Infectious genomic RNA of Rhopalosiphum padi virus transcribed in vitro from a full-length cDNA clone.
Availability of a cloned genome from which infectious RNA can be transcribed is essential for investigating RNA virus molecular mechanisms. To date, no such clones have been reported for the Dicistroviridae, an emerging family of invertebrate viruses. Previously we demonstrated baculovirus-driven expression of a cloned Rhopalosiphum padi virus (RhPV; Dicistroviridae) genome that was infectious to aphids, and we identified a cell line (GWSS-Z10) from the glassy-winged sharpshooter, that supports RhPV replication. Here we report that RNA transcribed from a full-length cDNA clone is infectious. Transfection of GWSS-Z10 cells with the RhPV transcript resulted in cytopathic effects, ultrastructural changes, and accumulation of progeny virions, consistent with virus infection. Virions from transcript-infected cells were infectious in aphids. This infectious transcript of a cloned RhPV genome provides a valuable tool, and a more tractable system without interference from baculovirus infection, for investigating replication and pathogenesis of dicistroviruses.
Boyapalle
Sandhya
S
Department of Entomology, Iowa State University, Ames, IA 50011, USA. sboyapalle@yahoo.com
Beckett
Randy J
RJ
Pal
Narinder
N
Miller
W Allen
WA
Bonning
Bryony C
BC
D000818:Animals; D001042:Aphids / Q000821:virology; D002460:Cell Line; D018076:DNA, Complementary / Q000235:genetics; D016679:Genome, Viral; D007304:Insect Viruses / Q000235:genetics* / Q000254:growth & development / Q000472:pathogenicity; D007313:Insecta; D012328:RNA Viruses / Q000235:genetics* / Q000254:growth & development / Q000472:pathogenicity; D014158:Transcription, Genetic; D014162:Transfection; D014771:Virion / Q000235:genetics* / Q000254:growth & development / Q000472:pathogenicity; D014774:Virulence
D018076:DNA, Complementary