Rhizopus microsporus
is a species in the order Mucorales that is known to cause mucormycosis, but it is poorly understood as a host of viruses. Here, we examined 25 clinical strains of
R. microsporus
for viral infection with a conventional double-stranded RNA (dsRNA) assay using agarose gel electrophoresis (AGE) and the recently established fragmented and primer-ligated dsRNA sequencing (FLDS) protocol. By AGE, five virus-infected strains were detected. Then, full-length genomic sequences of 12 novel RNA viruses were revealed by FLDS, which were related to the families
Mitoviridae, Narnaviridae,
and
Endornaviridae,
ill-defined groups of single-stranded RNA (ssRNA) viruses with similarity to the established families
Virgaviridae
and
Phasmaviridae,
and the proposed family “Ambiguiviridae.” All the characterized viruses, except a potential phasmavirid with a negative-sense RNA genome, had positive-sense RNA genomes. One virus belonged to a previously established species within the family
Mitoviridae
, whereas the other 11 viruses represented new species or even new genera. These results show that the fungal pathogen
R. microsporus
harbors diverse RNA viruses and extend our understanding of the diversity of RNA viruses in the fungal order Mucorales, division Mucoromycota. Identifying RNA viruses from clinical isolates of
R. microsporus
may expand the repertoire of natural therapeutic agents for mucormycosis in the future.
IMPORTANCE
The diversity of mycoviruses in fungal hosts in the division Mucoromycota has been underestimated, mainly within the species
Rhizopus microsporus
. Only five positive-sense RNA genomes had previously been discovered in this species. Because current sequencing methods poorly complete the termini of genomes, we used fragmented and primer-ligated double-stranded RNA sequencing to acquire the full-length genomes. Eleven novel mycoviruses were detected in this study, including the first negative-sense RNA genome reported in
R. microsporus
. Our findings extend the understanding of the viral diversity in clinical strains of Mucoromycota, may provide insights into the pathogenesis and ecology of this fungus, and may offer therapeutic options.
