Rapid and Accurate Identification by Real-Time PCR of Biotoxin-Producing Dinoflagellates from the  Family Gymnodiniaceae

Smith, Kirsty F.; De Salas, Miguel; Adamson, Janet; Rhodes, Lesley L.

doi:10.3390/md12031361

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Rapid and Accurate Identification by Real-Time PCR of Biotoxin-Producing Dinoflagellates from the Family Gymnodiniaceae

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Cawthron Institute, 98 Halifax Street East, Private Bag 2, Nelson 7042, New Zealand

Tasmanian Herbarium, Tasmanian Museum and Art Gallery, Private Bag 4, Hobart, Tasmania 7001, Australia

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Mar. Drugs 2014, 12(3), 1361-1376; https://doi.org/10.3390/md12031361

Received: 10 October 2013 / Revised: 27 January 2014 / Accepted: 18 February 2014 / Published: 7 March 2014

(This article belongs to the Special Issue Advances and New Perspectives in Marine Biotechnology)

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Abstract

The identification of toxin-producing dinoflagellates for monitoring programmes and bio-compound discovery requires considerable taxonomic expertise. It can also be difficult to morphologically differentiate toxic and non-toxic species or strains. Various molecular methods have been used for dinoflagellate identification and detection, and this study describes the development of eight real-time polymerase chain reaction (PCR) assays targeting the large subunit ribosomal RNA (LSU rRNA) gene of species from the genera Gymnodinium, Karenia, Karlodinium, and Takayama. Assays proved to be highly specific and sensitive, and the assay for G. catenatum was further developed for quantification in response to a bloom in Manukau Harbour, New Zealand. The assay estimated cell densities from environmental samples as low as 0.07 cells per PCR reaction, which equated to three cells per litre. This assay not only enabled conclusive species identification but also detected the presence of cells below the limit of detection for light microscopy. This study demonstrates the usefulness of real-time PCR as a sensitive and rapid molecular technique for the detection and quantification of micro-algae from environmental samples. View Full-Text

Keywords: real-time PCR; large subunit ribosomal RNA (LSU rRNA) gene; dinoflagellate monitoring; biotoxin production

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Smith, K.F.; De Salas, M.; Adamson, J.; Rhodes, L.L. Rapid and Accurate Identification by Real-Time PCR of Biotoxin-Producing Dinoflagellates from the Family Gymnodiniaceae. Mar. Drugs 2014, 12, 1361-1376. https://doi.org/10.3390/md12031361

AMA Style

Smith KF, De Salas M, Adamson J, Rhodes LL. Rapid and Accurate Identification by Real-Time PCR of Biotoxin-Producing Dinoflagellates from the Family Gymnodiniaceae. Marine Drugs. 2014; 12(3):1361-1376. https://doi.org/10.3390/md12031361

Chicago/Turabian Style

Smith, Kirsty F., Miguel De Salas, Janet Adamson, and Lesley L. Rhodes. 2014. "Rapid and Accurate Identification by Real-Time PCR of Biotoxin-Producing Dinoflagellates from the Family Gymnodiniaceae" Marine Drugs 12, no. 3: 1361-1376. https://doi.org/10.3390/md12031361

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Rapid and Accurate Identification by Real-Time PCR of Biotoxin-Producing Dinoflagellates from the Family Gymnodiniaceae

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