title: Turf Grasses.
abstract: A reliable and efficient genetic transformation protocol for various turfgrass species and elite cultivars has been achieved using Agrobacterium tumefaciens. We describe a general protocol for the establishment of embryogenic cell cultures, Agrobacterium tumefaciens-mediated transformation, selection, and regeneration of transgenic turfgrass plants. Embryogenic callus is initiated from mature seeds, maintained by visual selection, and infected with an Agrobacterium tumefaciens strain (LBA4404) that contains either an herbicide-resistant bar gene or an antibiotic-resistant hyg gene driven either by a rice ubiquitin or CaMV35S promoter. Stable transformation efficiencies up to 43.3% were achieved. Southern blot and genetic analysis was used to confirm transgene integration in the turfgrass genomes and normal transmission and stable expression of the transgene in the T1 generation. We demonstrate herein that five elite cultivars of bentgrass can be genetically transformed using this single tissue culture media regime. Additionally, we report the successful Agrobacterium-mediated transformation of an elite tall fescue variety using minor variations in the same transformation protocol.
authors.affiliation: HybriGene Inc., West Kingston, RI, USA.
authors.forename: Chip
authors.initials: C
authors.lastname: Longo
authors.forename: Colin
authors.initials: C
authors.lastname: Lickwar
authors.forename: Qian
authors.initials: Q
authors.lastname: Hu
authors.forename: Kimberly
authors.initials: K
authors.lastname: Nelson-Vasilchik
authors.forename: David
authors.initials: D
authors.lastname: Viola
authors.forename: Joel
authors.initials: J
authors.lastname: Hague
authors.forename: Joel M
authors.initials: JM
authors.lastname: Chandlee
authors.forename: Hong
authors.initials: H
authors.lastname: Luo
authors.forename: Albert P
authors.initials: AP
authors.lastname: Kausch
mesh_terms: D016960:Agrobacterium tumefaciens / Q000166:cytology / Q000235:genetics*; D031703:Agrostis / Q000033:anatomy & histology / Q000235:genetics* / Q000254:growth & development; D018929:Cell Culture Techniques; D003470:Culture Media; D005819:Genetic Markers; D012639:Seeds / Q000235:genetics / Q000254:growth & development; D046509:Tissue Culture Techniques; D014170:Transformation, Genetic
chemical_list: D003470:Culture Media; D005819:Genetic Markers