 26S rDNA D1/D2 domain sequence of Y41T and the ITS sequences of Y41T and Y01 are respectively AY274823, AY274824 and AY274825.
A phylogenetic tree based on ITS sequences is available as supplementary material in IJSEM Online.
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, b, 2002a, b, 2003), encouraged us to investigate basidiomycetous yeast diversity in other areas of Yunnan. In January 2001, approximately 130 strains were isolated from 16 plant leaf samples collected in Xishuang Banna, the tropical rain forest area of Yunnan, using the improved ballistoconidia fall method (Nakase & Takashima, 1993). After morphological, physiological and biochemical characterization according to standard methods (Yarrow, 1998), five strains isolated from wilting leaves of Synsepalum dulcificum (Y01), Sterculia sp. (Y02), Arenga candata (Y11 and Y87) and Theobroma cacao (Y41T) were classified in a single group because of their similar phenotypic characters. They formed pink-coloured colonies and asymmetric ballistoconidia. The major ubiquinone of the representative strain, Y41T, was Q-10, determined using the method of Yamada & Kondo (1973). Sexual structures were not observed in mating tests between the strains studied. Conventional taxonomic study suggested that this group of strains belongs to the genus Sporobolomyces as defined by Boekhout & Nakase (1998). The carbon and nitrogen compound assimilation patterns of these strains were similar to those of taxa in the Sporobolomyces roseus complex.
Molecular phylogenetic analyses of the five strains were then performed using methods described previously (Bai et al., 2002b). The D1/D2 sequences of these strains were identical. Their internal transcribed spacer (ITS) sequences were either identical or differed in only one or two positions. The molecular data suggest that these strains are conspecific. In phylogenetic trees drawn from the D1/D2 and ITS sequences, the Banna strains clustered in the Johnsonii clade of the Sporidiobolus lineage (Scorzetti et al., 2002). In the D1/D2 tree, these strains formed a separate branch (Fig. 1), whereas, in the ITS tree, they clustered closely with Sporobolomyces blumeae (tree available as supplementary material in IJSEM Online).
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3 %) in the D1/D2 domain. They differed from other species in the same clade by more than 20 nt in this domain. In the ITS region, the strains differed from S. blumeae by 27–28 nt (8 %) and from other closely related species by 
43 nt. These results indicate that the five strains from Banna represent a novel Sporobolomyces species, for which the name Sporobolomyces bannaensis is proposed.
It is not easy to find clear phenotypic differences between S. bannaensis sp. nov. and S. roseus when compared with the standard description of the latter (Boekhout & Nakase, 1998). Though molecular phylogenetic analysis showed that S. bannaensis sp. nov. is most closely related to S. blumeae of the species in the Johnsonii clade, the novel species differed from the latter in assimilation of inulin, ethanol, glycerol, KNO3, NaNO2, L-lysine and cadaverine when compared with the original description of S. blumeae (Takashima & Nakase, 2000).
Latin diagnosis of Sporobolomyces bannaensis Bai et Zhao sp. nov.
In liquido malti post dies 3 ad 20 °C, cellulae vegetativae, ovoideae, ellipsoideae vel elongatae, 2·5–5·0x5·0–10·0 µm, singulae aut binae. Post unum mensem ad 20 °C, annulus, pelliculum et sedimentum formantur. In agaro malti post unum mensem ad 20 °C, cultura rosea, glabra, butyracea vel viscida, margine glabra. In agaro farinae zeae pseudomycelium non formantur. Ballistosporae ellipsoidales vel reniformes, 2·5–5·0x4·0–7·0 µm. Fermentatio nulla. Glucosum, galactosum (variabile), L-sorbosum, saccharosum, maltosum, cellobiosum, trehalosum, raffinosum, melezitosum, inulin, amylum solubile, ethanolum, glycerolum, D-mannitolum, D-glucitolum, methyl 
-D-glucosidum et acidum succinicum assimilantur at non lactosum, melibiosum, D-xylosum, L-arabinosum, D-arabinosum, D-ribosum, L-rhamnosum, D-glucosaminum, methanolum, erythritolum, ribitolum (vel exigue), galactitolum, salicinum (vel exigue), acidum DL-lacticum, acidum citricum, inositolum nec hexadecanum. Kalium nitricum et natrum nitrosum assimilantur at non L-lysinum, ethylaminum nec cadaverinum. Maxima temperatura crescentiae: 33–34 °C. Materia amyloidea iodophila non formantur. Urea finditur. Ad crescentiam vitaminum non necessarium est. Diazonium caeruleum B positivum. Ubiquinonum majus: Q-10. Typus: Y41T, isolatus ex folio Theobroma cacao L. AS 2.2285T (=CBS 9204T) depositus in collectione China General Microbiological Culture Collection Center, Academia Sinica, Beijing.
Description of Sporobolomyces bannaensis Bai & Zhao sp. nov.
Sporobolomyces bannaensis (ban.na.en'sis. N.L. adj. bannaensis of Banna, referring to the geographical origin of the strains).
In malt extract, after 3 days at 20 °C, cells are ovoid, ellipsoidal to elongate, 2·5–5·0x5·0–10·0 µm, single or in pairs (Fig. 2a). A ring, pellicle and sediment are formed. After 1 month at 20 °C, a ring, pellicle and sediment are present. On malt extract agar, after 1 month at 20 °C, the streak culture is butyrous to viscous, pink to orange-red, smooth and glistening with an entire margin. Pseudohyphae are not formed in Dalmau plate culture on corn meal agar. On corn meal agar, ballistoconidia are formed on short sterigmata and are asymmetric, ellipsoidal to reniform, 2·5–5·0x4·0–7·0 µm (Fig. 2b). Fermentation of glucose is negative. The following carbon compounds are assimilated: glucose, galactose (variable), L-sorbose, sucrose, maltose, cellobiose, trehalose, raffinose, melezitose, inulin, soluble starch, ethanol, glycerol, D-mannitol, glucitol, methyl -D-glucoside and succinic acid. The following are not assimilated: lactose, melibiose, D-xylose, L-arabinose, D-arabinose, D-ribose, L-rhamnose, D-glucosamine, methanol, erythritol, ribitol (or weak), galactitol, salicin (or weak), DL-lactic acid, citric acid, inositol and hexadecane. KNO3 and NaNO2 are utilized as sole sources of nitrogen; L-lysine, ethylamine and cadaverine are not utilized. Growth in vitamin-free medium is positive. Maximum growth temperature is 33–34 °C. Starch-like compounds are not produced. Urease activity is positive. Diazonium blue B reaction is positive. The major ubiquinone is Q-10.
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ACKNOWLEDGEMENTS |
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REFERENCES |
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TOPABSTRACTMAIN TEXTREFERENCES |
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Bai, F.-Y., Takashima, M., Hamamoto, M. & Nakase, T. (2001b). Sporobolomyces yunnanensis sp. nov., a Q-10(H2)-containing yeast species with a close phylogenetic relationship to Erythrobasidium hasegawianum. Int J Syst Evol Microbiol 51, 231–235.[Abstract]
Bai, F.-Y., Takashima, M., Jia, J.-H. & Nakase, T. (2002a). Dioszegia zsoltii sp. nov., a new ballistoconidium-forming yeast species with two varieties. J Gen Appl Microbiol 48, 17–23.
Bai, F.-Y., Zhao, J.-H., Takashima, M., Jia, J.-H., Boekhout, T. & Nakase, T. (2002b). Reclassification of the Sporobolomyces roseus and Sporidiobolus pararoseus complexes, with the description of Sporobolomyces phaffii sp. nov. Int J Syst Evol Microbiol 52, 2309–2314.[Abstract]
Bai, F.-Y., Takashima, M., Zhao, J.-H., Jia, J.-H. & Nakase, T. (2003). Bullera anomala sp. nov. and Bullera pseudovariabilis sp. nov., two new ballistoconidium-forming yeast species from Yunnan, China. Antonie van Leeuwenhoek 83, 257–263.[Medline]
Boekhout, T. & Nakase, T. (1998). Sporobolomyces Kluyver & van Niel. In The Yeasts, a Taxonomic Study, 4th edn, pp. 828–843. Edited by C. P. Kurtzman & J. W. Fell. Amsterdam: Elsevier.
Nakase, T. & Takashima, M. (1993). A simple procedure for the high frequency isolation of new taxa of ballistosporous yeasts living on the surfaces of plants. RIKEN Rev 3, 33–34.
Scorzetti, G., Fell, J. W., Fonseca, A. & Statzell-Tallman, A. (2002). Systematics of basidiomycetous yeasts: a comparison of large subunit D1/D2 and internal transcribed spacer rDNA regions. FEMS Yeast Res 2, 495–517.[Medline]
Takashima, M. & Nakase, T. (2000). Four new species of the genus Sporobolomyces isolated from leaves in Thailand. Mycoscience 41, 357–369.[CrossRef]
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.
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.
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