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Background  Mycoviruses were recently discovered in the white pine blister rust (WPBR) fungus  Cronartium ribicola  (J.C. Fisch.). Detection and characterization of their double stranded RNA (dsRNA) would facilitate understanding of pathogen virulence and disease pathogenesis in WPBR systems.    Methods  Full-length cDNAs were cloned from the dsRNAs purified from viral-infected  C. ribicola , and their cDNA sequences were determined by DNA sequencing. Evolutionary relationships of the dsRNAs with related mycoviruses were determined by phylogenetic analysis. Dynamic distributions of the viral RNAs within samples of their fungal host  C. ribicola  were investigated by measurement of viral genome prevalence and viral gene expression.    Results  In this study we identified and characterized five novel dsRNAs from  C. ribicola , designated as  Cronartium ribicola  totivirus 1–5 (CrTV1 to CrTV5). These dsRNA sequences encode capsid protein and RNA-dependent RNA polymerase with significant homologies to dsRNA viruses of the family  Totiviridae . Phylogenetic analysis showed that the CrTVs were grouped into two distinct clades. CrTV2 through CrTV5 clustered within the genus  Totivirus . CrTV1 along with a few un-assigned dsRNAs constituted a distinct phyletic clade that is genetically distant from presently known genera in the  Totiviridae  family, indicating that CrTV1 represents a novel genus in the  Totiviridae  family. The CrTVs were prevalent in fungal samples obtained from infected western white pine, whitebark pine, and limber pines. Viral RNAs were generally expressed at higher levels during  in planta  mycelium growth than in aeciospores and urediniospores. CrTV4 was significantly associated with  C. ribicola  virulent pathotype and specific  C. ribicola  host tree species, suggesting dsRNAs as potential tools for dissection of pathogenic mechanisms of  C. ribicola  and diagnosis of  C. ribicola  pathotypes.    Conclusion  Phylogenetic and expression analyses of viruses in the WPBR pathogen,  C. ribicola,  have enchanced our understanding of virus diversity in the family  Totiviridae , and provided a potential strategy to utilize pathotype-associated mycoviruses to control fungal forest diseases.

Characterization of Cronartium ribicola dsRNAs reveals novel members of the family Totiviridae and viral association with fungal virulence