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Applied and Environmental Microbiology, April 2007, p. 2552-2560, Vol. 73, No. 8
0099-2240/07/$08.00+0     doi:10.1128/AEM.02389-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.

Development of a Real-Time PCR Probe for Quantification of the Heterotrophic Dinoflagellate Cryptoperidiniopsis brodyi (Dinophyceae) in Environmental Samples{triangledown}

Tae-Gyu Park,1 Miguel F. de Salas,1 Christopher J. S. Bolch,2 and Gustaaf M. Hallegraeff1*

School of Plant Science, University of Tasmania, Private Bag 55, Hobart, Tasmania 7001, Australia,1 School of Aquaculture, University of Tasmania, Locked Bag 1-370, Launceston, Tasmania 7250, Australia2

Received 10 October 2006/ Accepted 12 February 2007

A TaqMan format real-time PCR probe was developed against the internal transcribed spacer 2 ribosomal DNA region for the specific detection and quantification of Cryptoperidiniopsis brodyi in environmental samples. The assay specificity was confirmed by testing against related dinoflagellates and verified by sequencing PCR amplicons from natural water samples. Phylogenetic analysis of the sequenced environmental samples also showed that this assay is specific to C. brodyi. The C. brodyi-specific assay was used in conjunction with Pfiesteria piscicida- and Pfiesteria shumwayae-specific real-time PCR assays to investigate the temporal variations of C. brodyi, P. piscicida, and P. shumwayae abundance in the Derwent estuary, Tasmania. The 18-month field survey from November 2004 to April 2006 revealed that C. brodyi occurred in all seasons at very low densities, mostly below 25 cells liter–1, with higher abundance (maximum, 112 cells liter–1) in April and May. P. piscicida was detected only once, in May 2005 at 60 cells liter–1. P. shumwayae was not detected during the survey.

* Corresponding author. Mailing address: School of Plant Science, University of Tasmania, Private Bag 55, Churchill Avenue, Sandy Bay, Hobart, Tasmania 7001, Australia. Phone: (61) 3 6226-2623. Fax: (61) 3 6226-2698. E-mail: Hallegraeff{at}utas.edu.au

{triangledown} Published ahead of print on 23 February 2007.

Applied and Environmental Microbiology, April 2007, p. 2552-2560, Vol. 73, No. 8
0099-2240/07/$08.00+0     doi:10.1128/AEM.02389-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Park, T.-G., Bolch, C. J. S., Hallegraeff, G. M. (2007). Larval Crassostrea bivalve and Artemia brine shrimp bioassays to assess toxicity and micropredation by the heterotrophic dinoflagellates Cryptoperidiniopsis brodyi and Pfiesteria piscicida from Australian waters. J PLANKTON RES 29: 791-801 [Abstract] [Full Text]  


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