Composite potato plants with transgenic roots on non-transgenic shoots: a model system for studying gene silencing in roots

Plant Cell Rep. 2014 Dec;33(12):1977-92. doi: 10.1007/s00299-014-1672-x. Epub 2014 Sep 3.

Abstract

Composite potato plants offer an extremely fast, effective and reliable system for studies on gene functions in roots using antisense or inverted-repeat but not sense constructs for gene inactivation. Composite plants, with transgenic roots on a non-transgenic shoot, can be obtained by shoot explant transformation with Agrobacterium rhizogenes. The aim of this study was to generate composite potato plants (Solanum tuberosum) to be used as a model system in future studies on root-pathogen interactions and gene silencing in the roots. The proportion of transgenic roots among the roots induced was high (80-100%) in the four potato cultivars tested (Albatros, Desirée, Sabina and Saturna). No wild-type adventitious roots were formed at mock inoculation site. All strains of A. rhizogenes tested induced phenotypically normal roots which, however, showed a reduced response to cytokinin as compared with non-transgenic roots. Nevertheless, both types of roots were infected to a similar high rate with the zoospores of Spongospora subterranea, a soilborne potato pathogen. The transgenic roots of composite potato plants expressed significantly higher amounts of β-glucuronidase (GUS) than the roots of a GUS-transgenic potato line event. Silencing of the uidA transgene (GUS) was tested by inducing roots on the GUS-transgenic cv. Albatros event with strains of A. rhizogenes over-expressing either the uidA sense or antisense transcripts, or inverted-repeat or hairpin uidA RNA. The three last mentioned constructs caused 2.5-4.0 fold reduction in the uidA mRNA expression. In contrast, over-expression of uidA resulted in over 3-fold increase in the uidA mRNA and GUS expression, indicating that sense-mediated silencing (co-suppression) was not functional in roots. The results suggest that composite plants offer a useful experimental system for potato research, which has gained little previous attention.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Agrobacterium / drug effects
  • Agrobacterium / metabolism
  • Benzyl Compounds / pharmacology
  • Gene Silencing*
  • Genes, Plant
  • Glucuronidase / metabolism
  • Models, Biological*
  • Phenotype
  • Plant Growth Regulators / pharmacology
  • Plant Roots / drug effects
  • Plant Roots / genetics*
  • Plant Roots / growth & development
  • Plant Shoots / drug effects
  • Plant Shoots / physiology*
  • Plants, Genetically Modified
  • Plasmodiophorida / drug effects
  • Purines / pharmacology
  • RNA, Messenger / genetics
  • RNA, Messenger / metabolism
  • Solanum tuberosum / drug effects
  • Solanum tuberosum / genetics*
  • Solanum tuberosum / parasitology
  • Transformation, Genetic / genetics
  • Transgenes

Substances

  • Benzyl Compounds
  • Plant Growth Regulators
  • Purines
  • RNA, Messenger
  • Glucuronidase
  • benzylaminopurine