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Molecular probes and microarrays for the detection of toxic algae in the genera Dinophysis and Phalacroma (Dinophyta)

机译:用于检测Dinophysis和Phalacroma(Dinophyta)属中有毒藻类的分子探针和微阵列

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摘要

Dinophysis and Phalacroma species containing diarrheic shellfish toxins and pectenotoxins occur in coastal temperate waters all year round and prevent the harvesting of mussels during several months each year in regions in Europe, Chile, Japan, and New Zealand. Toxicity varies among morphologically similar species, and a precise identification is needed for early warning systems. Molecular techniques using ribosomal DNA sequences offer a means to identify and detect precisely the potentially toxic species. We designed molecular probes targeting the 18S rDNA at the family and genus levels for Dinophysis and Phalacroma and at the species level for Dinophysis acuminata, Dinophysis acuta, and Dinophysis norvegica, the most commonly occurring, potentially toxic species of these genera in Western European waters. Dot blot hybridizations with polymerase chain reaction (PCR)-amplified rDNA from 17 microalgae were used to demonstrate probe specificity. The probes were modified along with other published fluorescence in situ hybridization and PCR probes and tested for a microarray platform within the MIDTAL project (). The microarray was applied to field samples from Norway and Spain and compared to microscopic cell counts. These probes may be useful for early warning systems and monitoring and can also be used in population dynamic studies to distinguish species and life cycle stages, such as cysts, and their distribution in time and space.Electronic supplementary materialThe online version of this article (doi:10.1007/s11356-012-1403-1) contains supplementary material, which is available to authorized users.
机译:含有腹泻性贝类毒素和果胶毒素的恐龙和Ph类常年在沿海温带水域中生长,每年在欧洲,智利,日本和新西兰的地区,几个月中贻贝的捕捞都无法进行。毒性在形态相似的物种之间有所不同,因此预警系统需要精确识别。使用核糖体DNA序列的分子技术提供了一种方法,可以准确地识别和检测潜在的毒性物种。我们设计了针对18S rDNA的分子探针,该探针针对恐龙和Ph的家族和属水平,以及针对恐龙Dinophysis acuminata,Dinophysis acuta和Dinophysis norvegica的物种水平,这是这些属在欧洲西部水域中最常见,潜在有毒的物种。使用来自17个微藻的聚合酶链反应(PCR)扩增的rDNA进行斑点印迹杂交来证明探针特异性。这些探针与其他已发表的荧光原位杂交和PCR探针一起进行了修饰,并在MIDTAL项目()中测试了微阵列平台。将该微阵列应用于挪威和西班牙的野外样品,并与显微细胞计数进行比较。这些探针可能对预警系统和监测有用,也可以用于种群动态研究中以区分物种和生命周期阶段,例如囊肿及其在时间和空间上的分布。电子补充材料本文的在线版本(doi :10.1007 / s11356-012-1403-1)包含补充材料,授权用户可以使用。

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