New Microviridae isolated from Sulfitobacter reveals two cosmopolitan subfamilies of ssDNA phages infecting marine and terrestrial Alphaproteobacteria

The Microviridae family represents one of the major clades of ssDNA phages. Their cultivated members are lytic and infect Proteobacteria, Bacteroidetes , and Chlamydiae . Prophages have been predicted in genomes from Bacteroidales, Hyphomicrobiales , and Enterobacteraceae and cluster within the “Alpavirinae”, “Amoyvirinae” and Gokushovirinae . We have isolated “Ascunsovirus oldenburgi” ICBM5, a novel phage distantly related to known Microviridae . It infects Sulfitobacter dubius SH24-1b and uses both a lytic and a carrier-state life strategy. Using ICBM5 proteins as a query, we uncovered in publicly available resources 65 new microviridae prophages and episomes in bacterial genomes and retrieved 47 environmental viral genomes (EVGs) from various viromes. Genome clustering based on protein content and phylogenetic analysis showed that ICBM5, together with Rhizobium phages, new prophages, episomes, and EVGs cluster within two new phylogenetic clades, here tentatively assigned the rank of subfamily and named “Tainavirinae” and “Occultatumvirinae”. They both infect Rhodobacterales . Occultatumviruses also infect Hyphomicrobiales , including nitrogen-fixing endosymbionts from cosmopolitan legumes. A biogeographical assessment showed that tainaviruses and occultatumviruses are spread worldwide, in terrestrial and marine environments. The new phage isolated here shed light onto new and diverse branches of the Microviridae tree, suggesting that much of the ssDNA phage diversity remains in the dark. ### Competing Interest Statement The authors have declared no competing interest.