title: Citrus transformation using mature tissue explants.
abstract: Mature tissue protocol for production of transgenic Citrus plants via Agrobacterium-mediated transformation uses explants derived from branches of mature, fruit-bearing trees. Through the multiple cleaning steps consisting of grafting of apical tip meristems on rootstock plants grown under sanitary conditions, "mother" plants are produced that will serve as a source of budding material. These buds are grafted onto rootstock plants grown under the same, highly sanitary conditions. Newly obtained, one meter tall, young grafted plants serve as a source of explants for co-incubation experiments with Agrobacterium. Following successful transformation with Agrobacterium, selected transgenic shoots are micrografted onto rootstock plants in vitro where they are allowed to grow for a couple of months. Grafted transgenic plantlet together with the associated rootstock plant is taken out of culture tubes, severed from the root, and regrafted in terra on a 1-year-old rootstock plant. With the application of proper horticultural techniques, such a plant will yield first fruit about 12-15 months later.
authors.affiliation: Citrus Research and Education Center, University of Florida/IFAS, 700 Experiment Station Road, Room 103, Lake Alfred, FL, 33850, USA, orbovic@ufl.edu.
authors.forename: Vladimir
authors.initials: V
authors.lastname: Orbović
authors.forename: Alka
authors.initials: A
authors.lastname: Shankar
authors.forename: Michael E
authors.initials: ME
authors.lastname: Peeples
authors.forename: Calvin
authors.initials: C
authors.lastname: Hubbard
authors.forename: Janice
authors.initials: J
authors.lastname: Zale
mesh_terms: D016960:Agrobacterium tumefaciens / Q000235:genetics / Q000254:growth & development; D002957:Citrus / Q000166:cytology / Q000235:genetics* / Q000254:growth & development*; D018920:Coculture Techniques; D005818:Genetic Engineering / Q000379:methods*; D005966:Glucuronidase / Q000235:genetics; D014170:Transformation, Genetic
chemical_list: D005966:Glucuronidase