| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
|
|
|
|||||||
|
|||||||||
1 Coastal Marine Laboratory/Department of Biology, The Hong Kong University of Science and Technology, Clear Water Bay, Kowloon, Hong Kong SAR, People's Republic of China
2 Division of Environmental Science and Engineering, The National University of Singapore, Singapore
3 Department of Biology, The Chinese University of Hong Kong, Shatin, NT, Hong Kong SAR, People's Republic of China
Correspondence
Pei-Yuan Qian
boqianpy{at}ust.hk
 |
ABSTRACT |
|---|
 TOP ABSTRACT MAIN TEXTREFERENCES |
|---|
7c (altogether representing 69.0 % of the total fatty acids). MK-6 was the only respiratory quinone detected. Phylogenetic analysis based on 16S rRNA gene sequences indicated that the closest relatives of UST050418-085T were members of the genus Gillisia, with sequence similarities of 93.2–96.6 %. Strain UST050418-085T differed from its closest relatives by 11 to 18 phenotypic traits. Molecular evidence and phenotypic characteristics suggest that strain UST050418-085T represents a novel species within the genus Gillisia. The name Gillisia myxillae sp. nov. is proposed, with UST050418-085T (=JCM 13546T=NRRL B-41416T) as the type strain.
A scanning electron micrograph of a cell of strain UST050418-085T is available as supplementary material in IJSEM Online.
|
MAIN TEXT |
|---|
|
TOPABSTRACTMAIN TEXTREFERENCES |
|---|
). Gillisia is a recently established genus within this family (Van Trappen et al., 2004) and forms a phylogenetic cluster with the genera Mesonia, Salegentibacter and Psychroflexus (Nedashkovskaya et al., 2005). Currently, there are five species in the genus Gillisia, all of which were isolated from marine environments, including a microbial mat and sea-ice algae in Antarctica, as well as seawater in the Sea of Japan (Van Trappen et al., 2004; Bowman & Nichols, 2005; Nedashkovskaya et al., 2005). In the present study, a novel member of the genus Gillisia, isolated from the surface of a marine sponge, is described. During the characterization of bacteria isolated from the surface of the sponge Myxilla incrustans collected from Friday Harbor, San Juan Island, WA, USA, in April 2005, strain UST050418-085T was isolated on an agar medium consisting of 3 g yeast extract l–1, 5 g peptone l–1 and 0.22-µm-filtered seawater (referred to as marine agar hereafter) after 48 h incubation at 24 °C. Unless otherwise specified, all characteristics described are those of cultures grown on marine agar under these conditions. UST050418-085T appeared as yellow, convex, circular colonies (1–2 mm in diameter) with an entire edge and a smooth surface. No diffusible pigment was observed.
The nearly complete 16S rRNA gene sequence of UST050418-085T (1411 bp) was obtained bidirectionally with three replications as described by Lau et al. (2004). Phylogenetic analysis based on this nearly complete 16S rRNA gene sequence revealed that strain UST050418-085T was a member of the family Flavobacteriaceae. Its closest relatives were Gillisia mitskevichiae KMM 6034T (Nedashkovskaya et al., 2005), Gillisia limnaea LMG 21470T (Van Trappen et al., 2004) and Gillisia hiemivivida IC154T (Bowman & Nichols, 2005), with 96.6, 96.1 and 95.8 % 16S rRNA gene sequence similarity, respectively. A neighbour-joining (NJ) phylogenetic tree (Fig. 1) constructed using the ARB software package (Ludwig et al., 2004) placed strain UST050418-085T within the cluster comprising the five species of the genus Gillisia and showed that the most closely related strain was G. limnaea LMG 21470T. Sequence similarity shows the similarity between two sequences based on a comparison of their nucleotide sequences in overlapping regions nucleotide-by-nucleotide. However, construction of NJ trees uses a phenetic method, is distance-based using a clustering algorithm and takes other factors (e.g. type of mismatch, mutation, gap, etc.) into account to generate a similarity matrix using ARB software (Ludwig et al., 2004). Therefore, the closest phylogenetic neighbour observed in the NJ tree is not necessarily the one with the highest sequence similarity because these comparisons are based on two different methods of calculating similarity. In fact, this phenomenon is common and found in other recent publications in the same genus or family (Van Trappen et al., 2004; Lau et al., 2005). Two other trees based on cladistic methods (i.e. character-based) were also constructed. One was made using the maximum-likelihood method, which selects the tree that is most likely to have produced the observed data; the other one was based on the maximum-parsimony method, which selects the tree that requires fewer evolutionary changes (Ludwig et al., 2004). Both trees showed essentially the same topography for all species of the genus Gillisia (not shown). These results suggest that UST050418-085T represents a novel species within the genus Gillisia.
|
, was 34.6±0.1 mol% (three replicates). This value is within the range of G+C contents observed for members of the genus Gillisia (i.e. 32.0–37.8 mol%). The cellular fatty acids of UST050418-085T were determined using the Sherlock Microbial Identification System (MIDI) according to the manufacturer's protocol. The dominant fatty acids were i15 : 0, a15 : 0, i15 : 1, i16 : 0, i17 : 0 3-OH, 17 : 0 2-OH and summed feature 3, comprising i15 : 0 2-OH and/or 16 : 17c, which altogether represent 69.0 % of the total fatty acids (Table 1). This fatty acid profile is similar to those of G. limnaea LMG 21470T and G. mitskevichiae KMM 6034T, confirming the close phylogenetic relationships observed for these bacteria (Fig. 1
). MK-6 was the only respiratory quinone detected in UST050418-085T, as determined using an HPLC method according to Collins (1994). Menaquinones extracted from Cellulophaga lytica (Nakagawa & Yamasato, 1993) and Pedobacter heparinus (Steyn et al., 1998) served as references for MK-6 and MK-7, respectively.
|
). Colony and cell morphology were examined using light (x40 magnification) and scanning electron microscopy (6700F; JEOL) according to Neu et al. (2001) (see Supplementary Fig. S1 available in IJSEM Online). The reaction to Gram stain was determined using light microscopy according to Smibert & Krieg (1994). Gliding motility was observed under a phase-contrast light microscope (Olympia) after growth of the strain on quarter-strength marine 2216 medium (Oxoid) solidified with 1 % agar according to Bowman (2000). Susceptibility to antibiotics was tested according to Acar (1980). Flexirubin-type pigment production and carboxymethylcellulose hydrolysis were determined according to Bowman (2000). Casein hydrolysis was determined according to the procedures described by Norris et al. (1985); hydrolysis of Tweens 20, 40 and 80 and chitin was determined as described by Baumann & Baumann (1981), whereas hydrolysis of agar, DNA and starch was tested according to Smibert & Krieg (1994). Oxidase and catalase activities were determined as described by Smibert & Krieg (1994). Other enzyme activities, substrate utilization patterns, nitrate reduction and production of H2S, indole and acetoin were tested using the commercial systems API 20E, API 20NE, API 50CH and API ZYM (bioMérieux) and MicroLog 3 (Biolog). Cells for inoculating the API test systems were suspended in a sterile solution of seawater mixture at 22 
salinity (MacDonell et al., 1982). Growth on glycerol, D-glucose, sucrose, D-mannitol, D-galactose, starch, D-sorbitol, D-arabinose and D-melibiose as sole carbon sources was also tested using a medium containing 0.2 g NaNO3, 0.2 g NH4Cl, 0.05 g yeast extract and 4 % (w/v) carbon source in 1 l solution of seawater mixture at 35 salinity (Nedashkovskaya et al., 2003).
The morphological, physiological and biochemical characteristics of UST050418-085T are listed in the species description. Strain UST050418-085T differed from its three closest relatives by 11 to 18 traits, indicated in Table 2. Molecular evidence, together with phenotypic characteristics presented in this study, suggest that UST050418-085T constitutes a novel species within the genus Gillisia. The name Gillisia myxillae sp. nov. is proposed.
|
Cells are Gram-negative, short rods (1.3–2.0 µm in length and 0.5 µm in width), strictly aerobic, devoid of gliding and flagellar motility. Upon cultivation on marine agar, colonies are yellow in colour, 2–4 mm in diameter, circular and convex with a smooth surface and an entire edge. Does not produce flexirubin-type or diffusible pigments. MK-6 is the only respiratory quinone. Growth occurs between pH 5.0 and 10.0 (pH 7.0–9.0 optimum) and between 4.0 and 28.0 °C (12.0–20.0 °C optimum), but not at 36.0 °C or higher. Requires NaCl (2.0–10.0 %; 4.0–6.0 % optimum) for growth. Predominant fatty acids are i15 : 0, a15 : 0, i15 : 1, i16 : 0, i17 : 0 3-OH, 17 : 0 2-OH and summed feature 3 (comprising i15 : 0 2-OH and/or 16 : 17c), together representing 69.0 % of the total. Susceptible to benzylpenicillin (1.0 µg), chloramphenicol (1.0 µg), ampicillin (1.0 µg) and tetracycline (10.0 µg), but resistant to streptomycin and kanamycin (tested up to 100.0 µg). Acetoin, indole and H2S are not produced. Nitrate is not reduced. Citrate is not utilized. Casein, starch, and Tweens 20 and 40 are degraded, but agar, Tween 80, chitin and carboxymethylcellulose are not. Positive for oxidase, DNase, alkaline phosphatase, esterase (C4), esterase lipase (C8), lipase (C14), leucine arylamidase, valine arylamidase, cystine arylamidase, acid phosphatase, naphthol-AS-BI-phosphohydrolase and tryptophan deaminase activities. Negative for gelatinase, urease, trypsin, 
-chymotrypsin, -galactosidase, 
-galactosidase, -glucuronidase, -glucosidase, -glucosidase, N-acetyl--glucosaminidase, -mannosidase, -fucosidase, arginine dihydrolase, lysine decarboxylase and ornithine decarboxylase activities. Utilization of glycerol, D-glucose, sucrose, D-mannitol, D-galactose, starch, D-sorbitol, D-arabinose and D-melibiose as sole carbon sources is observed on agar medium supplemented with 4 % (w/v) carbon source and utilization of D-galacturonic acid, L-proline and putrescine is observed in the MicroLog 3 system. However, none of the carbon sources included in the API 20NE and 50CH systems are utilized. Acid is produced from L-arabinose, D-fructose, D-mannose, D-maltose, starch and glycogen in the API 50CH system, but no acid production is observed from the carbon sources in the API 20E system.
The type strain is UST050418-085T (=JCM 13546T=NRRL B-41416T), isolated from the surface of the marine sponge Myxilla incrustans at Friday Harbor, San Juan Island, WA, USA. The DNA G+C content of strain UST050418-085T is 34.6 mol%.
|
ACKNOWLEDGEMENTS |
|---|
|
REFERENCES |
|---|
|
TOPABSTRACTMAIN TEXTREFERENCES |
|---|
Baumann, P. & Baumann, L. (1981). The marine gram-negative eubacteria: genera Photobacterium, Beneckea, Alteromonas, Pseudomonas and Alcaligenes. In The Prokaryotes, vol. 1, pp. 1302–1331. Edited by M. P. Starr, H. Stolp, H. G. Trüper, A. Balows & H. Schlegel. Berlin: Springer.
Bowman, J. P. (2000). Description of Cellulophaga algicola sp. nov., isolated from the surfaces of Antarctic algae, and reclassification of Cytophaga uliginosa (ZoBell and Upham 1944) Reichenbach 1989 as Cellulophaga uliginosa comb. nov. Int J Syst Evol Microbiol 50, 1861–1868.[Abstract]
Bowman, J. P. (2004). Psychrophilic prokaryote structural-functional relationships, biogeography and evolution within marine sediment. Cell Mol Biol 50, 503–515.[Medline]
Bowman, J. P. & Nichols, D. S. (2005). Novel members of the family Flavobacteriaceae from Antarctic maritime habitats including Subsaximicrobium wynnwilliamsii gen. nov., sp. nov., Subsaximicrobium saxinquilinus sp. nov., Subsaxibacter broadyi gen. nov., sp. nov., Lacinutrix copepodicola gen. nov., sp. nov., and novel species of the genera Bizionia, Gelidibacter and Gillisia. Int J Syst Evol Microbiol 55, 1471–1486.
Collins, M. D. (1994). Isoprenoid quinones. In Chemical Methods in Prokaryotic Systematics, pp. 265–310. Edited by M. Goodfellow & A. G. O'Donnell. Chichester: Wiley.
Isnansetyo, A. & Kamei, Y. (2003). Pseudoalteromonas phenolica sp. nov., a novel marine bacterium that produces phenolic anti-methicillin-resistant Staphylococcus aureus substances. Int J Syst Evol Microbiol 53, 583–588.
Lau, S. C. K., Tsoi, M. M. Y., Li, X., Plakhotnikova, I., Wu, M., Wong, P. K. & Qian, P. Y. (2004). Loktanella hongkongensis sp. nov., a novel member of the -Proteobacteria originating from marine biofilms in Hong Kong waters. Int J Syst Evol Microbiol 54, 2281–2284.
Lau, S. C. K., Tsoi, M. M. Y., Li, X. & 7 other authors (2005). Winogradskyella poriferorum sp. nov., a novel member of the family Flavobacteriaceae isolated from a sponge in the Bahamas. Int J Syst Evol Microbiol 55, 1589–1592.
Ludwig, W., Strunk, O., Westram, R. & 29 other authors (2004). ARB: a software environment for sequence data. Nucleic Acids Res 32, 1363–1371.
MacDonell, M. T., Singleton, F. L. & Hood, M. A. (1982). Diluent composition for use of API 20E in characterizing marine and estuarine bacteria. Appl Environ Microbiol 44, 423–427.
Mesbah, M., Premachandran, U. & Whitman, W. B. (1989). Precise measurement of the G+C content of deoxyribonucleic acid by high-performance liquid chromatography. Int J Syst Bacteriol 39, 159–167.
Nakagawa, Y. & Yamasato, K. (1993). Phylogenetic diversity of the genus Cytophaga revealed by 16S rRNA sequencing and menaquinone analysis. J Gen Microbiol 139, 1155–1161.
Nedashkovskaya, O. I., Kim, S. B., Hans, S. K. & 7 other authors (2003). Mesonia algae gen. nov., sp. nov., a novel marine bacterium of the family Flavobacteriaceae isolated from the green alga Acrosiphonia sonderi (Kütz) Kornm. Int J Syst Evol Microbiol 53, 1967–1971.
Nedashkovskaya, O. I., Kim, S. B., Lee, K. H., Mikhailov, V. V. & Bae, K. S. (2005). Gillisia mitskevichiae sp. nov., a novel bacterium of the family Flavobacteriaceae, isolated from sea water. Int J Syst Evol Microbiol 55, 321–323.
Neu, B., Voigt, A., Mitlohner, R. & 7 other authors (2001). Biological cells as templates for hollow microcapsules. J Microencapsul 18, 385–395.[CrossRef][Medline]
Norris, J. R., Ribbons, D. W. & Varma, A. K. (editors) (1985). Methods in Microbiology, vol. 18. London: Academic Press.
Smibert, R. M. & Krieg, N. R. (1994). Phenotypic characteristics. In Methods for General and Molecular Biology, pp. 607–654. Edited by P. Gerhardt, R. G. E. Murray, W. A. Wood & N. R. Krieg. Washington, DC: American Society for Microbiology.
Steyn, P. L., Segers, P., Vancanneyt, M., Sandra, P., Kersters, K. & Joubert, J. J. (1998). Classification of heparinolytic bacteria into a new genus, Pedobacter, comprising four species: Pedobacter heparinus comb. nov., Pedobacter piscium comb. nov., Pedobacter africanus sp. nov. and Pedobacter saltans sp. nov. Proposal of the family Sphingobacteriaceae. Int J Syst Bacteriol 48, 165–177.
Van Trappen, S., Vandecandelaere, I., Mergaert, J. & Swings, J. (2004). Gillisia limnaea gen. nov., sp. nov., a new member of the family Flavobacteriaceae isolated from a microbial mat in Lake Fryxell, Antarctica. Int J Syst Evol Microbiol 54, 445–448.
This article has been cited by other articles:
|
|
 |
|
  O. O. Lee, Y. H. Wong, and P.-Y. Qian Inter- and Intraspecific Variations of Bacterial Communities Associated with Marine Sponges from San Juan Island, Washington Appl. Envir. Microbiol., June 1, 2009; 75(11): 3513 - 3521. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 M. W. Taylor, R. Radax, D. Steger, and M. Wagner Sponge-Associated Microorganisms: Evolution, Ecology, and Biotechnological Potential Microbiol. Mol. Biol. Rev., June 1, 2007; 71(2): 295 - 347. [Abstract] [Full Text] [PDF]
|
|
| ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
| INT J SYST EVOL MICROBIOL | MICROBIOLOGY | J GEN VIROL |
| J MED MICROBIOL | ALL SGM JOURNALS | |
