Antonie van Leeuwenhoek (2008) 93:391–399 DOI 10.1007/s10482-007-9216-9
ORIGINAL PAPER
Diversity of basidiomycetous phylloplane yeasts belonging to the genus Dioszegia (Tremellales) and description of Dioszegia athyri sp. nov., Dioszegia butyracea sp. nov. and Dioszegia xingshanensis sp. nov. Qi-Ming Wang Æ Jian-Hua Jia Æ Feng-Yan Bai
Received: 9 September 2007 / Accepted: 29 November 2007 / Published online: 9 December 2007 Ó Springer Science+Business Media B.V. 2007
Abstract From approximately 200 basidiomycetous yeast isolates forming orange or orange-red colonies isolated from senescent leaves collected in different regions of China, 29 representative strains varying in their geographic distribution and ballistoconidium forming ability were selected for further phenotypic and molecular taxonomic studies. Sequence analysis of the large subunit (26S) rDNA D1/D2 domain and the internal transcribed spacer (ITS) region including 5.8S rRNA from the strains resulted in the recognition of seven Dioszegia species, including four described species, namely D. aurantiaca, D. fristingensis, D. hungarica and D. zsoltii var. zsoltii and D. zsoltii var. yunnanensis, and three undescribed species. The three new species are described as Dioszegia athyri sp. nov. (type strain: CB 159T = AS 2.2559T = CBS 10119T), Dioszegia butyracea sp. nov. (type strain: CB 261T = AS 2.2600T = CBS 10122T) and Dioszegia xingshanensis sp. nov. (type strain: HB 1.4T = AS 2.2481T = CBS 10120T) in the present study.
Q.-M. Wang (&) J.-H. Jia F.-Y. Bai Systematic Mycology and Lichenology Laboratory, Institute of Microbiology, Chinese Academy of Sciences, Beijing 100101, China e-mail:
[email protected]
Keywords Basidiomycetous yeasts Dioszegia Species diversity Molecular phylogeny
Introduction The genus Dioszegia Zsolt in the Tremellales was revived and emended by Takashima et al. (2001) to accommodate both ballistoconidium and non ballistoconidium-forming basidiomycetous yeast species which usually form orange-coloured colonies. Three species were originally included in the emended genus. Since then, the number of species in the genus has increased to ten (Bai et al. 2002a; Ina´cio et al. 2005; Wang et al. 2003). Recent studies on yeast isolates from plant leaves collected in different regions in China indicated that the natural distribution of Dioszegia species in the phylloplane is common. Approximately 200 basidiomycetous yeasts forming orange or orange-red colonies were isolated from senescent leaves collected directly from plants in different regions of China. On the basis of morphological grouping, 29 representative strains isolated from different plants or different places were selected for rDNA sequence analysis and physiological characterization. Seven Dioszegia species including four described and three undescribed taxa were recognized from the strains studied. The three new species are described as Dioszegia athyri sp. nov., Dioszegia butyracea sp. nov. and Dioszegia xingshanensis sp. nov. in the present study.
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Morphological, physiological and biochemical characterization
Materials and methods Yeast strains The strains studied are listed in Table 1. They were isolated from senescent plant leaves using the improved ballistoconidia-fall method as described by Nakase and Takashima (1993). The leaves were collected from August to October during the years 1998–2004. All were still attached to the plants when collected.
Most of the morphological, physiological and biochemical characteristics were examined according to standard methods (Yarrow 1998). Extraction, purification and identification of ubiquinones were carried out according to Yamada and Kondo (1973). Assimilation of nitrogen compounds was investigated on solid media with starved inoculum (Nakase and Suzuki 1986).
Table 1 Yeast strains studied Species
Strain
Source
D. aurantiaca
CB 166 CB 170
Brachybotrys paridiformis, Changbai Mountain, Jilin, northeast China Brachybotrys paridiformis, Changbai Mountain, Jilin, northeast China
CB 323
Leptolepidium sp., Changbai Mountain, Jilin, northeast China
HX 4.4
Ranunculus sp., Xingshan, Hubei, central China
HX 6.2
Smilax stans, Xingshan, Hubei, central China
D. fristingensis D. hungarica
D. zsoltii var. zsoltii
HX 9.2
Viburnum betulifolium, Xingshan, Hubei, central China
HX 12.13
Lindera communis, Xingshan, Hubei, central China
HX 13.13
Decaisnea fargesii, Xingshan, Hubei, central China
HX 20.10
Porana henryi, Xingshan, Hubei, central China
HX 21.6
Artemisia sp., Xingshan, Hubei, central China
SH 67
Ligustrum sp., Taigu, Shanxi, north China
SH 47
Rhododendron oroedoxa, Taigu County, Shanxi, north China
XJ 4E3
Rubus sachalinensis, Xinjiang, northwest China
CB 145
Tilia amurensis, Changbai Mountain, Jilin, northeast China
CB 247
Dryopteris sp., Changbai Mountain, Jilin, northeast China
HX 21.3 SH 24
Artemisia sp., Xingshan County, Hubei, central China Salix yanbianica, Taigu County, Shanxi, north China
BH 4
Deutzia sp., Baihua Mountain, Beijing, north China
BH 19
Betula platyphylla, Baihua Mountain, Beijing, north China
CB 167
Brachybotrys paridiformis, Changbai Mountain, Jilin, northeast China
ST 5.13
Zanthoxylum sp., Taibai Mountain, Shaanxi, northwest China
SH 12
Vitex negundo, Taigu, Shanxi, north China
D. zsoltii var. yunnanensis
BH 3
Deutzia sp., Baihua Mountain, Beijing, north China
D. athyri sp. nov.
CB 159T
Athyrium sp., Changbai Mountain, Jilin, northeast China
D. butyracea sp. nov.
CB 261T
Betula ermanii, Changbai Mountain, Jilin, northeast China
CB 320
Leptolepidium sp., Changbai Mountain, Jilin, northeast China
SB 4.7
Euonymus verrucosoides, Taibai County, Shaanxi, northwest China
D. xingshanensis sp. nov.
123
XJ 7E2
Betula platyphylla, Xinjiang, northwest China
HB 1.4T
Artemisia sp., Xingshan County, Hubei, central China
Antonie van Leeuwenhoek (2008) 93:391–399
Sequencing and molecular phylogenetic analysis Nuclear DNA was extracted by using the method of Makimura et al. (1994). ITS (including 5.8S rDNA) and 26 rDNA D1/D2 domain sequences were determined as described previously (Bai et al. 2002b). Sequences were aligned with the Clustal X program (Thompson et al. 1997). Phylogenetic trees were constructed from the evolutionary distance data calculated from Kimura’s two parameter model (Kimura 1980) by using the neighbor-joining method (Saitou and Nei 1987). Bootstrap analyses (Felsenstein 1985) were performed from 1,000 random resamplings.
Results and discussion Seven species belonging to the genus Dioszegia were recognized among the 29 representative strains on the basis of D1/D2 domain and ITS region sequence analysis (Table 1). Eleven isolates from north, northeast and central China were assigned to D. aurantiaca. Their sequences matched that of the type (JCM 2956) of the species in the ITS region and differed by zero to two substitutions in the D1/D2 domain. All isolates formed ballistoconida. The present study shows that D. aurantiaca is widespread in nature and that its geographic distribution may not be restricted to colder regions as suggested by Ina´cio et al. (2005). The species was frequently isolated in Xingshan, Hubei Province, a subtropical area of central China (Table 1). Strains SH 47 and XJ 4E3 isolated in north and northwest China were assigned to D. fristingensis. This species was described recently based on a single isolate from Germany (Ina´cio et al. 2005). Strains SH 47 and XJ 4E3 differed from each other by three substitutions in each of the D1/D2 and ITS region. They differed from the type (PYCC 5861) by one or two substitutions in the D1/D2 and two or five mismatches in the ITS region. Whereas D. fristingensis was originally described as producing ballistoconidia abundantly (Ina´cio et al. 2005), the two Chinese isolates only rarely formed ballistoconidia. The species was not found among phylloplane yeasts identified in Portugal (Ina´cio et al. 2005). Four isolates from central, north and northeast China belonged to D. hungarica. They were identical to the type in the D1/D2 domain but differed by two substitutions in the ITS region. This species contains
393
both ballistoconidium and non ballistoconidium-forming strains (Takashima et al. 2001). Among the four Chinese representatives of D. hungarica, ballistoconidia were observed only in strain HX 21.3 on artificial media. Six strains isolated from north, northeast and northwest China were assigned to D. zsoltii based on their sequence identity to the type in the D1/D2 domain. This species was originally described based on six isolates from the subtropical part of Yunnan Province in southwest China (Bai et al. 2002a). Two varieties that differed by three substitutions in the ITS regions were recognized based on their intermediate DNA– DNA relatedness values (45–55%). Of the six strains of D. zsoltii identified in the present study, strain HB 3 was assigned to D. zsoltii var. yunnanensis based on a matching ITS sequence. The other five strains were assigned to D. zsoltii var. zsoltii. Three (BH 4, CB 167 and SH 12) had identical ITS sequences to the type, whereas strains HB 19 and ST 5.13 differed by one substitution in that region. This species was also identified among phylloplane yeasts in Portugal (Ina´cio et al. 2005). D. zsoltii therefore appears to be widely distributed across areas with different climate types. Analysis of D1/D2 and ITS sequences (Fig. 1) placed strain CB 159 in a clade that included also D. takashimae, D. catarinonii, D. zsoltii, an unnamed strain Dioszegia sp. TY-217, and ‘Cryptococcus hungaricus’ CBS 6576 with strong bootstrap support. In the D1/D2 domain, strain CB 159 differed from ‘C. hungaricus’ CBS 6576 by 28 mismatches and from the other four taxa by three to six substitutions. Differences of 13–20 mismatches were found with the latter in the ITS region. On this basis, we regard strain CB 159 as representative of a novel species, for which the name D. athyri sp. nov. is proposed. The four strains CB 261, CB 320, SB 4.7, and XJ 7E2 possessed identical ITS sequences and the first two, from northeast China, differed from the last two, from northwest China, by one substitution in the D1/D2 region. We interpret this as evidence that the four strains are conspecific and distinct from D. statzelliae and D. fristingensis (Fig. 1). The isolates differ from the types of the two described species by five and nine mismatches in the D1/D2 and by 13 and 26 mismatches in the ITS regions, respectively. We propose the name D. butyracea sp. nov. for this new species. Strain HB 1.4 from Hubei Province, central China, occupied a basal position (Fig. 1) with respect to
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Fig. 1 Phylogenetic tree drawn from neighborjoining analysis of the combined sequences of the ITS/5.8S and D1/D2 LSU rDNAs, depicting the relationships of the new Dioszegia species with closely related taxa. Bootstrap percentages over 50% from 1,000 bootstrap replicates are shown. Reference sequences were retrieved from GenBank under the accession numbers indicated
100
0.02 Knuc
100 67
95 100
D. crocea, D. aurantiaca and D. changbaiensis, with which it formed a well supported clade in the tree drawn from ITS sequences (bootstrap = 94%, not shown). The strain differed from D. crocea, D. aurantiaca, and D. changbaiensis by 11, 13 and 23 mismatches in the D1/D2 domain, respectively, and by 24, 23 and 18 mismatches in the ITS region. These data support our proposal of a new species, which we name D. xingshanensis sp. nov.
Latin diagnosis of Dioszegia athyri Q. M. Wang & F. Y. Bai sp. nov. In YM (Difco) liquido post dies 7 ad 17°C, cellulae vegetativae ellipsoideae, ovoideae, 2.5–6 9 5–10 lm,
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Dioszegia aurantiaca CB 166 (EU266497/EU266509) Dioszegia aurantiaca CB 170 (EU266498/EU266510) Dioszegia aurantiaca CB 323 (EU266499/EU266511) Dioszegia aurantiaca HX 4.4 (EU286789/EU266512) Dioszegia aurantiaca HX 6.2 (EU286790/EU266513) Dioszegia aurantiaca HX 9.2 (EU266500/EU266514) Dioszegia aurantiaca HX 20.10 (EU286791/EU266516) Dioszegia aurantiaca HX 21.6 (EU286792/EU266517) 94 Dioszegia aurantiaca SH 67 (EU286793/EU266518) Dioszegia aurantiaca HX 12.13 (EU286788/EU070934) 79 Dioszegia aurantiaca HX 13.13 (EU286787/EU266515) Dioszegia aurantiaca JCM 2956T (AB049613/AB104689) Dioszegia crocea CBS 6714T (AB022932/AF075508) Dioszegia xingshanensis sp. nov. HB 1.4T (EU070923/EU070928) Dioszegia changbaiensis AS 2.2309T (AY242818/AY242819) Dioszegia butyracea sp. nov. SB 4.7 (EU266507/EU070930) 55 99 Dioszegia butyracea sp. nov. XJ 7E2 (EU266508/EU266530) Dioszegia butyracea sp. nov. CB 261T (EU070924/EU070929) 90 Dioszegia butyracea sp. nov. CB 320 (EU266506/EU266529) Dioszegia statzelliae CBS 8926T (AY029344/AY029342) 85 Dioszegia fristingensis PYCC 5861T (AY562146/AY562158) Dioszegia fristingensis XJ 4E3 (EU070927/EU070933) 100 Dioszegia fristingensis SH 47 (EU070925/EU070932) Dioszegia hungarica CB 145 (EU286794/EU266519) Dioszegia hungarica CB 247 (EU266501/EU266520) 59 Dioszegia hungarica HX 21.3 (EU286795/EU266521) 100 Dioszegia hungarica SH 24 (EU286796/EU266522) 69 Dioszegia hungarica CBS 4214T (AB049614/AF075503) ‘Cryptococcus hungaricus’ CBS 5124 (AF272667/AF314231) ‘Cryptococcus hungaricus’ CBS 6953 (AF272668/AF314240) 100 Dioszegia buhagiarii PYCC 5866T (AY885687/AY562151) Dioszegia zsoltii var. yunnanensis BH 3 (EU266505/EU266528) Dioszegia zsoltii var. yunnanensis AS 2.2091T (AF385450/AF544246) Dioszegia zsoltii var. zsoltii BH 4 (EU266502/EU266523) 94 Dioszegia zsoltii var. zsoltii CB 167 (EU266503/EU266525) Dioszegia zsoltii var. zsoltii SH 12 (EU266504/EU266527) Dioszegia zsoltii var. zsoltii BH 19 (EU070921/EU266524) 59 Dioszegia zsoltii var. zsoltii ST 5.13 (EU070922/EU266526) 78 Dioszegia zsoltii var. zsoltii AS 2.2089T (AF385445/AF544245) Dioszegia catarinonii PYCC 5857T (AY562154/AY562142) ‘Cryptococcus hungaricus’ CBS 6576 (AF272667/AF314238) 53 Dioszegia takashimae PYCC 5864T (AY562160/AY562149) Dioszegia athyrii sp. nov. CB 159T (EU070926/EU070931) 100 Dioszegia sp. TY-217 (AY313016/AY313018) Cryptococcus luteolus CBS 943T (AF444323/AF075482) Bullera sinensis var. sinensis CBS 7238T (AF314989/AF189884) Bullera coprosmaensis CBS 8284T (AF314985/AF363660) Bullera mrakii JCM 8934T (AB022932/AB118871) Bullera huiaensis JCM 8933T (AB022931/AB118870) Bullera waltii AS 2.2414T (AB022935/AY487569) Bullera variabilis CBS 7347T (AF444403/AF179855)
singulae aut binae. Sedimentum formatur. Post unum mensem ad 17°C, annulus et sedimentum formantur. In agaro YM post unum mensem ad 17°C, cultura aurantiaca, glabra, nitida, butyracea, margine glabra. Pseudomycelium non formantur. Ballistosporae ellipsoideae, ampulliformes, 4–6 9 5–10 lm. Fermentatio nulla. Glucosum, galactosum, saccharosum, maltosum, cellobiosum, trehalosum, melibiosum, raffinosum, melezitosum, inulin (lente et exigue), amylum solubile (lente et exigue), D-xylosum, L-arabinosum, D-arabinosum (lente et exigue), D-ribosum (lente et exigue), L-rhamnosum (lente et exigue), galactitolum(exigue), D-glucitolum (lente et exigue), methyl a-D-glucosidum (lente et exigue) et salicinum (lente et exigue) assimilantur at non L-sorbosum, lactosum, D-glucosaminum, methanolum, ethanolum, glycerolum, erithritolum,
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ribitolum, D-mannitolum, acidum DL-lacticum, acidum succinicum, acidum citricum, inositolum nec hexdecanum. Ammonium sulfatum, natrum nitrosum et L-lysinum assimilantur at non kalium nitricum, ethylaminum nec cadaverinum. Vitamina externa ad crescentiam necessaria sunt. Maxima temperatura crescentiae: 29°C. Materia amyloidea iodophila non formatur. Urea finditur. Diazonium caeruleum B positivum. Ubiquinonum majus: Q-10. Typus: Isolatus ex folio Athyrium sp., CB 159T, depositus in collectione China General Microbiological Culture Collection Center, Academia Sinica (AS 2.2559T = CBS 10119T).
Description of Dioszegia athyri Q. M. Wang & F. Y. Bai sp. nov. In YM broth, after 7 days at 17°C, cells are ovoid, ellipsoidal, 2.5–6 9 5–10 lm and single or pairs, budding is polar, sediment is formed (Fig. 2). After 1 month at 17°C, a ring and sediment are present. On YM agar, after 1 month at 17°C, the streak culture is orange, butyrous, smooth, shining. The margin is entire. In Dalmau plate culture on corn meal agar, pseudohyphyae are not formed. Ballistoconidia are napiform or ellipsoidal, 4–6 9 5–10 lm. Fermentation is negative. Glucose, galactose, sucrose, maltose,
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cellobiose, trehalose, melibiose, raffinose, melezitose, inulin (delayed and weak), soluble starch (delayed and weak), D-xylose, L-arabinose, D-arabinose (delayed and weak), D-ribose (delayed and weak), L-rhamnose (delayed and weak), galactitol (weak), D-glucitol (delayed and weak), methyl a-D-glucoside (delayed and weak) and salicin (delayed and weak) are assimilated. L-sorbose, lactose, D-glucosamine, methanol, ethanol, glycerol, erythritol, ribitol, D-mannitol, DL-lactic acid, succinic acid, critic acid, inositol and hexdecane are not assimilated. Ammonium sulfate, sodium nitrite and L-lysine are assimilated. Potassium nitrate, ethylamine hydrochloride and cadaverine dihydrochloride are not assimilated. Maximum growth temperature is 29°C. Growth in vitamin-free medium is negative. Starch-like substances are not produced. Growth on 50% (w/w) glucose-yeast extract agar is negative. Urease activity is positive. Diazonium Blue B reaction is positive. The major ubiquinone is Q-10. The type strain of Dioszegia, CB 159T, was isolated from a senescent leaf of Athyrium sp. collected in Changbai Mountain, Jilin province, China in October 1998. This strain has been deposited in the China General Microbiological Culture Collection Center (CGMCC), Academia Sinica, Beijing, China, as AS 2.2559T, and in the Centraalbureau voor Schimmelcultures, Utrecht, The Netherlands, as CBS 10119T.
Fig. 2 Dioszegia athyri sp. nov. CB 159T vegetative cells grown in YM broth for 7 days at 17°C (left) and ballistoconidia produced on corn meal agar after 14 days at 17°C (right). Bars, 10 lm
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396
Etymology: the specific epithet athyri refers to plant genus Athyrium from which the type strain of the species was isolated. Physiologically, D. athyri sp. nov. differs from its phylogenetically close relatives D. catarinonii, D. takashimae, and D. zsoltii by its inability to assimilate succinic acid, DL-lactate and inositol.
Latin diagnosis of Dioszegia butyracea Q. M. Wang & F. Y. Bai sp. nov. In YM (Difco) liquido post dies 7 ad 17°C, cellulae vegetativae ellipsoideae, ovoideae, 3–6 9 5–12 lm, singulae or binae. Annulus et sedimentum formantur. Post unum mensem ad 17°C, annulus, pellicula et sedimentum formantur. In agaro YM post unum mensem ad 17°C, cultura aurantiaca, glabra, nitida, butyracea, margine glabra. Mycelium formantur vel non. Ballistosporae ellipsoideae ad ovoideae, 4–5 9 5–10 lm. Fermentatio nulla. Glucosum, galactosum, saccharosum (exigue), maltosum, cellobiosum, trehalosum, melibiosum, raffinosum, melezitosum, inulin, D-xylosum, L-arabinosum, D-arabinosum, L-rhamnosum (exigue), D-glucosaminum (lente et exigue), galactitolum, D-mannitolum et methyl a-D-glucosidum (exigue) assimilantur at non L-sorbosum, lactosum, amylum solubile, D-ribosum,
Fig. 3 Dioszegia butyracea sp. nov. CB 261T vegetative cells grown in YM broth for 7 days at 17°C (left) and ballistoconidia produced on corn meal agar after 7 days at 17°C (right). Bars, 10 lm
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methanolum, ethanolum, glycerolum, erithritolum, ribitolum, D-glucitolum, salicinum, acidum DL-lacticum, acidum succinicum, acidum citricum, inositolum nec hexdecanum. Ammonium sulfatum, natrum nitrosum et L-lysinum assimilantur at non kalium nitricum, ethylaminum nec cadaverinum. Vitamina externa ad crescentiam necessaria sunt. Maxima temperatura crescentiae: 25°C. Materia amyloidea iodophila formatur. Urea finditur. Diazonium caeruleum B positivum. Ubiquinonum majus: Q-10. Typus: Isolatus ex folio Betula ermanii Cham., CB 261T, depositus in collectione China General Microbiological Culture Collection Center, Academia Sinica (AS 2.2600T = CBS 10122T).
Description of Dioszegia butyracea Q. M. Wang & F. Y. Bai sp. nov. In YM broth, after 7 days at 17°C, cells are ovoid, ellipsoidal, 3–6 9 5–12 lm and single or pairs, budding is polar, a ring and sediment is formed (Fig. 3). After 1 month at 17°C, a ring, pellicle and sediment are present. On YM agar, after 1 month at 17°C, the streak culture is orange, butyrous, smooth, shining. The margin is entire. In Dalmau plate culture on corn meal agar, true hyphae are formed or not. Ballistoconidia are ellipsoidal, ovoid. 4–5 9 5–10 lm. Fermentation is
Antonie van Leeuwenhoek (2008) 93:391–399
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negative. Glucose, galactose, sucrose (weak), maltose, cellobiose, trehalose, melibiose, raffinose, melezitose, inulin, D-xylose, L-arabinose, D-arabinose, L-rhamnose (weak), D-glucosamine (delayed and weak), galactitol, D-mannitol and methyl a-D-glucoside (weak) are assimilated. L-sorbose, lactose, soluble starch, D-ribose, methanol, ethanol, glycerol, erythritol, ribitol, D-glucitol, salicin, DL-lactic acid, succinic acid, critic acid, inositol and hexdecane are not assimilated. Ammonium sulfate, sodium nitrite and L-lysine are assimilated. Potassium nitrate, ethylamine hydrochloride and cadaverine dihydrochloride are not assimilated. Maximum growth temperature is 25°C. Growth in vitamin-free medium is negative. Starch-like substances are produced. Growth on 50% (w/w) glucose-yeast extract agar is negative. Urease activity is positive. Diazonium Blue B reaction is positive. The major ubiquinone is Q-10. The type strain, CB 261T, was isolated from a senescent leaf of Betula ermanii Cham. collected in Changbai Mountain, Jilin province, China, in October 1998. This strain has been deposited in the China General Microbiological Culture Collection Center (CGMCC), Academia Sinica, Beijing, China, as AS 2.2600T, and in the Centraalbureau voor Schimmelcultures, Utrecht, The Netherlands, as CBS 10122T.
Etymology: the specific epithet butyracea refers to the butyrous colony texture of the strains of this species. Phenotypically, Dioszegia butyracea sp. nov. differs from its closest relative D. statzalliae by its ability to assimilate inulin and its inability to assimilate D-glucitol and DL-lactate. The inulin utilizing ability can also differentiate the new species from all the other described Dioszegia species (Table 2).
Latin diagnosis of Dioszegia xingshanensis Q. M. Wang & F. Y. Bai sp. nov. In YM (Difco) liquido post dies 7 ad 17°C, cellulae vegetativae ellipsoideae, ovoideae, 2–5 9 4–10 lm, singulae or binae. Sedimentum formatur. Post unum mensem ad 17°C, annulus et sedimentum formantur. In agaro YM post unum mensem ad 17°C, cultura aurantiaca, glabra, nitida, butyracea, margine glabra. Pseudomycelium non formantur. Ballistoconidia napiforma aut ellipsoidea, 4–6 9 5–10 lm. Fermentatio nulla. Glucosum, galactosum (exigue), saccharosum, maltosum, cellobiosum, trehalosum, melibiosum, raffinosum, melezitosum, D-xylosum, L-arabinosum,
Table 2 Diagnostic physiological characters of taxa in the genus Dioszegia Taxon
Assimilation of 1
2
3
St 4
5
6
7
8
9
10
Bc
25°C
11
D. athyri sp. nov.
+
lw
lw
lw
-
-
lw
lw
-
-
-
-
+
+
D. butyracea sp. nov.
+
+
-
-
-
+
-
-
-
-
-
+
+
+
D. xingshanensis sp. nov.
+
-
-
-
-
+
-
w
-
-
w
+
+
+
D. aurantiacaa
+
-
l
w
-
+
+
l
w
+
-
+
+
-
D. buhagiariib
-
-
-
+
l
+
l
+
-
+
-
+
-
+
D. catarinoniib
+
-
-
+,l
-
-,l
-
+
+,l
+
-,l
+
+,-
+
D. changbaiensisc D. croceaa
+ +
-
+ w
+
+
l +
l +
+ +
+
+ +
-
+ +
+
+ +
D. fristingensisb
+
-
-
l
l
+
+
l
l
+
-
+
+
-
D. hungaricaa
-
-
-
-
+
+
+
+
-
+
+
+
+,-
+
w
-
-
w
w
+
+
w
+
w
-
+
-
-
D. statzelliaed D. takashimae
b
+
-
-
+,l
-
+,l
-
+
+,l
+
l
+
+
+
D. zsoltii var. zsoltiie
+
-
+
-
-
+(w)
-
+,-
-
+,w
-
w
+
+
D. zsoltii var. yunnanensise
+
-
+
-
-
-
-
+,-
-(w)
+,w
-
w
+
+
a
b
c
d
e
Data from Takashima et al. (2001); Ina´cio et al. (2005); Wang et al. (2003); Thomas-Hall et al. (2002); and Bai et al. (2002a) Abbreviations: 1, melibiose; 2, inulin; 3, soluble starch; 4, Ribose; 5, ribitol; 6, D-mannitol; 7, D-glucitol; 8, salicin; 9, lactate; 10, succinate; 11, inositol; St, starch-like substance formation; Bc, ballistoconidia formation; 25°C, growth at 25°C; +, positive; l, delayed positive; w, weakly positive; lw, delayed and weakly positive
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398 D-arabinosum, L-rhamnosum,
galactitolum (lente et exigue), D-mannitolum (exigue), methyl a-D-glucosidum (exigue), salicinum (exigue) et inositolum (exigue) assimilantur at non L-sorbosum, lactosum, inulin, amylum solubile, D-ribosum, D-glucosaminum, methanolum, ethanolum, glycerolum, erithritolum, ribitolum, D-glucitolum, acidum DL-lacticum, acidum succinicum, acidum citricum nec hexdecanum. Ammonium sulfatum, natrum nitrosum et L-lysinum assimilantur at non kalium nitricum, ethylaminum nec cadaverinum. Ad crescentiam vitamina non necessaria sunt. Maxima temperatura crescentiae: 26°C. Materia amyloidea iodophila formatur. Urea finditur. Diazonium caeruleum B positivum. Ubiquinonum majus: Q-10. Typus: Isolatus ex folio Artemisia sp., HB 1.4T, depositus in collectione China General Microbiological Culture Collection Center, Academia Sinica (AS 2.2481T = CBS 10120T).
Description of Dioszegia xingshanensis Q. M. Wang & F. Y. Bai sp. nov. In YM broth, after 7 days at 17°C, cells are ovoid, ellipsoidal, 2–5 9 4–10 lm and single or pairs, budding is polar or bipolar, sediment is formed (Fig. 4). After 1 month at 17°C, a ring and sediment are present. On YM agar, after 1 month at 17°C, the streak culture
Fig. 4 Dioszegia xingshanensis sp. nov. HB 1.4T vegetative cells grown in YM broth for 7 days at 17°C (left) and ballistoconidia produced on corn meal agar after 14 days at 17°C (right). Bars, 10 lm
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Antonie van Leeuwenhoek (2008) 93:391–399
is orange, butyrous, smooth, shining. The margin is entire. In Dalmau plate culture on corn meal agar, true or pseudohyphae are not formed. Ballistoconidia are ellipsoidal, napiform, 4–6 9 5–10 lm. Fermentation is negative. Glucose, galactose (weak), sucrose, maltose, cellobiose, trehalose, melibiose, raffinose, melezitose, D-xylose, L-arabinose, D-arabinose, L-rhamnose, galactitol (delayed and weak), D-mannitol (weak), methyl a-D-glucoside (weak), salicin (weak) and inositol (weak) are assimilated. L-sorbose, lactose, inulin, soluble starch, D-ribose, D-glucosamine, methanol, ethanol, glycerol, erythritol, ribitol, D-glucitol, DL-lactic acid, succinic acid, critic acid and hexdecane are not assimilated. Ammonium sulfate, sodium nitrite and L-lysine are assimilated. Potassium nitrate, ethylamine hydrochloride and cadaverine dihydrochloride are not assimilated. Maximum growth temperature is 26°C. Growth in vitamin-free medium is positive (delayed and weak). Starch-like substances are produced. Growth on 50% (w/w) glucose-yeast extract agar is negative. Urease activity is positive. Diazonium Blue B reaction is positive. The major ubiquinone is Q-10. The type strain, HB 1.4T, was isolated from a senescent leaf of Artemisia sp. collected in Hubei province, China in October 2002. This strain has been deposited in the China General Microbiological Culture Collection Center (CGMCC), Academia Sinica, Beijing, China, as AS 2.2481T and in the
Antonie van Leeuwenhoek (2008) 93:391–399
Centraalbureau voor Schimmelcultures, Utrecht, The Netherlands, as CBS 10120T. Etymology: the specific epithet xingshanensis refers to the geographic origin of the type strain of this species. D. xingshanensis sp. nov. is physiologically similar to D. butyracea sp. nov., however, the former can be distinguished from the latter by its inability to assimilate inulin (Table 2). Acknowledgements This study was supported by grant no. 30470005 from the National Natural Science Foundation of China (NSFC) and the Youth Association of the Institute of Microbiology (IMYA), Chinese Academy of Sciences.
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