title: Evasion of short interfering RNA-directed antiviral silencing in Musa acuminata persistently infected with six distinct banana streak pararetroviruses.
abstract: Vegetatively propagated crop plants often suffer from infections with persistent RNA and DNA viruses. Such viruses appear to evade the plant defenses that normally restrict viral replication and spread. The major antiviral defense mechanism is based on RNA silencing generating viral short interfering RNAs (siRNAs) that can potentially repress viral genes posttranscriptionally through RNA cleavage and transcriptionally through DNA cytosine methylation. Here we examined the RNA silencing machinery of banana plants persistently infected with six pararetroviruses after many years of vegetative propagation. Using deep sequencing, we reconstructed consensus master genomes of the viruses and characterized virus-derived and endogenous small RNAs. Consistent with the presence of endogenous siRNAs that can potentially establish and maintain DNA methylation, the banana genomic DNA was extensively methylated in both healthy and virus-infected plants. A novel class of abundant 20-nucleotide (nt) endogenous small RNAs with 5'-terminal guanosine was identified. In all virus-infected plants, 21- to 24-nt viral siRNAs accumulated at relatively high levels (up to 22% of the total small RNA population) and covered the entire circular viral DNA genomes in both orientations. The hotspots of 21-nt and 22-nt siRNAs occurred within open reading frame (ORF) I and II and the 5' portion of ORF III, while 24-nt siRNAs were more evenly distributed along the viral genome. Despite the presence of abundant viral siRNAs of different size classes, the viral DNA was largely free of cytosine methylation. Thus, the virus is able to evade siRNA-directed DNA methylation and thereby avoid transcriptional silencing. This evasion of silencing likely contributes to the persistence of pararetroviruses in banana plants.
  
  
  OBJECTIVE
  We report that DNA pararetroviruses in Musa acuminata banana plants are able to evade DNA cytosine methylation and transcriptional gene silencing, despite being targeted by the host silencing machinery generating abundant 21- to 24-nucleotide short interfering RNAs. At the same time, the banana genomic DNA is extensively methylated in both healthy and virus-infected plants. Our findings shed light on the siRNA-generating gene silencing machinery of banana and provide a possible explanation why episomal pararetroviruses can persist in plants whereas true retroviruses with an obligatory genome-integration step in their replication cycle do not exist in plants.
authors: University of Basel, Department of Environmental Sciences, Botany, Basel, Switzerland.
authors: Rajendran
authors: R
authors: Rajeswaran
authors: University of Basel, Department of Environmental Sciences, Botany, Basel, Switzerland FASTERIS SA, Plan-les-Ouates, Switzerland.
authors: Jonathan
authors: J
authors: Seguin
authors: CIRAD, UMR BGPI, Montpellier, France.
authors: Matthieu
authors: M
authors: Chabannes
authors: CIRAD, UMR BGPI, Montpellier, France.
authors: Pierre-Olivier
authors: PO
authors: Duroy
authors: CIRAD, UMR BGPI, Montpellier, France.
authors: Nathalie
authors: N
authors: Laboureau
authors: FASTERIS SA, Plan-les-Ouates, Switzerland.
authors: Laurent
authors: L
authors: Farinelli
authors: CIRAD, UMR BGPI, Montpellier, France.
authors: Marie-Line
authors: ML
authors: Iskra-Caruana
authors: University of Basel, Department of Environmental Sciences, Botany, Basel, Switzerland Mikhail.Pooggin@unibas.ch.
authors: Mikhail M
authors: MM
authors: Pooggin
mesh_terms: D019175:DNA Methylation; D018506:Gene Expression Regulation, Plant / Q000276:immunology; D015967:Gene Expression Regulation, Viral; D020868:Gene Silencing; D016679:Genome, Viral; D057131:Immune Evasion / Q000235:genetics*; D028521:Musa / Q000235:genetics* / Q000276:immunology / Q000821:virology; D010935:Plant Diseases / Q000235:genetics / Q000276:immunology / Q000821:virology; D057865:Plant Immunity / Q000235:genetics; D010942:Plant Viruses / Q000235:genetics* / Q000472:pathogenicity; D034741:RNA, Small Interfering / Q000235:genetics / Q000276:immunology*; D012367:RNA, Viral / Q000235:genetics / Q000276:immunology; D012190:Retroviridae / Q000235:genetics* / Q000472:pathogenicity; D014158:Transcription, Genetic
chemical_list: D034741:RNA, Small Interfering; D012367:RNA, Viral