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Nitrogen and phosphorus budgets of the Changjiang River estuary

ISSN号：0254-4059
期刊名称：《中国海洋湖沼学报：英文版》
时间：0
分类：Q949.24[生物学—植物学] Q943[生物学—植物学]
作者机构：[1]Key Laboratory of Marine Chemistry Theory and Technology, Ministry of Education, Ocean University of China, Qingdao 266100, China, [2]Key Laboratory of Marine Ecology and Environmental Sciences, Institute of Oceanology, Chinese Academy of Sciences, Qingdao 266071, China
相关基金：Supported by the National Natural Science Foundation of China （No. 41006082）, the Postdoctoral Science Foundation （No. 20090461272）, and the National Natural Science Foundation of China for Creative Research Groups （No. 40821004）

作者：李祥安[1,2], 俞志明[1], 宋秀贤[1], 曹西华[1], 袁涌铨[1]

关键词：微小亚历山大藻, 荧光原位杂交, 基因探针, RRNA, 有毒甲藻, 快速检测, 荧光标记探针, GENBANK, fuorescence in situ hybridization （FISH）, Alexandrium minutum, rRNA probe

中文摘要：

dinoflagellate Alexandrium minutum 经常与有害海藻的花蕾(HAB ) 被联系。这种由处于他们生产毒素，但是有类似的形态学的能力不同的许多紧张组成，使鉴定困难。在这研究，种类特定的 rRNA 探针在 situ 杂交(鱼) 为整个房间的荧光被设计区分 A。从二种系发生的 clades 的 minutum。我们获得了完全的 SSU 到 11 Alexandrium 紧张的 LSU rDNA 序列(GenBank 就职数字 JF906989-JF906999 ) 并且使用了这些设计指向的 oligonucleotide 探查的 rRNA。三根 ribotype 特定的探针， M-GC-1， M-PC-2，和 M-PC-3，被设计。前者为 A 的 GC clade (全球 clade ) 是特定的。minutum，的多数被发现无毒，并且二是的后者为生产 PC clade (和平的 clade ) 的 PSP (毒害的麻痹贝壳) 特定。这三根探针的特性被鱼证实。当探针和目标种类在优化的鱼条件下面被孵化时，看法的观察领域里的所有房间荧光灯地被标记。然而，在核糖体的 rRNA 分子的可接近性在探查绑定位置之中变化了。因此，在在在核和 cytosol 以内的标记的房间的积极信号的分发有变化(M-GC-1， M-PC-3 ) ，或就核(M-PC-2 ) 。我们的结果为学习生物地理学和 A 的人口动力学提供一个方法学的基础。minutum，并且提供有毒的 HAB 的一个早警告。

英文摘要：

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.