Genetic variability of microcystin biosynthesis genes in Planktothrix as elucidated from samples preserved by heat desiccation during three decades.

Historic samples of phytoplankton can provide information on the abundance of the toxigenic genotypes of cyanobacteria in dependence on increased or decreased eutrophication. The analysis of a time-series from preserved phytoplankton samples by quantitative PCR (qPCR) extends observation periods con...

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Main Authors: Veronika Ostermaier, Guntram Christiansen, Ferdinand Schanz, Rainer Kurmayer
Format: Article
Language:English
Published: Public Library of Science (PLoS) 2013-01-01
Series:PLoS ONE
Online Access:https://doi.org/10.1371/journal.pone.0080177
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author Veronika Ostermaier
Guntram Christiansen
Ferdinand Schanz
Rainer Kurmayer
author_facet Veronika Ostermaier
Guntram Christiansen
Ferdinand Schanz
Rainer Kurmayer
author_sort Veronika Ostermaier
collection DOAJ
description Historic samples of phytoplankton can provide information on the abundance of the toxigenic genotypes of cyanobacteria in dependence on increased or decreased eutrophication. The analysis of a time-series from preserved phytoplankton samples by quantitative PCR (qPCR) extends observation periods considerably. The analysis of DNA from heat-desiccated samples by qPCR can be aggravated by point substitutions or the fragmentation of DNA introduced by the high temperature. In this study, we analyzed whether the heat desiccation of the cellular material of the cyanobacterium Planktothrix sp. introduced potential errors to the template DNA that is used for qPCR within (i) 16S rDNA and phycocyanin genes and (ii) the mcyA gene indicative of the incorporation of either dehydrobutyrine (Dhb) or N-methyl-dehydroalanine (Mdha) in position 7, and (ii) the mcyB gene, which is indicative of homotyrosine (Hty) in position 2 of the microcystin (MC) molecule. Due to high temperature desiccation, the deterioration of the DNA template quality was rather due to fragmentation than due to nucleotide substitutions. By using the heat-desiccated samples of Lake Zürich, Switzerland the abundance of the Dhb, Mdha and Hty genotypes was determined during three decades (1977-2008). Despite major changes in the trophic state of the lake resulting in a major increase of the total Planktothrix population density, the proportion of these genotypes encoding the synthesis of different MC congeners showed high stability. Nevertheless, a decline of the most abundant mcyA genotype indicative of the synthesis of Dhb in position 7 of the MC molecule was observed. This decline could be related to the gradual incline in the proportion of a mutant genotype carrying a 1.8kbp deletion of this gene region. The increase of this mcyA (Dhb) gene deletion mutant has been minor so far, however, and likely did not affect the overall toxicity of the population.
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spelling doaj-art-67c36a0214ac49af9679eba35f9119782025-08-20T02:34:14ZengPublic Library of Science (PLoS)PLoS ONE1932-62032013-01-01811e8017710.1371/journal.pone.0080177Genetic variability of microcystin biosynthesis genes in Planktothrix as elucidated from samples preserved by heat desiccation during three decades.Veronika OstermaierGuntram ChristiansenFerdinand SchanzRainer KurmayerHistoric samples of phytoplankton can provide information on the abundance of the toxigenic genotypes of cyanobacteria in dependence on increased or decreased eutrophication. The analysis of a time-series from preserved phytoplankton samples by quantitative PCR (qPCR) extends observation periods considerably. The analysis of DNA from heat-desiccated samples by qPCR can be aggravated by point substitutions or the fragmentation of DNA introduced by the high temperature. In this study, we analyzed whether the heat desiccation of the cellular material of the cyanobacterium Planktothrix sp. introduced potential errors to the template DNA that is used for qPCR within (i) 16S rDNA and phycocyanin genes and (ii) the mcyA gene indicative of the incorporation of either dehydrobutyrine (Dhb) or N-methyl-dehydroalanine (Mdha) in position 7, and (ii) the mcyB gene, which is indicative of homotyrosine (Hty) in position 2 of the microcystin (MC) molecule. Due to high temperature desiccation, the deterioration of the DNA template quality was rather due to fragmentation than due to nucleotide substitutions. By using the heat-desiccated samples of Lake Zürich, Switzerland the abundance of the Dhb, Mdha and Hty genotypes was determined during three decades (1977-2008). Despite major changes in the trophic state of the lake resulting in a major increase of the total Planktothrix population density, the proportion of these genotypes encoding the synthesis of different MC congeners showed high stability. Nevertheless, a decline of the most abundant mcyA genotype indicative of the synthesis of Dhb in position 7 of the MC molecule was observed. This decline could be related to the gradual incline in the proportion of a mutant genotype carrying a 1.8kbp deletion of this gene region. The increase of this mcyA (Dhb) gene deletion mutant has been minor so far, however, and likely did not affect the overall toxicity of the population.https://doi.org/10.1371/journal.pone.0080177
spellingShingle Veronika Ostermaier
Guntram Christiansen
Ferdinand Schanz
Rainer Kurmayer
Genetic variability of microcystin biosynthesis genes in Planktothrix as elucidated from samples preserved by heat desiccation during three decades.
PLoS ONE
title Genetic variability of microcystin biosynthesis genes in Planktothrix as elucidated from samples preserved by heat desiccation during three decades.
title_full Genetic variability of microcystin biosynthesis genes in Planktothrix as elucidated from samples preserved by heat desiccation during three decades.
title_fullStr Genetic variability of microcystin biosynthesis genes in Planktothrix as elucidated from samples preserved by heat desiccation during three decades.
title_full_unstemmed Genetic variability of microcystin biosynthesis genes in Planktothrix as elucidated from samples preserved by heat desiccation during three decades.
title_short Genetic variability of microcystin biosynthesis genes in Planktothrix as elucidated from samples preserved by heat desiccation during three decades.
title_sort genetic variability of microcystin biosynthesis genes in planktothrix as elucidated from samples preserved by heat desiccation during three decades
url https://doi.org/10.1371/journal.pone.0080177
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