The RNA silencing pathway mediated by small interfering RNAs (siRNAs) plays an important antiviral role in eukaryotes. To counteract this defense barrier, a large number of plant viruses express proteins with RNA silencing suppression activity. Recently, it was reported that the ipomovirus Cucumber vein yellowing virus (CVYV), which lacks the typical silencing suppressor of members of the family Potyviridae, i.e., HCPro, has a duplicated P1 coding sequence and that the downstream P1 copy, named P1b, has silencing suppression activity. In this study, we provide experimental evidence that P1b is a serine protease that self-cleaves at its C terminus but that its proteolytic activity is not essential for silencing suppression. In contrast, a putative zinc finger and a conserved basic motif in the N-terminal region of the protein are required for efficient silencing suppression. In vitro gel filtration-fast protein liquid chromatography and in vivo bimolecular fluorescence complementation assays showed that P1b binds itself to form oligomeric structures and that the zinc finger-like motif is essential for the self interaction. Moreover, we observed that CVYV P1b forms complexes with synthetic siRNAs, and this ability correlated with both silencing suppression activity and enhancement of Potato virus X pathogenicity in a mutational analysis. Together, these results suggest that CVYV P1b resembles potyviral HCPro and other viral proteins in interfering RNA silencing by preventing siRNA loading into the RNAinduced silencing complex.

A virus causing chlorotic mottling symptoms on sunflower was found in various locations in Argentina. Symptoms were small chlorotic spots, yellow blotches on leaves, and plant stunting. Virus transmission efficiency by mechanical inoculation was 73 to 100%, and by Myzus persicae was 31 to 49%. The host range included members of the Amaranthaceae, Asteraceae, Chenopodiaceae, and Solanaceae families. Electron microscopy of leaf dips from infected plants revealed flexuous particles 17 nm wide and 770 nm long. Cytoplasmic laminar aggregates and pinwheel inclusions were observed in ultrathin sections. Purified virus preparations analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis resolved a capsid protein of 33 kDa. A monoclonal antibody to aphid-transmitted potyviruses reacted with the capsid protein of this virus. In dot blot immunoassays, a polyclonal antiserum (early bleeding) reacted with infected sunflowers and weakly with Bidens mottle potyvirus, but not with either maize dwarf mosaic potyvirus or potato virus Y. The evidence suggests that a potyvirus is infecting sunflower, and a partial characterization of the causal agent is reported.

We have sequenced 1873 nucleotides from the 3'-end region of a sunflower potyvirus genome including the 3'-NIb protein coding region (813 nucleotides), the entire coat protein coding region (807 nucleotides) and 3'-NCR (253 nucleotides), excluding the poly (A) tail. Amino acids identity of the whole CP between the sunflower virus and Potyvirus members ranged from 49.5% (SCMV) to 81.5% (PVY-NsNr), and the core ranged from 55% (TVMV) to 87% (PVY-NsNr; PepMoV). The 3'-NCR nucleotides showed 38.7% homology to PeSMV and 61% to PepMoV-C. The sequence of 3' end region and analysis of phylogenetic relationships suggest this sunflower virus could belong to PVY subgroup and the name of "sunflower chlorotic mottle virus" (SuCMoV) is proposed. This is the first report on the partial nucleotide sequence of a potyvirus infecting sunflower.