title: Replication of Lettuce chlorosis virus (LCV), a crinivirus in the family Closteroviridae, is accompanied by the production of LCV RNA 1-derived novel RNAs.
abstract: Cloned infectious complementary DNAs of the bipartite genomic RNAs of Lettuce chlorosis virus (LCV) were constructed. Inoculation of tobacco protoplasts with the in vitro produced RNAs 1 and 2 transcripts, or with RNA 1 transcript alone, resulted in viral replication accompanied by the production of novel LCV RNA 1-derived RNAs. They included the abundantly accumulating LM-LCVR1-1 (~0.38 kb) and LM-LCVR1-2 (~0.3 kb), and the lowly accumulating HM-LCVR1-1 (~8.0 kb) and HM-LCVR1-2 (~6.6 kb), all of which reacted with riboprobes specific to the 5' end of RNA 1 in Northern blot analysis. LM-LCVR1-1 and HM-LCVR1-2 accumulated as positive-stranded RNAs that lacked complementary negative strands, while HM-LCVR1-1 and LM-LCVR1-2 accumulated in both polarities. Additional Northern blot, reverse transcription-polymerase chain reaction, cloning, and sequence analyses revealed LM-LCVR1-2 to be an authentic RNA 1-derived defective (D)RNA, suggesting that its synthesis and maintenance are supported in trans by an RNA 1 encoded replication machinery.
authors.affiliation: Department of Plant Pathology and Microbiology, University of California, Riverside, CA 92521, USA.
authors.forename: Chawin
authors.initials: C
authors.lastname: Mongkolsiriwattana
authors.forename: Angel Y S
authors.initials: AY
authors.lastname: Chen
authors.forename: James C K
authors.initials: JC
authors.lastname: Ng
mesh_terms: D001483:Base Sequence; D015152:Blotting, Northern; D003001:Cloning, Molecular; D029383:Crinivirus / Q000235:genetics* / Q000502:physiology*; D018076:DNA, Complementary / Q000235:genetics; D016679:Genome, Viral; D018545:Lactuca / Q000821:virology; D010935:Plant Diseases / Q000821:virology; D012367:RNA, Viral / Q000096:biosynthesis* / Q000235:genetics*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D017423:Sequence Analysis, RNA; D014026:Nicotiana / Q000821:virology; D014158:Transcription, Genetic; D014779:Virus Replication / Q000235:genetics*
chemical_list: D018076:DNA, Complementary; D012367:RNA, Viral