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PCR detection of dinoflagellate cysts in field sediment samples from tropic and temperate environments
Department of Fishery Microbiology, College of Fisheries, University of Agricultural Sciences, Mangalore.
Kristianstad University, School of Education and Environment, Avdelningen för Naturvetenskap. Kristianstad University, Forskningsmiljön Man and Biosphere Health (MABH).ORCID iD: 0000-0002-8059-0156
Department of Fishery Microbiology, College of Fisheries, University of Agricultural Sciences, Mangalore.
Department of Fishery Microbiology, College of Fisheries, University of Agricultural Sciences, Mangalore.
2002 (English)In: Harmful Algae, ISSN 1568-9883, E-ISSN 1878-1470, Vol. 1, no 4, p. 361-373Article in journal (Refereed) Published
Abstract [en]

Species-specific primers were constructed for Scrippsiella trochoidea, Protoceratium reticulatum and Lingulodinium polyedrum, which all are common cosmopolitan cyst forming dinoflagellates. The designed primers amplified a product of expected size from cultured planktonic cells of the three species, and did not yield any product with a wide range of other algal species used as negative controls. The PCR method for detection and identification of dinoflagellate cysts from the three species was applied on field samples. Undisturbed surface sediment was collected along the southwest coast of India and the west coast of Sweden. DNA extract from sediment including DNA from dinoflagellate cysts could be obtained after repeated grinding with mortar and pestle under liquid nitrogen followed by microwave boiling. All sediment samples that contained any of the target species as confirmed by microscopy, were also positive for PCR. Field samples negative for any of the target species by microscopy, were also negative by PCR. Restriction enzyme digestion and/or DNA sequencing confirmed the specificity of all the PCR products from field samples. The yield of DNA from sediment extraction was low, and therefore nested PCR was necessary for accurate species-specific detection of the three species in most of the field samples.

Place, publisher, year, edition, pages
2002. Vol. 1, no 4, p. 361-373
Keywords [en]
Cyst, Dinoflagellate, Dinophyceae, Field samples, PCR, Species-specific primers
National Category
Biological Sciences
Identifiers
URN: urn:nbn:se:hkr:diva-10972DOI: 10.1016/S1568-9883(02)00053-7OAI: oai:DiVA.org:hkr-10972DiVA, id: diva2:642040
Available from: 2013-08-20 Created: 2013-08-20 Last updated: 2017-12-06Bibliographically approved

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Rehnstam-Holm, Ann-Sofi

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Avdelningen för NaturvetenskapForskningsmiljön Man and Biosphere Health (MABH)
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