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Gene Silencing in Medicago truncatula Roots Using RNAi

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Legume Genomics

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1069))

Abstract

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.

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Acknowledgments

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.

Author information

Authors and Affiliations

  1. Boyce Thompson Institute for Plant Research, Ithaca, NY, USA

    Daniela S. Floss, Alexa M. Schmitz & Maria J. Harrison

  2. Department of Plant Biology, University of Minnesota, St. Paul, MN, USA

    Colby G. Starker & J. Stephen Gantt

  3. Department of Genetics, Cell Biology, and Development University of Minnesota, Minneapolis, MN, USA

    Colby G. Starker

  4. Center for Genome Engineering, University of Minnesota, Minneapolis, MN, USA

    Colby G. Starker

Authors
  1. Daniela S. Floss
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  2. Alexa M. Schmitz
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  3. Colby G. Starker
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  4. J. Stephen Gantt
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  5. Maria J. Harrison
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Editor information

Editors and Affiliations

  1. CILR, The University of Newcastle Environmental & Life Sciences, Callaghan, New South Wales, Australia

    Ray J. Rose

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© 2013 Springer Science+Business Media, LLC

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Floss, D.S., Schmitz, A.M., Starker, C.G., Gantt, J.S., Harrison, M.J. (2013). Gene Silencing in Medicago truncatula Roots Using RNAi. In: Rose, R. (eds) Legume Genomics. Methods in Molecular Biology, vol 1069. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-613-9_12

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  • DOI: https://doi.org/10.1007/978-1-62703-613-9_12

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  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-62703-612-2

  • Online ISBN: 978-1-62703-613-9

  • eBook Packages: Springer Protocols

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