Gene Silencing in Medicago truncatula Roots Using RNAi

  • Daniela S. Floss
  • Alexa M. Schmitz
  • Colby G. Starker
  • J. Stephen Gantt
  • Maria J. Harrison
Part of the Methods in Molecular Biology book series (MIMB, volume 1069)


Medicago truncatula is used widely as a model system for studies of root symbioses, interactions with parasitic nematodes and fungal pathogens, as well as studies of development and secondary metabolism. In Medicago truncatula as well as other legumes, RNA interference (RNAi) coupled with Agrobacterium rhizogenes-mediated root transformation, has been used very successfully for analyses of gene function in roots. One of the major advantages of this approach is the ease and relative speed with which transgenic roots can be generated. There are several methods, both for the generation of the RNAi constructs and the root transformation. Here we provide details of an RNAi and root transformation protocol that has been used successfully in M. truncatula and which can be scaled up to enable the analysis of several hundred constructs.

Key words

Legume Gene knockdown Functional genomics 



The authors thank Armando Bravo and Sergey Ivanov for their reviews and useful comments on the manuscript. Financial support for research was provided by the US National Science Foundation, grants DBI-0421676 and IOS-1127155.


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Copyright information

© Springer Science+Business Media, LLC 2013

Authors and Affiliations

  • Daniela S. Floss
    • 1
  • Alexa M. Schmitz
    • 1
  • Colby G. Starker
    • 2
    • 3
    • 4
  • J. Stephen Gantt
    • 2
  • Maria J. Harrison
    • 1
  1. 1.Boyce Thompson Institute for Plant ResearchIthacaUSA
  2. 2.Department of Plant BiologyUniversity of MinnesotaSt. PaulUSA
  3. 3.Department of GeneticsCell Biology, and Development University of MinnesotaMinneapolisUSA
  4. 4.Center for Genome EngineeringUniversity of MinnesotaMinneapolisUSA

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