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Development of a quantitative PCR method to explore the historical occurrence of a nuisance microalga under expansion

机译：定量PCR方法的开发，以探索扩展下令人讨厌的微藻的历史发生

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A number of marine and freshwater harmful algal bloom (HAB) species have colonized new areas and expanded their habitat range in recent years. Nevertheless it is notoriously difficult to establish when colonization first occurred, what the dispersal routes are, and to separate recent invasion from increases in existent but small populations. The freshwater raphidophyte Gonyostomum semen is a nuisance species that has expanded its habitat range and increased in abundance in northern Europe during the past decades. To evaluate to what extent sediments can be used for determining historic occurrence of G. semen, a quantitative real-time PCR method for detecting cysts of this algae was developed. This paper presents a qPCR protocol with a set of primers that are specific to Gonyostomum and with PCR conditions optimized for sediment samples from humic lakes, which are the common habitat of G. semen. With this sensitive method as few as 1.6 cysts per PCR reaction could be reliably quantified, corresponding to 320 cysts per g wet weight sediment Cysts were present in sediments with ages ranging from years to decades and their persistence allows detection of historic populations up to at least 50 years old. With this qPCR assay it will be possible to trace the presence of G. semen in environments prior to the onset of algae-specific monitoring programs as well as for quantification in water column samples. (C) 2016 The Authors. Published by Elsevier B.V.
机译：近年来，许多海洋和淡水有害藻华（HAB）物种已定居在新地区，并扩大了其栖息地范围。然而，众所周知，很难确定什么时候首次发生定殖，什么传播途径，以及将最近的入侵与现有但数量很少的种群的增加区分开来。在过去的几十年中，淡水菱角藻Gonyostomum精液是一个令人讨厌的物种，它扩大了其栖息地范围，并增加了北欧的数量。为了评估在何种程度上可以使用沉积物确定G.精液的历史发生，开发了一种定量实时PCR方法来检测这种藻类的囊肿。本文介绍了一种qPCR方案，该方案具有一套针对灵芝的引物，并优化了针对腐殖质湖常见的腐殖质湖中沉积物样品的PCR条件。通过这种灵敏的方法，每个PCR反应中只有1.6个囊肿可以可靠地定量，相当于每g湿重沉积物有320个囊肿。囊肿存在于年龄范围从数年到数十年的沉积物中，并且它们的持久性使得至少可以检测历史种群50岁。通过此qPCR分析，有可能在藻类特异性监测程序开始之前以及在水柱样品中进行定量之前，先在环境中追踪精液的存在。 （C）2016作者。由Elsevier B.V.发布

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