Development of Molecular Identification Method for

Authors: ChoongJae Kim ChangHoon Kim Yoshihiko Sako

Publish Date: 2005/05/17

Volume: 7, Issue: 3, Pages: 215-222

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Abstract

We have developed a method to identify species in the genus Alexandrium using wholecell fluorescent in situ hybridization with FITClabeled oligonucleotide probes that target large subunit ribosomal rRNA molecules The probes were designed based on the sequence of the rDNA D1D2 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 crossreactivity to noncorresponding Alexandrium species In addition no crossreactivity 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 speciesThis work was supported by a grant R02200000226 from the Basic Research Program of the Korea Science and Engineering Foundation and supported in part by a grantinaid for scientific research 13556033 from the Ministry of Education Culture Sports Science and Technology of JAPAN

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