Journal of Microbiology and Biotechnology

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
권호 표지
DOI QR코드

DOI QR Code

Establishment of an Axenic Culture of Microcystin-Producing Microcystis aeruginosa Isolated from a Korean Reservoir

  • Han, Ah-Won (Department of Environmental Engineering, Chungbuk National University) ;
  • Oh, Kyoung-Hee (Department of Environmental Engineering, Chungbuk National University) ;
  • Jheong, Weon-Hwa (National Institute of Environmental Research) ;
  • Cho, Young-Cheol (Department of Environmental Engineering, Chungbuk National University)
  • Received : 2010.03.17
  • Accepted : 2010.04.03
  • Published : 2010.07.28
Download PDF
⟨ Previous Next ⟩

Abstract

In order to establish an axenic (bacteria-free) culture of Microcystis aeruginosa NIER 10039 isolated from a Korean reservoir, the culture was subjected to sequential treatment, including ultrasonication, washing, and addition of antibiotics. Three broad-spectrum antibiotics, namely, kanamycin, ampicillin, and imipenem, were applied separately in that order. Axenicity of the culture was confirmed by cultivation on bacterial media and observation under epifluorescence and scanning electron microscopes. We are the first to establish an axenic culture of a Microcystis strain isolated from Korean reservoirs and can be used in physiological and molecular studies to control toxic Microcystis blooms.

Keywords

References

  1. Ahn, C.-Y., H.-S. Kim, B.-D. Yoon, and H.-M. Oh. 2003. Influence of rainfall on cyanobacterial bloom in Daechung Reservoir. Korean J. Limnol. 36: 413-419.
  2. Bolch, C. J. S. and S. I. Blackburn. 1996. Isolation and purification of Australian isolates of the toxic cyanobacterium Microcystis aeruginosa Kutz. J. Appl. Phycol. 8: 5-13. https://doi.org/10.1007/BF02186215
  3. Cadel-Six, S., C. Dauga, A. M. Castets, R. Rippka, C. Bouchier, N. T. de Marsac, and M. Welker. 2008. Halogenase genes in nonribosomal peptide synthetase gene clusters of Microcystis (cyanobacteria): Sporadic distribution and evolution. Mol. Biol. Evol. 25: 2031-2041 https://doi.org/10.1093/molbev/msn150
  4. Carmichael, W. W. 1996. Toxic Microcystis and the environment, pp. 1-11. In M. F. Watanabe, K. Harada, W. W. Carmichael, and K. Fujii (eds.). Toxic Microcystis. CRC Press, Boca Raton, FL, U.S.A.
  5. Choi, G.-H., M.-S. Bae, C.-Y. Ahn, and H.-M. Oh. 2008. Induction of axenic culture of Arthrospira (Spirulina) platensis based on antibiotic sensitivity of contaminating bacteria. Biotechnol. Lett. 30: 87-92.
  6. Ferris, M. J. and C. F. Hirsch. 1991. Method for isolation and purification of cyanobacteria. Appl. Environ. Microbiol. 57: 1448-1452.
  7. Hisbergues, M., G. Christiansen, L. Rouhiainen, K. Sivonen, and T. Borner. 2003. PCR-based identification of microcystinproducing genotypes of different cyanobacterial genera. Arch. Microbiol. 180: 402-410. https://doi.org/10.1007/s00203-003-0605-9
  8. Huang, W.-J., C.-H. Lai, and Y.-L. Cheng. 2007. Evaluation of extracellular products and mutagenicity in cyanobacteria cultures separated from a eutrophic reservoir. Sci. Tot. Environ. 377: 214-223. https://doi.org/10.1016/j.scitotenv.2007.01.075
  9. Kim, J.-S., Y.-H. Park, B.-D. Yoon, and H.-M. Oh. 1999. Establishment of axenic cultures of Anabeana flos-aquae and Aphanothece nidulans (cyanobacteria) by lysozyme treatment. J. Phycol. 35: 865-869. https://doi.org/10.1046/j.1529-8817.1999.3540865.x
  10. Kondo, R., T. Yoshida, Y. Yuki, and S. Hiroishi. 2000. DNADNA reassociation among a bloomforming cyanobacterial genus, Microcystis. Int. J. Sys. Evol. Microb. 50: 767-770. https://doi.org/10.1099/00207713-50-2-767
  11. Lee, H.-S., K.-H. Oh, and Y.-C. Cho. 2008. Isolation of cyanobacteria producing microcystin from lakes. Kor. J. Microbiol. 44: 251-257.
  12. Lee, K. L., W. H. Jheong, J. M. Kim, and H. S. Kim. 2007. Detection of toxigenicity of cyanobacteria by molecular method. Korean J. Limnol. 40: 149-154.
  13. NCCLS. 2001. Performance standards for antimicrobial disk susceptibility tests. Approved standard. National Committee for Clinical Laboratory Standards, Wayne, PA, U.S.A.
  14. Ohtake, A., M. Shirai, T. Aida, N. Mori, K.-I. Harada, K. Matsuura, M. Suzuki, and M. Nakano. 1989. Toxicity of Microcystis species isolated from natural blooms and purification of the toxin. Appl. Environ. Microbiol. 55: 3202-3207.
  15. Park, H. D., B. Kim, E. Kim, and T. Okino. 1998. Hepatotoxic microcystins and neurotoxic anatoxin-a in cyanobacterial blooms from Korean lakes. Environ. Toxicol. Water Qual. 13: 225-234. https://doi.org/10.1002/(SICI)1098-2256(1998)13:3<225::AID-TOX4>3.0.CO;2-9
  16. Pyle, B. H., S. C. Broadaway, and G. A. McFeters. 1995. Factors affecting the determination of respiratory activity on the basis of cyanoditolyl tetrazolium chloride reduction with membrane filtration. Appl. Environ. Microbiol. 61: 4304-4309.
  17. Rinta-Kanto, J. M. and S. W. Wilhelm. 2006. Diversity of microcystin-producing cyanobacteria in spatially isolated regions of Lake Erie. Appl. Environ. Microbiol. 72: 5083-5085. https://doi.org/10.1128/AEM.00312-06
  18. Saker, M. L., M. Vale, D. Kramer, and V. M. Vasconcelos. 2007. Molecular techniques for the early warning of toxic cyanobacteria blooms in freshwater lakes and rivers. Appl. Microbiol. Biotechnol. 75: 441-449. https://doi.org/10.1007/s00253-006-0813-8
  19. Shirai, M., K. Matsumara, A. Ohotake, Y. Takamura, T. Aida, and M. Nakano. 1989. Development of a solid medium for growth and isolation of axenic microcystin strain (cyanobacteria). App. Environ. Microbiol. 55: 2569-2571.
  20. Tamura, K., J. Dudley, M. Nei, and S. Kumar. 2007. MEGA4: Molecular Evolutionary Genetics Analysis (MEGA) software version 4.0. Mol. Biol. Evol. 24: 1596-1599. https://doi.org/10.1093/molbev/msm092
  21. Vazquez-Martinez, G., M. H. Rodriguez, F. Hernandez-Hernandez, and J. E. Ibarra. 2004. Strategy to obtain axenic cultures from field-collected samples of the cyanobacterium Phormidium animalis. J. Microbiol. Methods 57: 115-121. https://doi.org/10.1016/j.mimet.2003.12.003
  22. Vezie, C., J. Rapala, J. Vaitomaa, J. Seitsonen, and K. Sivonen, K. 2002. Effect of nitrogen and phosphorus on growth of toxic and nontoxic Microcystis strains and on intracellular microcystin concentrations. Microb. Ecol. 43: 443-454. https://doi.org/10.1007/s00248-001-0041-9

Cited by

  1. Simultaneous Quantification of Cyanobacteria and Microcystis spp. Using Real-Time PCR vol.22, pp.2, 2010, https://doi.org/10.4014/jmb.1109.09051
  2. Quantification of toxigenic Microcystis spp. in freshwaters by quantitative real-time PCR based on the microcystin synthetase A gene vol.51, pp.1, 2010, https://doi.org/10.1007/s12275-013-2354-z