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Microbial Genomics and Bioprocessing Research Unit, National Center for Agricultural Utilization Research, Agricultural Research Service, US Department of Agriculture, 1815 N. University St, Peoria, IL 61604, USA
Correspondence
Cletus P. Kurtzman
kurtzman{at}ncaur.usda.gov
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ABSTRACT |
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 TOP ABSTRACT MAIN TEXTREFERENCES |
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The GenBank/EMBL/DDBJ accession numbers for the LSU and mitochondrial SSU rDNA sequences, ITS sequences and COXII sequences of the proposed type strains and related strains are listed in Table 1.
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TOPABSTRACTMAIN TEXTREFERENCES |
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), but phylogenetic analysis of domains D1/D2 of the large-subunit (LSU) rDNA demonstrated that assigned species are actually anamorphic members of the Saccharomycetales (Kurtzman & Robnett, 1995, 1998). The D1/D2 LSU rDNA analysis also showed close relatedness between Blastobotrys, Arxula, Sympodiomyces and several Candida species. More recently, Kurtzman & Robnett (2007) re-examined relationships among these taxa with a more robust dataset consisting of nucleotide sequences from the nearly complete LSU rDNA, mitochondrial small-subunit (SSU) rDNA and the cytochrome oxidase II (COXII) gene. This multigene analysis demonstrated Blastobotrys, Arxula, Sympodiomyces and the above-noted Candida species to be members of the same clade, which was interpreted as a single genus. Of described genera, Blastobotrys (von Klopotek, 1967) has taxonomic priority for this clade over Arxula (van der Walt et al., 1990) and Sympodiomyces (Fell & Statzell, 1971), resulting in the transfer of species assigned to the latter two genera to Blastobotrys. The multigene sequence analysis also showed Trichomonascus to represent the ascosporic state of this clade. During the course of that study, six novel species of Blastobotrys were detected based on their unique nucleotide sequences in LSU rDNA, ITS rDNA, mitochondrial SSU rDNA and the COXII gene. In this report, descriptions are given for these six novel species.
Strains of the proposed novel species and reference taxa are given in Table 1 along with GenBank accession numbers for the genes sequenced. The strains are maintained in the Agricultural Research Service Culture Collection (NRRL), National Center for Agricultural Utilization Research, Peoria, IL, USA. Sources of the novel species are given in each species description. The composition of culture media used in this study, as well as the methods for fermentation and assimilation tests, was given previously (Yarrow, 1998).
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, 2003, 2007). Both strands of the DNAs analysed were sequenced with the ABI BigDye Terminator Cycle Sequencing kit (Applied Biosystems) using either an ABI 3100 or an ABI 3730 automated DNA sequencer according to the manufacturer's instructions. For phylogenetic analysis, sequences were aligned visually and regions of uncertain alignment were removed. Estimates of phylogenetic relatedness among species were determined using the maximum-parsimony and neighbour-joining programs of PAUP* 4.063a (Swofford, 1998). Bootstrap support for phylogenetic trees was determined from 1000 replications.
Maximum-parsimony analysis of concatenated nucleotide sequences from the nearly entire LSU rDNA, mitochondrial SSU rDNA and COXII gene demonstrated genetic separation of the six proposed novel species from previously described species in the Trichomonascus/Blastobotrys clade (Fig. 1). Results were the same when the dataset was analysed by neighbour-joining with Kimura's two-parameter correction. Two of the proposed novel species are closely related to each other and to Blastobotrys mokoenaii, and one of the proposed novel species is closely related to Blastobotrys adeninivorans. Genetic separation of these species was further examined from ITS1–5.8S–ITS2 sequence analysis as discussed in the species descriptions. Sequences for LSU rDNA, mitochondrial SSU rDNA and the COXII gene were not determined for the recently described Sympodiomyces attinorum (Carreiro et al., 2004) in the present study but, because D1/D2 LSU rDNA sequence analysis showed it to be a sister species of Blastobotrys (Sympodiomyces) parvus, Kurtzman & Robnett (2007) transferred it to Blastobotrys along with other species of Sympodiomyces.
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) and further suggests that these taxa are separate species, because the divergence is greater than that seen between strains of a species in closely related clades (Kurtzman & Robnett, 2007). Noteworthy in comparisons of these particular species pairs is that many of the sequence differences in ITS and mitochondrial SSU rDNA result from indels, whereas differences in D1/D2 LSU rDNA and the COXII gene are exclusively from nucleotide substitutions.
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reported that three of seven known species formed setae, i.e. Blastobotrys aristata, Blastobotrys capitulata and Blastobotrys proliferans. In the present study, two of the six novel species, B. illinoisensis and B. malaysiensis, were found to produce setae, as did the recently described B. mokoenaii (Mokwena et al., 2000). Seta-forming species are not distributed uniformly within Blastobotrys and occur in only two subclades, but not all members of the B. proliferans subclade produce setae. All presently known ascosporogenous species of the Trichomonascus/Blastobotrys clade are heterothallic (Kurtzman, 2004; Smith & de Hoog, 1998), which raises the possibility that anamorphic species in the clade may represent mating types.
Latin diagnosis of Blastobotrys americana Kurtzman sp. nov.
In agaro malti post dies 3 ad 25 °C, cellulae vegetativae globosae (2.3–4.1 µm) aut elongatae (1.3–3.5x2.0–12.0 µm), singulae, binae et in racemis brevibus. In agaro morphologico post dies 7 ad 25 °C, incrementum fuscum pallidum, hebes, filamentosum; margo glabra vel undulata. Pseudohyphae et hyphae verae fiunt. Ascosporae non fiunt. Glucosum et trehalosum (infirme) fermentantur. Sucrosum, raffinosum, galactosum, lactosum et maltosum non fermentantur. Assimilantur glucosum, raffinosum, melibiosum, galactosum, trehalosum, methyl-
-D-glucosidum, cellobiosum, salicinum, L-sorbosum, D-xylosum, L-arabinosum, D-arabinosum, D-ribosum, ethanolum, glycerolum, erythritolum, ribitolum, galactitolum, D-mannitolum, D-glucitolum, inositolum, DL-acidum lacticum (infirme), acidum succinicum, D-gluconatum, D-glucosaminum, N-acetyl-D-glucosaminum, hexadecanum, 5-keto-D-gluconatum et cadaverinum. Non assimilantur inulinum, sucrosum, lactosum, maltosum, melezitosum, amylum solubile, L-rhamnosum, methanolum, acidum citricum, potassii nitras, 2-keto-D-gluconatum et saccharatum. Amylum non formatur. Vitamina externa ad crescentiam necessaria sunt. Gelatinum non liquescit; esteres non fiunt; pellicula fiunt. Crescit in medio 100 µg cycloheximido ml–1 addito, et in medio 10 % sodii chloridium/5 % glucosum. Augmentum non fiunt in temperatura 37 °C. Species nova a speciebus aliis sequentibus nucleotiditis nucleus submonas largus rRNA gene, mitochondrial submonas parvus rRNA gene et COXII gene distinguenda. Typus: NRRL Y-6844T (CBS 10337T) designat stirpem typicam. Isolata e substrato incognito, Kansas, USA. Depositata in Collectione Culturarum ARS (NRRL), Peoria, IL, USA.
Description of Blastobotrys americana Kurtzman sp. nov.
Blastobotrys americana (a.me.ri.ca'na. N.L. nom. fem. adj. americana from America, i.e. the USA).
After 3 days growth on 5 % malt extract (ME) agar at 25 °C, budding is multilateral and cells are occasionally spherical, 2.3–4.1 µm, but are generally elongate, 1.3–3.5x2.0–12.0 µm, single, in pairs and in short chains (Fig. 2a). Colony growth is restricted, dull, tannish-white in colour and mycelial. Growth under the cover glass of a Dalmau plate culture with yeast morphology (YMo) agar is quite restricted and is composed of pseudohyphae bearing infrequent blastoconidia after 7 days at 25 °C (Fig. 2b). Aerobic growth on this medium is also restricted, dull, mycelial and tannish-white. The colony margin is entire to lobed. After 7–10 days, growth begins to show true hyphae, often with tip cells that are inflated and globose. However, these inflated hyphal cells do not appear to be asci. Occasionally, intercalary hyphal cells become enlarged and spherical. Cultures were examined weekly for ascospore formation on yeast extract, malt extract, peptone, glucose (YM), 5 % ME, restricted growth (RG) and McClary's acetate agar media incubated at 15 and 25 °C. Ascospores were not detected after 3 months. Reactions for fermentation, assimilation and other diagnostic tests are given in Table 3. Dry, climbing pellicles are formed on stationary liquid media.
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Latin diagnosis of Blastobotrys illinoisensis Kurtzman sp. nov.
In agaro malti post dies 3 ad 25 °C, cellulae vegetativae globosae (2.0–4.0 µm) aut elongatae (2.0–3.0x2.5–8.0 µm) et ovoideae cellulae cum saetis (1.0–1.3x18–125 µm), singulae et binae. In agaro morphologico post dies 7 ad 25 °C, incrementum fuscum pallidum, hebes, filamentosum; margo glabra vel undulata. Pseudohyphae et hyphae verae fiunt. Ascosporae non fiunt. Glucosum, sucrosum (infirme), galactosum, trehalosum et maltosum fermentantur. Raffinosum et lactosum non fermentantur. Assimilantur glucosum, sucrosum, galactosum, maltosum, methyl--D-glucosidum, amylum solubile, cellobiosum, salicinum, L-sorbosum, L-rhamnosum, D-xylosum, L-arabinosum, D-arabinosum, ethanolum, glycerolum, erythritolum, ribitolum, galactitolum, D-mannitolum, D-glucitolum, inositolum, acidum succinicum, acidum citricum, D-gluconatum, D-glucosaminum, N-acetyl-D-glucosaminum, hexadecanum, 2-keto-D-gluconatum, 5-keto-D-gluconatum et cadaverinum. Non assimilantur inulinum, raffinosum, melibiosum, lactosum, trehalosum, melezitosum, D-ribosum, methanolum, DL-acidum lacticum, potassii nitras et saccharatum. Amylum non formatur. Vitamina externa ad crescentiam necessaria non sunt. Gelatinum liquescit; esteres non fiunt; pellicula fiunt. Crescit in medio 100 µg cycloheximido ml–1 addito, et in medio 10 % sodii chloridium/5 % glucosum. Augmentum fiunt in temperatura 37 °C. Species nova a speciebus aliis sequentibus nucleotiditis nucleus submonas largus rRNA gene, ITS1–5.8S–ITS2 rRNA genes, mitochondrial submonas parvus rRNA gene et COXII gene distinguenda. Typus: NRRL YB-1343T (CBS 10339T) designat stirpem typicam. Isolata detrita a arbor mortua, Marion, IL, USA. Depositata in Collectione Culturarum ARS (NRRL), Peoria, IL, USA.
Description of Blastobotrys illinoisensis Kurtzman sp. nov.
Blastobotrys illinoisensis (il.lin.ois.en'sis. N.L. nom. fem. adj. illinoisensis from the State of Illinois, USA).
After 3 days growth on 5 % ME agar at 25 °C, cells are spherical, 2.0–4.0 µm, to elongate, 2.0–3.0x2.5–8.0 µm, and often teardrop-shaped. Cells form by multilateral budding and may be single or in pairs. Not infrequently, cells 4–5x6–8 µm have a tapered seta-like extension that has a basal diameter of 1.0–1.3 µm and a length of 18–125 µm (Fig. 2c
). Seta-bearing cells often produce buds. Colony growth on 5 % ME agar is white with an almost powdery appearance and a prominent mycelial fringe. After 7 days at 25 °C, growth under the cover glass of a Dalmau plate culture with YMo agar shows pseudohyphae and abundant true hyphae with side branches having highly denticulate tips bearing blastoconidia. Hyphal cells also form denticles with blastoconidia (Fig. 2d). Aerobic colonies are white, mycelial and somewhat powdery with an entire to lobate margin. Reactions for fermentation, assimilation and other diagnostic tests are given in Table 3. Moderately thick, climbing pellicles are formed on stationary liquid media. Cultures grown on YM, 5 % ME and RG agar media at 15 and 25 °C produced no ascospores after 6 weeks. The type strain differs from B. mokoenaii NRRL Y-27120T and B. malaysiensis sp. nov. NRRL Y-6417T by just two nucleotide substitutions in the D1/D2 domain of the LSU rDNA. This small difference raised the possibility that the taxa may be conspecific. However, greater divergence was found in the nucleotide sequences of ITS1–5.8S–ITS2 rDNA, mitochondrial SSU rDNA and the COXII gene, providing additional evidence that the three taxa are closely related but separate species (Table 2). In further tests, cultures of the three species were mixed on YM, 5 % ME and RG media, incubated at 25 °C and observed over a 6-week period. Neither conjugation between cells nor ascospores was detected. Cultures of B. mokoenaii, especially on 5 % ME agar, occasionally formed spherical endospores in hyphal cells.
NRRL YB-1343T (=CBS 10339T) is the designated type strain and is preserved as a lyophilized preparation in the ARS Culture Collection (NRRL), Peoria, IL, USA. The strain was isolated by L. J. Wickerham in 1949, from debris surrounding the base of a standing, dead deciduous tree near Marion, IL, USA.
Latin diagnosis of Blastobotrys malaysiensis Kurtzman sp. nov.
In agaro malti post dies 3 ad 25 °C, cellulae vegetativae globosae (3.0–5.5 µm) aut ellipsoideae (2.5–4.0x3.5–6.0 µm) et ovoideae cellulae cum saetis, singulae et binae. In agaro morphologico post dies 7 ad 25 °C, incrementum fuscum pallidum, hebes et filamentosum; margo glabra vel undulata. Pseudohyphae et hyphae verae fiunt. Ascosporae non fiunt. Glucosum, sucrosum (infirme), galactosum, trehalosum et maltosum fermentantur. Raffinosum et lactosum non fermentantur. Assimilantur glucosum, sucrosum, galactosum, lactosum, trehalosum, maltosum, melezitosum (infirme), methyl--D-glucosidum, amylum solubile, cellobiosum, salicinum, L-sorbosum, L-rhamnosum, D-xylosum, L-arabinosum, D-arabinosum, D-ribosum, ethanolum, glycerolum, erythritolum, ribitolum, D-mannitolum, D-glucitolum, inositolum, DL-acidum lacticum, acidum succinicum, acidum citricum, D-glucosaminum, N-acetyl-D-glucosaminum, 2-keto-D-gluconatum, 5-keto-D-gluconatum et cadaverinum. Non assimilantur inulinum, raffinosum, melibiosum, methanolum, galactitolum, D-gluconatum, hexadecanum, potassii nitras et saccharatum. Amylum non formatur. Vitamina externa ad crescentiam necessaria non sunt. Gelatinum liquescit; esteres non fiunt; pellicula fiunt. Crescit in medio 100 µg cycloheximido ml–1 addito, et in medio 10 % sodii chloridium/5 % glucosum. Augmentum fiunt in temperatura 37 °C. Species nova a speciebus aliis sequentibus nucleotiditis nucleus submonas largus rRNA gene, ITS1–5.8S–ITS2 rDNA genes, mitochondrial submonas parvus rRNA gene et COXII gene distinguenda. Typus: NRRL Y-6417T (CBS 10336T) designat stirpem typicam. Isolata soli, Malaysia. Depositata in Collectione Culturarum ARS (NRRL), Peoria, IL, USA.
Description of Blastobotrys malaysiensis Kurtzman sp. nov.
Blastobotrys malaysiensis (ma.lay.si.en'sis. N.L. nom. fem. adj. malaysiensis from the country of Malaysia).
After 3 days growth on 5 % ME agar at 25 °C, yeast cells are spherical, 3.0–5.5 µm, to ellipsoidal, 2.5–4.0x3.5–6.0 µm, form by multilateral budding and occur singly or in pairs (Fig. 2e). In addition, seta-bearing cells are also present, as seen for B. illinoisensis and B. mokoenaii (observation from this study), but lengths of setae seldom exceed 25 µm. Growth on 5 % ME agar is tannish-white, dull, somewhat butyrous and with a prominent mycelial fringe. Growth under the cover glass of a Dalmau plate culture with YMo agar shows mostly true hyphae with a few interspersed pseudohyphae after 7 days at 25 °C. Aerobic growth is white, mycelial and almost powdery, with radial furrows and with margins that are broadly lobate. Hyphae on various media often show a spherical, inflated terminal cell that may produce buds (Fig. 2f). Blastoconidia often form on short denticles on the hyphal cells (Fig. 2g). Cultures grown on YM, 5 % ME and RG agar media incubated at 15 and 25 °C produced no ascospores after 6 weeks and, as noted above, there was no mating reaction or ascosporulation in mixtures with the closely related species B. illinoisensis and B. mokoenaii. Reactions for fermentation, assimilation and other diagnostic tests are given in Table 3. Dry, climbing pellicles are formed on stationary liquid media.
NRRL Y-6417T (=CBS 10336T) is the designated type strain and is preserved as a lyophilized preparation in the ARS Culture Collection (NRRL), Peoria, IL, USA. The strain was received by the ARS Culture Collection in 1963 from Chester W. Emmons, who reported that the strain had been isolated from soil in a cave in Malaysia.
Latin diagnosis of Blastobotrys muscicola Kurtzman sp. nov.
In agaro malti post dies 3 ad 25 °C, cellulae vegetativae globosae (2.0–4.1 µm) aut elongatae et fusiformae (1.5–2.5x8.0–22.0 µm), singulae et binae. In agaro morphologico post dies 7 ad 25 °C, incrementum fuscum pallidum, hebes, filamentosum; margo undulata. Pseudohyphae et hyphae verae fiunt. Ascosporae non fiunt. Saccharas non fermentantur. Assimilantur glucosum, sucrosum, raffinosum (variabile), melibiosum (variabile), galactosum, lactosum, trehalosum (variabile), maltosum, melezitosum, methyl--D-glucosidum (variabile), amylum solubile, cellobiosum, salicinum (variabile), glycerolum, D-mannitolum, acidum succinicum, acidum citricum, D-glucosaminum (variabile), N-acetyl-D-glucosaminum et cadaverinum. Non assimilantur inulinum, L-sorbosum, L-rhamnosum, D-xylosum, L-arabinosum, D-arabinosum, D-ribosum, methanolum, ethanolum, erythritolum, ribitolum, galactitolum, D-glucitolum, inositolum, DL-acidum lacticum, D-gluconatum, hexadecanum, potassii nitras, 2-keto-D-gluconatum, 5-keto-D-gluconatum et saccharatum. Amylum non formatur. Vitamina externa ad crescentiam necessaria non sunt (variabile). Gelatinum liquescit; esteres non fiunt; pellicula fiunt. Crescit in medio 100 µg cycloheximido ml–1 addito. Non crescit in medio 10 % sodii chloridium/5 % glucosum. Augmentum fiunt in temperatura 37 °C. Species nova a speciebus aliis sequentibus nucleotiditis nucleus submonas largus rRNA gene, mitochondrial submonas parvus rRNA gene et COXII gene distinguenda. Typus: NRRL Y-7993T (CBS 10338T) designat stirpem typicam. Isolata musca, propinqua New Orleans, LA, USA. Depositata in Collectione Culturarum ARS (NRRL), Peoria, IL, USA.
Description of Blastobotrys muscicola Kurtzman sp. nov.
Blastobotrys muscicola (mus.ci'co.la. N.L. nom. fem. n. muscicola moss-dweller).
After 3 days growth on 5 % ME agar at 25 °C, budding is multilateral and cells are single or in pairs, infrequently spherical, 2.0–4.1 µm, but the majority are elongate and often spindle-shaped, 1.5–2.5x8.0–22.0 µm (Fig. 2h). Larger elongated cells often form terminal denticles that produce blastoconidia. Colony growth is dull, white and mycelial. Growth under the cover glass of a Dalmau plate culture with YMo agar shows both true hyphae and pseudohyphae after 7 days at 25 °C. Aerobic growth on this medium is dull, white and mycelial with a lobate margin. Cultures were examined weekly for ascospore formation on YM, 5 % ME, RG and McClary's acetate agar media incubated at 15 and 25 °C, but ascospores were not detected after 3 months. Hyphae on various media develop inflated spherical cells borne on short denticles (Fig. 2i), but these cells do not form ascospores and do not appear to be asci. Reactions for fermentation, assimilation and other diagnostic tests are given in Table 3. Dry, climbing pellicles are formed on stationary liquid media.
NRRL Y-7993T (=CBS 10338T) is the designated type strain and is preserved as a lyophilized preparation in the ARS Culture Collection (NRRL), Peoria, IL, USA. The strain was isolated by the author in 1976 from an unidentified moss growing on a fallen log in a wilderness area in St Tammany Parish, near New Orleans, LA, USA.
Latin diagnosis of Blastobotrys peoriensis Kurtzman sp. nov.
In agaro malti post dies 3 ad 25 °C, cellulae vegetativae globosae (2.0–5.0 µm) aut elongatae (2.0–4.0x2.5–12.0 µm), singulae et binae. In agaro morphologico post dies 7 ad 25 °C, incrementum fuscum pallidum, hebes, butyrosum; margo glabra vel serrata. Pseudohyphae et hyphae verae fiunt. Ascosporae non fiunt. Glucosum, galactosum et trehalosum fermentantur. Sucrosum, raffinosum, lactosum et maltosum non fermentantur. Assimilantur glucosum, sucrosum, raffinosum, melibiosum, galactosum, lactosum, maltosum, melezitosum, methyl--D-glucosidum, amylum solubile (infirme), cellobiosum, salicinum, L-sorbosum, D-xylosum, L-arabinosum, D-arabinosum, D-ribosum, ethanolum, glycerolum, erythritolum, ribitolum, galactitolum, D-mannitolum, D-glucitolum, inositolum, acidum succinicum, D-gluconatum, D-glucosaminum, N-acetyl-D-glucosaminum, hexadecanum, potassii nitras, 2-keto-D-gluconatum, 5-keto-D-gluconatum et cadaverinum. Non assimilantur inulinum, trehalosum, L-rhamnosum, methanolum, DL-acidum lacticum, acidum citricum et saccharatum. Amylum non formatur. Vitamina externa ad crescentiam necessaria sunt. Gelatinum non liquescit; esteres non fiunt; pellicula fiunt. Crescit in medio 100 µg cycloheximido ml–1 addito, et in medio 10 % sodii chloridium/5 % glucosum. Augmentum non fiunt in temperatura 37 °C. Species nova a speciebus aliis sequentibus nucleotiditis nucleus submonas largus rRNA gene, mitochondrial submonas parvus rRNA gene et COXII gene distinguenda. Typus: NRRL YB-2290T (CBS 10340T) designat stirpem typicam. Isolatus ex fungus incognitus, Peoria, IL, USA. Depositata in Collectione Culturarum ARS (NRRL), Peoria, IL, USA.
Description of Blastobotrys peoriensis Kurtzman sp. nov.
Blastobotrys peoriensis (pe.o.ri.en'sis. N.L. nom. fem. adj. peoriensis from Peoria, IL, USA, the collection site of the type strain).
After 3 days growth on 5 % ME agar at 25 °C, cells are spherical, 2.0–5.0 µm, ovoid and elongate, 2.0–4.0x2.5–12.0 µm, occur singly or in pairs and divide by multilateral budding (Fig. 2j, k). Colony growth is tannish-white and dull and ranges from butyrous to mycelial. Growth under the cover glass of a Dalmau plate culture with YMo agar shows abundant pseudohyphae with clusters of blastoconidia after 7 days at 25 °C. True hyphae are infrequent but become more common in older cultures. Both hyphae and individual cells form rachis-like extensions that produce blastoconidia (Fig. 2j, k). Aerobic colony growth on the Dalmau plate is white to tannish-white, dull with a convoluted surface and butyrous in texture. Ascosporulation was not observed in cultures grown on YM, 5 % ME, RG and McClary's acetate agar media incubated at 15 and 25 °C for 3 months. Reactions for fermentation, assimilation and other diagnostic tests are given in Table 3. Thick pellicles are formed on stationary liquid media.
NRRL YB-2290T (=CBS 10340T) is the designated type strain and is preserved as a lyophilized preparation in the ARS Culture Collection (NRRL), Peoria, IL, USA. The strain was isolated in 1950 by L. J. Wickerham from unidentified fungal growth on a rotted log, Peoria, IL, USA.
Latin diagnosis of Blastobotrys raffinosifermentans Kurtzman sp. nov.
In agaro malti post dies 3 ad 25 °C, cellulae vegetativae globosae (2.0–4.0 µm) aut elongatae (2.0–3.5x3.0–13.0 µm), singulae, binae et fasciculatae. In agaro morphologico post dies 7 ad 25 °C, incrementum fuscum pallidum, hebes et butyrosum; margo glabra vel undulata. Pseudohyphae fiunt; hyphae verae non fiunt. Ascosporae non fiunt. Glucosum, sucrosum (infirme), raffinosum, galactosum, trehalosum et maltosum fermentantur. Lactosum non fermentatur. Assimilantur glucosum, inulinum, sucrosum, raffinosum, melibiosum, galactosum, maltosum, melezitosum, methyl--D-glucosidum, amylum solubile, cellobiosum, salicinum, L-sorbosum, D-xylosum, L-arabinosum, D-arabinosum, D-ribosum, ethanolum, glycerolum, erythritolum, ribitolum, galactitolum, D-mannitolum, D-glucitolum, inositolum, DL-acidum lacticum, acidum succinicum, acidum citricum, D-gluconatum, D-glucosaminum, N-acetyl-D-glucosaminum, hexadecanum, potassii nitras, 2-keto-D-gluconatum, 5-keto-D-gluconatum et cadaverinum. Non assimilantur lactosum, trehalosum, L-rhamnosum, methanolum et saccharatum. Amylum non formatur. Vitamina externa ad crescentiam necessaria sunt. Gelatinum non liquescit; esteres non fiunt; pellicula fiunt. Crescit in medio 100 µg cycloheximido ml–1 addito, et in medio 10 % sodii chloridium/5 % glucosum. Augmentum fiunt in temperatura 37 °C. Species nova a speciebus aliis sequentibus nucleotiditis nucleus submonas largus rRNA gene, mitochondrial submonas parvus rRNA gene et COXII gene distinguenda. Typus: NRRL Y-27150T (CBS 6800T) designat stirpem typicam. Isolata e substrato incognito. Depositata in Collectione Culturarum ARS (NRRL), Peoria, IL, USA.
Description of Blastobotrys raffinosifermentans Kurtzman sp. nov.
Blastobotrys raffinosifermentans (raf'fi.no.si.fer.men'tans. N.L. part. adj. raffinosifermentans fermenting raffinose, a property rare among species of the Trichomonascus/Blastobotrys clade).
After 3 days growth on 5 % ME agar at 25 °C, cells are spherical, 2.0–4.0 µm, to elongate, 2.0–3.5x3.0–13.0 µm, and occur singly, in pairs and in small clusters (Fig. 2l). Budding is multilateral. Some cells form short denticles that produce blastoconidia. Colony growth is white, dull and butyrous. Growth under the cover glass of a Dalmau plate culture with YMo agar shows abundant well-branched pseudohyphae bearing blastoconidia after 7 days at 25 °C (Fig. 2m), but true hyphae are not detected. Aerobic colony growth is tannish-white, smooth, dull, with a cupulate centre and margins with infrequent small lobes. Ascosporulation was not observed in cultures grown on YM, 5 % ME and RG agar media at 15 and 25 °C for 2 months. Reactions for fermentation, assimilation and other diagnostic tests are given in Table 3. Moderately thick, climbing pellicles form on stationary liquid media.
NRRL Y-27150T (=CBS 6800T) is the designated type strain and is preserved as a lyophilized preparation in the ARS Culture Collection (NRRL), Peoria, IL, USA. The strain was received at the CBS Culture Collection from J. Shennan, but the source is unknown.
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ACKNOWLEDGEMENTS |
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REFERENCES |
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TOPABSTRACTMAIN TEXTREFERENCES |
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de Hoog, G. S. & Smith, M. Th. (1998). Blastobotrys von Klopotek. In The Yeasts, a Taxonomic Study, 4th edn, pp. 443–448. Edited by C. P. Kurtzman & J. W. Fell. Amsterdam: Elsevier.
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50 % are given at nodes. Trigonopsis variabilis NRRL Y-1579T was the designated outgroup for the analysis. ARS Culture Collection strain numbers follow the names of taxa; GenBank accession numbers for the gene sequences are given in Table 1