Expression of a synthetic rust fungal virus cDNA in XXXX .

@entity3196 @entity71934 @entity66 @entity71935 @entity1460 @entity375
UNASSIGNED: Mycoviruses are viruses that infect fungi. Recently, mycovirus-like RNAs were sequenced from the fungus @entity71934 , the causal agent of @entity1460 rust. One of the RNAs appeared to represent a novel mycovirus and was designated @entity71934 virus 2383 (PpV2383). The genome of PpV2383 resembles @entity71935 , a double-stranded (ds) RNA mycovirus of @entity375 . PpV2383 encodes two major, overlapping open reading frames with similarity to gag (capsid protein) and pol (RNA-dependent RNA polymerase), and a -1 ribosomal frameshift is necessary for the translation of a gag-pol fusion protein. Phylogenetic analysis of pol relates PpV2383 to members of the family Totiviridae, including L-A. Because the obligate biotrophic nature of @entity71934 makes it genetically intractable for in vivo analysis and because PpV2383 is similar to L-A, we synthesized a DNA clone of PpV2383 and tested its infectivity in @entity375 cells. PpV2383 RNA was successfully expressed in @entity375 , and mass spectrometry confirmed the translation of gag and gag-pol fusion proteins. There was, however, no production of PpV2383 dsRNA, the evidence of viral replication. Neither the presence of endogenous L-A nor the substitution of the 5' and 3' untranslated regions with those from @entity3196 was sufficient to rescue replication of PpV2383. Nevertheless, the proof of transcription and translation from the clone in vivo are steps toward confirming that PpV2383 is a mycovirus. Further development of a surrogate biological system for the study of rust mycoviruses is necessary, and such research may facilitate biological control of @entity66 .
