Agrobacterium rhizogenes-mediated transformation of Prunus as an alternative for gene functional analysis in hairy-roots and composite plants

Bosselut Nathalie, Van Ghelder Cyril, Claverie Michel, Voisin Roger, Onesto Jean-Paul, Rosso Marie-Noëlle, Esmenjaud Daniel. 2011. Agrobacterium rhizogenes-mediated transformation of Prunus as an alternative for gene functional analysis in hairy-roots and composite plants. Plant Cell Reports, 30 (7) : 1313-1326.

Quartile : Q2, Sujet : PLANT SCIENCES

Résumé : Resistant rootstocks offer an alternative to pesticides for the control of soil pests. In Prunus spp., resistance loci to root-knot nematodes (RKN) have been mapped and a transformation method is needed to validate candidate genes. Our efforts have focused on the generation of transformed hairy-roots and composite plants appropriate for nematode infection assays. An efficient and reliable method using the A4R strain of Agrobacterium rhizogenes for the transformation of Prunus roots with an Egfp reporter gene is given. The rooting efficiency, depending on the genotypes, was maximal for the interspecific hybrid 253 (Myrobalan plum × almond-peach), susceptible to RKN, that was retained for subsequent studies. From the agro-inoculated cuttings, 72% produced roots, mainly at the basal section of the stem. Transformed roots were screened by microscope detection of Egfp fluorescence and molecular analyses of the integration of the transgene. The absence of residual agrobacteria in the plants was checked by the non-amplification of the chromosomal gene chvH. Egfp was expressed visually in 76% of the rooted plants. Isolated hairy roots in Petri dishes and composite plants (transformed roots and non-transformed aerial part) in soil containers were inoculated with the RKN Meloidogyne incognita. In both cases, root transformation did not affect the ability of the nematodes to develop in the root tissues. Our results showed that isolated hairy-roots can be used to validate candidate genes and the conditions in which composite plants offer a complementary system for studying the function of root genes in physiological conditions of whole plants are discussed.

Mots-clés Agrovoc : Prunus, Agrobacterium rhizogenes, porte greffe, système racinaire, poil végétal, transformation génétique, plante transgénique, génie génétique, résistance aux organismes nuisibles, nématode des plantes

Mots-clés géographiques Agrovoc : France

Classification Agris : H10 - Ravageurs des plantes
F30 - Génétique et amélioration des plantes

Champ stratégique Cirad : Axe 1 (2005-2013) - Intensification écologique

Auteurs et affiliations

• Bosselut Nathalie, INRA (FRA)
• Van Ghelder Cyril, INRA (FRA)
• Claverie Michel, CIRAD-BIOS-UMR DAP (FRA)
• Voisin Roger, INRA (FRA)
• Onesto Jean-Paul, INRA (FRA)
• Rosso Marie-Noëlle, INRA (FRA)
• Esmenjaud Daniel, INRA (FRA)

Source : Cirad - Agritrop (https://agritrop.cirad.fr/562641/)

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