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Environmental DNA metabarcoding for fish community analysis in backwater lakes: A comparison of capture methods

机译:在回水湖中用于鱼类群落分析的环境DNA元条形码:捕获方法的比较

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摘要

The use of environmental DNA (eDNA) methods for community analysis has recently been developed. High-throughput parallel DNA sequencing (HTS), called eDNA metabarcoding, has been increasingly used in eDNA studies to examine multiple species. However, eDNA metabarcoding methodology requires validation based on traditional methods in all natural ecosystems before a reliable method can be established. To date, relatively few studies have performed eDNA metabarcoding of fishes in aquatic environments where fish communities were intensively surveyed using multiple traditional methods. Here, we have compared fish communities’ data from eDNA metabarcoding with seven conventional multiple capture methods in 31 backwater lakes in Hokkaido, Japan. We found that capture and field surveys of fishes were often interrupted by macrophytes and muddy sediments in the 31 lakes. We sampled 1 L of the surface water and analyzed eDNA using HTS. We also surveyed the fish communities using seven different capture methods, including various types of nets and electrofishing. At some sites, we could not detect any eDNA, presumably because of the polymerase chain reaction (PCR) inhibition. We also detected the marine fish species as sewage-derived eDNA. Comparisons of eDNA metabarcoding and capture methods showed that the detected fish communities were similar between the two methods, with an overlap of 70%. Thus, our study suggests that to detect fish communities in backwater lakes, the performance of eDNA metabarcoding with the use of 1 L surface water sampling is similar to that of capturing methods. Therefore, eDNA metabarcoding can be used for fish community analysis but environmental factors that can cause PCR inhibition, should be considered in eDNA applications.
机译:最近开发了使用环境DNA(eDNA)方法进行社区分析的方法。高通量并行DNA测序(HTS),称为eDNA元条形码,已越来越多地用于eDNA研究中以检查多种物种。但是,eDNA元条形码技术需要在所有自然生态系统中基于传统方法进行验证,然后才能建立可靠的方法。迄今为止,在使用多种传统方法对鱼类群落进行密集调查的水生环境中,对鱼类进行eDNA元条形码的研究相对较少。在这里,我们比较了日本北海道31个死水湖泊中eDNA元条形码与7种常规多重捕获方法中鱼类群落的数据。我们发现,在31个湖泊中,鱼类捕获和现场调查通常被大型植物和泥泞的沉积物打断。我们采样了1 L的地表水,并使用HTS分析了eDNA。我们还使用七种不同的捕获方法(包括各种类型的网和电钓鱼)对鱼类群落进行了调查。在某些位置,我们可能无法检测到任何eDNA,这可能是由于聚合酶链反应(PCR)受到抑制。我们还检测到海水鱼类是污水来源的eDNA。 eDNA元条形码和捕获方法的比较表明,两种方法之间检测到的鱼类群落相似,重叠率为70%。因此,我们的研究表明,要检测回水湖中的鱼类群落,使用1 L地表水采样的eDNA元条形码的性能类似于捕获方法。因此,eDNA元条形码可用于鱼类群落分析,但在eDNA应用中应考虑可能引起PCR抑制的环境因素。

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