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PCR-based identification of microcystin-producing genotypes of different cyanobacterial genera

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Abstract

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.

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Fig. 1 A
Fig. 2A–C
Fig. 3
Fig. 4 A

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Abbreviations

RFLP :

Restriction fragment length polymorphism

MALDI-TOF :

Matrix-assisted laser desorption/ionization-time of flight spectrometry

HPLC :

High performance liquid chromatography

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Acknowledgements

This study was supported by grants from the European Commission (TOPIC project FMRX-CT97-0097 and CYANOTOX project ENV4-CT98-0892) to T.B. and K.S. We are grateful to Dr. Chantal Vezie and Prof. Vitor M. Vasconcelos for allowing use of Lake Grand Lieu Microcystis strains and the Portuguese strain Microcystis IZANCYA5, respectively, in our study, and to Lorena Via-Ordorika for providing the environmental samples. We thank Dr. Jutta Fastner for HPLC analyses and Dr. Marcel Erhard for MALDI-TOF data from strain NIVA-CYA34. Jana Müller and Matti Wahlsten are thanked for their excellent technical assistance, and Prof. Elke Dittmann for critical reading of the manuscript.

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  1. Michael Hisbergues

    Present address: Institute for Biology of Lille, Department of Microbiology of Ecosystems, 1 rue du Professeur Calmette, 59019 , Lille cedex, France

Authors and Affiliations

  1. Department of Biology (Genetics), Humboldt University, Chausseestrasse 117, 10115 , Berlin, Germany

    Michael Hisbergues, Guntram Christiansen & Thomas Börner

  2. Department of Applied Chemistry and Microbiology, Helsinki University, Biocenter Viikki, P. O. Box 56, 00014 , Helsinki, Finland

    Leo Rouhiainen & Kaarina Sivonen

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  1. Michael Hisbergues
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  2. Guntram Christiansen
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  4. Kaarina Sivonen
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Correspondence to Thomas Börner.

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Hisbergues, M., Christiansen, G., Rouhiainen, L. et al. PCR-based identification of microcystin-producing genotypes of different cyanobacterial genera. Arch Microbiol 180, 402–410 (2003). https://doi.org/10.1007/s00203-003-0605-9

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