Agrobacterium-mediated transformation of apricot (Prunus armeniaca L.) leaf explants

Plant Cell Rep. 2008 Aug;27(8):1317-24. doi: 10.1007/s00299-008-0550-9. Epub 2008 May 1.

Abstract

A protocol for Agrobacterium-mediated stable transformation for scored, whole leaf explants of the apricot (Prunus armeniaca) cultivar Helena was developed. Regenerated shoots were selected using a two-step increased concentrations of paromomycin sulphate. Different factors affecting survival of transformed buds, including possible toxicity of green fluorescent protein (GFP) and time of exposure to high cytokine concentration in the regeneration medium, were examined. Transformation efficiency, based on PCR analysis of individual putative transformed shoots from independent lines was 5.6%, when optimal conditions for bud survival were provided. Southern blot analysis on four randomly chosen PCR-positive shoots confirmed the presence of the nptII transgene. This is the first time that stable transformation of an apricot cultivar is reported and constitutes also one of the few reports on the transformation of Prunus cultivars.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Blotting, Southern
  • Culture Media / pharmacology
  • Green Fluorescent Proteins / genetics
  • Green Fluorescent Proteins / metabolism
  • Paromomycin / pharmacology
  • Plant Leaves / drug effects
  • Plant Leaves / genetics*
  • Plant Leaves / physiology
  • Plants, Genetically Modified / drug effects
  • Plants, Genetically Modified / genetics
  • Plants, Genetically Modified / physiology
  • Polymerase Chain Reaction
  • Prunus / drug effects
  • Prunus / genetics*
  • Prunus / physiology
  • Regeneration / drug effects
  • Regeneration / genetics
  • Regeneration / physiology
  • Rhizobium / genetics*
  • Transformation, Genetic*

Substances

  • Culture Media
  • Green Fluorescent Proteins
  • Paromomycin