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

IOCAS-IR

	Application of rRNA probes and fluorescence in situ hybridization for rapid detection of the toxic dinoflagellate Alexandrium minutum
	Tang XH(唐祥海); Yu RC(于仁成); Zhou MJ(周名江); Yu ZG(于志刚)
发表期刊	Chinese Journal of Oceanology and Limnology
ISSN	0254-4059
	2012-03-15
出版年	2012
卷号	v.30 期号:02 页码:66-73
文献类型	CNKI期刊论文
摘要	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.
关键词	fluorescence in situ hybridization (FISH) Alexandrium minutum rRNA probe
CNKI专辑号	A;B;
CNKI专辑名称	基础科学;工程科技Ⅰ辑;
CNKI专题号	B027;
CNKI专题名称	环境科学与资源利用;
分类号	X835
收录类别	中科院核心
语种	英文;
资助项目(基金）	Supported by the National Natural Science Foundation of China(No.41006082) ; the Postdoctoral Science Foundation(No.20090461272) ; the National Natural Science Foundation of China for Creative Research Groups(No.40821004)
文献类型	CNKI期刊论文
条目标识符	http://ir.qdio.ac.cn/handle/337002/191331
专题	中国科学院海洋研究所
作者单位	1.KeyLaboratoryofMarineChemistryTheoryandTechnology,MinistryofEducation,OceanUniversityofChina 2.KeyLaboratoryofMarineEcologyandEnvironmentalSciences,InstituteofOceanology,ChineseAcademyofSciences
推荐引用方式 GB/T 7714	Tang XH,Yu RC,Zhou MJ,et al. Application of rRNA probes and fluorescence in situ hybridization for rapid detection of the toxic dinoflagellate Alexandrium minutum. 2012.