A simple shoot multiplication procedure using internode explants, and its application for particle bombardment and Agrobacterium-mediated transformation in watercress

J Plant Res. 2009 Jul;122(4):455-63. doi: 10.1007/s10265-009-0226-8. Epub 2009 Mar 24.

Abstract

A shoot multiplication system derived from internode explants was investigated with the aim of improving genetic characteristics of watercress (Nasturtium officinale R. Br.). Internodes of ca. 1 cm excised from in vitro stock shoot culture were placed on half-strength Murashige and Skoog (MS) medium supplemented with 3 muM 2,4-dichlorophenoxyacetic acid as a pre-treatment. Laser scanning microscopy indicated clearly that the first sign of meristematic cell division could be seen after 1-2 days of pre-culture, and meristematic tissues multiplied along the vascular cambium of the internode segment during 7 days of culture. Multiple shoots could be obtained from more than 90% of the pre-treated explants when they were subsequently transferred to MS medium supplemented with 1 muM thidiazuron for 3 weeks. These findings indicate that pre-treatment of the internodes for 7 days promoted their capacity for organogenesis. Using this pre-treatment, frequent generation of transgenic watercress plants was achieved by adapting particle bombardment and Agrobacterium-mediated transformation techniques with a construct expressing a synthetic green florescent protein gene.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Biolistics / methods*
  • Brassicaceae / cytology
  • Brassicaceae / genetics*
  • Brassicaceae / microbiology*
  • Gene Expression Profiling
  • Green Fluorescent Proteins / genetics
  • Onions / genetics
  • Organogenesis / drug effects
  • Plant Growth Regulators / pharmacology
  • Plant Shoots / cytology
  • Plant Shoots / drug effects
  • Plant Shoots / embryology
  • Plant Shoots / growth & development*
  • Plant Stems / cytology
  • Plant Stems / drug effects
  • Plant Stems / embryology
  • Plant Stems / growth & development*
  • Plants, Genetically Modified
  • Rhizobium / metabolism*
  • Transformation, Genetic* / drug effects

Substances

  • Plant Growth Regulators
  • Green Fluorescent Proteins