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1 Biological Resource Center, KRIBB, Daejeon 305-806, Korea
2 Fudan University, Shanghai 200433, China
3 Yeungnam University, Gyeongsangbuk-do 712-749, Korea
4 University of Science and Technology, Daejeon 305-333, Korea
5 Environmental Biotechnology National Core Research Center, Gyeongsang National University, Jinju 660-701, Korea
Correspondence
Jin-Woo Bae
baejw{at}kribb.re.kr
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; Ivanova et al., 2004a) currently comprises 34 recognized species, with Pseudoalteromonas haloplanktis as the type species. Many Pseudoalteromonas species have been isolated from the marine environment and show interstrain 16S rRNA gene sequence similarity values ranging from 90 to 99.9 %. To better describe the Pseudoalteromonas community associated with the tidal flats along the Korean coast (Kim et al., 2004), marine sediment was sampled and its bacterial diversity was investigated. Here, we describe two novel Pseudoalteromonas-like strains, which were determined to belong to the genus Pseudoalteromonas on the basis of their 16S rRNA gene sequences. Accordingly, the objective of the present work was to elucidate the taxonomic position of these newly isolated strains, designated mano4T and mano6, via phenotypic, genetic and chemo-taxonomic analyses. An additional aim was to determine whether the current taxonomic status of Pseudoalteromonas sagamiensis is appropriate.
Strains mano4T and mano6 were isolated from a tidal flat area of Dae-Chun, Chung-Nam, Korea (36° 17' 45.2'' N 126° 31' 9.5'' E) using the dilution plating technique on marine agar 2216 (MA; Difco). The two strains were grown routinely at 25 °C for 3 days. Their closest relatives, as judged by 16S rRNA gene similarity, were Pseudoalteromonas undina DSM 6065T, Pseudoalteromonas translucida DSM 14402T and Pseudoalteromonas aliena DSM 16473T. These three strains were obtained from DSMZ, Germany, and were grown under the same conditions and used as reference strains. Cultures of the isolates and the reference strains were stored at –80 °C in marine broth (MB) containing 20 % glycerol. For morphological and physiological characterization, strains mano4T and mano6 and the reference strains were generally cultivated in MB with shaking at 25 °C. API 20NE and API ZYM test strips (bioMérieux) were used to analyse the biochemical and physiological traits of these bacterial strains. Strips were inoculated with a heavy bacterial suspension in ASW or AUX medium (bioMérieux), supplemented with 2 % (w/v) sea salts. Other biochemical tests were performed using the methods and media described by Gordon et al. (1973). Catalase activity was determined by bubble production in a 3 % (v/v) hydrogen peroxide solution. Growth under anaerobic conditions was determined microscopically (Nikon E600) after incubation for 7 days in anaerobic Gaspak jars (BBL) containing an atmosphere of 80 % N2, 10 % CO2 and 10 % H2 (by vol.). Growth at various NaCl concentrations and at various temperatures and pH values was measured in MB. Cellular morphology and the presence of spores were also determined microscopically. Cellular motility for the novel isolate was examined using fresh wet-mounts of young bacterial cultures in MB. For TEM, cells from exponentially growing cultures were negatively stained with 1 % (w/v) phosphotungstic acid. After air-drying, the grid was examined using a model H-7600 transmission electron microscope (Hitachi). Results from the biochemical and physiological tests are given in Table 1 and in the species description. The new isolates could be readily differentiated from other closely related species by several phenotypic properties, as shown in Table 1.
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). Chromosomal DNA was extracted and purified as described by Sambrook et al. (1989). DNA G+C content was assessed by using the methods described by Tamaoka & Komagata (1984). DNA was hydrolysed and the resultant nucleotides were analysed via HPLC using a reversed-phase column (Supelcosil LC-18-S; Supelco). Amplification, sequencing and phylogenetic analysis of the 16S rRNA gene were performed according to the methods described by Ivanova et al. (2002c). DNA–DNA hybridization was performed fluorometrically by using the method of Bae et al. (2005) with Cy5-labelled DNA probes and microarrays.
Phylogenetic trees based on 16S rRNA gene sequences of members of the genus Pseudoalteromonas showed that strains mano4T and mano6 fall within the cluster of Pseudoalteromonas species (Fig. 1). Strains mano4T and mano6 exhibited 16S rRNA gene sequence similarities of 92.8–98.4 % to the type strains of 34 other Pseudoalteromonas species.
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Ivanova et al. (2000, 2002a, b, c, d, e, 2004b) have extensively explored the diversity and systematics of the genus Pseudoalteromonas. In successive studies, they observed that Pseudoalteromonas bacteriolytica Sawabe et al. 1998 (basonym of Algicola bacteriolytica) branched deeply in the phylogenetic tree of the genus and lacked a signature sequence (Ivanova et al., 2004a). Thus, the genus Algicola has been newly proposed to resolve the phylogenetic relationships among the marine Alteromonas-like proteobacteria. However, Pseudoalteromonas sagamiensis (Kobayashi et al., 2003), which is the species most closely related to Algicola bacteriolytica, was not included in the study. The 16S rRNA gene sequences of members of the genera Pseudoalteromonas and Algicola and a related species were retrieved from the NCBI database. DNA sequence alignment was conducted using CLUSTAL_X software (Thompson et al., 1997). Phylogenetic trees were constructed using the Fitch & Margoliash (Fitch & Margoliash, 1967) and neighbour-joining (Saitou & Nei, 1987) methods. The resultant unrooted tree topology was evaluated by bootstrap analysis based on 1000 resamplings (Felsenstein, 1985), using the neighbour-joining method. From the unrooted evolutionary tree shown in Fig. 1
, we concluded that P. sagamiensis forms a branch with Algicola bacteriolytica that is distinct from other Pseudoalteromonas species. The relationship is based on data from different tree-making algorithms and the bootstrap value of 100 %. P. sagamiensis showed very low sequence similarity (90.4 %) to Algicola bacteriolytica. Despite this low similarity, Kobayashi et al. (2003) did not classify P. sagamiensis as representing a new genus because the phenotypic differences between P. sagamiensis and currently known related genera seemed too small to warrant generic separation.
P. sagamiensis can also be clearly differentiated by phenotypic characteristics determined in previous studies (Ivanova et al., 2004a; Kobayashi et al., 2003; Sawabe et al., 1998). As shown in Table 2, P. sagamiensis and Algicola bacteriolytica showed low halotolerance, in contrast to the other Pseudoalteromonas species. They also shared the characteristic of lack of growth at 4 and 37 °C. However, some phenotypic properties, such as bacteriolytic activity and utilization of D-mannose, D-fructose and sucrose, differentiated P. sagamiensis from Algicola bacteriolytica.
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) and the data presented here, we propose that Pseudoalteromonas sagamiensis Kobayashi et al. 2003 be reassigned to the genus Algicola as Algicola sagamiensis comb. nov.
Description of Pseudoalteromonas marina sp. nov.
Pseudoalteromonas marina (ma.ri'na. L. fem. adj. marina of the sea, marine).
Cells are Gram-negative, rod-shaped on MA (measuring 0.5–0.7x2.1–3.0 µm) and motile. Cells do not form endospores. Colonies are pale yellow in colour, 0.2–0.5 mm in diameter, smooth and circular to slightly irregular in shape after 3 days of culture on MA. Growth occurs at 4–37 °C and at pH 5.3–8.8, but not at pH values lower than 4.1 nor higher than 9.3. Growth occurs in the presence of 3–12 % NaCl, but not in the absence of NaCl or in 15 % NaCl. Growth is not observed under anaerobic conditions. Catalase-positive and Voges–Proskauer test negative. Casein and starch are hydrolysed, but nitrate is not reduced to nitrite. Produce amylase, caseinase, alginase, DNase, but not agarase or chitinase, and utilize glucose, maltose and melibiose as a sole carbon and energy source. The following substrates are not utilized: D-arabinose, L-arginine, D-galactose, lactose, D-mannitol, sorbitol, citrate and xylose. Major fatty acids are C15 : 0 (6.8±0.4 %), C16 : 0 (21.3±0.7 %) and C16 : 1
7c (24.7±1.2 %).
The DNA G+C content of the type strain is 41.2 mol%. The type strain, mano4T (=KCTC 12242T=DSM 17587T), was isolated from a tidal flat area of Dae-Chun, Chung-Nam, Korea.
Description of Algicola sagamiensis Kobayashi et al. 2003 comb. nov.
Algicola sagamiensis (sa.ga.mi.en'sis. N.L. fem. adj. sagamiensis referring to Sagami Bay, the place of isolation).
Basonym: Pseudoalteromonas sagamiensis Kobayashi et al. 2003. The description is identical to that given by Kobayashi et al. (2003).
The type strain is B-10-31T (=DSM 14643T=JCM 11461T).
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ACKNOWLEDGEMENTS |
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REFERENCES |
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Egan, S., Holmström, C. & Kjelleberg, S. (2001). Pseudoalteromonas ulvae sp. nov., a bacterium with antifouling activities isolated from the surface of a marine alga. Int J Syst Evol Microbiol 51, 1499–1504.[Abstract]
Felsenstein, J. (1985). Confidence limits on phylogenies: an approach using the bootstrap. Evolution 39, 783–791.[CrossRef]
Fitch, W. M. & Margoliash, E. (1967). Construction of phylogenetic trees. Science 155, 279–284.
Gauthier, G., Gauthier, M. & Christen, R. (1995). Phylogenetic analysis of the genera Alteromonas, Shewanella, and Moritella using genes coding for small-subunit rRNA sequences and division of the genus Alteromonas into two genera, Alteromonas (emended) and Pseudoalteromonas gen. nov., and proposal of twelve new species combinations. Int J Syst Bacteriol 45, 755–761.
Gordon, R. E., Haynes, W. C. & Pang, C. H. (1973). The Genus Bacillus. Washington, DC: USDA.
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.
Ivanova, E. P., Chun, J., Romanenko, L. A., Matte, M. E., Mikhailov, V. V., Frolova, G. M., Huq, A. & Colwell, R. R. (2000). Reclassification of Alteromonas distincta Romanenko et al. 1995 as Pseudoalteromonas distincta comb. nov. Int J Syst Evol Microbiol 50, 141–144.[Abstract]
Ivanova, E. P., Gorshkova, N. M., Sawabe, T., Hayashi, K., Kalinovskaya, N. I., Lysenko, A. M., Zhukova, N. V., Nicolau, D. V., Kuznetsova, T. A. & other authors (2002a). Pseudomonas extremorientalis sp. nov., isolated from a drinking water reservoir. Int J Syst Evol Microbiol 52, 2113–2120.[Abstract]
Ivanova, E. P., Sawabe, T., Alexeeva, Y. V., Lysenko, A. M., Gorshkova, N. M., Hayashi, K., Zukova, N. V., Christen, R. & Mikhailov, V. V. (2002b). Pseudoalteromonas issachenkonii sp. nov., a bacterium that degrades the thallus of the brown alga Fucus evanescens. Int J Syst Evol Microbiol 52, 229–234.[Abstract]
Ivanova, E. P., Sawabe, T., Lysenko, A. M., Gorshkova, N. M., Hayashi, K., Zhukova, N. V., Nicolau, D. V., Christen, R. & Mikhailov, V. V. (2002c). Pseudoalteromonas translucida sp. nov. and Pseudoalteromonas paragorgicola sp. nov., and emended description of the genus. Int J Syst Evol Microbiol 52, 1759–1766.[Abstract]
Ivanova, E. P., Sawabe, T., Lysenko, A. M., Gorshkova, N. M., Svetashev, V. I., Nicolau, D. V., Yumoto, N., Taguchi, T., Yoshikawa, S. & other authors (2002d). Pseudoalteromonas ruthenica sp. nov., isolated from marine invertebrates. Int J Syst Evol Microbiol 52, 235–240.[Abstract]
Ivanova, E. P., Shevchenko, L. S., Sawabe, T., Lysenko, A. M., Svetashev, V. I., Gorshkova, N. M., Satomi, M., Christen, R. & Mikhailov, V. V. (2002e). Pseudoalteromonas maricaloris sp. nov., isolated from an Australian sponge, and reclassification of [Pseudoalteromonas aurantia] NCIMB 2033 as Pseudoalteromonas flavipulchra sp. nov. Int J Syst Evol Microbiol 52, 263–271.[Abstract]
Ivanova, E. P., Flavier, S. & Christen, R. (2004a). Phylogenetic relationships among marine Alteromonas-like proteobacteria: emended description of the family Alteromonadaceae and proposal of Pseudoalteromonadaceae fam. nov., Colwelliaceae fam. nov., Shewanellaceae fam. nov., Moritellaceae fam. nov., Ferrimonadaceae fam. nov., Idiomarinaceae fam. nov. and Psychromonadaceae fam. nov. Int J Syst Evol Microbiol 54, 1773–1788.
Ivanova, E. P., Gorshkova, N. M., Zhukova, N. V., Lysenko, A. M., Zelepuga, E. A., Prokof'eva, N. G., Mikhailov, V. V., Nicolau, D. V. & Christen, R. (2004b). Characterization of Pseudoalteromonas distincta-like sea-water isolates and description of Pseudoalteromonas aliena sp. nov. Int J Syst Evol Microbiol 54, 1431–1437.
Kim, B. S., Oh, H. M., Kang, H., Park, S. S. & Chun, J. (2004). Remarkable bacterial diversity in the tidal flat sediment as revealed by 16S rDNA analysis. J Microbiol Biotechnol 14, 205–211.
Kobayashi, T., Imada, C., Hiraishi, A., Tsujibo, H., Miyamoto, K., Inamori, Y., Hamada, N. & Watanabe, E. (2003). Pseudoalteromonas sagamiensis sp. nov., a marine bacterium that produces protease inhibitors. Int J Syst Evol Microbiol 53, 1807–1811.
Lau, S. C. K., Tsoi, M. M. Y., Li, X., Dobretsov, S., Plakhotnikova, Y., Wong, P.-K. & Qian, P.-Y. (2005). Pseudoalteromonas spongiae sp. nov., a novel member of the 
-Proteobacteria isolated from the sponge Mycale adhaerens in Hong Kong waters. Int J Syst Evol Microbiol 55, 1593–1596.
Romanenko, L. A., Mikhailov, V. V., Lysenko, A. M. & Stepanenko, V. I. (1995). A new species of melanin-producing bacteria of the genus Alteromonas. Mikrobiologiia 64, 74–77 (in Russian).
Romanenko, L. A., Zhukova, N. V., Lysenko, A. M., Mikhailov, V. V. & Stackebrandt, E. (2003a). Assignment of ‘Alteromonas marinoglutinosa’ NCIMB 1770 to Pseudoalteromonas mariniglutinosa sp. nov., nom. rev., comb. nov. Int J Syst Evol Microbiol 53, 1105–1109.
Romanenko, L. A., Zhukova, N. V., Rohde, M., Lysenko, A. M., Mikhailov, V. V. & Stackebrandt, E. (2003b). Pseudoalteromonas agarivorans sp. nov., a novel marine agarolytic bacterium. Int J Syst Evol Microbiol 53, 125–131.
Saitou, N. & Nei, M. (1987). The neighbour-joining method: a new method for reconstructing phylogenetic trees. Mol Biol Evol 4, 406–425.[Abstract]
Sambrook, J., Fritsch, E. F. & Maniatis, T. (1989). Molecular Cloning: a Laboratory Manual, 2nd edn. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory.
Sasser, M. (1990). Identification of bacteria by gas chromatography of cellular fatty acids, MIDI Technical Note 101. Newark, DE: MIDI Inc.
Sawabe, T., Makino, H., Tatsumi, M., Nakano, K., Tajima, K., Iqbal, M. M., Yumoto, I., Ezura, Y. & Christen, R. (1998). Pseudoalteromonas bacteriolytica sp. nov., a marine bacterium that is the causative agent of red spot disease of Laminaria japonica. Int J Syst Bacteriol 48, 769–774.
Tamaoka & Komagata (1984). Determination of DNA base composition by reverse-phase high-performance liquid chromatography. FEMS Microbiol Lett 25, 125–128.
Thompson, J. D., Gibson, T. J., Plewniak, F., Jeanmougin, F. & Higgins, D. G. (1997). The CLUSTAL_X windows interface: flexible strategies for multiple sequence alignment aided by quality analysis tools. Nucleic Acids Res 25, 4876–4882.
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