Accumulation of transgene-derived siRNAs is not sufficient for RNAi-mediated protection against Citrus tristeza virus in transgenic Mexican lime

Mol Plant Pathol. 2010 Jan;11(1):33-41. doi: 10.1111/j.1364-3703.2009.00566.x.

Abstract

Mexican lime plants transformed with the 3'-terminal 549 nucleotides of the Citrus tristeza virus (CTV) genome in sense, antisense and intron-hairpin formats were analysed for transgene-derived transcript and short interfering RNA (siRNA) accumulation, and for CTV resistance. Propagations from all sense, antisense and empty-vector transgenic lines were susceptible to CTV, except for a single sense-line plant with a complex transgene integration pattern that showed transgene-derived siRNAs in association with low levels of the transgene-derived transcript. In contrast, nine of 30 intron-hairpin lines showed CTV resistance, with 9%-56% of bud-propagated plants, depending on the line, remaining uninfected on graft inoculation, and the others being susceptible. Although resistance was always associated with the presence of transgene-derived siRNAs, their level in different sense and intron-hairpin transformants was variable irrespective of the response to CTV infection. In intron-hairpin lines with single transgene integration, CTV resistance was correlated with low accumulation of the transgene-derived transcript rather than with high accumulation of transgene-derived siRNAs.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • 3' Untranslated Regions
  • Base Sequence
  • Citrus / virology*
  • DNA Primers
  • DNA, Complementary
  • Introns
  • Open Reading Frames
  • Plant Viruses / physiology*
  • Plants, Genetically Modified
  • RNA Interference*
  • RNA, Small Interfering*
  • Transformation, Genetic
  • Transgenes*

Substances

  • 3' Untranslated Regions
  • DNA Primers
  • DNA, Complementary
  • RNA, Small Interfering