离子注入法将外源DNA直接导入小麦的研究  

STUDIES ON INTRODUCING EXOGENOUS DNA INTOWHEAT BY ION IMPLANTATION

作　　者：吴丽芳[1] 尹若春[2] 谷运红[1] 吴李君[1] 余增亮[1] 

机构地区：[1]中国科学院等离子体物理研究所离子束生物工程学实验室,安徽合肥230031 [2]安徽大学生命科学学院生物技术系,安徽合肥230039

出　　处：《生物物理学报》2001年第4期724-730,共7页Acta Biophysica Sinica

基　　金：国家重点科技攻关项目 (96 -538 -02 -01)资助课题;安徽省自然科学基金资助项目 (99047640)

摘　　要：通过离子束介导法将外源GUS基因直接导入小麦成熟种子。组织化学染色结果表明 :GUS基因的瞬时表达率可以达到70 %以上。当代(R0)PCR分析结果表明 ,阳性植株的频率与离子注入剂量有关 ,适宜的注入剂量为7×1016 离子/cm2。PCR -Southern和SouthernBlot分析结果表明外源基因已整合到小麦基因组中 ,说明离子束介导外源DNA直接导入小麦是可行的。The exogenous GUS gene was introduced into wheat mature seeds by ion implantation. Histochemical staining showed the temporary expression rate of GUS gene reached more than 70%. PCR assays of R0 generation displayed that the positive plant rate was related to dosage of implanted ions, and the suitable dosage for transformation was 7×1016ions/cm2. Further PCR-Southern and Southern blot assays proved that exogenous gene had integrated into wheat genome. The experiments provided an effective method for DNA delivery into wheat and showed a good prospect of introducing exogenous genomic DNA with helpful gene into wheat.

关 键 词：离子注入 GUS基因 小麦 直接导入 成熟种子 外源DNA 

分 类 号：S512.103[农业科学—作物学] S188