根癌农杆菌介导甲型肝炎病毒(HAV)衣壳蛋白融合基因转化柑桔成年材料的研究.PDFVIP

分子植物育种，2009 年，第7 卷，第1 期，第51-55 页 Molecular Plant Breeding, 2009, Vol.7, No.1, 51-55 研究报告 Research Report 根癌农杆菌介导甲型肝炎病毒(HAV)衣壳蛋白融合基因转化柑桔成年 材料的研究 1,3 1,3 1,2,3 1,3 1,3 1,3 1,2,3 1,2,3* 彭爱红 彭祝春 何永睿 许兰珍 雷天刚 刘小丰 姚利晓 陈善春 1 中国农业科学院柑桔研究所, 重庆, 400712; 2 国家柑桔工程技术研究中心, 重庆, 400712; 3 国家柑桔品种改良中心, 重庆, 400712 * 通讯作者, scchen2004@ 摘 要 本研究以柑桔优良品种不知火(shinanui)为试料，通过携带质粒pBI121-A 的根癌农杆菌转化成年 树腋芽，获得转甲型肝炎病毒(HAV)衣壳蛋白融合基因不知火植株。研究结果表明，锦橙试管砧木培养15 d 时不知火成年树腋芽的嫁接成活率最高，达到90% ；不知火不同成熟度的枝条应采用不同的消毒时间，木 质化的枝条消毒时间要长些，半木质化的枝条消毒时间要短些。对获得的拟转化植株进行PCR 分析以及 对PCR 阳性植株进行测序，结果表明HAV 衣壳蛋白融合基因已整合到不知火的基因组中。 关键词 柑桔, 根癌农杆菌, 甲型肝炎病毒衣壳蛋白融合基因, 转化 Agrobacterium -mediated Transformation of Citrus Mature Explants with Hepatitis a Virus Capsid Protein Fusion Gene Peng Aihong 1,3 Peng Zhuchun 1,3 He Yongrui 1,2,3 Xu Lanzhen 1,3 Lei Tiangang 1,3 Liu Xiaofeng 1,3 Yao Lixiao 1,2,3 Chen Shanchun 1,2,3* 1 Citrus Research Institute, Chinese Academy of Agricultural Sciences, Chongqing, 400712; 2 Engineering Research Center of National Citrus, Chongqing, 400712; 3 National Citrus Cultivar Improvement Center of China, Chongqing, 400712 * Corresponding author, scchen2004@ Abstract With the mature axillary buds of citrus excellent variety Shinanui ( Citrus Reticulate×Citrus sinensis) severed as the explant, The plantlets which carried the hepatitis a virus capsid protein fusion gene were obtained through Agrobacterium -mediated transformation. In this study, survival in graft of mature axillary buds of shinanui was the highest, up to 90%, when the orange rootstocks were cultured 15 days. Sterilization time was effected on maturity of shinanui branches. The more maturity of branches, the longer of sterilization time was. The putative transgenic plantlets was selected by PCR, PCR amplification product of PCR positive plantlets were sequenced, and