Single-cell genome assembly of marine bacterial communities metabolising plastic waste Robert Sugar 2014

Plastics in Oceans 725 million tons of plastic produced each year 10% ends up in the oceans Polluting, dangerous to wildlife, an endocrine toxin

Pictures of Plastic

Plastic-eating bacteria Plastic levels are steady from the 80’s on – where does it go? Plastic metabolism has been seen in landfills (Pseudomonas) Recently discovered in seawater (Vibrio)* 10 μm * Zettler, E. R. et al. Microbial Communities on Plastic Marine Debris (2013)

Why? ~90% of bacterial species are unculturable Single-cell sequencing is a unique window into understanding these organisms

Fighting Contamination with Low Volumes

Whole Genome Amplification (WGA) with Multiple displacement amplification (MDA)

Steps Collect samples from the plastisphere Cell-sort them by physical properties Single-cell DNA sequencing, and initial genome assembly Cluster cell by sequence similarity Co-assemble highly similar samples (likely clonal or same species) to close gaps, and manage coverage differences Downstream analysis of genomes to look for pathways relevant to plastic metabolism