基因枪微弹轰击烟草叶绿体遗传转化体系的建立  

Establishment of Tobacco Plastid Genetic Transformation Via Bombardment Microprojectiles

作　　者：任延国[1] 邹竹荣[1] 张中林[1] 于晓兰[1] 山松[1] 沈桂芳[1] 

机构地区：[1]中国农业科学院生物技术研究中心

出　　处：《农业生物技术学报》1997年第1期47-53,共7页Journal of Agricultural Biotechnology

基　　金：国家自然科学基金

摘　　要：以质粒ｐＺＳ１９７ＤＮＡ制备金粉子弹，通过基因枪轰击烟草叶片，经壮观霉素筛选获得了分化愈伤组织和抗性植株。进一步对烟草叶绿体转化植株进行点杂交、Ｓｏｕｔｈｅｒｎ杂交和ＰＣＲ扩增等分子检测，结果证明２－０１植株中外源ａａｄＡ（氨基糖苷３＇－腺苷酸转移酶）基因通过同源重组已经整合到它的叶绿体基因组上，并在１６ＳｒＲＮＡ启动子下得到表达而产生壮观霉素（Ｓｐｃ）抗性；同时以２－０１植株为母本的杂交及自交后代种子的萌发子叶均抗壮观霉素，呈现母系遗传方式。Spectinomycin resistant callus and shoots were obtained from tobacco leaves bombarded with golden particles coated with pZS197 DNA. Further molecular screening on the transformed tobacco plants such as dot blot, Southern blot and PCR amplifying were performed, which indicated that foreign aadA gene had been integrated into the plastid genome of No.2 01 transformant and expressed under the control of 16SrRNA promoter with the resistance to spectinomycin. Moreover, seedlings of the hybrid of No.2 01 transgenic tobacco plant acted as the female showed the resistance to spectinomycin as well as those of its self pollinated progeny with the mode of maternal heredity. All of above results showed that stable tobacco plastid genetic transformation system had been established, which provided a new pathway for plant engineering.

关 键 词：基因枪 叶绿体 遗传转化 烟草 

分 类 号：S572.035.3[农业科学—烟草工业]