2024-03-29T09:47:28Zhttp://digital.csic.es/dspace-oai/requestoai:digital.csic.es:10261/266872020-11-12T09:43:51Zcom_10261_123com_10261_8col_10261_376
00925njm 22002777a 4500
dc
Penna, Antonella
author
Bertozzini, Elena
author
Battocchi, Cecilia
author
Galluzzi, Luca
author
Giacobbe, Maria Grazia
author
Vila, Magda
author
Garcés, Esther
author
Lugliè, Antonella
author
Magnani, Mauro
author
2007-01
A qualitative and semi-quantitative polymerase chain reaction (PCR)-based assay was developed for the detection of several potentially Harmful Algal Bloom (HAB) species and genera belonging to Dinophyceae, Bacillariophyceae and Raphydophyceae. Oligonucleotide primers were designed based on Internal Transcribed Spacer (ITS)–5.8S ribosomal DNA (rDNA) sequences available in public database or identified in this study. The specificity and sensitivity of the PCR assay were validated using clonal cultures and then natural seawater samples, as well as the known copy number of plasmids containing the target ITS–5.8S rDNA regions. A filter system for collecting mixed phytoplankton cells coupled to a target species-specific PCR assay was performed on spatial and temporal series of net and surface seawater samples during coastal water monitoring carried out in several localities of the Mediterranean Sea. The application of PCR allowed a rapid detection of various genera and species-specific potential HAB taxa in all examined natural samples. Qualitative and semi-quantitative PCR results obtained from field samples were compared with microscopic [light microscope (LM)] examinations. The two methods gave comparable results, and the molecular assay was able to detect HAB target cells at concentrations not detectable by microscopy or those of uncertain identity. The highest values of positive detection of potential HAB taxon presence obtained by PCR assay compared with the microscopic examination ranged from 67 to 8.0%
Journal of Plankton Research 29(1): 19-38 (2007)
0142-7873
http://hdl.handle.net/10261/26687
10.1093/plankt/fbl053
1464-3774
Monitoring of HAB species in the Mediterranean Sea through molecular methods