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http://dx.doi.org/10.5657/fas.2004.7.4.175

Molecular Identification of the Toxic Alexandrium tamiyavanichii (Dinophyceae) by the Whole-cell FISH Method  

Kim Choong-Jae (Korea Research Institute of Bioscience and Biotechnology, Environmental Biotechnology Laboratory)
Yoshimatsu Sada-Akfi (Akashowo Research Institute of Kagawa Prefecture)
Sako Yoshihiko (Department of Applied Biosciences, Graduate School of Agriculture, Kyoto University)
Kim Chang-Hoon (Department of Aquaculture, Pukyoung National University)
Publication Information
Fisheries and Aquatic Sciences / v.7, no.4, 2004 , pp. 175-183 More about this Journal
Abstract
The dinoflagellate Alexandrium tamiyavanichii Balech, a producer of toxins causing paralytic shellfish poisoning (PSP), has recently been considered as one of main organisms responsible for toxication of shellfish in Japan. In this study, A. tamiyavanichii was subjected to a molecular phylogenetic analysis inferred from 28S rDNA D1-D2 sequences and a species-specific LSU rRNA-targeted oligonucleotide DNA probe was designed to identify A. tamiyavanichii using the whole cell-FISH (fluorescence in situ hybridization). The sequences of the 28S rDNA D1-D2 region of A. tamiyavanichii showed no difference from A. cohorticular AF1746l4 (present name A. tamiyavanichii) and formed a distinct clade from the 'tamarensis species complex'. The probe, TAMID2, reacted specifically with A. tamiyavanichii cultured cells, without any cross-reaction with other species belonging to the same genus, including A. tamarense, A. catenella, A. affine, A. fraterculus, A. insuetum and A. pseudogonyaulax. In a test of cross-reactivity with a field sample, TAMID2 reacted consistently with only A. tamiyavanichii, indicating that the present protocol involving the TAMID2 probe might be useful for detecting toxic A. tamiyavanichii in a simple and rapid manner.
Keywords
Whole cell-FISH (fluorescence in situ hybridization); DNA probe; Alexandrium tamiyavanichii;
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