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A simple Agrobacterium tumefaciens-mediated transformation method for rapid transgene expression in Medicago truncatula root hairs

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Abstract

Medicago truncatula is widely used as a model legume for symbiotic and pathogenic microbial interaction studies. Although a number of Agrobacterium-mediated transformation methods have been developed for M. truncatula, a rapid root transformation system was not yet available for this model plant. Here, we describe an easy method for rapid transgene expression in root hairs of M. truncatula, using young seedlings co-cultivated with the disarmed hypervirulent A. tumefaciens strain AGL1. This method leads to efficient expression of various GUS and fluorescent reporters in M. truncatula root hairs. We showed that transgene expression is detected as soon as 2 days following co-culture, in root hairs of a particular responsive zone lying 0.5–2 cm behind the root tip. This method can be used with a variety of M. truncatula genotypes, and is particularly useful for rapid investigation of the sub-cellular localization of fluorescent fusion proteins. Moreover, combining distinct Agrobacterium strains during the initial co-culture step efficiently generates co-transformed root hairs, suitable for co-localization of different fluorescent fusion proteins in the same cell.

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Acknowledgements

We would like to thank Christophe Jacquet (LRSV, Toulouse) for providing F83.005-5 seeds, Sandra Bensmihen (LIPM, Toulouse) for the pLeEXT-GUS and pCO2-GUS constructs, the FRAIB imaging platform for technical support on confocal imaging, Marie-Françoise Jardinaud (LIPM, Toulouse) for helpful advices on statistical analyses, David G. Barker (LIPM, Toulouse) for helpful discussions and the reviewers for valuable comments. This work was supported by the French Laboratory of Excellence project “TULIP” (ANR-10-LABX-41) and the ANR COME-IN (ANR- 14–CE35-0007).

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Authors and Affiliations

  1. LIPM, Université de Toulouse, INRA, CNRS, Castanet-Tolosan, France

    Céline Remblière, Joëlle Fournier, Fernanda de Carvalho-Niebel & Mireille Chabaud

Authors
  1. Céline Remblière
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  2. Joëlle Fournier
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  3. Fernanda de Carvalho-Niebel
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  4. Mireille Chabaud
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Contributions

MC and F de CN conceived and designed research. MC and CR conducted experiments and analyzed the data. JF and F de CN contributed with microscopy analyses. MC made statistical analyses. MC, JF, and F de CN wrote the paper. All authors read and approved the final manuscript.

Corresponding authors

Correspondence to Fernanda de Carvalho-Niebel or Mireille Chabaud.

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The authors declare that they have no competing interests.

Additional information

Communicated by Sergio J. Ochatt.

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Remblière, C., Fournier, J., de Carvalho-Niebel, F. et al. A simple Agrobacterium tumefaciens-mediated transformation method for rapid transgene expression in Medicago truncatula root hairs. Plant Cell Tiss Organ Cult 132, 181–190 (2018). https://doi.org/10.1007/s11240-017-1323-3

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  • DOI: https://doi.org/10.1007/s11240-017-1323-3

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