Virus recovery is induced in Brome mosaic virus p2 transgenic plants showing synchronous complementation and RNA-2-specific silencing

Mol Plant Microbe Interact. 2000 Mar;13(3):247-58. doi: 10.1094/MPMI.2000.13.3.247.

Abstract

Nicotiana benthamiana plants expressing Brome mosaic virus (BMV) p2 protein complemented replication of RNAs1 + 3 but, surprisingly, supported little or no replication of RNA-2. Despite this, the p2 transgenic plants were able to support systemic migration of RNAs-1 and -3. Kinetic analyses showed identical degradation rates for RNAs-2 and -3, greatly detracting from the concept of an induction of an RNA-2-specific degradation system. Deletion analysis identified a 200-nucleotide sequence that may contribute to silencing in a context-specific manner. When R1 progeny of a severely silencing p2 transgenic line were tested for virus resistance, three different classes of reactions were observed. In class 1 and class 3 plants, the virus moved systemically and showed various extents of RNA-2 silencing. However, in class 2 plants, there was a stochastic onset of post-transcriptional silencing in the systemic leaves that was reminiscent of virus recovery. Plants showing recovery tended to have a greater number of transgene loci than did those exhibiting component-specific silencing. The induction of silencing did not appear to be dependent solely on the combined steady state levels of the transgene and viral RNA. Some plants transformed with a p2 frameshift construct showed a complete silencing phenotype, but none showed RNA-2-specific silencing. While the relationship between the two types of silencing remains unclear, we speculate that our observations reflect early events in the induction of virus recovery.

MeSH terms

  • Bromovirus / genetics
  • Bromovirus / physiology*
  • Frameshift Mutation
  • Gene Silencing*
  • Molecular Probe Techniques
  • Nicotiana / genetics
  • Nicotiana / virology*
  • Plants, Genetically Modified
  • Plants, Toxic*
  • Polymerase Chain Reaction
  • RNA Processing, Post-Transcriptional
  • RNA, Viral / biosynthesis
  • RNA, Viral / genetics*
  • RNA, Viral / metabolism
  • Transfection
  • Viral Proteins / genetics*
  • Virus Replication

Substances

  • RNA, Viral
  • Viral Proteins