柑桔栽培品种高效基因转化新技术研究

采用试管实生苗去芽创伤－农杆菌感染法，将人工合成的抗菌肽基因转入柑桔栽培品种，利用后期筛选和Ｐｅｒａｆｉｌｍ包缠法，提高了转化效率。拟转化植株的叶圆片抗Ｋａｎ能力分析结果表明，锦橙、新会橙和沙田袖的转化效率分别达到４６．３％，３９．５％和４３．７％。根据对 Ｋａｎ阳性植株的ＰＣＲ分析，证明有抗菌肽基因出现。结果表明，这是一种简便、高效、重复性好的实用基因转化方法。

New Gene Transformation Technique for Commercial Citrus Cultivars

An efficient 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 transformation efficiency 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 genes had already been integrated into genomes of the citrus transgenic plants. This approach was a simple.efficient and easily repeated method for the gene transformation of a wide range of woody fruit species.