DNA amplification fingerprinting as a tool for checking genetic purity of strains in the cyanobacterial inoculum Summary
The electrophoretic patterns for 17 different cyanobacterial cultures derived from 6 different decamer primers were analysed to provide diagnostic fingerprints for each culture and their genetic distances based on RAPD markers.The primer OPB 09 produced a maximum of 24 amplified products. The primers OPB 09, OPG 04 and OPAH 02 generated markers specific for
cultures. Westiellopsis was found to be distinct from other cyanobacterial cultures in the RAPD profile obtained with the primer OPAH 02. The primer OPF 03 generated specific markers for Tolypothrix tenuis. Fischerella cultures could be identified with the primers OPB 09, OPAG 03 and OPF 05. The study revealed that these RAPD markers could be further used to identify and establish the genetic purity of the strains in the cyanobacterial inoculum. There was a similarity of 60–90% within Westiellopsis cultures. Nostoc
cultures shared 50–80% similarity with Westiellopsis cultures. Anabaenacultures were similar to Westiellopsiscultures by 60–70. The markers produced for each culture were also applied to phylogenetic analysis to infer genetic relatedness in this group of prokaryotes. The dendrogram analysis clearly revealed that free-living cyanobacterial cultures are closely related to each other and are distinct from the symbiotic forms. Keywords Cyanobacteria cluster analysis genetic purity RAPD–PCR profile strain identification References Clark, M.S. 1997Plant Molecular Biology – A laboratory manual Springer-Verlag New York Google Scholar Doers, M.P., Paker, D.L. 1988 Properties of Microcystis aeruginosa and M. flosaquae in culture: taxonomic implications Journal of Phycology 24 502 508 Google Scholar Eskew, D.L., Caetano-Anolles, G., Bassam, B.J., Gresshoff, P.M. 1993 DNA amplification fingerprinting of the Azolla–Anabaena symbiosis Plant Molecular Biology 21 263 373 CrossRef Google Scholar Giovannoni, S.J., Turner, S., Olsen, G.J., Barns, S., Lane, D.J., Pace, N.R. 1988 Evolutionary relationship among cyanobacteria and green chloroplasts Journal of Bacteriology 170 3584 3592 PubMed Google Scholar Kannaiyan, S. 1993Biofertilizers for Rice Tamil Nadu Agricultural University Coimbatore: Tamil Nadu, India 60 Google Scholar Kannaiyan, S. 1985Studies on the Algal Application for Low Land Rice Crop Tamil Nadu Agricultural University Coimbatore: Tamil Nadu, India 24 Google Scholar Kato, T., Watanabe, M.F., Watanabe, M. 1991 Allozyme divergence in Microcystis and its taxonomic inference Archives of Hydrobiology Suppl. Bd 64 129 140 Google Scholar Kelly, M.L., Cowie, D.B. 1972 DNA-DNA hybridization studies of blue-green algae Carnegie Institute of Washington Yearbook 71 276 287 Google Scholar Kumar, K., Annapoorna, N., Kannaiyan, S. 2000Determination of genetic purity of the strains in the cyanobacterial inoculants by RAPD-PCR techniques Paper presented in the Asian Pacific Conference on Plant Tissue Culture and Agribiotechnology (APCPTCA) National University of Singapore Singapore 229 Google Scholar Neilan, B.A. 1995 Identification and phylogenic analysis of toxigenic cyanobacteria by multiple randomly amplified polymorphic DNA-PCR Applied and Environmental Microbiology 61 2286 2291 Google Scholar Nelissen, B., De Baere, R., Wilmotte, A., De Wachter, R. 1996 Phylogenetic relationships of non-axenic filamentous cyanobacterial strains based on 16S rRNA sequence analysis Journal of Molecular Evolution 42 194 200 PubMed Google Scholar Nierzwicki-Bauer, S.A., Haselkorn, R. 1986 Differences in mRNA levels in Anabaena living freely or in symbiotic association with Azolla EMBO Journal 5 29 35 PubMed Google Scholar Nishihara, H., Miwa, H., Watanabe, M., Nagashima, M., Yagi, O., Takamura, Y. 1997 Random Amplified Polymorphic DNA (RAPD) analysis for discriminating genotypes of Microcystis cyanobacteria Bioscience Biotechnology and Biochemistry 61 1067 1072 Google Scholar Rasmussen, U., Svenning, M.M. 1998 Fingerprinting of cyanobacteria based on PCR with primers derived from short and long tandemly repeated repetitive sequences Applied and Environmental Microbiology 64 265 272 Google Scholar Rippka, R., Deruelles, J., Waterbury, J.B., Herdman, M., Stanier, M. 1979 Genetic assignments, strain histories and properties of pure culture of cyanobacteria Journal of General Microbiology 111 1 61 Google Scholar Roger, P.A., Watanabe, I. 1986 Technologies for utilizing biological nitrogen fixation in Wetland rice Potentialities current usage and limiting factors Dutta, S.K. Patrick, N.H., eds. Nitrogen Economy in Flooded Rice Soils. Jr. Martinus Nijhoff Publishers Dordrecht The Netherlands 37 77 Google Scholar
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