Persistent co-infection mediates inter-virus and host-virus gene exchange in marine protists

Genomics and metagenomics have transformed our understanding of the diversity of large DNA viruses infecting eukaryotic microorganisms. Using long-read sequencing, we discovered ubiquitous, co-culturing nucleocytoviruses in several thraustochytrid protists. These viruses are closely related to SmDNAV, an 'orphan' virus of the thraustochytrid Sicyoidochytrium minutum. Transcriptomics, proteomics and electron microscopy demonstrate viral gene expression and particle formation in these cultures without obvious defects in culture growth. We also found SmDNAV-type viruses associated with previously sequenced thraustochytrids. Phylogenetic analysis reveals that the SmDNAV and SmDNAV-like genomes belong to a hitherto unrecognized order of Nucleocytoviricota, here named "Skiavirales". All fifteen identified skiavirus genomes lack numerous viral hallmark genes including DNA-dependent RNA polymerase; all but three also lack DNA polymerase family B (PolB). PolB phylogeny suggests a specific relationship between Skiavirales and the recently discovered mirusviruses, which belong to a different viral realm. Gene exchange between skiaviruses, co-occurring mirusviruses, and host nuclear genomes shows that persistent infection by diverse large DNA viruses provides an opportunity for virus-virus and virus-host co-evolution in thraustochytrids and, perhaps, other microbial eukaryotes.