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Applied and Environmental Microbiology, January 2006, p. 117-123, Vol. 72, No. 1
0099-2240/06/$08.00+0     doi:10.1128/AEM.72.1.117-123.2006
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

Transposons Inactivate Biosynthesis of the Nonribosomal Peptide Microcystin in Naturally Occurring Planktothrix spp.{dagger}

Guntram Christiansen,1,3 Rainer Kurmayer,2* Qian Liu,2 and Thomas Börner3

Department of Chemistry, University of Hawaii at Manoa, 2545 McCarthy Mall, Honolulu, Hawaii 96822,1 Institute for Limnology, Austrian Academy of Sciences, Mondseestrasse 9, 5310 Mondsee, Austria,2 Institute for Biology (Genetics), Humboldt University, Chausseestrasse 117, D-10115 Berlin, Germany3

Received 2 August 2005/ Accepted 7 October 2005

The filamentous cyanobacteria Planktothrix spp. occur in the temperate region of the Northern hemisphere. The red-pigmented Planktothrix rubescens bacteria occur in deep, physically stratified, and less eutrophic lakes. Planktothrix is a known producer of the toxic heptapeptide microcystin (MC), which is produced nonribosomally by a large enzyme complex consisting of peptide synthetases and polyketide synthases encoded by a total of nine genes (mcy genes). Planktothrix spp. differ in their cellular MC contents as well as the production of MC variants; however, the mechanisms favoring this diversity are not understood. Recently, the occurrence of Planktothrix strains containing all mcy genes but lacking MC has been reported. In this study, 29 such strains were analyzed to find out if mutations of the mcy genes lead to the inability to synthesize MC. Two deletions, spanning 400 bp (in mcyB; one strain) and 1,869 bp (in mcyHA; three strains), and three insertions (IS), spanning 1,429 bp (in mcyD; eight strains), 1,433 bp (in mcyEG; one strain), and 1,433 bp (in mcyA; one strain), were identified. Though found in different genes and different isolates and transcribed in opposite directions, IS were found to be identical and contained conserved domains assigned to transposable elements. Using mutation-specific primers, two insertions (in mcyD and mcyA) and one deletion (in mcyHA) were found regularly in populations of P. rubescens in different lakes. The results demonstrate for the first time that different mutations resulting in inactivation of MC synthesis do occur frequently and make up a stable proportion of the mcy gene pool in Planktothrix populations over several years.

* Corresponding author. Mailing address: Austrian Academy of Sciences, Institute for Limnology, Mondseestrasse 9, 5310 Mondsee, Austria. Phone: 43-6232-312532. Fax: 43-6232-3578. E-mail: rainer.kurmayer{at}oeaw.ac.at

{dagger} Supplemental material for this article may be found at http://aem.asm.org/.

Applied and Environmental Microbiology, January 2006, p. 117-123, Vol. 72, No. 1
0099-2240/06/$08.00+0     doi:10.1128/AEM.72.1.117-123.2006
Copyright © 2006, American Society for Microbiology. All Rights Reserved.



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