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Agrobacterium rhizogenes-dependent production of transformed roots from foliar explants of pepper (Capsicum annuum): a new and efficient tool for functional analysis of genes

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Abstract

Pepper is known to be a recalcitrant species to genetic transformation via Agrobacterium tumefaciens. A. rhizogenes-mediated transformation offers an alternative and rapid possibility to study gene functions in roots. In our study, we developed a new and efficient system for A. rhizogenes transformation of the cultivated species Capsicum annuum. Hypocotyls and foliar organs (true leaves and cotyledons) of Yolo Wonder (YW) and Criollo de Morelos 334 (CM334) pepper cultivars were inoculated with the two constructs pBIN-gus and pHKN29-gfp of A. rhizogenes strain A4RS. Foliar explants of both pepper genotypes infected by A4RS-pBIN-gus or A4RS-pHKN29-gfp produced transformed roots. Optimal results were obtained using the combination of the foliar explants with A4RS-pHKN29-gfp. 20.5% of YW foliar explants and 14.6% of CM334 foliar explants inoculated with A4RS-pHKN29-gfp produced at least one root expressing uniform green fluorescent protein. We confirmed by polymerase chain reaction the presence of the rolB and gfp genes in the co-transformed roots ensuring that they integrated both the T-DNA from the Ri plasmid and the reporter gene. We also demonstrated that co-transformed roots of YW and CM334 displayed the same resistance response to Phytophthora capsici than the corresponding untransformed roots. Our novel procedure to produce C. annuum hairy roots will thus support the functional analysis of potential resistance genes involved in pepper P. capsici interaction.

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Acknowledgments

We thank Hervé Etienne (CIRAD, Montpellier, France) and Claudine Franche (IRD, Montpellier, France) for the generous gift of the A. rhizogenes A4RS strain carrying pBIN-gus and pHKN29-gfp, respectively. This work was financially supported by the Genoplante Project PhytoSol-2 that resides under the ANR call. Stéphanie Mallard received a fellowship funded by the ANR, Patricio Castro received a fellowship funded by the Secreteria Nacional de Cienca y Tecnologia del Ecuador (SENACYT) from Ecuador.

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Authors and Affiliations

  1. INRA Avignon, UR 1052, Unité de Génétique et Amélioration des Fruits et Légumes, BP 94, 84143, Montfavet Cedex, France

    J. Aarrouf, P. Castro-Quezada, S. Mallard, B. Caromel & V. Lefebvre

  2. UAPV Avignon, Laboratoire de Physiologie des Fruits et des Légumes, Pôle Agrosciences, BP 21239, 84916, Avignon Cedex 9, France

    J. Aarrouf & Y. Lizzi

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  1. J. Aarrouf
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  2. P. Castro-Quezada
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  4. B. Caromel
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  5. Y. Lizzi
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  6. V. Lefebvre
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Correspondence to J. Aarrouf or V. Lefebvre.

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Communicated by P. Ozias-Akins.

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Aarrouf, J., Castro-Quezada, P., Mallard, S. et al. Agrobacterium rhizogenes-dependent production of transformed roots from foliar explants of pepper (Capsicum annuum): a new and efficient tool for functional analysis of genes. Plant Cell Rep 31, 391–401 (2012). https://doi.org/10.1007/s00299-011-1174-z

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  • DOI: https://doi.org/10.1007/s00299-011-1174-z

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