Abstract
Pseudanabaena species are poorly known filamentous bloom-forming cyanobacteria closely related to Limnothrix. We isolated 28 Pseudanabaena strains from the Baltic Sea (BS) and the Albufera de Valencia (AV; Spain). By combining phenotypic and genotypic approaches, the phylogeny, diversity and evolutionary diversification of these isolates were explored. Analysis of the in vivo absorption spectra of the Pseudanabaena strains revealed two coexisting pigmentation phenotypes: (i) phycocyanin-rich (PC-rich) strains and (ii) strains containing both PC and phycoerythrin (PE). Strains of the latter phenotype were all capable of complementary chromatic adaptation (CCA). About 65 kb of the Pseudanabaena genomes were sequenced through a multilocus sequencing approach including the sequencing of the16 and 23S rRNA genes, the ribosomal intergenic spacer (IGS), internal transcribed spacer 1 (ITS-1), the cpcBA operon encoding PC and the IGS between cpcA and cpcB. In addition, the presence of nifH, one of the structural genes of nitrogenase, was investigated. Sequence analysis of ITS and cpcBA-IGS allowed the differentiation between Pseudanabaena isolates exhibiting high levels of microdiversity. This multilocus sequencing approach revealed specific clusters for the BS, the AV and a mixed cluster with strains from both ecosystems. The latter comprised exclusively CCA phenotypes. The phylogenies of the 16 and 23S rRNA genes are consistent, but analysis of other loci indicated the loss of substructure, suggesting that the recombination between these loci has occurred. Our preliminary results on population genetic analyses of the PC genes suggest an evolutionary diversification of Pseudanabaena through purifying selection.
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Acknowledgements
We are grateful to Marije Doeleman for her assistance in sequencing and to Ute Wollenzien for isolation, culturing and maintenance of the Pseudanabaena strains. We thank Maayke Stomp and the crew of the RV Aranda for their assistance during sampling of the Baltic Sea. Special thanks to Caroline Jenkins for providing us the 0.45 μm filters during the Cyano-cruise in 2004 and Dr. Beatríz Díez for sharing her unpublished data with us. Research of JH was supported by the Earth and Life Sciences Foundation (ALW), which was subsidized by the Netherlands Organization for Scientific Research (NWO). SGA was supported in the framework of the Flemish-Dutch collaboration for Sea Research (VLANEZO), TH and LJS acknowledge the support from the European Commission through the project MIRACLE (EVK3-CT-2002–00087). This is publication 4365 of NIOO-KNAW.
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Acinas, S., Haverkamp, T., Huisman, J. et al. Phenotypic and genetic diversification of Pseudanabaena spp. (cyanobacteria). ISME J 3, 31–46 (2009). https://doi.org/10.1038/ismej.2008.78
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