A universal expression/silencing vector in plants

Plant Physiol. 2007 Dec;145(4):1251-63. doi: 10.1104/pp.107.108217. Epub 2007 Sep 28.

Abstract

A universal vector (IL-60 and auxiliary constructs), expressing or silencing genes in every plant tested to date, is described. Plants that have been successfully manipulated by the IL-60 system include hard-to-manipulate species such as wheat (Triticum duram), pepper (Capsicum annuum), grapevine (Vitis vinifera), citrus, and olive (Olea europaea). Expression or silencing develops within a few days in tomato (Solanum lycopersicum), wheat, and most herbaceous plants and in up to 3 weeks in woody trees. Expression, as tested in tomato, is durable and persists throughout the life span of the plant. The vector is, in fact, a disarmed form of Tomato yellow leaf curl virus, which is applied as a double-stranded DNA and replicates as such. However, the disarmed virus does not support rolling-circle replication, and therefore viral progeny single-stranded DNA is not produced. IL-60 does not integrate into the plant's genome, and the construct, including the expressed gene, is not heritable. IL-60 is not transmitted by the Tomato yellow leaf curl virus's natural insect vector. In addition, artificial satellites were constructed that require a helper virus for replication, movement, and expression. With IL-60 as the disarmed helper "virus," transactivation occurs, resulting in an inducible expressing/silencing system. The system's potential is demonstrated by IL-60-derived suppression of a viral-silencing suppressor of Grapevine virus A, resulting in Grapevine virus A-resistant/tolerant plants.

MeSH terms

  • Animals
  • DNA, Satellite
  • DNA, Single-Stranded
  • Escherichia coli / genetics
  • Flexiviridae
  • Geminiviridae / genetics*
  • Gene Expression
  • Gene Silencing*
  • Genetic Engineering*
  • Genetic Vectors*
  • Genome, Plant
  • Hemiptera / virology
  • Nicotiana / virology
  • Plants / genetics*
  • Plants / metabolism
  • Plants / virology
  • Plasmids
  • RNA, Double-Stranded
  • Transcriptional Activation

Substances

  • DNA, Satellite
  • DNA, Single-Stranded
  • RNA, Double-Stranded