Identification of a Novel Mycovirus Belonging to the "flexivirus"-related Family with Icosahedral Virion

The order currently comprises five viral families with positive-sense RNA [(+)RNA] genomes that infect plants, fungi, and insects. Virion morphologies within the order differ between families, with icosahedral virions in the and filamentous virions in the other families. Despite their different morphologies, these viruses are placed in the same order based on phylogenetic analyses of replicase-associated polyproteins. However, one of the families in the , , is considered to be capsidless because there have been no published reports of virion isolation. Here, we report that a new "flexivirus"-related (+)RNA virus, prospectively named Fusarium oxysporum icosahedral virus 1 (FoIV1), is icosahedral and that most deltaflexiviruses may have icosahedral virions. Phylogenetic analyses based on replicase-associated polyproteins indicated that FoIV1 forms a distinct group in the with some viruses originally assigned to the . Electron microscopy, protein analysis, and protein structure predictions indicate that FoIV1 open reading frame 4 encodes a single jelly-roll (SJR)-like coat protein (CP) that constitutes the icosahedral virions. Results of clustering analyses based on amino acid sequences and predicted CP structures suggested that most of the deltaflexiviruses have icosahedral virions composed of SJR-like CPs as in FoIV1, rather than having filamentous virions or capsidless. These results challenge the conventional understanding of viruses in the order , with important implications for revising its taxonomic framework and providing insights into the evolutionary relationships within this diverse and broad host range group of (+)RNA viruses.