| SS4.02 Ecology and Physiology of Marine Organisms: Insights from Genes, Genomes, and Proteomes |
| Huang, B, Q, Marine Environmental Laboratory, Xiamen, China, bqhuang@jingxian.xmu.edu.cn |
| Chen, J, X, Marine Environmental Laboratory, Xiamen, China, smmd@sohu.com |
| Jiao, N, Z, Marine Environmental Laboratory, Xiamen, China, jiao@xmu.edu.cn |
| Hong, J, S, Marine Environmental Laboratory, Xiamen, China, hshong@xmu.edu.cn |
| Chen, F, , Center of Marine Biotechnology, Baltimore, USA, chenf@umbi.umd.edu |
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| FLUORESCENT IN SITU HYBRIDIZATION WITH OLIGONUCLEOTIDE PROBES TO IDENTIFY ALEXANDRIUM TAMARENSE AND GENUS ALEXANDRIUM |
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| This study was laid out to develop oligonucleotide probes that can be coupled with fluorescent in situ hybridization (FISH) to specifically detect the bloom-forming Alexandrium spp. and the species A. tamarense, respectively. Multiple probes were designed based on the sequences of 5S, 28S rDNA, and 18-28S internal transcribed spacer region, respectively. Three complementary probes (Tama28s01, Tama5s01, Tamaits01) specific for A. tamarense and one (Alex24s01) specific for genus Alexandrium, labeled with fluorescein, were tested on several different algal species using FISH method. The hybridization results were detected by both flow cytometry (FCM) and epifluorescence microscopy (EFM). The three A. tamarense-specific probes could discriminate A. tamarense from other species. The hybridization signal of each probe was affected by the growth stage of algae. Probes Tama28s01 and Tama5s01 yielded much brighter hybridization signal than probe Tamaits01. Targeted cells and negative control cells were well separated using FCM. Three auxiliary probes were designed to enhance the efficiency of hybridization. The probes and protocols developed here can be used to monitor in situ distribution of Alexandrium spp. and specific species of A. tamarenser. |
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