低能离子束介导水稻遗传转化的研究

STUDY ON RICE TRANSFORMATION MEDIATED BY LOW ENERGY ION BEAM IMPLANTATION

为研究影响低能离子束介导水稻遗传转化效率的各种因素 ,以粳稻 0 2 42 8、花培 94 鉴 0 9、籼稻明恢 63这 3个水稻品种为实验材料 ,分析了离子的种类、能量、剂量、剂量率等参数对遗传转化的影响 ,建立了有利于遗传转化的组织培养条件和筛选程序。分子生物学检测证明 ,外源GUS报告基因已经整合到水稻基因组中 ,3个水稻品种获得抗性愈伤组织的转化率分别为 1 1 %、1 1 4%和 7 1 % ,获得再生试管苗的转化率分别为 1 5 2 %、1 87%和 1 1 3 %。为应用离子束介导法进行其它对农杆菌不敏感的禾谷类作物转基因提供了方法学的参照 ,并为深入研究离子束与生物体相互作用机制打下了基础。

Delivery of foreign DNA into rice via ion beam was first reported in 1994. In recent years we have aimed to set up efficient transformation system mediated by low energy ion beam. The factors that influence the transformation including type of ion, parameters of ion energy, dose and dose rate, plant genotype, composition of media, concentration of hormones and antibiotics were carefully investigated. Treated with 25keV Ar +, the transformation efficiencies of the mature embryos of rice variety 02428, Huapei94\|jian\|09 and Minghui63 reached 11%, 11 4% and 7 1% measured by produced antibiotic resistant callus and 1 52%, 1 87% and 1 13% measured by regenerated plants respectively. PCR detection and Southern blot analysis showed that GUS report gene had inserted in rice genome. Low energy ion beam mediated gene transfer will be extended to other cereal recalcitrant to Agrobacterium tumefaciens as soon as methodological parameters were optimized.