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Applied and Environmental Microbiology, June 2009, p. 3598-3604, Vol. 75, No. 11
0099-2240/09/$08.00+0     doi:10.1128/AEM.02448-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.

Identifying the Source of Unknown Microcystin Genes and Predicting Microcystin Variants by Comparing Genes within Uncultured Cyanobacterial Cells{triangledown}

Christopher J. Allender,1 Gary R. LeCleir,1 Johanna M. Rinta-Kanto,2 Randall L. Small,3 Michael F. Satchwell,4 Gregory L. Boyer,4 and Steven W. Wilhelm1*

Department of Microbiology, University of Tennessee, Knoxville, Tennessee 37996,1 Department of Marine Sciences, University of Georgia, Athens, Georgia 30602,2 Department of Ecology and Evolutionary Biology, University of Tennessee, Knoxville, Tennessee 37996,3 Department of Biochemistry, State University of New York College of Environmental Science and Forestry, Syracuse, New York 132104

Received 24 October 2008/ Accepted 1 April 2009

While multiple phylogenetic markers have been used in the culture-independent study of microcystin-producing cyanobacteria, in only a few instances have multiple markers been studied within individual cells, and in all cases these studies have been conducted with cultured isolates. Here, we isolate and evaluate large DNA fragments (>6 kb) encompassing two genes involved in microcystin biosynthesis (mcyA2 and mcyB1) and use them to identify the source of gene fragments found in water samples. Further investigation of these gene loci from individual cyanobacterial cells allowed for improved analysis of the genetic diversity within microcystin producers as well as a method to predict microcystin variants for individuals. These efforts have also identified the source of the novel mcyA genotype previously termed Microcystis-like that is pervasive in the Laurentian Great Lakes and they predict the microcystin variant(s) that it produces.

* Corresponding author. Mailing address: Department of Microbiology, The University of Tennessee, Knoxville, TN 37996-0845. Phone: (865) 974-0665. Fax: (865) 974-4007. E-mail: Wilhelm{at}utk.edu

{triangledown} Published ahead of print on 10 April 2009.

Applied and Environmental Microbiology, June 2009, p. 3598-3604, Vol. 75, No. 11
0099-2240/09/$08.00+0     doi:10.1128/AEM.02448-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.





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