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Departamento de Microbiología y Genética, Edificio Departamental, Campus Miguel de Unamuno, Universidad de Salamanca, Spain
Correspondence
Martha E. Trujillo
mett{at}usal.es
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ABSTRACT |
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 TOP ABSTRACT MAIN TEXTREFERENCES |
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The GenBank/EMBL/DDBJ accession number for the 16S rDNA sequence of strain XIL11T is AY303668.
A fuller phylogenetic tree is available as supplementary material in IJSEM Online.
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MAIN TEXT |
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TOPABSTRACTMAIN TEXTREFERENCES |
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; Funke et al., 1995; Collins & Pascual, 2000; Elberson et al., 2000; Stackebrandt et al., 2002). Further study of this strain, based on a polyphasic approach that included chemotaxonomic, physiological and DNA–DNA hybridization analyses, confirmed its position as a representative of a novel species within the genus Cellulomonas.
Strain XIL11T was isolated on XED medium at 28 °C from decayed wood of an elm tree, as described previously (Rivas et al., 2003). Extraction and amplification of genomic DNA for 16S rDNA sequence analysis were carried out as described previously (Rivas et al., 2001, 2003). An almost-complete 16S rDNA sequence was obtained and aligned against 16S rDNA sequences that were available from public databases. Pairwise evolutionary distances were computed by using the correction algorithm of Jukes & Cantor (1969). The least-squares distance method of DeSoete (1983) was used in the construction of the phylogenetic dendrogram from distance matrices. The MEGA2 package (Kumar et al., 2001) was used for all analyses.
Isolate XIL11T was observed through a phase-contrast microscope after 48 h growth in YED medium (0·4 % yeast extract, 0·7 % dextrose, 1·8 % agar) to check for cell shape and motility. Cells were also stained according to the classical Gram procedure that was described by Doetsch (1981).
Physiological and biochemical tests were determined by using API 20NE, API 20E and API 50 CH strips (bioMérieux) according to the manufacturer's instructions. Enzyme activities for amylase, casein, catalase, cellulase and oxidase were determined as described previously (Rivas et al., 2003). A temperature range for growth on YED medium of 4–45 °C was determined.
Amino acid and sugar analyses of whole-cell hydrolysates were performed according to procedures described by Staneck & Roberts (1974). Peptidoglycan type was determined as described by Schleifer & Kandler (1972) and Schleifer (1985). Menaquinone and cellular fatty acid compositions were determined as described by Zimmermann et al. (1998). Polar lipids were extracted and examined by two-dimensional TLC (Minnikin et al., 1984).
The G+C content of genomic DNA was determined by using the thermal denaturation method (Mandel & Marmur, 1968).
DNA–DNA hybridization was performed [in 2x SSC+10 % DMSO (v/v) at 68 °C] between strain XIL11T and Cellulomonas humilata ATCC 25174T. DNA was isolated as described by Cashion et al. (1977). Renaturation rates were calculated by using the TRANSFER.BAS program (Jahnke, 1992).
Strain XIL11T was characterized morphologically as a Gram-positive, non-filamentous actinomycete that grew as lemon-yellow colonies on XED and YED media. Cells were non-motile and coccoid in shape (2·4x1·4 µm). Details of various differentiating characteristics of strain XIL11T and phylogenetically related species are shown in Table 1. Other characteristics determined are given in the species description. Isolate XIL11T grew at 4–37 °C. No growth was recorded at 45 °C.
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), which was supported by a high bootstrap value (100 %) based on 1000 resamplings. A fuller phylogenetic tree is available as supplementary material in IJSEM Online.
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), except for the nucleotides at positions 614–626 (A–U) and 1438–1463 (A–U), which were identical to those found in the sequence of C. humilata, but differed from those of the rest of the species in the genus Cellulomonas.
The peptidoglycan composition of strain XIL11T corresponded to type A4
; it contained D-ornithine–D-glutamic acid. This composition is reported for most members of the genus Cellulomonas and has been emphasized as an important feature for delineation at genus level in actinobacteria (Stackebrandt & Schumann, 2000). In the case of C. humilata ATCC 25174T, Gledhill & Casida (1969) found that the peptidoglycan of this strain contained lysine and ornithine, whereas Stackebrandt et al. (2002) reported L-ornithine–D-glutamic acid; the latter composition is found in most Cellulomonas species, including strain XIL11T. If the peptidoglycan results obtained by Stackebrandt et al. (2002) are confirmed, the genus Cellulomonas would then embrace a homogeneous group with respect to peptidoglycan structure, with L-ornithine in the third position of the peptide side-chain and D-aspartic acid or D-glutamic acid in the interpeptide bridge.
The cell-wall sugars in strain XIL11T were rhamnose, mannose and traces of fucose. The major fatty acids were anteiso-C15 : 0 (41·66 %), iso-C16 : 0 (13·37 %), iso-C18 : 0 (12·83 %) and anteiso-C17 : 0 (6·87 %). The main difference was the fairly large amount of C18 : 0 in the new isolate with respect to other Cellulomonas species (Schumann et al., 2001).
HPLC analysis of menaquinones revealed two peaks: the main peak corresponded to MK-9(H4) and the smaller one to MK-8(H4). MK-9(H4) is the major menaquinone found in members of the family Cellulomonadaceae. Polar lipids detected were diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylinositol and phosphatidylinositol mannosides.
The results of DNA–DNA hybridization showed 36·6 % relatedness between strain XIL11T and C. humilata ATCC 25174T, suggesting that isolate XIL11T should be classified as a novel species, in view of the recommendations of Wayne et al. (1987) for species delineation. The G+C content of the DNA of this strain is 73 mol%, a value that falls within the range reported for other members of the genus. Therefore, on the basis of chemotaxonomic, phylogenetic and physiological data, we propose that isolate XIL11T should be classified as the type strain of a novel species, Cellulomonas xylanilytica sp. nov.
Description of Cellulomonas xylanilytica sp. nov.
Cellulomonas xylanilytica (xy.la.ni.ly'ti.ca. N.L. n. xylan xylan, a plant polymer; N.L. adj. lyticus from Gr. adj. lytos dissolving; N.L. fem. adj. xylanilytica xylan-dissolving).
Gram-positive, non-flagellated, non-spore-forming, coccoid or rod-shaped cells (2·4x1·4 µm). Aerobic or facultatively anaerobic and chemo-organotrophic. Colonies on YED medium are smooth, yellow and usually 1–3 mm in diameter within 7 days at 28 °C. Aerial mycelium is not formed. Optimal growth temperature is 30 °C. Optimal growth pH is 7. Oxidase- and catalase-positive. Phylogenetically related to members of the family Cellulomonadaceae. Utilizes arabinose, cellulose, gentiobiose, maltose, mannose, N-acetylglucosamine, starch and xylan as sole carbon sources. By contrast, no growth occurs in acetate, adipate, caprate, citrate, malate, mannitol, phenylacetate or ribose. Acid is produced from amygdalin, arbutin, cellobiose, D-fructose, galactose, glucose, glycerol, glycogen, inulin, lactose, D-lyxose, maltose, D-mannose, melezitose, melibiose, D-raffinose, rhamnose, salicin, L-sorbose, sucrose, trehalose, xylitol, D-xylose, methyl 
-D-glucoside, methyl -D-mannoside, N-acetylglucosamine and -gentiobiose. Amylases, cellulases, -galactosidase and xylanases are actively produced. Aesculin, casein and gelatin are hydrolysed. Nitrate is reduced to nitrite. Arginine dehydrolase, indole, tryptophan deaminase and urease are not produced. Major fatty acids are ai-C15 : 0, C16 : 0 and C18 : 0. Major isoprenoid quinones are MK-9(H4) and MK-8(H4). Peptidoglycan contains L-ornithine–L-glutamic acid (type A4). Cell-wall sugars are rhamnose, mannose and fucose.
The type strain is XIL11T (=LMG 21723T=CECT 5729T).
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ACKNOWLEDGEMENTS |
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REFERENCES |
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TOPABSTRACTMAIN TEXTREFERENCES |
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Collins, M. D. & Pascual, C. (2000). Reclassification of Actinomyces humiferus (Gledhill and Casida) as Cellulomonas humilata nom. corrig., comb. nov. Int J Syst Evol Microbiol 50, 661–663.[Abstract]
DeSoete, G. (1983). A least squares algorithm for fitting additive trees to proximity data. Psychometrika 48, 621–626.[CrossRef]
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Elberson, M. A., Malekzadeh, F., Yazdi, M. T., Kameranpour, N., Noori-Daloii, M. R., Matte, M. H., Shahamat, M., Colwell, R. R. & Sowers, K. R. (2000). Cellulomonas persica sp. nov. and Cellulomonas iranensis sp. nov., mesophilic cellulose-degrading bacteria isolated from forest soils. Int J Syst Evol Microbiol 50, 993–996.[Abstract]
Funke, G., Pascual Ramos, C. & Collins, M. D. (1995). Identification of some clinical strains of CDC coryneform group A-3 and A-4 bacteria as Cellulomonas species and proposal of Cellulomonas hominis sp. nov. for some group A-3 strains. J Clin Microbiol 33, 2091–2097.[Abstract]
Gledhill, W. E. & Casida, L. E., Jr (1969). Predominant catalase-negative soil bacteria. II. Occurrence and characterization of Actinomyces humiferus, sp. N. Appl Microbiol 18, 114–121.[Medline]
Jahnke, K.-D. (1992). Basic computer program for evaluation of spectroscopic DNA renaturation data from GILFORD system 2600 spectrometer on a PC/XT/AT type personal computer. J Microbiol Methods 15, 61–73.
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Mandel, M. & Marmur, J. (1968). Use of ultraviolet absorbance temperature profile for determining the guanine plus cytosine content of DNA. Methods Enzymol 12B, 195–206.
Minnikin, D. E., O'Donnell, A. G., Goodfellow, M., Alderson, G., Athalye, M., Schaal, K. & Parlett, J. H. (1984). An integrated procedure for the extraction of isoprenoid quinones and polar lipids. J Microbiol Methods 2, 233–241.[CrossRef]
Rivas, R., Velázquez, E., Valverde, A., Mateos, P. F. & Martínez-Molina, E. (2001). A two primers random amplified polymorphic DNA procedure to obtain polymerase chain reaction fingerprints of bacterial species. Electrophoresis 22, 1086–1089.[CrossRef][Medline]
Rivas, R., Sánchez, M., Trujillo, M. E., Zurdo-Piñeiro, J. L., Mateos, P. F., Martínez-Molina, E. & Velázquez, E. (2003). Xylanimonas cellulosilytica gen. nov., sp. nov., a xylanolytic bacterium isolated from a decayed tree (Ulmus nigra). Int J Syst Evol Microbiol 53, 99–103.
Schleifer, K. H. (1985). Analysis of the chemical composition and primary structure of murein. Methods Microbiol 18, 123–156.
Schleifer, K. H. & Kandler, O. (1972). Peptidoglycan types of bacterial cell walls and their taxonomic implications. Bacteriol Rev 36, 407–477.
Schumann, P., Weiss, N. & Stackebrandt, E. (2001). Reclassification of Cellulomonas cellulans (Stackebrandt and Keddie 1986) as Cellulosimicrobium cellulans gen. nov., comb. nov. Int J Syst Evol Microbiol 51, 1007–1010.[Abstract]
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Stackebrandt, E. & Schumann, P. (2000). Description of Bogoriellaceae fam. nov., Dermacoccaceae fam. nov., Rarobacteraceae fam. nov. and Sanguibacteraceae fam. nov. and emendation of some families of the suborder Micrococcineae. Int J Syst Evol Microbiol 50, 1279–1285.[Abstract]
Stackebrandt, E., Breymann, S., Steiner, U., Prauser, H., Weiss, N. & Schumann, P. (2002). Re-evaluation of the status of the genus Oerskovia, reclassification of Promicromonospora enterophila (Jáger et al. 1983) as Oerskovia enterophila comb. nov. and description of Oerskovia jenensis sp. nov. and Oerskovia paurometabola sp. nov. Int J Syst Evol Microbiol 52, 1105–1111.[Abstract]
Staneck, J. L. & Roberts, G. D. (1974). Simplified approach to identification of aerobic actinomycetes by thin-layer chromatography. Appl Microbiol 28, 226–231.[Medline]
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