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Comparison of gene delivery methods for transformation ofRobinia pseudoacacia L. by Ti-plasmid
Authors:Ren Chen  Koichiro Gyokusen  Akira Saito
Institution:(1) Department of Forestry, Faculty of Agriculture, Kyushu University, 812-8581 Fukuoka, Japan
Abstract:The binary vector plasmid pIG121-Hm carrying the chimeric neomycin phosphotransferase (npt II), β-glucuronidase (gusA) and hygromycin phosphotransferase (hpt) genes was delivered intoRobinia pseudoacacia L. callus using theAgrobacterium tumefaciens-mediated transformation method and particle gun transformation method. It was determined that adding 10 mg/L of acetosyringone toAgrobacterium infection medium and callus-Agrobacterium co-culture medium could increaseAgrobacterium-mediated transformation efficiency. In contrast, particle gun transformation could successfully deliver plasmid DNA into the callus, but its transformation efficiency was lower and gene expression was transient in comparison with theAgrobacterium-mediated method, suggesting that substantially stronger promoters might be required.
Keywords:Agrobacterium tumefaciens            callus culture  particle gun            Robinia pseudoacacia L    Ti-plasmid transformation
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