A single-stranded DNA virus replicates in the mitochondria of a marine oomycete

Abstract A virus with a circular single-stranded DNA genome, HthCRESSV1, was discovered in an oomycete, Halophytophthora thermoambigua, isolated from brackish waters off the Algarve coast in southern Portugal. Phylogenetic analyses place this virus outside of all currently classified families within the phylum Cressdnaviricota, suggesting it represents a distinct lineage. Cellular fractionation, mitochondrial marker co-enrichment, and Southern blot analyses indicate that HthCRESSV1 is associated with mitochondria and replicates episomally. The virus is stably transmitted through zoospores and is associated with a reduced host mycelial growth. Homologous sequences corresponding to both the replication-associated protein and a membrane-associated hypothetical protein were identified in the mitochondrial genomes of several oomycete species, suggesting recurrent endogenization of HthCRESSV1-like viruses in oomycete mitochondria. Together, these findings expand the known diversity of oomycete-associated DNA viruses, identify mitochondria as an unexpected niche for DNA virus replication, and highlight marine environments as underexplored reservoirs of DNA viruses in stramenopiles.