School of Engineering Sciences in Chemistry, Biotechnology and Health, Department of Gene Technology, Science for Life Laboratory, KTH Royal Institute of Technology, Stockholm, Sweden.
Centre for Ecology and Evolution in Microbial Model Systems, EEMiS, Linnaeus University, Kalmar, Sweden.
Microbiome. 2018 Sep 28;6(1):173. doi: 10.1186/s40168-018-0550-0.
BACKGROUND: Prokaryotes dominate the biosphere and regulate biogeochemical processes essential to all life. Yet, our knowledge about their biology is for the most part limited to the minority that has been successfully cultured. Molecular techniques now allow for obtaining genome sequences of uncultivated prokaryotic taxa, facilitating in-depth analyses that may ultimately improve our understanding of these key organisms. RESULTS: We compared results from two culture-independent strategies for recovering bacterial genomes: single-amplified genomes and metagenome-assembled genomes. Single-amplified genomes were obtained from samples collected at an offshore station in the Baltic Sea Proper and compared to previously obtained metagenome-assembled genomes from a time series at the same station. Among 16 single-amplified genomes analyzed, seven were found to match metagenome-assembled genomes, affiliated with a diverse set of taxa. Notably, genome pairs between the two approaches were nearly identical (average 99.51% sequence identity; range 98.77-99.84%) across overlapping regions (30-80% of each genome). Within matching pairs, the single-amplified genomes were consistently smaller and less complete, whereas the genetic functional profiles were maintained. For the metagenome-assembled genomes, only on average 3.6% of the bases were estimated to be missing from the genomes due to wrongly binned contigs. CONCLUSIONS: The strong agreement between the single-amplified and metagenome-assembled genomes emphasizes that both methods generate accurate genome information from uncultivated bacteria. Importantly, this implies that the research questions and the available resources are allowed to determine the selection of genomics approach for microbiome studies.
背景:原核生物主宰着生物圈,并调节着所有生命都必不可少的生物地球化学过程。然而,我们对它们的生物学的了解在很大程度上仅限于那些已经成功培养的少数原核生物。分子技术现在可以获得未培养原核生物类群的基因组序列,从而促进深入分析,最终可能提高我们对这些关键生物的理解。
结果:我们比较了两种从近海站采集的样本中获取细菌基因组的非培养依赖策略的结果:单扩增基因组和宏基因组组装基因组。单扩增基因组与来自同一站时间序列的先前获得的宏基因组组装基因组进行了比较。在分析的 16 个单扩增基因组中,有 7 个与宏基因组组装基因组匹配,与多种分类群相关。值得注意的是,两种方法之间的基因组对几乎完全相同(重叠区域平均序列同一性为 99.51%;范围为 98.77-99.84%)。在匹配的对中,单扩增基因组始终更小且更不完整,而遗传功能谱得以维持。对于宏基因组组装基因组,只有平均 3.6%的碱基由于错误分类的连续体而被估计从基因组中丢失。
结论:单扩增和宏基因组组装基因组之间的强烈一致性强调了这两种方法都可以从未培养的细菌中生成准确的基因组信息。重要的是,这意味着研究问题和可用资源允许确定用于微生物组研究的基因组学方法的选择。
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