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Development of Molecular Identification Method for Genus Alexandrium (Dinophyceae) Using Whole-Cell FISH

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Abstract

We have developed a method to identify species in the genus Alexandrium using whole-cell fluorescent in situ hybridization with FITC-labeled oligonucleotide probes that target large subunit ribosomal rRNA molecules. The probes were designed based on the sequence of the rDNA D1-D2 region of Alexandrium species. DNA probes specific for toxic A. tamarense and A. catenella and nontoxic A. affine, A. fraterculus, A. insuetum, and A. pseudogonyaulax, respectively, were applied to vegetative cells of all above Alexandrium species to test the sensitivity of the probes. Each DNA probe hybridized specifically with vegetative cells of the corresponding Alexandrium species and showed no cross-reactivity to noncorresponding Alexandrium species. In addition, no cross-reactivity of the probes was observed in experiments using concentrated natural seawater samples. The TAMAD2 probe, which is highly specific to A. tamarense, a common toxic species in Korean coastal waters, provides a simple and reliable molecular tool for identification of toxic Alexandrium species.

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Acknowledgments

This work was supported by a grant (R02-2000-00226) from the Basic Research Program of the Korea Science and Engineering Foundation, and supported in part by a grant-in-aid for scientific research (13556033) from the Ministry of Education, Culture, Sports, Science and Technology of JAPAN.

We declare that the experiments performed comply with the current laws of the country in which the experiments were performed.

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Authors and Affiliations

  1. Environmental Biotechnology Laboratory, Korea Research Institute of Bioscience & Biotechnology, 305-333, Yuseong-gu, Daejeon, Korea

    Choong-Jae Kim

  2. Department of Aquaculture, Pukyong National University, 599-1 Daeyeon-3-dong, 608-737 Nam Gu, Busan, South Korea

    Chang-Hoon Kim

  3. Department of Applied Biosciences, Graduate School of Agriculture, Kyoto University, 606-8502, Kyoto, Japan

    Yoshihiko Sako

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  1. Choong-Jae Kim
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Correspondence to Choong-Jae Kim.

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Kim, CJ., Kim, CH. & Sako, Y. Development of Molecular Identification Method for Genus Alexandrium (Dinophyceae) Using Whole-Cell FISH. Mar Biotechnol 7, 215–222 (2005). https://doi.org/10.1007/s10126-004-0424-2

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  • DOI: https://doi.org/10.1007/s10126-004-0424-2

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