LOCALIZATION OF MICROCYSTIN SYNTHETASE GENES IN COLONIES OF THE CYANOBACTERIUM MICROCYSTIS USING FLUORESCENCE IN SITU HYBRIDIZATION(1)

James S Metcalf; Marianne Reilly; Fiona M Young; Geoffrey A Codd

doi:10.1111/j.1529-8817.2009.00750.x

LOCALIZATION OF MICROCYSTIN SYNTHETASE GENES IN COLONIES OF THE CYANOBACTERIUM MICROCYSTIS USING FLUORESCENCE IN SITU HYBRIDIZATION(1)

J Phycol. 2009 Dec;45(6):1400-4. doi: 10.1111/j.1529-8817.2009.00750.x. Epub 2009 Sep 29.

Authors

James S Metcalf¹, Marianne Reilly¹, Fiona M Young¹, Geoffrey A Codd¹

Affiliation

¹ Division of Molecular Microbiology, College of Life Sciences, University of Dundee, Dundee, DD1 4HN, UK.

PMID: 27032597
DOI: 10.1111/j.1529-8817.2009.00750.x

Abstract

To better understand the production of microcystins (MCs) in Microcystis colonies, fluorescence in situ hybridization (FISH) methods were developed to detect DNA involved in the synthesis of these cyanobacterial hepatotoxins. Using colonies of Microcystis aeruginosa (Kütz.) Kütz. isolated from environmental blooms of cyanobacteria and from a colony-forming, MC-producing laboratory strain of Microcystis, amplified PCR products were observed, coincident with positive controls. The total MC content of individual colonies of Microcystis, determined by ELISA, showed a positive correlation with colony cross-sectional area. FISH analysis of Microcystis colonies gave high fluorescence in comparison to negative controls, indicating the presence of MC synthetase DNA (mcyA) in situ. FISH analysis for MC synthetase genes has the potential to be developed into an effective early warning tool for drinking and recreational water management.

Keywords: DNA; ELISA; Microcystis; PCR; fluorescence in situ hybridization; microcystin.