Validation of the application of gel beads-based single-cell genome sequencing platform to soil and seawater

Single-cell genomics is applied to environmental samples as a method to solve the problems of current metagenomics. However, in the fluorescence-activated cell sorting-based cell isolation and subsequent whole genome amplification, the sorting efficiency and the sequence quality are greatly affected by the type of target environment, limiting its adaptability. Here, we developed an improved single-cell genomics platform, named SAG-gel, which utilizes gel beads for single-cell isolation, lysis, and whole genome amplification. To validate the versatility of SAG-gel, single-cell genome sequencing was performed with model bacteria and microbial samples collected from eight environmental sites, including soil and seawater. Gel beads enabled multiple lysis treatments. The genome coverage with model bacteria was improved by 9.1-25%. A total of 734 single amplified genomes were collected from the diverse environmental samples, and almost full-length 16S rRNA genes were recovered from 57.8% of them. We also revealed two marine Rhodobacter strains harboring nearly identical 16S rRNA genes but having different genome contents. In addition, searching for viral sequences elucidated the virus-host linkage over the sampling sites, revealing the geographic distribution and diverse host range of viruses.

Automated summary

An improved single-cell genomics platform, SAG-gel, was developed to overcome the limitations of current metagenomics. By using gel beads, SAG-gel achieved efficient single-cell isolation, lysis, and whole genome amplification. The versatility of SAG-gel was validated through single-cell genome sequencing of model bacteria and environmental samples. The results showed improved genome coverage and revealed virus-host linkages.