Division of Biological and Environmental Science and Engineering (BESE), Red Sea Research Center, King Abdullah University of Science and Technology (KAUST), Thuwal, Saudi Arabia.
Center for Genomics and Systems Biology, New York University Abu Dhabi, Abu Dhabi, UAE.
Mol Ecol Resour. 2019 Jul;19(4):1063-1080. doi: 10.1111/1755-0998.13004. Epub 2019 Apr 26.
We present SymPortal (SymPortal.org), a novel analytical framework and platform for genetically resolving the algal symbionts of reef corals using next-generation sequencing (NGS) data of the ITS2 rDNA. Although the ITS2 marker is widely used to genetically characterize taxa within the family Symbiodiniaceae (formerly the genus Symbiodinium), the multicopy nature of the marker complicates its use. Commonly, the intragenomic diversity resultant from this multicopy nature is collapsed by analytical approaches, thereby focusing on only the most abundant sequences. In contrast, SymPortal employs logic to identify within-sample informative intragenomic sequences, which we have termed 'defining intragenomic variants' (DIVs), to identify ITS2-type profiles representative of putative Symbiodiniaceae taxa. By making use of this intragenomic ITS2 diversity, SymPortal is able to resolve genetic delineations using the ITS2 marker at a level that was previously only possible by using additional genetic markers. We demonstrate this by comparing this novel approach to the most commonly used alternative approach for NGS ITS2 data, the 97% similarity clustering to operational taxonomic units (OTUs). The SymPortal platform accepts NGS raw sequencing data as input to provide an easy-to-use, standardization-enforced, and community-driven framework that integrates with a database to gain resolving power with increased use. We consider that SymPortal, in conjunction with ongoing large-scale sampling and sequencing efforts, should play an instrumental role in making future sampling efforts more comparable and in maximizing their efficacy in working towards the classification of the global Symbiodiniaceae diversity.
我们提出了 SymPortal(SymPortal.org),这是一种新颖的分析框架和平台,可使用下一代测序(NGS)ITS2 rDNA 数据对珊瑚藻共生体进行基因解析。虽然 ITS2 标记广泛用于对 Symbiodiniaceae 科(前身为 Symbiodinium 属)内的分类群进行基因特征描述,但该标记的多拷贝性质使其使用变得复杂。通常,由于这种多拷贝性质,基因组内的多样性会被分析方法所简化,从而只关注最丰富的序列。相比之下,SymPortal 采用逻辑方法来识别样本内的信息性基因组内序列,我们称之为“定义基因组内变体”(DIV),以识别代表假定的 Symbiodiniaceae 分类群的 ITS2 型谱。通过利用这种基因组内 ITS2 多样性,SymPortal 能够在以前只能通过使用其他遗传标记才能实现的水平上,使用 ITS2 标记来解析遗传界限。我们通过将这种新方法与最常用的 NGS ITS2 数据替代方法(97%相似性聚类到操作分类单元(OTUs))进行比较,证明了这一点。SymPortal 平台接受 NGS 原始测序数据作为输入,提供了一个易于使用、标准化执行且社区驱动的框架,该框架与数据库集成,可随着使用的增加而提高分辨率。我们认为,SymPortal 应与正在进行的大规模采样和测序工作一起,在使未来的采样工作更具可比性和最大程度地提高其在全球 Symbiodiniaceae 多样性分类工作中的效率方面发挥重要作用。