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柑桔栽培品种高效基因转化新技术研究

New Gene Transformation Technique for Commercial Citrus Cultivars

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【作者】 洪勇何永睿陈善春唐益雄张进仁贾士荣

【Author】 Hong Yong, He Yongrui, Chen Shanchun, Tang Yixiong, Zhang Jinren, Jia Shirong ( Citrus Research Institute, Chinese Academy of Agricultural Sciences, Chongqing 400712; Biotechnology Research Center, CAAS, Beijing 100081)

【机构】 中国农业科学院柑桔研究所!重庆400712中国农业科学院生物技术中心!北京100081

【摘要】 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。

【Abstract】 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.

【基金】 本课题为“九五”农业部重点“高新技术与基础研究”项目!(编号:95农-15-03-12)
  • 【文献出处】 热带作物学报 ,CHINESE JOURNAL OF TROPICAL CROPS , 编辑部邮箱 ,2000年02期
  • 【分类号】S666
  • 【被引频次】49
  • 【下载频次】137
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