Transformation of Medicago truncatula via infiltration of seedlings or flowering plants with Agrobacterium

Plant J. 2000 Jun;22(6):531-41. doi: 10.1046/j.1365-313x.2000.00757.x.

Abstract

Two rapid and simple in planta transformation methods have been developed for the model legume Medicago truncatula. The first approach is based on a method developed for transformation of Arabidopsis thaliana and involves infiltration of flowering plants with a suspension of Agrobacterium. The second method involves infiltration of young seedlings with Agrobacterium. In both cases a proportion of the progeny of the infiltrated plants is transformed. The transformation frequency ranges from 4.7 to 76% for the flower infiltration method, and from 2.9 to 27.6% for the seedling infiltration method. Both procedures resulted in a mixture of independent transformants and sibling transformants. The transformants were genetically stable, and analysis of the T2 generation indicates that the transgenes are inherited in a Mendelian fashion. These transformation systems will increase the utility of M. truncatula as a model system and enable large-scale insertional mutagenesis. T-DNA tagging and the many adaptations of this approach provide a wide range of opportunities for the analysis of the unique aspects of legumes.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Blotting, Southern
  • DNA, Bacterial / genetics
  • DNA, Plant / analysis
  • Gene Transfer Techniques*
  • Medicago sativa / genetics*
  • Plants / genetics
  • Plants, Genetically Modified
  • Rhizobium / genetics*
  • Transformation, Genetic

Substances

  • DNA, Bacterial
  • DNA, Plant
  • T-DNA