Chinese Journal of Oceanology and Limnology

, Volume 30, Issue 2, pp 256–263

Application of rRNA probes and fluorescence in situ hybridization for rapid detection of the toxic dinoflagellate Alexandrium minutum

  • Xianghai Tang唐祥海
  • Rencheng Yu于仁成
  • Mingjiang Zhou周名江
  • Zhigang Yu于志刚
Biology

DOI: 10.1007/s00343-012-1142-7

Cite this article as:
Tang, X., Yu, R., Zhou, M. et al. Chin. J. Ocean. Limnol. (2012) 30: 256. doi:10.1007/s00343-012-1142-7

Abstract

The dinoflagellate Alexandrium minutum is often associated with harmful algal blooms (HABs). This species consists of many strains that differ in their ability to produce toxins but have similar morphology, making identification difficult. In this study, species-specific rRNA probes were designed for whole-cell fluorescence in situ hybridization (FISH) to distinguish A. minutum from two phylogenetic clades. We acquired the complete SSU to LSU rDNA sequences (GenBank accession numbers JF906989-JF906999) of 11 Alexandrium strains and used these to design rRNA targeted oligonucleotide probes. Three ribotype-specific probes, M-GC-1, M-PC-2, and M-PC-3, were designed. The former is specific for the GC clade (“Global clade”) of A. minutum, the majority of which have been found non-toxic, and the latter two are specific for the PSP (paralytic shellfish poisoning)-producing PC clade (“Pacific clade”). The specificity of these three probes was confirmed by FISH. All cells in observed fields of view were fluorescently labeled when probes and target species were incubated under optimized FISH conditions. However, the accessibility of rRNA molecules in ribosomes varied among the probe binding positions. Thus, there was variation in the distribution of positive signals in labeled cells within nucleolus and cytosol (M-GC-1, M-PC-3), or just nucleolus (M-PC-2). Our results provide a methodological basis for studying the biogeography and population dynamics of A. minutum, and providing an early warning of toxic HABs.

Keyword

fluorescence in situ hybridization (FISH)Alexandrium minutumrRNA probe

Copyright information

© Chinese Society for Oceanology and Limnology, Science Press and Springer-Verlag Berlin Heidelberg 2012

Authors and Affiliations

  • Xianghai Tang唐祥海
    • 1
  • Rencheng Yu于仁成
    • 2
  • Mingjiang Zhou周名江
    • 2
  • Zhigang Yu于志刚
    • 1
  1. 1.Key Laboratory of Marine Chemistry Theory and Technology, Ministry of EducationOcean University of ChinaQingdaoChina
  2. 2.Key Laboratory of Marine Ecology and Environmental Sciences, Institute of OceanologyChinese Academy of SciencesQingdaoChina