title: Initiation and transformation of grapevine embryogenic cultures.
abstract: Protocols for the production and transformation of grapevine embryogenic cultures are described. Embryogenic cultures are initiated from leaves or stamens and pistils and transformed with Agrobacterium containing an enhanced green fluorescent protein/neomycin phosphotransferase II (egfp/nptII) fusion gene. Cultures are transferred to induction medium in the dark for callus formation and proliferation. Resulting cultures are transferred to somatic embryo development medium to induce secondary embryogenesis and formation of transgenic somatic embryos. Transgenic embryos identified on the basis on GFP fluorescence and kanamycin resistance are transferred to germination medium to regenerate transgenic plants. The presence of transgenes in independent plant lines is confirmed by PCR.
authors.affiliation: Mid-Florida Research and Education Center, University of Florida/IFAS, Apopka, FL, USA.
authors.forename: Sadanand A
authors.initials: SA
authors.lastname: Dhekney
authors.forename: Zhijian T
authors.initials: ZT
authors.lastname: Li
authors.forename: Manjul
authors.initials: M
authors.lastname: Dutt
authors.forename: Dennis J
authors.initials: DJ
authors.lastname: Gray
mesh_terms: D060054:Agrobacterium; D004269:DNA, Bacterial / Q000235:genetics; D018744:DNA, Plant / Q000235:genetics; D035264:Flowers / Q000196:embryology; D018014:Gene Transfer Techniques; D049452:Green Fluorescent Proteins / Q000235:genetics; D007612:Kanamycin / Q000494:pharmacology; D019868:Kanamycin Kinase / Q000235:genetics*; D007613:Kanamycin Resistance / Q000235:genetics; D018515:Plant Leaves / Q000196:embryology; D030821:Plants, Genetically Modified / Q000187:drug effects / Q000235:genetics / Q000254:growth & development; D011993:Recombinant Fusion Proteins; D046509:Tissue Culture Techniques; D014170:Transformation, Genetic; D027843:Vitis / Q000196:embryology / Q000235:genetics* / Q000254:growth & development
chemical_list: D004269:DNA, Bacterial; D018744:DNA, Plant; D011993:Recombinant Fusion Proteins; C036483:T-DNA; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins; D007612:Kanamycin; D019868:Kanamycin Kinase