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

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

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

Content Provider	Springer Nature Link
Author	Bosselut, Nathalie Ghelder, Cyril Claverie, Michel Voisin, Roger Onesto, Jean Paul Rosso, Marie ëlle Esmenjaud, Daniel
Copyright Year	2011
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.
Starting Page	1313
Ending Page	1326
Page Count	14
File Format	PDF
ISSN	07217714
Journal	Plant Cell Reports
Volume Number	30
Issue Number	7
e-ISSN	1432203X
Language	English
Publisher	Springer-Verlag
Publisher Date	2011-03-17
Publisher Place	Berlin, Heidelberg
Access Restriction	One Nation One Subscription (ONOS)
Subject Keyword	Prunus Agrobacterium rhizogenes Transgenic roots Genetic engineering Acclimation Nematode Cell Biology Plant Sciences Plant Biochemistry Biotechnology
Content Type	Text
Resource Type	Article
Subject	Plant Science Medicine Agronomy and Crop Science

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