title: Identification of cyanobacteria and their toxigenicity in environmental samples by rapid molecular analysis.
abstract: We report molecular analyses which identify cyanobacterial strains present in environmental samples. These analyses do not require the isolation and culture of strains. Identification of cyanobacteria used the polymerase chain reaction (PCR), based on the phycocyanin operon. Differentiation was either by restriction endonuclease digestion (restriction fragment length polymorphisms) or sequencing of the PCR products. Identification was based on sequence homology of the intergenic spacer region (IGS) between the beta- and alpha-phycocyanin subunits (PC-IGS) with database records. We have found that the length and sequence of the PC-IGS is capable of predicting the genus accurately, but not the species. Toxigenicity was determined with oligonucleotide probes for key steps in the microcystin toxin synthesis pathway. We have shown that it is possible to easily and routinely obtain PCR amplification products and differentiate the strains in bloom samples. The methods can detect even minor components in bloom samples, which may not be apparent on microscopic examination. Genetic probes for microcystin toxigenicity are effective on environmental samples, eliminating the need for isolation and culture of the organisms. The use of a suite of tests described here will allow water managers to determine the presence and the type of cyanobacteria and their microcystin toxigenicity.
authors.affiliation: School of Biological Sciences, University of New England, Armidale, NSW 2351, Australia.
authors.forename: J A
authors.initials: JA
authors.lastname: Baker
authors.forename: B A
authors.initials: BA
authors.lastname: Neilan
authors.forename: B
authors.initials: B
authors.lastname: Entsch
authors.forename: D B
authors.initials: DB
authors.lastname: McKay
mesh_terms: D000818:Animals; D001315:Australia; D001483:Base Sequence; D000458:Cyanobacteria / Q000235:genetics* / Q000502:physiology; D015342:DNA Probes; D004262:DNA Restriction Enzymes; D003625:Data Collection; D051379:Mice; D052998:Microcystins; D008969:Molecular Sequence Data; D009152:Mutagenicity Tests; D010456:Peptides, Cyclic / Q000633:toxicity; D010798:Phycocyanin / Q000235:genetics / Q000378:metabolism; D016133:Polymerase Chain Reaction; D012150:Polymorphism, Restriction Fragment Length; D012689:Sequence Homology, Nucleic Acid; D014873:Water Pollutants / Q000032:analysis*
chemical_list: D015342:DNA Probes; D052998:Microcystins; D010456:Peptides, Cyclic; D014873:Water Pollutants; D010798:Phycocyanin; C078588:microcystin; D004262:DNA Restriction Enzymes