Rapid and Accurate Identification by Real-Time PCR of Biotoxin-Producing Dinoflagellates from the Family Gymnodiniaceae
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 dinofla...
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doaj-8561c9fb9b704bc9be791e597a1b411b2020-11-25T01:06:27ZengMDPI AGMarine Drugs1660-33972014-03-011231361137610.3390/md12031361md12031361Rapid and Accurate Identification by Real-Time PCR of Biotoxin-Producing Dinoflagellates from the Family GymnodiniaceaeKirsty F. Smith0Miguel de Salas1Janet Adamson2Lesley L. Rhodes3Cawthron Institute, 98 Halifax Street East, Private Bag 2, Nelson 7042, New ZealandTasmanian Herbarium, Tasmanian Museum and Art Gallery, Private Bag 4, Hobart, Tasmania 7001, AustraliaCawthron Institute, 98 Halifax Street East, Private Bag 2, Nelson 7042, New ZealandCawthron Institute, 98 Halifax Street East, Private Bag 2, Nelson 7042, New ZealandThe 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.http://www.mdpi.com/1660-3397/12/3/1361real-time PCRlarge subunit ribosomal RNA (LSU rRNA) genedinoflagellate monitoringbiotoxin production |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Kirsty F. Smith Miguel de Salas Janet Adamson Lesley L. Rhodes |
spellingShingle |
Kirsty F. Smith Miguel de Salas Janet Adamson Lesley L. Rhodes Rapid and Accurate Identification by Real-Time PCR of Biotoxin-Producing Dinoflagellates from the Family Gymnodiniaceae Marine Drugs real-time PCR large subunit ribosomal RNA (LSU rRNA) gene dinoflagellate monitoring biotoxin production |
author_facet |
Kirsty F. Smith Miguel de Salas Janet Adamson Lesley L. Rhodes |
author_sort |
Kirsty F. Smith |
title |
Rapid and Accurate Identification by Real-Time PCR of Biotoxin-Producing Dinoflagellates from the Family Gymnodiniaceae |
title_short |
Rapid and Accurate Identification by Real-Time PCR of Biotoxin-Producing Dinoflagellates from the Family Gymnodiniaceae |
title_full |
Rapid and Accurate Identification by Real-Time PCR of Biotoxin-Producing Dinoflagellates from the Family Gymnodiniaceae |
title_fullStr |
Rapid and Accurate Identification by Real-Time PCR of Biotoxin-Producing Dinoflagellates from the Family Gymnodiniaceae |
title_full_unstemmed |
Rapid and Accurate Identification by Real-Time PCR of Biotoxin-Producing Dinoflagellates from the Family Gymnodiniaceae |
title_sort |
rapid and accurate identification by real-time pcr of biotoxin-producing dinoflagellates from the family gymnodiniaceae |
publisher |
MDPI AG |
series |
Marine Drugs |
issn |
1660-3397 |
publishDate |
2014-03-01 |
description |
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. |
topic |
real-time PCR large subunit ribosomal RNA (LSU rRNA) gene dinoflagellate monitoring biotoxin production |
url |
http://www.mdpi.com/1660-3397/12/3/1361 |
work_keys_str_mv |
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