Journal of Environmental Science International (한국환경과학회지)

The Korean Environmental Sciences Society (한국환경과학회)
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Strain Identification and Comparative Analysis of Toxigenic Cyanobacteria Determined by PCR

  • Jung Jong-Mun (Busan Water Quality Institute, Water Authority) ;
  • Jung Eun-Young (Busan Water Quality Institute, Water Authority) ;
  • LEE You-Jung (Busan Water Quality Institute, Water Authority) ;
  • Park Hong-Ki (Busan Water Quality Institute, Water Authority) ;
  • Jung Mi-Eun (Busan Water Quality Institute, Water Authority) ;
  • Ji Ki-Won (Busan Water Quality Institute, Water Authority) ;
  • Joo Gea-Jae (Department of Biology, Pusan National University)
  • Published : 2005.07.01
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Abstract

Microcystis aeruginosa is common form of cyanobacteria (blue-green algae) capable of producing toxic heptapeptide (microcystin) that cause illness or death. The comparison of molecular genetic method with the morphological characteristics of cyanobacteria was conducted. We have designed PCR primers (JJM98F, JJM1141R) for cyanobacterial 16S rRNA and phycocyanin intergenic spacer (PC-IGS) gene domain. To confirm the production of microcystins, PCR primers for the N-methyltransferase (NMT) domain of microcystin synthetase gene mcyA were designed using 21 cyanobacteria strains Most of isolated strains from the Nakdong River was classified as Microcystis aeruginosa and the similarities were $99\%$ with M. aeruginosa AF 139292. $38.1\%$ of isolated strains contained microcystin synthesis gene. NMT (N-methyltransferase) were not detected in isolated strain in several strains, which means non-toxic. However, the NMTs of the strains were detected during the cultivation.

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References

  1. Carmichael, W. W., 1997, The cyanotoxins. In: Callow, J. A. (ED.) Advances in Botanicak research, 27, Academic press, London, 211- 256pp
  2. Ha, K., H. W. Kim and G. J. Joo, 1998, The phytoplankton succession in the lower part of hypertrophic Nakdong River (Mulgum), South Korea, Hydrobiologia, 369/370, 217-227 https://doi.org/10.1023/A:1017067809089
  3. Oh, H. M., S. J. Lee, M. H. Jang and B. D. Yoon, 2000, Microcystin production by Microcystis aeruginosa in a phosphorus-limited chemostat, Appl. Environ. Microbiol., 66, 176-179 https://doi.org/10.1128/AEM.66.1.176-179.2000
  4. Park, H. D., E. B. 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
  5. Ben-Amotz, A., T. G. Tornabene and W. H. Thomas, 1985, chemical profile of selected species of microalgae with emphasis on lipids, J. Phycol, 21, 72-81 https://doi.org/10.1111/j.0022-3646.1985.00072.x
  6. Komarek, J., 1994, Current trends and species delimination in the cyanoprokaryote taxanomy, Archiv fur Hydrobiologie (suppliment: Algological Studies), 75, 11-29
  7. Tatsumi, K., M. F. Watanabe and M. Watanabe, 1991, Allozyme divergence in Anabaena(Cyanophyceae) and its taxonomic inference, Arch. Hydro bioI. Suppl., 92, Algological Studies, 6, 129-140
  8. Neilan, B. A., P. T. Cox, P. R. Hawkins and A. E. Goodman, 1994, 16S ribosomal DNA gene sequence and phylogeny of toxic Microcystis sp. (Cyanobacteria), DNA Sequence, 4:333-337 https://doi.org/10.3109/10425179409020861
  9. Nubel, D., F. Garcia-Pichel and G. Muyzer, 1997, PCR Primers to Amplify 16S rRNA Genes from Cyanobacteria, Appl. Environ. Microbiol., 63, 3327-3332
  10. Rudi, K., O. M. Skulberg and K. S. Jakobsen, 1998, Evolution of Cyanobacteria by Exchange of Genetic Material among Phyletically Related Strains, J. Bacteriol., 180, 3453-3461
  11. Neilan, B. A., D. Jacobs and A. E. Goodman, 1995, Genetic Diversity and Phylogeny of Toxic Cyanobacteria Determined by DNA polymorphisms within the phycocyanin Locus, Appl. Environ. Microbiol., 61, 3875-3883
  12. Wilson, K. M., M. A. Schembri, P. D. Baker and C. P. Saint, 2000, Molecular Characterization of Toxic Cyanobacterium Cylindrospermopsis raciborskii and Design of a Species-Specific PCR, Appl. Environ. Microbiol., 66, 332-338 https://doi.org/10.1128/AEM.66.1.332-338.2000
  13. Harada, K. I., K. Matsuura and M. Suzuki, 1988, Analysis and purification of toxic peptides from cyanobacteria by reversed-phase high-performance liquid chromatography, J. Chromatography, 448, 275-283 https://doi.org/10.1016/S0021-9673(01)84589-1
  14. Wilmotte, A., J. M. Neefs and R. D. Wachter, 1994, Evolutionary affiliation of the marine nitrogen-fixing cyanobacterium Trichodesmium sp. strain NIBB 1067, derived by 16S ribosomal RNA sequence analysis, Microbiology, 140, 2159-2164 https://doi.org/10.1099/13500872-140-8-2159
  15. Otsuka, S., S. Suda, R. Li, M. Watanabe, H. Oyaizu, S. Matsumoto and M. M. Watanabe, 1998, 16S rRNA sequence and phylogenetic analysis of Microcystis strains with and without phycoerythrin, FEMS Microbiol. Lett., 164, 119-124 https://doi.org/10.1111/j.1574-6968.1998.tb13076.x
  16. Otsuka, S., S. Suda, R. Li, M. Watanabe, H. Oyaizu, S. Matsumoto and M. M. Watanabe, 1999, Phylogenetic relationships between toxic and nontoxic strains of the genus Microcystis based on 16S to 23 S internal transcribed spacer sequence, FEMS Microbiol Lett., 172, 15-21 https://doi.org/10.1111/j.1574-6968.1999.tb13443.x
  17. Neilan, B. A., E. Dittmann, J. Rouhiainen, R. A. Bass, V. Schaub, K. Sivonen and T. Bomer, 1999, N onribosomal Peptide Synthesis and Toxigenicity of Cyanobacteria, J. Bacteriol., 181(13), 4089-4097
  18. Nishizawa, T., M. Asayama, K. Fujii, K. Harada and M. Shirai, 1999, Genetic analysis of the peptide synthetase genes for a cylic heptapeptide microcystin in Microcystis, spp. J. Biochem., 126, 520-529 https://doi.org/10.1093/oxfordjournals.jbchem.a022481