Browse > Article

Molecular Characterization of Marine Cyanobacteria from the Indian Subcontinent Deduced from Sequence Analysis of the Phycocyanin Operon (cpcB-IGS-cpcA) and 16S-23S ITS Region  

Premanandh, Jagadeesan (National Facility for Marine Cyanobacteria, Bharathidasan University)
Priya, Balakrishnan (National Facility for Marine Cyanobacteria, Bharathidasan University)
Teneva, Ivanka (Department of Botany, Faculty of Biology, University of Plovdiv)
Dzhambazov, Balik (Department of Cell and Molecular Biology, Lund University)
Prabaharan, Dharmar (National Facility for Marine Cyanobacteria, Bharathidasan University)
Uma, Lakshmanan (National Facility for Marine Cyanobacteria, Bharathidasan University)
Publication Information
Journal of Microbiology / v.44, no.6, 2006 , pp. 607-616 More about this Journal
Abstract
Molecular characterization of ten marine cyanobacterial isolates belonging to the order Oscillatoriales was carried out using the phycocyanin locus (cpcBA-IGS) and the 16S-23S internally transcribed spacer region. DNA sequences from the phycocyanin operon discriminated ten genotypes, which corresponded to seven morphotypes identified by traditional microscopic analysis. The cpcB coding region revealed 17% nucleotide variation, while cpcA exhibited 29% variation across the studied species. Phylogenetic analyses support the conclusion that the Phormidium and Leptolyngbya genera are not monophyletic. The nucleotide variations were heterogeneously distributed with no or minimal informative nucleotides. Our results suggest that the discriminatory power of the phycocyanin region varies across the cyanobacterial species and strains. The DNA sequence analysis of the 16S-23S internally transcribed spacer region also supports the polyphyletic nature of the studied oscillatorian cyanobacteria. This study demonstrated that morphologically very similar strains might differ genotypically. Thus, molecular approaches comprising different gene regions in combination with morphological criteria may provide better taxonomical resolution of the order Oscillatoriales.
Keywords
cyanobacteria; genetic diversity; IGS; ITS; Leptolyngbya; Phormidium; phycocyanin operon; phylogeny; taxonomy;
Citations & Related Records

Times Cited By Web Of Science : 5  (Related Records In Web of Science)
Times Cited By SCOPUS : 2
연도 인용수 순위
1 Barker, G.L., P.K. Hayes, and S.L. O'Mahony. 1999. A molecular and phenotypic analysis of Nodularia (Cyanobacteria) from the Baltic Sea. J. Phycol. 35, 931-937   DOI
2 Barker, G.L., A. Konopka, B.A. Handley, and P.K. Hayes. 2000b. Genetic variation in Aphanizomenon (Cyanobacteria) colonies from the Baltic Sea and North America. J. Phycol. 36, 947-950   DOI   ScienceOn
3 Boyer, S.L., V.R. Flechtner, and J.R. Johansen. 2001. Is the 16S-23S rRNA internal transcribed spacer region a good tool for use in molecular systematics and population genetics? A case study in cyanobacteria. Mol. Biol. Evol. 18, 1057-1069   DOI   ScienceOn
4 Crosbie, N.D., M. Pöckl, and T. Weisse. 2003. Dispersal and phylogenetic diversity of nonmarine Picocyanobacteria, inferred from 16S rRNA gene and cpcBA-Intergenic spacer sequence analyses. Appl. Environ. Microbiol. 69, 5716-5721   DOI
5 Dyble, J. H.W. Paerl, and B.A. Neilan. 2002. Genetic characterization of Cylindrospermopsis raciborskii (Cyanobacteria) isolates from diverse geographic origins based on nifH and cpcBA-IGS nucleotide sequence analysis. Appl. Environ. Microbiol. 68, 2567-2571   DOI
6 Komárek, J. and K. Anagnostidis. 1989. Modern approach to the classification system of cyanophytes. 4: Nostocales. Arch. Hydrobiol. Suppl./Algological Studies 56, 247-345
7 Manen, J.F. and J. Falque. 2002. The cpcB-cpcA locus as a tool for the genetic characterization of the genus Arthrospira (Cyanobacteria): evidence for horizontal transfer. Int. J. System. Evol. Microbiol. 52, 861-867   DOI   ScienceOn
8 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
9 Orcutt, K.M., U. Rasmussen, E.A. Webb, J.B. Waterbury, K. Gundersen, and B. Bergman. 2002. Characterization of Trichodesmium spp. by genetic techniques. Appl. Environ. Microbiol. 68, 2236-2245   DOI
10 Payne, M.C., J.R. Johansen, and S.L. Boyer. 2001. Taxonomic resolution of Leptolyngbya (Cyanophyta) utilizing the 16S rRNA gene sequence. 55th Annual Meeting of the Phycological Society of America, June 23-28, 2001, Estes Park, CO, USA; J. Phycol. 37, 40
11 Tillett, D., D.L. Parker, and B.A. Neilan. 2001. Detection of toxigenicity by a probe for the microcystin synthetase A gene (mcyA) of the cyanobacterial genus Microcystis: comparison of toxicities with 16S rRNA and phycocyanin operon (phycocyanin intergenic spacer) phylogenies. Appl. Environ. Microbiol. 67, 2810-2818   DOI   ScienceOn
12 Wu, X., A. Zarka, and S. Boussiba. 2000. A simplified protocol for preparing DNA from filamentous cyanobacteria. Plant. Mol. Biol. Rep. 18, 385-392   DOI
13 Barker, G.L., B.A. Handley, P. Vacharapiyasophon, J.R. Stevens, and P.K. Hayes. 2000a. Allele-specific PCR shows that genetic exchange occurs among genetically diverse Nodularia (Cyanobacteria) filaments in the Baltic Sea. Microbiology 146, 2865-2875   DOI
14 Young, J.M., 2001. Implications of alternative classifications and horizontal gene transfer for bacterial taxonomy. Int. J. System. Evol. Microbiol. 51, 945-953   DOI   ScienceOn
15 Stanier, R.Y., R. Kunisawa, M. Mandel, and G. Cohen-Bazire. 1971. Purification and properties of unicellular blue-green algae (order Chroococcales). Bacteriol. Rev. 35, 171-205
16 Felsenstein, J. 2004. PHYLIP (Phylogeny Inference Package) version 3.6. Distributed by the author. Department of Genome Sciences, University of Washington, Seattle, USA
17 Lu, W. 1999. Application of molecular biological techniques to the identification of cyanobacteria. Master's thesis, Liverpool John Moores University, England
18 Margheri, M.C., R. Piccardi, S. Ventura, C. Viti, and L. Giovannetti. 2003. Genotypic diversity of oscillatoriacean strains belonging to the genera Geitlerinema and Spirulina determined by 16S rDNA restriction analysis. Curr. Microbiol. 46, 359-364   DOI
19 Rippka, R., J. Deruelles, J.B. Waterbury, M. Herdman, and R.Y. Stanier. 1979. Generic assignments, strain histories and properties of pure cultures of cyanobacteria. J. Gen. Microbiol. 111, 1-61   DOI
20 Saker, M.L., B.A. Neilan, and D.J. Griffiths. 1999. Two morphological forms of Cylindrospermopsis raciborskii (Cyanobacteria) isolated from Solomon Dam, Palm Island, Queensland. J. Phycol. 35, 599-606   DOI
21 Lu, W., E.H. Evans, S.M. McColl, and V.A. Saunders. 1997. Identification of cyanobacteria by polymorphisims of PCRamplified ribosomal DNA spacer regions. FEMS Microbiol. Lett. 153, 147-149
22 Salomon, P.S., S. Janson, and E. Granéli. 2003. Molecular identification of bacteria associated with filaments of Nodularia spumigena and their effect on the cyanobacterial growth. Harmful. Algae 2, 261-272   DOI   ScienceOn
23 Iteman, I., R. Rippka, N. Tandeau De Marsac, and M. Herdman. 2000. Comparison of conserved structural and regulatory domains within divergent 16S rRNA-23S rRNA spacer sequences of cyanobacteria. Microbiol. 146, 1275-1286   DOI
24 Litvaitis, M.K. 2002. A molecular test of cyanobacterial phylogeny: inferences from constraint analyses. Hydrobiologia 468, 135-145   DOI   ScienceOn
25 Komarek, J. and K. Anagnostidis. 2005. Susswasserflora von Mitteleuropa Bd.19/2 Cyanoprokaryota, Oscillatoriales, Verlag: Spektrum Akademischer Verlag; Urban and Fischer Bei Els, 759 p
26 Prabaharan, D. 1988. Partial characterization of the marine cyanobacterium Phormidium valderianum 30501 for biotechnological purposes. M. Phil Dissertation, Bharathidasan University, Tiruchirapalli, India
27 Nübel, U., F. Garcia-Pichel, and G. Muyzer. 1997. PCR primers to amplify 16S rRNA genes from cyanobacteria. Appl. Environ. Microbiol. 63, 3327-3332
28 Castenholz, R.W. and J.B. Waterbury. 1989. Cyanobacteria. In Stanley, J.T., M.P. Bryant, N. Pfenning, and J.H. Holt (Eds.). Bergy's Manual Systematic Bacteriology, Vol. 3. Section 19: Oxygenic photosynthetic bacteria. Williams and Wilkins, Baltimore, USA
29 Fox, G.E., J.D. Wisotzkey, and P. Jurtshuk, Jr. 1992. How close is close: 16S rRNA sequence identity may not be sufficient to guarantee species identity. Int. J. Syst. Bacteriol. 42, 166-170   DOI   ScienceOn
30 Garcia-Pichel, F., L. Prufert-Bebout, and G. Muyzer. 1996. Phenotypic and phylogenetic analysis show Microcoleus chthonoplastes to be a cosmopolitan cyanobacterium. Appl. Environ. Microbiol., 62, 3284-3291
31 Teneva, I., B. Dzhambazov, R. Mladenov, and K. Schirmer. 2005. Molecular and phylogenetic characterization of Phormidium species (Cyanoprokaryota) using the cpcBGS- cpcA locus. J. Phycol. 41, 188-194   DOI   ScienceOn
32 Ballot, A., P. Dadheech, and L. Krienitz. 2004. Phylogenetic relationship of Arthrospira, Phormidium, and Spirulina strains from Kenyan and Indian waterbodies. Arch. Hydrobiol. Suppl./Algological Studies 113, 37-56
33 Baurain, D., L. Renquin, S. Grubisic, P. Scheldeman, A. Belay, and A. Wilmotte. 2002. Remarkable conservation of internally transcribed spacer sequences of Arthrospira ('Spirulina') (Cyanophyceae, Cyanobacteria) strains from four continents and of recent and 30-years-old dried samples from Africa. J. Phycol. 38, 384-393   DOI   ScienceOn
34 Bolch, C.J.S., S.I. Blackburn, B.A. Neilan, and P.M. Grewe. 1996. Genetic characterization of strains of cyanobacteria using PCR-RFLP of the phycocyanin intergenic spacer and flanking regions. J. Phycol. 32, 445-451   DOI   ScienceOn
35 Ishida, T., A. Yokota, and J. Sugiyama. 1997. Phylogenetic relationships of filamentous cyanobacterial taxa inferred from 16S rDNA sequence divergence. J. Gen. Appl. Microbiol. 43, 237-241   DOI   ScienceOn
36 Janson, S. and E. Granéli. 2002. Phylogenetic analyses of nitrogen-fixing cyanobacteria from the Baltic Sea reveal sequence anomalies in the phycocyanin operon. Int. J. System. Evol. Microbiol. 52, 1397-1404   DOI   ScienceOn
37 Li, X. 2000. Phenotypic and genotypic characterization of Nostoc species from 6 different sites in the Mojave desert. Master's thesis, John Carroll University, Cleveland, Ohio
38 Scheldeman, P., D. Baurain, R. Bouhy, M. Scott, M. Mühling, B.A. Whitton, A. Belay, and A. Wilmotte. 1999. Arthrospira ('Spirulina') strains from four continents are resolved in only two clusters, based on amplified ribosomal DNA restriction analysis on the internally transcribed spacer. FEMS Microbiol. Lett. 172, 213-222   DOI
39 Robertson, B.R., N. Tezuka, and M.M. Watanabe. 2001. Phylogenetic analyses of Synechococcus strains (Cyanobacteria) using sequences of 16S rDNA and part of the phycocyanin operon reveal multiple evolutionary lines and reflect phycobilin content. Int. J. Syst. Evol. Microbiol. 51, 861-871   DOI   ScienceOn
40 Taton, A., S. Grubusic, E. Brambilla, R.D. Wit, and A. Wilmotte. 2003. Cyanobacterial diversity in natural and artificial microbial mats of lake fryxell (Mc Murdo dry valleys, Antartica): a morphological and molecular approach. Appl. Environ. Microbiol. 69, 5157-5169   DOI
41 Otsuka, S., S. Suda, R. Li, S. Matsumoto, and M. Watanabe. 2000. Morphological variability of colonies of Microcystis morphospecies in culture. J. Gen. Appl. Microbiol. 46, 39-50   DOI   ScienceOn
42 Anagnostidis, K. and J. Komarek. 1985. Modern approach to the classification system of cyanophytes 1-Introduction. Arch. Hydrobiol./ Suppl. 71, Algological. Studies 38 / 39, 291-302