转苏云金杆菌杀虫晶体蛋白基因抗虫水稻的GUS和PCR辅助选择

GUS and PCR assisted-selection of antipest rice of transferring Bacillus thuringiensis crystal protein genes

报道了转基因抗虫水稻克螟稻 1号 (Ts 9)与非转基因恢复系川恢 94 9、H97810 17及保持系 2 12B等杂交F3 代的GUS和PCR检测。GUS检测的结果 ,Ts 9与非转基因川恢 94 9、H97810 17杂交F3 代有较多的GUS阳性植株。GUS阴性植株与阳性植株的分离比约为 2∶1。用设计合成的一对引物进行PCR扩增的结果 ,大部分GUS检测呈阳性的植株及一个保持系 2 12B与Ts 9杂交F3 代植株获得了大小约 12 0 0bp的PCR产物 ,即cryIA (b)基因片段。试验结果表明PCR是比GUS叶片染色法更为灵敏、准确、方便的检测方法。

GUS and PCR detection of crossing F 3 progenies between transgenic antipest rice KMD1(Ts-9) and non-transgenic rice restoring line Chuanhui 949 and H97810-17,a maintainer line 212B,and etc.were reported in this paper.Resuls of the GUS detection showed that there were more positive plants in the F 3 progenies crossing between Ts-9 and Chuanhui 949 and H97810-17.Separation ratio of GUS negative plants and the GUS positive plants were about 2∶1.Results of the PCR detection with a pair of designed and synthesized primers showed that a size of 1200 bp PCR product was amplified in the majority of the GUS positive rice plants and a plant from crossing between Ts-9 and normal rice maintainer line 212B.The experimental results showed that the PCR detective method was more sensitive,accurate and easier than the GUS histochemical method.