Highly efficient transformation and plant regeneration of tall fescue mediated by Agrobacterium tumefaciens.
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.
Hu
Zhang-Hua
ZH
Institute for Virology and Biotechnology, Zhejiang Academy of Agricultural Sciences, Hangzhou 310021, China.
Chen
Jin-Qing
JQ
Wu
Guan-Ting
GT
Jin
Wei
W
Lang
Chun-Xiu
CX
Huang
Rui-Zhi
RZ
Wang
Fu-Lin
FL
Liu
Zhi-Hong
ZH
Chen
Xiao-Yun
XY
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*