Mende Daniel R, Aylward Frank O, Eppley John M, Nielsen Torben N, DeLong Edward F
Daniel K. Inouye Center for Microbial Oceanography Research and Education, University of Hawai'i at Manoa, Honolulu HI, USA.
Front Microbiol. 2016 Feb 11;7:143. doi: 10.3389/fmicb.2016.00143. eCollection 2016.
Assembling complete or near complete genomes from complex microbial communities remains a significant challenge in metagenomic studies. Recent developments in single cell amplified genomes (SAGs) have enabled the sequencing of individual draft genomes representative of uncultivated microbial populations. SAGs suffer from incomplete and uneven coverage due to artifacts that arise from multiple displacement amplification techniques. Conversely, metagenomic sequence data does not suffer from the same biases as SAGs, and significant improvements have been realized in the recovery of draft genomes from metagenomes. Nevertheless, the inherent genomic complexity of many microbial communities often obfuscates facile generation of population genome assemblies from metagenomic data. Here we describe a new method for metagenomic-guided SAG assembly that leverages the advantages of both methods and significantly improves the completeness of initial SAGs assemblies. We demonstrate that SAG assemblies of two cosmopolitan marine lineages-Marine Group 1 Thaumarchaeota and SAR324 clade bacterioplankton-were substantially improved using this approach. Moreover, the improved assemblies strengthened biological inferences. For example, the improved SAR324 clade genome assembly revealed the presence of many genes in phenylalanine catabolism and flagellar assembly that were absent in the original SAG.
从复杂的微生物群落中组装完整或近乎完整的基因组仍然是宏基因组学研究中的一项重大挑战。单细胞扩增基因组(SAG)的最新进展使得对代表未培养微生物种群的单个草图基因组进行测序成为可能。由于多重置换扩增技术产生的假象,SAG存在覆盖不完整和不均匀的问题。相反,宏基因组序列数据不会受到与SAG相同的偏差影响,并且在从宏基因组中恢复草图基因组方面已经取得了显著进展。然而,许多微生物群落固有的基因组复杂性常常使从宏基因组数据中轻松生成种群基因组组装变得模糊不清。在这里,我们描述了一种用于宏基因组引导的SAG组装的新方法,该方法利用了两种方法的优点,并显著提高了初始SAG组装的完整性。我们证明,使用这种方法,两个世界性海洋谱系——海洋类群I Thaumarchaeota和SAR324进化枝浮游细菌——的SAG组装得到了实质性改进。此外,改进后的组装加强了生物学推断。例如,改进后的SAR324进化枝基因组组装揭示了原始SAG中不存在的许多参与苯丙氨酸分解代谢和鞭毛组装的基因。