A chimeric satellite transgene sequence is inefficiently targeted by viroid-induced DNA methylation in tobacco

Plant Mol Biol. 2010 Jul;73(4-5):439-47. doi: 10.1007/s11103-010-9631-6. Epub 2010 Apr 3.

Abstract

In plants, transgenes containing Potato spindle tuber viroid (PSTVd) cDNA sequences were efficient targets of PSTVd infection-mediated RNA-directed DNA methylation. Here, we demonstrate that in PSTVd-infected tobacco plants, a 134 bp PSTVd fragment (PSTVd-134) did not become densely methylated when it was inserted into a chimeric Satellite tobacco mosaic virus (STMV) construct. Only about 4-5% of all cytosines (Cs) of the PSTVd-134 were methylated when flanked by satellite sequences. In the same plants, C methylation was approximately 92% when the PSTVd-134 was in a PSTVd full length sequence context and roughly 33% when flanked at its 3' end by a 19 bp PSTVd and at its 5' end by a short viroid-unrelated sequence. In addition, PSTVd small interfering RNAs (siRNAs) produced from the replicating viroid failed to target PSTVd-134-containing chimeric STMV RNA for degradation. Satellite RNAs appear to have adopted secondary structures that protect them against RNA interference (RNAi)-mediated degradation. Protection can be extended to short non-satellite sequences residing in satellite RNAs, rendering them poor targets for nuclear and cytoplasmic RNAi induced in trans.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Base Sequence
  • Blotting, Northern
  • Blotting, Southern
  • DNA Methylation / genetics*
  • DNA, Recombinant / genetics*
  • Gene Expression Regulation, Plant
  • Gene Silencing
  • Molecular Sequence Data
  • Nicotiana / genetics*
  • Nicotiana / virology*
  • Plant Viruses / genetics
  • RNA Stability / genetics
  • Sequence Analysis, DNA
  • Tobacco mosaic satellite virus / genetics*
  • Transgenes / genetics*
  • Viroids / genetics*

Substances

  • DNA, Recombinant