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Applied and Environmental Microbiology, April 2001, p. 1839-1845, Vol. 67, No. 4
0099-2240/01/$04.00+0   DOI: 10.1128/AEM.67.4.1839-1845.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Varied Diazotrophies, Morphologies, and Toxicities of Genetically Similar Isolates of Cylindrospermopsis raciborskii (Nostocales, Cyanophyceae) from Northern Australia

Martin L. Saker1,* and Brett A. Neilan2

Departamento de Zoologia e Antropologia, Faculdade de Ciências, Praça Gomes Teixeira, 4050 Porto, Portugal,1 and School of Microbiology and Immunology, University of New South Wales, Sydney, New South Wales 2051, Australia2

Received 6 November 2000/Accepted 31 January 2001

The potentially toxic freshwater cyanobacterium Cylindrospermopsis raciborskii has become increasingly prevalent in tropical and temperate water bodies worldwide. This paper investigates the effects of different nitrogen sources (NO3-, NH4+, and omission of a fixed form of nitrogen) on the growth rates, morphologies, and cylindrospermopsin (CYL) concentrations (expressed as a percentage of the freeze-dried weight) of seven C. raciborskii isolates obtained from a range of water bodies in northern Australia and grown in batch culture. In general, growth rates were lowest in the absence of a fixed-nitrogen source and highest with NH4+ as the nitrogen source. Conversely, the highest concentrations of CYL were recorded in cultures grown in the absence of a fixed-nitrogen source and the lowest were found in cultures supplied with NH4+. Cultures supplied with NO3- were intermediate with respect to both CYL concentration and growth rate. Different nitrogen sources resulted in significant differences in the morphology of C. raciborskii trichomes. Most notable were the loss of heterocysts and the tapering of end cells in cultures supplied with NH4+ and the statistically significant increase in vegetative cell length (nitrogen depleted < NO3- < NH4+). The morphological changes induced by different nitrogen sources were consistent for all isolates, despite measurable differences in vegetative-cell and heterocyst dimensions among isolates. Such induced morphological variation has implications for Cylindrospermopsis taxonomy, given that distinctions between species are based on minor and overlapping differences in cell lengths and widths. The close phylogenetic association among all seven isolates was confirmed by the high level (>99.8%) of similarity of their 16S rRNA gene sequences. Another genetic technique, analysis of the HIP1 octameric-palindrome repeated sequence, showed greater heterogeneity among the isolates and appears to be a useful method for distinguishing among isolates of C. raciborskii.

* Corresponding author. Mailing address: Departamento de Zoologia e Antropologia, Faculdade de Ciências, Praça Gomes Teixeira, 4050 Porto, Portugal. Phone: 351 22 340 1516. Fax: 351 22 340 1511. E-mail: msaker{at}fc.up.pt.

Applied and Environmental Microbiology, April 2001, p. 1839-1845, Vol. 67, No. 4
0099-2240/01/$04.00+0   DOI: 10.1128/AEM.67.4.1839-1845.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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