title: Highly efficient transformation and plant regeneration of tall fescue mediated by Agrobacterium tumefaciens.
abstract: An efficient and reproducible system has been developed for the production of transgenic plants in tall fescue (Festuca arundinacea Schreb.) using A. tumefaciens-mediated transformation. Two-months-old suspension cultures served as excellent explants for transformation. The explants were inoculated with an A. tumefaciens strain EHA105 carrying a plasmid pDBA121 containing genes for hygromycin phosphotransferase (hpt) and phosphinotricin acetyltransferase (bar). The commercial herbicide Basta was used as a selective agent. Inclusion of acetosyringone (ACS) 20 mg/L in the co-culture medium led to an increase in transformation efficiency. The efficiency of transformation was highly dependent on the genotype, the explant, the culture medium and selective agent used. Tall fescue transformation efficiency is 2.85-10.9 plants per gram fresh weight (FW) of suspension cultures. This is much higher than the corresponding values reported before (2-5 plants). So far more than 300 transgenic plants have been obtained, the fertility of some transgenic plants had decreased. Stable integration and high expression of the transgenes were confirmed by PCR analysis and Southern blot hybridization or herbicide Basta spraying test.
authors.affiliation: Institute for Virology and Biotechnology, Zhejiang Academy of Agricultural Sciences, Hangzhou 310021, China.
authors.forename: Zhang-Hua
authors.initials: ZH
authors.lastname: Hu
authors.forename: Jin-Qing
authors.initials: JQ
authors.lastname: Chen
authors.forename: Guan-Ting
authors.initials: GT
authors.lastname: Wu
authors.forename: Wei
authors.initials: W
authors.lastname: Jin
authors.forename: Chun-Xiu
authors.initials: CX
authors.lastname: Lang
authors.forename: Rui-Zhi
authors.initials: RZ
authors.lastname: Huang
authors.forename: Fu-Lin
authors.initials: FL
authors.lastname: Wang
authors.forename: Zhi-Hong
authors.initials: ZH
authors.lastname: Liu
authors.forename: Xiao-Yun
authors.initials: XY
authors.lastname: Chen
mesh_terms: D016960:Agrobacterium tumefaciens / Q000235:genetics*; D015139:Blotting, Southern; D031751:Festuca / Q000235:genetics* / Q000502:physiology; D030821:Plants, Genetically Modified / Q000235:genetics* / Q000502:physiology; D016133:Polymerase Chain Reaction; D012038:Regeneration; D014170:Transformation, Genetic / Q000235:genetics*