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DNA metabarcoding of benthic algae and associated eukaryotes from Lake Baikal in the face of rapid environmental changes

机译:面对快速环境变化贝加尔湖底栖藻类和相关真核生物的 DNA 元条形码

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摘要

Here we report new data describing the biodiversity of phytobenthic communities based on DNA-metabarcoding using the 18S rDNA marker and the Illumina MiSeq system. The study was initiated due to the blooming of f ilamentous algae (mainly of the genus Spirogyra) and cyanobacteria in the coastal zone of Lake Baikal under climate change and anthropogenic impact. The composition and taxonomic diversity of algae and other organisms associated with them on different sites of Lake Baikal (near Bolshoi Ushkaniy Island, in Listvennichny Bay) and in the Kaya (within the city of Irkutsk, located in the same drainage basin as Lake Baikal) were determined using DNAmetabarcoding. About 15 thousand reads of the 18S rRNA marker were obtained by applying NGS (next-generation sequencing). The species of algae dominating in the number of reads, as well as the diff icult-to-identify taxa (Stramenopiles, Alveolata, Euglenozoa, Chromista, Rhizaria, Amoebozoa, etc.), which play an important role in the functioning and formation of the structure of algal communities, were revealed. The Shannon index of the communities studied ranges from 1.56 to 2.72. The advantages and weaknesses of using DNA-metabarcoding based on the 18S rRNA gene fragment for studying the structure of algal communities are shown. The advantage of this method is the possibility to more fully determine the diversity of eukaryotes taxa, which are diff icult to identify by morphology, without involving a large number of specialists, while the disadvantage of the method is the distortion that may occur during the PCR. Here, ways of solving this problem are proposed. The results of the study show that the analysis of the minor component of the eukaryotic community in samples (organisms with low biomass) consisting of a mixture of multicellular and unicellular organisms requires a read-depths of at least 100,000 sequences per sample. In general, the DNA-metabarcoding method is recommended for studying the structure of algal communities and eukaryotes associated with them.
机译:在这里,我们报告了使用 18S rDNA 标记和 Illumina MiSeq 系统基于 DNA 宏条形码描述底栖植物群落生物多样性的新数据。这项研究是由于在气候变化和人为影响下,贝加尔湖沿岸地区的野生藻类(主要是 Spirogyra 属)和蓝藻的大量繁殖而启动的。使用 DNA 超条形码确定了贝加尔湖(靠近 Bolshoi Ushkaniy 岛,位于 Listvennichny 湾)和 Kaya(在伊尔库茨克市内,与贝加尔湖位于同一流域)不同地点的藻类和其他相关生物的组成和分类多样性。通过应用 NGS(下一代测序)获得了大约 15,000 个 18S rRNA 标记物。揭示了在读取次数中占主导地位的藻类种类,以及难以识别的分类群(Stramenopiles、Alveolata、Euglenozoa、Chromista、Rhizaria、Amoebozoa 等),它们在藻类群落结构的功能和形成中起着重要作用。所研究社区的香农指数范围为 1.56 至 2.72。显示了使用基于 18S rRNA 基因片段的 DNA 宏条形码研究藻类群落结构的优缺点。这种方法的优点是可以更全面地确定真核生物分类群的多样性,这些分类群很难通过形态学来识别,而无需大量专家参与,而该方法的缺点是在 PCR 过程中可能发生的扭曲。在这里,提出了解决这个问题的方法。研究结果表明,分析由多细胞和单细胞生物混合物组成的样品(生物量低的生物体)中真核生物群落的次要成分需要每个样品至少 100,000 个序列的读深。一般来说,推荐使用 DNA 元条形码方法研究藻类群落和与之相关的真核生物的结构。

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