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基因组解析长读长测序扩展了陆地生境中已知的微生物多样性。

Genome-resolved long-read sequencing expands known microbial diversity across terrestrial habitats.

作者信息

Sereika Mantas, Mussig Aaron James, Jiang Chenjing, Knudsen Kalinka Sand, Jensen Thomas Bygh Nymann, Petriglieri Francesca, Yang Yu, Jørgensen Vibeke Rudkjøbing, Delogu Francesco, Sørensen Emil Aarre, Nielsen Per Halkjær, Singleton Caitlin Margaret, Hugenholtz Philip, Albertsen Mads

机构信息

Center for Microbial Communities, Aalborg University, Aalborg, Denmark.

Australian Centre for Ecogenomics, School of Chemistry and Molecular Biosciences, The University of Queensland, Brisbane, Queensland, Australia.

出版信息

Nat Microbiol. 2025 Jul 24. doi: 10.1038/s41564-025-02062-z.

Abstract

The emergence of high-throughput, long-read DNA sequencing has enabled recovery of microbial genomes from environmental samples at scale. However, expanding the terrestrial microbial genome catalogue has been challenging due to the enormous complexity of these environments. Here we performed deep, long-read Nanopore sequencing of 154 soil and sediment samples collected during the Microflora Danica project, yielding genomes of 15,314 previously undescribed microbial species, recovered using our custom mmlong2 workflow. The recovered microbial genomes span 1,086 previously uncharacterized genera and expand the phylogenetic diversity of the prokaryotic tree of life by 8%. The long-read assemblies also enabled the recovery of thousands of complete ribosomal RNA operons, biosynthetic gene clusters and CRISPR-Cas systems. Furthermore, the incorporation of the recovered genomes into public genomic databases substantially improved species-level classification rates for soil and sediment metagenomic datasets. These findings demonstrate that long-read sequencing allows cost-effective recovery of high-quality microbial genomes from highly complex ecosystems, which remain an untapped source of biodiversity.

摘要

高通量、长读长DNA测序技术的出现,使得大规模从环境样本中获取微生物基因组成为可能。然而,由于这些环境极其复杂,扩充陆地微生物基因组目录一直具有挑战性。在此,我们对在丹麦微生物区系项目中收集的154份土壤和沉积物样本进行了深度、长读长的纳米孔测序,使用我们定制的mmlong2工作流程,获得了15314个此前未被描述的微生物物种的基因组。所获得的微生物基因组涵盖了1086个此前未被表征的属,将原核生物生命树的系统发育多样性扩展了8%。长读长组装还使得数千个完整的核糖体RNA操纵子、生物合成基因簇和CRISPR-Cas系统得以恢复。此外,将所获得的基因组纳入公共基因组数据库,显著提高了土壤和沉积物宏基因组数据集的物种水平分类率。这些发现表明,长读长测序能够以具有成本效益的方式,从高度复杂的生态系统中获取高质量的微生物基因组,而这些生态系统仍是未被开发的生物多样性来源。

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