Abstract

Giant viruses have large genomes, often within the size range of cellular organisms. This distinguishes them from most other viruses and demands additional effort for the successful recovery of their genomes from environmental sequence data. Here, we tested the performance of genome-resolved metagenomics on a recently isolated giant virus, Fadolivirus, by spiking it into an environmental sample from which two other giant viruses were isolated. At high spike-in levels, metagenome assembly and binning led to the successful genomic recovery of Fadolivirus from the sample. A complementary survey of the major capsid protein indicated the presence of other giant viruses in the sample matrix but did not detect the two isolated from this sample. Our results indicate that genome-resolved metagenomics is a valid approach for the recovery of near-complete giant virus genomes given that sufficient clonal particles are present. However, our data also underline that a vast majority of giant viruses remain currently undetected, even in an era of terabase-scale metagenomics.IMPORTANCE The discovery of large and giant nucleocytoplasmic large DNA viruses (NCLDV) with genomes in the megabase range and equipped with a wide variety of features typically associated with cellular organisms was one of the most unexpected, intriguing, and spectacular breakthroughs in virology. Recent studies suggest that these viruses are highly abundant in the oceans, freshwater, and soil, impact the biology and ecology of their eukaryotic hosts, and ultimately affect global nutrient cycles. Genome-resolved metagenomics is becoming an increasingly popular tool to assess the diversity and coding potential of giant viruses, but this approach is currently lacking validation.

Highlights

  • Giant viruses have large genomes, often within the size range of cellular organisms

  • For giant virus cocultivation experiments, a sample of wastewater was collected from a treatment plant in Toulon, France, and particles within the sample were sorted by flow cytometry into microplates containing host cells

  • Giant virus identification by flow cytometry characteristics showed 2 different populations; the first population corresponded to Mimivirus, and the second population was unidentified

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Summary

Introduction

Giant viruses have large genomes, often within the size range of cellular organisms. This distinguishes them from most other viruses and demands additional effort for the successful recovery of their genomes from environmental sequence data. Most viral genomes are small, and when found in metagenomic data, they are readily present on a single contig and often considered complete or nearly complete [7] This is in stark contrast to genomes of large and giant viruses of the nucleocytoplasmic large DNA viruses (NCLDV), which can be up to several megabases [8, 9]. The genomes from the uncultivated Klosneuvirinae revealed that they encoded comprehensive translation system components [14], subsequently found in isolated tupanviruses [27] Taken together, these studies indicate that metagenomics is of profound value in deriving genomes of giant viruses from the environment, enabling important novel insights into their predicted biology, ecology, and evolutionary history. Spiking viral particles into a wastewater sample, we tested the performance of commonly used assembly and binning tools, as well as the ability to detect giant virus genomic information in metagenomes

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