PCR-based identification of microcystin-producing genotypes of different cyanobacterial genera.
Microcystins are harmful hepatotoxins produced by many, but not all strains of the cyanobacterial genera Anabaena, Microcystis, Anabaena, Planktothrix, and Nostoc. Waterbodies have to be monitored for the mass development of toxic cyanobacteria; however, because of the close genetic relationship of microcystin-producing and non-producing strains within a genus, identification of microcystin-producers by morphological criteria is not possible. The genomes of microcystin-producing cells contain mcy genes coding for the microcystin synthetase complex. Based on the sequence information of mcy genes from Microcystis and Planktothrix, a primer pair for PCR amplification of a mcyA gene fragment was designed. PCR with this primer pair is a powerful means to identify microcystin-producing strains of the genera Anabaena, Microcystis, and Planktothrix. Moreover, subsequent RFLP analysis of the PCR products generated genus-specific fragments and allowed the genus of the toxin producer to be identified. The assay can be used with DNA from field samples.
Hisbergues
Michael
M
Department of Biology, Humboldt University, Chausseestrasse 117, 10115 Berlin, Germany.
Christiansen
Guntram
G
Rouhiainen
Leo
L
Sivonen
Kaarina
K
Börner
Thomas
T
D017033:Anabaena / Q000145:classification / Q000235:genetics / Q000378:metabolism; D001426:Bacterial Proteins; D001427:Bacterial Toxins / Q000096:biosynthesis / Q000235:genetics; D001483:Base Sequence; D000458:Cyanobacteria / Q000145:classification* / Q000235:genetics* / Q000378:metabolism; D016172:DNA Fingerprinting; D004269:DNA, Bacterial / Q000737:chemistry / Q000302:isolation & purification; D005798:Genes, Bacterial; D052998:Microcystins; D008969:Molecular Sequence Data; D010453:Peptide Synthases / Q000235:genetics*; D010456:Peptides, Cyclic / Q000096:biosynthesis* / Q000235:genetics*; D016133:Polymerase Chain Reaction; D012150:Polymorphism, Restriction Fragment Length; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA; D012689:Sequence Homology, Nucleic Acid; D014871:Water Microbiology
D001426:Bacterial Proteins; D001427:Bacterial Toxins; D004269:DNA, Bacterial; D052998:Microcystins; D010456:Peptides, Cyclic; C078588:microcystin; D010453:Peptide Synthases; C432283:microcystin synthetase A