根癌农杆菌介导的虎杖茎尖转化研究  

Agrobacterium tumefaciens-mediated Transformation of Shoot Tip of Polygonum cuspidatum

作　　者：柳忠玉[1] 赵树进[2] 

机构地区：[1]长江大学生命科学学院,荆州434025 [2]广州军区广州总医院,广州510010

出　　处：《生物技术通报》2014年第2期79-84,共6页Biotechnology Bulletin

基　　金：广东省自然科学基金(10151001002000012);长江大学博士启动基金项目(801100010122)

摘　　要：为了建立根癌农杆菌介导的虎杖茎尖遗传转化体系,以虎杖的茎尖为转化受体,研究了携带白藜芦醇合酶基因(PcRS)的根癌农杆菌载体介导的虎杖遗传转化若干因素对转化效果的影响。结果显示,较适宜的转化系统为预培养2 d,农杆菌菌液(OD600值为0.6)侵染10 min,共培养3 d,在含8 mg/L潮霉素的培养基上诱导不定芽。利用该体系从300块茎尖外植体中共转化获得15株抗性再生植株,经PCR和Southern杂交检测,有6株虎杖的基因组中已整合进了目的基因。In order to establish Agrobacterium tumefaciens-mediated transformation system of Polygonum cuspidatum shoot tip, some key factors that affect transformation frequency of P. cuspidatum shoot tip were studied. The shoot tips of P. cuspidatum was infected by Agrobacterium tumefaciens EHA105 with the plasmid which contains PcRS gene. The superior transformation system was as following, precultured 2 days and then infected 10 minutes in suspension（OD 600 =0.6）, co-cultured 3 days, transgenic tissues and shoots were selected with 8 mg/L hygromycin. Six transgenic plants from 300 explants were obtained and the intergration of transgene was confirmed by PCR and Southern blot analysis.

关 键 词：虎杖 茎尖 根癌农杆菌 遗传转化 

分 类 号：S567.239[农业科学—中草药栽培]