柑桔栽培品种高效基因转化新技术研究

An efficent Agrobacterium-mediated in vivo transformation system was established by infecting wounded in vitro seedlings whose shoot-tops were excised with the wound wrapped with Parafilm. The two synthetic antibacterial peptide genes (Cecropin B gene and Shiva A gene ) were simultaneously introduced into the commercial citrus cultivars, and the transgenic plants were obtained. Evaluation of resistance to kanamycin of leaf disc from Kan-resistant plantlets indicated that the transfromation efficency of Jincheng orange (Citrus sinensis Osb ), Xinhui orange (C. sinensis Osb) and Shatian pummelo (C. grandis ) was up to 46.3%, 39.5%, and 43.7% respectively. PCR analysis of the Kan-resistant plantlets showed that both antibacterial peptide gene had already been integrated into genomes of the citrus transgenic plants. This approach was a simple efficent and easily repeated method fro the gene transfromation of a wide range of woody fruit species.