Department of Marine Biology and Oceanography, Institute of Marine Sciences (ICM)-CSIC, Pg. Marítim de la Barceloneta, 37-49, Barcelona E-08003, Spain.
CEA, Institut de Génomique, Génoscope, 2 Rue Gaston Crémieux, Evry F-91000, France.
Sci Rep. 2017 Jan 27;7:41498. doi: 10.1038/srep41498.
Pico-sized eukaryotes play key roles in the functioning of marine ecosystems, but we still have a limited knowledge on their ecology and evolution. The MAST-4 lineage is of particular interest, since it is widespread in surface oceans, presents ecotypic differentiation and has defied culturing efforts so far. Single cell genomics (SCG) are promising tools to retrieve genomic information from these uncultured organisms. However, SCG are based on whole genome amplification, which normally introduces amplification biases that limit the amount of genomic data retrieved from a single cell. Here, we increase the recovery of genomic information from two MAST-4 lineages by co-assembling short reads from multiple Single Amplified Genomes (SAGs) belonging to evolutionary closely related cells. We found that complementary genomic information is retrieved from different SAGs, generating co-assembly that features >74% of genome recovery, against about 20% when assembled individually. Even though this approach is not aimed at generating high-quality draft genomes, it allows accessing to the genomic information of microbes that would otherwise remain unreachable. Since most of the picoeukaryotes still remain uncultured, our work serves as a proof-of-concept that can be applied to other taxa in order to extract genomic data and address new ecological and evolutionary questions.
真核微型生物在海洋生态系统的功能中起着关键作用,但我们对它们的生态学和进化仍然知之甚少。MAST-4 谱系尤其引人注目,因为它广泛存在于海洋表面,具有生态型分化,并且迄今为止一直抵制培养。单细胞基因组学(SCG)是从这些未培养的生物体中获取基因组信息的有前途的工具。然而,SCG 基于全基因组扩增,这通常会引入扩增偏差,从而限制从单个细胞中获取的基因组数据量。在这里,我们通过共组装来自进化上密切相关的细胞的多个单个扩增基因组(SAG)的短读长,从两个 MAST-4 谱系中增加了基因组信息的回收。我们发现,不同的 SAG 中会检索到互补的基因组信息,生成的共组装特征是基因组恢复的>74%,而单独组装时约为 20%。即使这种方法不是为了生成高质量的草图基因组,它也允许访问否则无法获得的微生物的基因组信息。由于大多数微型真核生物仍然未被培养,我们的工作可以作为一个概念验证,应用于其他分类群,以提取基因组数据并解决新的生态和进化问题。