Hidrológiai Közlöny, 2015 (95. évfolyam)
2015 / 5-6. különszám - LVI. Hidrobiológus Napok előadásai
31 ségben sikerült tenyésztésbe vonnunk, valószínűleg az alkalmazott egyedi táptalajoknak is köszönhetően. A bíbor színű réteg mikroszkópos vizsgálata során ostorral rendelkező, görbült pálcika alakú, fototaxist mutató pirosas színű baktériumok tömegét láttuk. Irodalmi a- datok (Imhoff, 1995; Imhoff, 2006) szerint az obiigát anaerob bíbor kénbaktérium, az Ectothiorhodospira nemzetség tagjaira a megfigyelt morfológiai tulajdonságok ráillenek. Köszönetnyilvánítás A kutatást az OTKA PD 105407 és PD 112449 pályázat támogatta. A kutatás során használt műszerek beszerzését a KM- OP-4.2.1/B-10-2011-0002 és TÁMOP-4.2.2/B-10/1-2010-0030 pályázatok támogatták. Somogyi Boglárka és Felföldi Tamás munkáját a Magyar Tudományos Akadémia Bolyai János kutatói ösztöndíja segítette. A szerzők köszönetüket fejezik ki Németh Balázsnak a mintavételbeli, valamint Balogh Lajosné Anikónak technikai segítségéért. Irodalom Biel, A.J. 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Fanjing, K„ Qinxian, J., Jia, E„ Mianping, Z. (2009) Characterization of a eukaryotic picoplankton alga, strain DGN-Z1, isolated from a soda lake in Inner Mongolia, China. - Natural Res. Environ. Iss. 15: article 38. Fuerst, J.A., Hawkins, J.A., Holmes, A. Sly, L.I., Moore, C.J., Stackebrandt, E. (1993) Porphyrobacter neustonensis gen. nov., sp. nov., an aerobic bacteriochlorophyll-synthesizing budding bacterium from fresh water. - Int. J. Syst. Bacteriol. 43:125-134. Imhoff, J.F. (1995) Taxonomy and Physiology of Phototrophic Purple Bacteria and Green Sulfur Bacteria. In Anoxygenic Photosynthetic Bacteria. Kluwer Academic Publishers, pp. 1-15. Imhoff, J.F. (2006) The family Ectothiorhodospiraceae. In The Prokaryotes. Springer, pp. 874-886. Klindworth, A., Pruesse, E., Schweer, T, Peplies, J., Quast, C., Horn, M., Glöckner, F.O. (2013) Evaluation of general 16S ribosomal R- NA gene PCR primers for classical and next-generation sequencing-based diversity studies. - Nucleic Acids Res. 41: el. Kim, O.S., Cho, Y.J., Lee, K, Yoon, S.H., Kim, M„ Na, H, Park, S. C„ Jeon, Y.S., Lee J.H., Yi, H, Won, S„ Chun, J. (2012) Introducing EzTaxon-e: a prokaryotic 16S rRNA gene sequence database with phylotypes that represent uncultured species. - Int. J. Syst. Evol. Microbiol. 62: 716-721. Lane, D.J. (1991) 16S/23S rRNA sequencing. In Nucleic acid techniques in bacterial systematics. Wiley, pp. 125-175. Madigan, M.T., Martinka, J.M., Stahl, D., Clark, D.P. (2010) Brock Biology of Microorganisms (13th Ed). Pearson, San Francisco. Ollivier, B., Caumette, P., Garcia, J.L., Mah, R.A. (1994) Anaerobic bacteria from hypersaline environments. — Microbiol. Rev. 58: 27- 38. Pálffy, K, Felföldi, T, Mentes, A., Horváth, H., Márialigeti, K, Boros, E„ Vörös, L„ Somogyi, B. (2014) Unique picoeukaryotic algal community under multiple environmental stress conditions in a shallow, alkaline pan. - Extremophiles 18:111-119. Polz, M.F., Cavanaugh, C.M. (1998) Bias in template-to-product ratios in multitemplate PCR. — Appl. Environ. Microbiol. 64: 3724-3730. Pruesse, E., Peplies, J., Glöckner, F.O. (2012) SINA: accurate high- throughput multiple sequence alignment of ribosomal RNA genes. — Bioinformatics 28: 1823-1829. Puspita, I.D., Kamagata, Y, Tanaka, M., Asano, K., Nakatsu, C.H. (2012) Are Uncultivated Bacteria Really Uncultivable? - Microbes Environ. 27:356-366. Quast, C., Pruesse, E., Yilmaz, P„ Gerken, J., Schweer, T, Yarza, P., Peplies, G„ Glöckner, F. O. (2013) The SILVA ribosomal RNA gene database project: improved data processing and web-based tools. — Nucleic Acids Res. 41: D590-D596. Schloss, P.D., Westcott, S.L., Ryabin, T, Hall, J.R., Hartmann, M., Hollister, E.B., Lesniewski, R.A., Oakley, B.B., Parks, D.H., Robinson, C.J., Sahl, J.W., Stres, B., Thallinger, G.G., Van Horn, D.J., Weber, C.F. (2009) Introducing mothur: open-source, platform-independent, community-supported software for describing and comparing microbial communities. - Appl. Environ. Microbiol. 75: 7537-7541. Somogyi, B., Felföldi, T, Vanyovszki, J., Agyi, A., Márialigeti, K., Vörös, L. (2009) Winter bloom of picoeukaryotes in Hungarian shallow turbid soda pans and the role of light and temperature. - Aqu- at. Ecol. 43: 735-744. Sorokin, D.Y., Gorlenko, V.M., Namsaraev, B.B., Namsaraev, Z.B., Lysenko, A.M., Eshinimaev, B. T., Khmelenina, V.N., Trotsenko, Y.A., Kuenen, J.G. (2004) Prokaryotic communities of the north-eastern Mongolian soda lakes. - Hydrobiologia 522: 235-248. Tindall, B. J., Rosselló-Móra, R., Busse, H.-J., Ludwig, W., Kämpfer, P. (2010) Notes on the characterization of prokaryote strains for taxonomic purposes. - Int. J. Syst. Evol. Microbiol. 60:249-266. Van Trappen, S., Mergaert, J., Swings, J. (2004) Loktanella salsilacus gen. nov., sp. nov., Loktanella fryxellensis sp. nov. and Loktanella vestfoldensis sp. nov., new members of the Rhodobacter group, i- solated from microbial mats in Antarctic lakes. - Int. J. Syst. Evol. Microbiol. 54: 1263-1269. Wellburn, A.R. (1994) The spectral determination of chlorophylls a and b, as well as total carotenoids, using various solvents with spectrophotometers of different resolution. - J. Plant. Physiol. 144: 307- 313. Determination of purple bacterium community composition combining next-generation DNA sequencing and cultivation techniques Korponai, K., Somogyi, B., Szabó, A., Boros, E., Vörös, L. and Felföldi, T. Abstract: In April 2014 a spectacular, dual bloom occurred in a shallow alkaline soda pan near the village Fülöpszállás (Danube-Tisza Interfluve, Hungary). The upper three cm water layer was inhabited by a dense population of the planktonic green alga, Oocystis submarina; and under that, there was a separate purple layer formed by rod-shaped purple bacteria with flagella and showing phototaxis. Identification of these bacteria was carried out using cultivation and high-throughput DNA-sequencing methods. As a result of next-generation sequencing (Roche GS Junior platform) 3131 sequences were obtained, of which more than half were identified as members of the genera Ectothiorhodospira and Rhodobaca. Members from many dominant groups were successfully cultivated. Three different solid media were used during aerobic and anaerobic cultivation, two of them were designed particularly for this work. During aerobic cultivation, 1.7-35 x 106 CF- U/mL plate counts were obtained after 19 days of incubation, while under anaerobic conditions highly dissimilar values were recorded on different culture media. Half of the isolated bacterial strains had reddish coloration, a few of them showed irisation. Among the bacterial strains, members of phyla Proteobacteria and Bacteroidetes dominated, and the strains showed the highest similarity with the genera Halo- monas, Pseudomonas, Nitrincola, Vibrio, Alkalimonas, Porphyrobacter, Loktanella, Paracoccus, Roseicitreum, Roseinatronobacter, Rhodobaca, Belliella and Aquiflexum. Several strains represent potentially new species, their taxonomic description will be carried out in the near future. Keywords: soda pan, purple bacteria, bloom, cultivation, next-generation DNA sequencing.
