Zhao Dandan, Salas-Leiva Dayana E, Williams Shelby K, Dunn Katherine A, Shao Jason D, Roger Andrew J
Institute for Comparative Genomics, Dalhousie University, Halifax, Nova Scotia, Canada.
Department of Biochemistry and Molecular Biology, Dalhousie University, Halifax, Nova Scotia, Canada.
mBio. 2025 May 14;16(5):e0069925. doi: 10.1128/mbio.00699-25. Epub 2025 Apr 10.
UNLABELLED: Whole-genome shotgun (WGS) metagenomic sequencing of microbial communities enables the discovery of the functions, physiologies, and evolutionary histories of prokaryotic and eukaryotic microbes. However, metagenomic studies of microbial eukaryotes lag due to challenges in identifying and assembling high-quality genomes from WGS data. To address this problem, we developed Eukfinder, a bioinformatics pipeline that identifies potential eukaryotic sequences from WGS metagenomic data, with a complementary binning workflow for recovering nuclear and mitochondrial genomes. Eukfinder uses two specialized databases for read/contig classification, customizable to specific data sets or environments. We tested Eukfinder on simulated gut microbiome data sets which included varying numbers of reads from the protist , a human gut commensal. We also applied Eukfinder to previously published human gut microbiome WGS metagenomic data to recover new genomes of . Compared to other workflows, Eukfinder offers the potential to recover high-quality, near-complete genomes of diverse eukaryotes, including different subtypes, without relying on a reference genome. With sufficient sequencing depth, Eukfinder outperforms similar tools for recovering eukaryotic genomes from metagenomic data. Eukfinder is a valuable tool for reference-independent and cultivation-free studies of eukaryotic microbial genomes from environmental WGS metagenomic samples. IMPORTANCE: Advancements in next-generation sequencing have made whole-genome shotgun (WGS) metagenomic sequencing an efficient method for reconstruction of microbial genomes from various environments. Thousands of new prokaryotic genomes have been characterized; however, the large size and complexity of protistan genomes have hindered the use of WGS metagenomics to sample microbial eukaryotic diversity. Eukfinder enables the recovery of eukaryotic microbial genomes from environmental WGS metagenomic samples. Retrieval of high-quality protistan genomes from diverse metagenomic samples increases the number of reference genomes available. This aids future metagenomic investigations into the functions, physiologies, and evolutionary histories of eukaryotic microbes in the gut microbiome and other ecosystems.
未标注:对微生物群落进行全基因组鸟枪法(WGS)宏基因组测序能够发现原核和真核微生物的功能、生理特征及进化历史。然而,由于从WGS数据中识别和组装高质量基因组存在挑战,对真核微生物的宏基因组研究进展滞后。为解决这一问题,我们开发了Eukfinder,这是一种生物信息学流程,可从WGS宏基因组数据中识别潜在的真核序列,并通过互补的分箱工作流程来恢复核基因组和线粒体基因组。Eukfinder使用两个专门的数据库进行读段/重叠群分类,可根据特定数据集或环境进行定制。我们在模拟肠道微生物组数据集上测试了Eukfinder,该数据集包含来自原生生物(一种人类肠道共生菌)的不同数量的读段。我们还将Eukfinder应用于先前发表的人类肠道微生物组WGS宏基因组数据,以恢复新的基因组。与其他工作流程相比,Eukfinder有潜力在不依赖参考基因组的情况下,恢复包括不同亚型在内的多种真核生物的高质量、近乎完整的基因组。在有足够测序深度的情况下,Eukfinder在从宏基因组数据中恢复真核基因组方面优于类似工具。Eukfinder是一种有价值的工具,可用于对来自环境WGS宏基因组样本的真核微生物基因组进行不依赖参考基因组和免培养的研究。 重要性:下一代测序技术的进步使全基因组鸟枪法(WGS)宏基因组测序成为从各种环境中重建微生物基因组的有效方法。数千个新的原核基因组已得到表征;然而,原生生物基因组的巨大规模和复杂性阻碍了WGS宏基因组学用于对真核微生物多样性进行采样。Eukfinder能够从环境WGS宏基因组样本中恢复真核微生物基因组。从不同宏基因组样本中检索高质量的原生生物基因组会增加可用参考基因组的数量。这有助于未来对肠道微生物组和其他生态系统中真核微生物的功能、生理特征及进化历史进行宏基因组研究。
Microbiome. 2018-9-15
Brief Bioinform. 2023-9-22
mSystems. 2022-8-30
Microbiome. 2019-2-8
Bioinformatics. 2022-1-3
Genome Biol. 2019-11-28