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

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.

Author(s): 
Bosselut, Nathalie
Van Ghelder, Cyril
Claverie, Michel
Voisin, Roger
Onesto, Jean-Paul
Rosso, Marie-Noëlle
Esmenjaud, Daniel
Item Type: 
Journal Article
Publication Title: 
Plant Cell Reports
Journal Abbreviation: 
Plant Cell Rep.
Publication Date: 
2011-07
Publication Year: 
2011
Pages: 
1313-1326
Volume: 
30
Issue: 
7
ISSN: 
1432-203X
DOI: 
10.1007/s00299-011-1043-9
Library Catalog: 
PubMed
Extra: 
PMID: 21409553

Turabian/Chicago Citation

Nathalie Bosselut, Cyril Van Ghelder, Michel Claverie, Roger Voisin, Jean-Paul Onesto, Marie-Noëlle Rosso and Daniel Esmenjaud. 2011-07. "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. 10.1007/s00299-011-1043-9.

Wikipedia Citation

<ref> {{Cite journal | doi = 10.1007/s00299-011-1043-9 | issn = 1432-203X | volume = 30 | pages = 1313-1326 | last = Bosselut | first = Nathalie | coauthors = Van Ghelder, Cyril, Claverie, Michel, Voisin, Roger, Onesto, Jean-Paul, Rosso, Marie-Noëlle, Esmenjaud, Daniel | title = Agrobacterium rhizogenes-mediated transformation of Prunus as an alternative for gene functional analysis in hairy-roots and composite plants | journal = Plant Cell Reports | date = 2011-07 | pmid = | pmc = }} </ref>