Skip to main content
Applied and Environmental Microbiology logoLink to Applied and Environmental Microbiology
. 1995 Nov;61(11):3875–3883. doi: 10.1128/aem.61.11.3875-3883.1995

Genetic diversity and phylogeny of toxic cyanobacteria determined by DNA polymorphisms within the phycocyanin locus.

B A Neilan 1, D Jacobs 1, A E Goodman 1
PMCID: PMC167692  PMID: 8526499

Abstract

Cyanobacteria are a highly diverse group in relation to form, function, and habitat. Current cyanobacterial systematics relies on the observation of minor and plastic morphological characters. Accurate and reliable delineation of toxic and bloom-forming strains of cyanobacteria has not been possible by traditional methods. We have designed general primers to the phycocyanin operon (cpc gene) and developed a PCR which allows the amplification of a region of this gene, including a variable intergenic spacer sequence. Because of the specificity of this PCR for cyanobacterial isolates, the assay is appropriate for the rapid and reliable identification of strains in freshwater samples. Successive restriction endonuclease digestion of this amplification product, with a total of nine enzymes, yielded many identifying DNA profiles specific to the various taxonomic levels of cyanobacteria. The restriction enzyme profiles for MspI, RsaI, and TaqI were conserved for strains within each of the eight genera (40 strains) studied and clearly discriminated among these genera. Intrageneric delineation of strains was revealed by the enzymes AluI, CfoI, and HaeIII for members of the genus Microcystis, while strains of genus Anabaena were differentiated by the digestion patterns provided by AluI, CfoI, and ScrFI. Phenetic and cladistic analyses of the data were used to infer the genetic relatedness and evolution of toxic and bloom-forming cyanobacteria.

Full Text

The Full Text of this article is available as a PDF (491.6 KB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Akopyanz N., Bukanov N. O., Westblom T. U., Berg D. E. PCR-based RFLP analysis of DNA sequence diversity in the gastric pathogen Helicobacter pylori. Nucleic Acids Res. 1992 Dec 11;20(23):6221–6225. doi: 10.1093/nar/20.23.6221. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Altschul S. F., Gish W., Miller W., Myers E. W., Lipman D. J. Basic local alignment search tool. J Mol Biol. 1990 Oct 5;215(3):403–410. doi: 10.1016/S0022-2836(05)80360-2. [DOI] [PubMed] [Google Scholar]
  3. Belknap W. R., Haselkorn R. Cloning and light regulation of expression of the phycocyanin operon of the cyanobacterium Anabaena. EMBO J. 1987 Apr;6(4):871–884. doi: 10.1002/j.1460-2075.1987.tb04833.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Britschgi T. B., Giovannoni S. J. Phylogenetic analysis of a natural marine bacterioplankton population by rRNA gene cloning and sequencing. Appl Environ Microbiol. 1991 Jun;57(6):1707–1713. doi: 10.1128/aem.57.6.1707-1713.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Carmichael W. W. Cyanobacteria secondary metabolites--the cyanotoxins. J Appl Bacteriol. 1992 Jun;72(6):445–459. doi: 10.1111/j.1365-2672.1992.tb01858.x. [DOI] [PubMed] [Google Scholar]
  6. Caudales R., Wells J. M. Differentiation of free-living Anabaena and Nostoc cyanobacteria on the basis of fatty acid composition. Int J Syst Bacteriol. 1992 Apr;42(2):246–251. doi: 10.1099/00207713-42-2-246. [DOI] [PubMed] [Google Scholar]
  7. Darrasse A., Priou S., Kotoujansky A., Bertheau Y. PCR and restriction fragment length polymorphism of a pel gene as a tool to identify Erwinia carotovora in relation to potato diseases. Appl Environ Microbiol. 1994 May;60(5):1437–1443. doi: 10.1128/aem.60.5.1437-1443.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Dubbs J. M., Bryant D. A. Molecular cloning and transcriptional analysis of the cpeBA operon of the cyanobacterium Pseudanabaena species PCC7409. Mol Microbiol. 1991 Dec;5(12):3073–3085. doi: 10.1111/j.1365-2958.1991.tb01867.x. [DOI] [PubMed] [Google Scholar]
  9. Giovannoni S. J., Turner S., Olsen G. J., Barns S., Lane D. J., Pace N. R. Evolutionary relationships among cyanobacteria and green chloroplasts. J Bacteriol. 1988 Aug;170(8):3584–3592. doi: 10.1128/jb.170.8.3584-3592.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Glazer A. N. Light guides. Directional energy transfer in a photosynthetic antenna. J Biol Chem. 1989 Jan 5;264(1):1–4. [PubMed] [Google Scholar]
  11. Mullis K. B., Faloona F. A. Specific synthesis of DNA in vitro via a polymerase-catalyzed chain reaction. Methods Enzymol. 1987;155:335–350. doi: 10.1016/0076-6879(87)55023-6. [DOI] [PubMed] [Google Scholar]
  12. Nei M., Li W. H. Mathematical model for studying genetic variation in terms of restriction endonucleases. Proc Natl Acad Sci U S A. 1979 Oct;76(10):5269–5273. doi: 10.1073/pnas.76.10.5269. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Neilan B. A., Cox P. T., Hawkins P. R., Goodman A. E. 16S ribosomal RNA gene sequence and phylogeny of toxic Microcystis sp. (cyanobacteria). DNA Seq. 1994;4(5):333–337. doi: 10.3109/10425179409020861. [DOI] [PubMed] [Google Scholar]
  14. Neilan B. A., Gurvitz A., Leigh D. A., Lai L. Y., McDonald B. Rapid preparation of limited biological samples for small-volume PCR. PCR Methods Appl. 1993 Feb;2(3):261–262. doi: 10.1101/gr.2.3.261. [DOI] [PubMed] [Google Scholar]
  15. Neilan B. A. Identification and phylogenetic analysis of toxigenic cyanobacteria by multiplex randomly amplified polymorphic DNA PCR. Appl Environ Microbiol. 1995 Jun;61(6):2286–2291. doi: 10.1128/aem.61.6.2286-2291.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Padhy R. N., Hottat F. G., Coene M. M., Hoet P. P. Restriction analysis and quantitative estimation of methylated bases of filamentous and unicellular cyanobacterial DNAs. J Bacteriol. 1988 Apr;170(4):1934–1939. doi: 10.1128/jb.170.4.1934-1939.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Saitou N., Nei M. The neighbor-joining method: a new method for reconstructing phylogenetic trees. Mol Biol Evol. 1987 Jul;4(4):406–425. doi: 10.1093/oxfordjournals.molbev.a040454. [DOI] [PubMed] [Google Scholar]
  18. Schopf J. W., Packer B. M. Early Archean (3.3-billion to 3.5-billion-year-old) microfossils from Warrawoona Group, Australia. Science. 1987 Jul 3;237:70–73. doi: 10.1126/science.11539686. [DOI] [PubMed] [Google Scholar]
  19. Woese C. R. Bacterial evolution. Microbiol Rev. 1987 Jun;51(2):221–271. doi: 10.1128/mr.51.2.221-271.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. de Lorimier R., Bryant D. A., Porter R. D., Liu W. Y., Jay E., Stevens S. E., Jr Genes for the alpha and beta subunits of phycocyanin. Proc Natl Acad Sci U S A. 1984 Dec;81(24):7946–7950. doi: 10.1073/pnas.81.24.7946. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. de Lorimier R., Wilbanks S. M., Glazer A. N. Genes of the R-phycocyanin II locus of marine Synechococcus spp., and comparison of protein-chromophore interactions in phycocyanins differing in bilin composition. Plant Mol Biol. 1993 Jan;21(2):225–237. doi: 10.1007/BF00019939. [DOI] [PubMed] [Google Scholar]

Articles from Applied and Environmental Microbiology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES