农杆菌转化花粉愈伤组织获取纯合的转基因水稻植株  

Homozygous transgenic plants regenerated from rice pollen derived callus via Agrobacterium tumefaciens.

作　　者：傅亚萍 斯华敏 朱正歌 胡国成 孙宗修 

机构地区：[1]中国水稻研究所农业部水稻生物学重点实验室,浙江杭州310006

出　　处：《浙江大学学报（农业与生命科学版）》2001年第4期407-410,共4页Journal of Zhejiang University:Agriculture and Life Sciences

基　　金：国家自然科学基金重点资助项目 (397890 2 0 );国家 973计划资助项目

摘　　要：应用农杆菌介导法成功地将玉米转座子 Ds导入粳稻 (Oryza sativa L.subsp.japonica)中花 11的花粉愈伤组织中 ,基因转化以对除草剂 PPT有抗性的 Bar基因和潮霉素抗性基因 H pt为选择标记 .共获得了 18个独立的转基因绿苗 ,其中单倍体 11株 ,二倍体 7株 .转基因植株均抗除草剂 BASTA.考查了二倍体植株种子后代 (T1)对 BASTA的抗性 ,7个株系的所有植株均抗 BASTA,而未发生感抗分离 ,表明外源基因在转基因植株中是纯合的 .PCR和 Southern blotting的研究支持上述结果 .Maize transposable element Ds was inserted into pollen derived calli of Zhonghua 11(Oryza sativa L. subsp. japonica) mediated by Agrobacterium tumefaciens. The plasmid T DNA contained a hygromycin resistance gene (Hpt) and a PPT resistant gene (Bar) as the selection markers. Seventeen green transgenic plants were regenerated, including 10 haploid and 7 diploid plants. All of the transgenic plants showed the resistance to herbicide BASTA, and the T 1 lines from diploid plants also resisted to BASTA without segregation within each line, which indicated that foreign gene was homozygous in diploid transgenic lines. PCR and Southern blot analysis indicated that Bar gene did insert in genomic DNA of Zhonghua 11. The approaches to obtain transgenic homozygous lines are discussed.

关 键 词：水稻 花药培养 转基因稻 农杆菌转化 

分 类 号：S511.03[农业科学—作物学]