建立低能离子束介导小麦转基因方法并获得转GUS基因植株  

Establishment of a Method for GUS Gene Transferring into Wheat (Triticum astivum L.) Embryos by Low Energy Ion

作　　者：吴丽芳[1] 李红[1] 宋道君 冯慧云 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实验室,合肥230031

出　　处：《Acta Genetica Sinica》2000年第11期982-991,共10页

基　　金：国家自然科学基金重大项目!(119890300);国家重点科技攻关项目!(96-538-02-01)资助

摘　　要：研究了注入离子种类、能量、剂量等参数对于低能离子束介导的遗传转化的影响，建立了适于小麦成熟胚转化的组培条件和筛选程序。以携带ＧＵＳ基因的质粒为供体，进行了报告基因转化研究。分子生物学证据表明ＧＵＳ基因已整合到小麦基因组中。３个小麦品种的抗性愈伤转化率分别为 ９．５％、 １０．８％、 １１．２％，再生植株转化率分别为 １．４％、 ３．４％、 １７％。首次证明了离子束介导小麦遗传转化是可行的，为离子束介导小麦遗传转化体系的建立打下了基础。Physical parameters influencing transformation of wheat mediated by low energy ion beam, including type of ion, parameters of ion energy, dose and dose rate, were studied. Ar+ was regarded as suitable ions implanted in transformation. 20- 25keV of energy, 4.68 ×1016 ions/cm2 of dose, 2.6 ×1015 ions/cm2 of dose rate were chosen as appropriate implantation parameters. The suitable culture conditions for induction and growth of callus and the optimal selection scheme were established, After implantation and selection, resistant calli and hygromycin-resistant plantlets were obtained from three varieties. Molecular analysis data proved that GUS gene had integrated into the wheat genome. The mature embryo transformation efficiency of wherat variety Yangmai 5, Yangmai 158, Wanmai 32 reached 9.5%, 10.8%, 11.2% measured in produced hygromycin-resistant callus and l.4%, 3.4%, l.7% measured in regenerated plants, respectively, This experiment provides a basis for further investigation of wheat transformation system. Low energy ion beam mediated transformation can be extended to other plant recalcitrant to Agrobacterium tumefaciens as soon as methodological parameters are optimized.

关 键 词：低能离子束 小麦 遗传转化 GUS基因 基因表达 

分 类 号：Q943[生物学—植物学]