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

Authors

  • Anthony T. Trieu,

    1. The Samuel Roberts Noble Foundation, 2510 Sam Noble Parkway, Ardmore, Oklahoma 73402, USA, and
    Search for more papers by this author
    • Present address: Genomics Department, Dow AgroSciences, 9330 Zionsville Road, Indianapolis, IN 46268-1054, USA.

    • First co-authors.

  • Stephen H. Burleigh,

    1. The Samuel Roberts Noble Foundation, 2510 Sam Noble Parkway, Ardmore, Oklahoma 73402, USA, and
    Search for more papers by this author
    • Present address: Centre for Plant-Microbe Symbioses, Risoe National Laboratory, Box 49, DK4000, Roskilde, Denmark.

    • First co-authors.

  • Igor V. Kardailsky,

    1. The Samuel Roberts Noble Foundation, 2510 Sam Noble Parkway, Ardmore, Oklahoma 73402, USA, and
    2. The Salk Institute for Biological Studies, 10010 N. Torrey Pines Road, La Jolla, CA 92037, USA
    Search for more papers by this author
    • §

      Present address: Plant Gene Expression Centre/USDA, 800 Buchanan Street, Albany, CA 94710, USA.

    • First co-authors.

  • Ignacio E. Maldonado-Mendoza,

    1. The Samuel Roberts Noble Foundation, 2510 Sam Noble Parkway, Ardmore, Oklahoma 73402, USA, and
    Search for more papers by this author
  • Wayne K. Versaw,

    1. The Samuel Roberts Noble Foundation, 2510 Sam Noble Parkway, Ardmore, Oklahoma 73402, USA, and
    Search for more papers by this author
  • Laura A. Blaylock,

    1. The Samuel Roberts Noble Foundation, 2510 Sam Noble Parkway, Ardmore, Oklahoma 73402, USA, and
    Search for more papers by this author
  • Heungsop Shin,

    1. The Samuel Roberts Noble Foundation, 2510 Sam Noble Parkway, Ardmore, Oklahoma 73402, USA, and
    Search for more papers by this author
  • Tzyy-Jen Chiou,

    1. The Samuel Roberts Noble Foundation, 2510 Sam Noble Parkway, Ardmore, Oklahoma 73402, USA, and
    Search for more papers by this author
  • Hiroaki Katagi,

    1. The Samuel Roberts Noble Foundation, 2510 Sam Noble Parkway, Ardmore, Oklahoma 73402, USA, and
    Search for more papers by this author
  • Gary R. Dewbre,

    1. The Samuel Roberts Noble Foundation, 2510 Sam Noble Parkway, Ardmore, Oklahoma 73402, USA, and
    Search for more papers by this author
  • Detlef Weigel,

    1. The Salk Institute for Biological Studies, 10010 N. Torrey Pines Road, La Jolla, CA 92037, USA
    Search for more papers by this author
  • Maria J. Harrison

    Corresponding author
    1. The Samuel Roberts Noble Foundation, 2510 Sam Noble Parkway, Ardmore, Oklahoma 73402, USA, and
      For correspondence (fax +1 580 2217380; e-mail
    Search for more papers by this author

For correspondence (fax +1 580 2217380; e-mail mjharrison@noble.org).

Summary

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.

Ancillary