Journal
DEEP-SEA RESEARCH PART II-TOPICAL STUDIES IN OCEANOGRAPHY
Volume 57, Issue 3-4, Pages 288-300Publisher
PERGAMON-ELSEVIER SCIENCE LTD
DOI: 10.1016/j.dsr2.2009.09.010
Keywords
HAB species; Mediterranean Sea; PCR; Ribosomal genes; Resting stages; Sediments
Categories
Funding
- EU [GOCE-CT-2005-003875]
- Spanish Ministry of Science and Education
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A PCR-based assay was developed and applied to sediment and sediment trap samples for the detection of different cysts belonging to dinoflagellates and raphidophytes in European coastal areas. Oligonucleotide primers were designed based on the ITS-5.8S and LSU ribosomal gene sequences. The specificity and sensitivity of the PCR assay were assessed using genomic DNA from clonal cultures, plasmid copy number of cloned target sequences, as well as from sediment samples. Qualitative PCR determinations of different cysts in sediment and sediment trap samples were compared to taxonomic examinations by light microscopy. This molecular methodology permitted a fast and specific detection of target cysts in sediment samples. We also detected dinoflagellate and raphidophyte cysts at concentrations not detectable by microscopic methods or that are difficult to identify. The results given by molecular and microscopic methods were comparable. However, higher values of positive detection for target cysts were obtained by PCR than with microscopy. Some taxa were detected in 100% of the samples using PCR, while others were only found in 10% of the samples. The data obtained in this study showed that the PCR-based method is a valid tool for cyst identification in marine sediments. (C) 2009 Elsevier Ltd. All rights reserved.
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