title: Suppression of antiviral silencing by cucumber mosaic virus 2b protein in Arabidopsis is associated with drastically reduced accumulation of three classes of viral small interfering RNAs.
abstract: We investigated the genetic pathway in Arabidopsis thaliana targeted during infection by cucumber mosaic virus (CMV) 2b protein, known to suppress non-cell-autonomous transgene silencing and salicylic acid (SA)-mediated virus resistance. We show that 2b expressed from the CMV genome drastically reduced the accumulation of 21-, 22-, and 24-nucleotide classes of viral small interfering RNAs (siRNAs) produced by Dicer-like4 (DCL4), DCL2, and DCL3, respectively. The defect of a CMV 2b-deletion mutant (CMV-Delta2b) in plant infection was efficiently rescued in Arabidopsis mutants producing neither 21- nor 22-nucleotide viral siRNAs. Since genetic analysis further identifies a unique antiviral role for DCL3 upstream of DCL4, our data indicate that inhibition of the accumulation of distinct viral siRNAs plays a key role in 2b suppression of antiviral silencing. Strikingly, disease symptoms caused by CMV-Delta2b in Arabidopsis mutants defective in antiviral silencing were as severe as those caused by CMV, demonstrating an indirect role for the silencing suppressor activity in virus virulence. We found that production of CMV siRNAs without 2b interference depended largely on RNA-dependent RNA polymerase 1 (RDR1) inducible by SA. Given the known role of RDR6-dependent transgene siRNAs in non-cell-autonomous silencing, our results suggest a model in which 2b inhibits the production of RDR1-dependent viral siRNAs that confer SA-dependent virus resistance by directing non-cell-autonomous antiviral silencing.
authors.affiliation: Center for Plant Cell Biology, Department of Plant Pathology and Microbiology, University of California, Riverside, CA 92521, USA.
authors.forename: Juan A
authors.initials: JA
authors.lastname: Diaz-Pendon
authors.forename: Feng
authors.initials: F
authors.lastname: Li
authors.forename: Wan-Xiang
authors.initials: WX
authors.lastname: Li
authors.forename: Shou-Wei
authors.initials: SW
authors.lastname: Ding
mesh_terms: D017360:Arabidopsis / Q000378:metabolism* / Q000821:virology*; D029681:Arabidopsis Proteins / Q000378:metabolism; D018797:Cell Cycle Proteins / Q000378:metabolism; D020868:Gene Silencing; D010935:Plant Diseases / Q000821:virology; D018515:Plant Leaves / Q000821:virology; D010942:Plant Viruses / Q000235:genetics*; D034741:RNA, Small Interfering / Q000235:genetics / Q000378:metabolism*; D012367:RNA, Viral / Q000378:metabolism*; D043244:Ribonuclease III / Q000378:metabolism; D012260:Ribonucleases / Q000378:metabolism; D014764:Viral Proteins / Q000235:genetics / Q000378:metabolism*
chemical_list: C404227:2b protein, cucumber mosaic virus; D029681:Arabidopsis Proteins; D018797:Cell Cycle Proteins; D034741:RNA, Small Interfering; D012367:RNA, Viral; D014764:Viral Proteins; C504982:DCL4 protein, Arabidopsis; D012260:Ribonucleases; C511919:DCL2 protein, Arabidopsis; D043244:Ribonuclease III