GENETIC CHARACTERIZATION OF STRAINS OF CYANOBACTERIA USING PCR‐RFLP OF THE cpcBA INTERGENIC SPACER AND FLANKING REGIONS 1

Oligonucleotide primers, specific for conserved regions of the genes encoding the β‐ and α‐phycocyanin subunits of phycobilisomes (cpcB and cpcA) of cyanobacteria, were used to amplify a DNA fragment containing the intervening intergenic spacer region (cpcBA‐IGS) of 19 strains of three morphospecies of cyanobacteria. Six Australian strains were identified as Anabaena circinalis Rabenhorst, six strains were identified as Microcystis aeruginosa Kützing, and seven strains were identified as Nodularia spumigena Mertens. Restriction enzyme digestion of the amplification products from the strains revealed restriction fragment length polymorphism (RFLP) within all three morphospecies. Strains corresponding to M. aeruginosa were highly polymorphic: 11 of the 14 restriction enzymes used displayed RFLPs. The A. circinalis and N. spumigena strains were less variable: three of 14 enzymes and seven of 14 enzymes, respectively, showed RFLPs. The presence of genetic variation between strains within these three divergent morphospecies, which span two orders of cyanobacteria (Chroococcales Wettstein and Nostocales (Borzi) Geitler), show that the cpcBA‐ IGS fragment has broad application as a molecular marker for intrageneric studies of cyanobacteria systematics and genetics.

[1]  J. Sambrook,et al.  Molecular Cloning: A Laboratory Manual , 2001 .

[2]  F. Zechman,et al.  USE OF RIBOSOMAL DNA INTERNAL TRANSCRIBED SPACERS FOR PHYLOGENETIC STUDIES IN DIATOMS 1 , 1994 .

[3]  R. Gasser,et al.  Species identification of trichostrongyle nematodes by PCR-linked RFLP. , 1994, International journal for parasitology.

[4]  F. Martin,et al.  Identification of the ectomycorrhizal basidiomycete Tylospora fibrillosa Donk by RFLP analysis of the PCR-amplified ITS and IGS regions of ribosomal DNA. , 1994, The New phytologist.

[5]  E. Carpenter,et al.  GENOTYPIC RELATIONSHIPS IN TRICHODESMIUM (CYANOPHYCEAE) BASED ON nifH SEQUENCE COMPARISONS 1 , 1993 .

[6]  R. Castenholz,et al.  SPECIES USAGE, CONCEPT, AND EVOLUTION IN THE CYANOBACTERIA (BLUE‐GREEN ALGAE) , 1992 .

[7]  K. Kersters,et al.  DNA probes for Bordetella species and a colorimetric reverse hybridization assay for the detection of Bordetella pertussis. , 1992, Molecular and cellular probes.

[8]  R. Grumet,et al.  A quick and inexpensive method for removing polysaccharides from plant genomic DNA. , 1992, BioTechniques.

[9]  J. Komárek,et al.  A review of water bloom forming microcystis species with regard to populations from japan , 1991 .

[10]  J. Komárek,et al.  Modern approach to the classification system of cyanophytes 5. stigonematales , 1990 .

[11]  D. Townsend,et al.  DNA POLYMORPHISM WITHIN THE WH7803 SEROGROUP OF MARINE SYNECHOCOCCUS SPP. (CYANOBACTERIA) 1, 2 , 1990 .

[12]  D. L. Parker,et al.  PROPERTIES OF MICROCYSTIS AERUGINOSA AND M. FLOS‐AQUAE (CYANOPHYTA) IN CULTURE: TAXONOMIC IMPLICATIONS 1 , 1988 .

[13]  S. Douglas,et al.  Examination of Genetic Relatedness of Marine Synechococcus spp. by Using Restriction Fragment Length Polymorphisms , 1988, Applied and environmental microbiology.

[14]  S. R. Barnum,et al.  Rapid small-scale DNA isolation from filamentous cyanobacteria , 1988 .

[15]  D. Mazel,et al.  Green light induces transcription of the phycoerythrin operon in the cyanobacterium Calothrix 7601. , 1986, Nucleic acids research.

[16]  W. Stam,et al.  Genotypic relationships between strains of Anabaena (Cyanophyceae) and their correlation with morphological affinities , 1984 .

[17]  W. Sidler,et al.  Phycobilisome and Phycobiliprotein Structures , 1994 .

[18]  A. Wilmotte Molecular Evolution and Taxonomy of the Cyanobacteria , 1994 .

[19]  R. Rippka Isolation and purification of cyanobacteria. , 1988, Methods in enzymology.

[20]  K. Anagnostidis Modern approach to the classification system of cyanophytes 1-Introduction. , 1985 .