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Agrobacterium rhizogenes mediated transformation of grapevine (Vitis vinifera L.)
Authors:Véronique Guellec  Chantal David  Michel Branchard  Jacques Tempé
Institution:(1) Laboratoire d'AAmélioration des Plantes, Université de Paris-Sud, UA 115, Bt, 360, 91405 Orsay cedex, France;(2) Groupe de Recherche sur les Interactions entre Plantes et Microorganismes, Université de Paris-Sud, Bt. 409, 91405 Orsay cedex, France;(3) Institut des Sciences Végétales, CNRS, Av. de la Terrasse, 91198 Gif sur Yvette cedex, France
Abstract:Genetically transformed grapevine (Vitis vinifera L.) roots were obtained after inocultation of in vitro grown whole plants (cv. Grenache) with Agrobacterium rhizogenes. The strain used contains two plasmids: the wild-type Ri plasmid pRi 15834 and a Ti-derived plasmid which carries a chimaeric neomycin phosphotrans-ferase gene (NPT II) and the nopaline synthase gene. Expression of the NPT II gene can confer kanamycin resistance to transformed plant cells. Slowly growing axenic root cultures derived from single root tips were obtained. Opine analysis indicated the presence of agropine and/or nopaline in established root cultures. For one culture, the presence of T-DNA was confirmed by dot-blot hybridization with pRi 15834 TL-DNA. Callogenesis was induced by subculturing root fragments on medium supplemented with benzylaminopurine and indoleacetic acid.Transformation of in vitro cultured grapevine cells has recently been reported (baribault T.J. et al., Plant Cell Rep (1989) 8: 137–140). In contrast with the results presented here, expession of the NPT II gene Conferred kanamycin resistance to Vitis vinifera calli that was sufficient for selection of trasformed cells.Abbreviations BAP benzylaminopurine - IAA indoleacetic acid - NAA naphtaleneacetic acid - NPT II neomycin phosphostransferase II - EDTA ethylenediaminetetraacetic acid
Keywords:Agrobacterium rhizogenes  grapevine  NPT II gene  opine  T-DNA
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