大麦转化中潮霉素和草丁膦的筛选效果研究  

COMPARISON OF THE HYGROMYCIN AND BIALAPHOS AS SELECTIVE AGENTS IN BARLEY TRANSFORMATION

作　　者：韩秀丽[1] 王文良[1] 田田[1] 张陵云[2] 吴佳洁[1] 

机构地区：[1]作物生物学国家重点实验室,山东省作物生物学重点实验室,山东泰安271018 [2]东营职业学院,山东东营257000

出　　处：《山东农业大学学报（自然科学版）》2013年第4期525-532,共8页Journal of Shandong Agricultural University：Natural Science Edition

基　　金：国家自然基金重大国际合作项目(31110103917);山东省自然科学杰出青年基金(JQ201107);国家转基因专项任务(2013ZX08009-003-001-006)

摘　　要：本研究以大麦品种"Golden Promise"幼胚为受体材料,建立了农杆菌介导的大麦遗传转化方法。实验利用携带潮霉素磷酸转移酶基因HptII(潮霉素抗性)和草丁膦乙酰转移酶基因Bar(草丁膦抗性)的载体PC551,研究了潮霉素及草丁膦对转基因幼胚的愈伤组织诱导、植株再生、生根等过程的影响,结果表明潮霉素的筛选效果优于草丁膦。通过除草剂Rely280涂抹、PCR检测以及黄色荧光观察等多种方法验证转基因后代,结果表明以潮霉素为筛选剂得到再生苗的阳性率显著高于以草丁膦为筛选剂的阳性率。因此在大麦遗传转化体系中,潮霉素更适合作为筛选剂。Selective agents play important roles in generating transgenic plants. In this study, a binary construct PC551 was prepared containing two selective matrices of the hygromycin phosphotransferase gene （HptII, Hygromycin resistance） and the phosphinothricin acetyhransferase gene （Bar, Bialaphos resistance）. PC551 was used to study the efficiency of hygromycin and bialaphos in screening of transgenic plants in cultivated barley ＂ Golden Promise＂. It was shown that hygromycin performed as a superior selective agent during the course of callus induction, shoot regeneration, and root regeneration. Herbicide painting, PCR analysis, and fluorescence - detection were further used to confirm transgenic To plants. It was demonstrated that hygromycin was associated with a higher transformation efficiency than bialaphos. Therefore, hygromycin is a preferable selective agent in genetic transformation of barley.

关 键 词：大麦 遗传转化 潮霉素 草丁膦 Rely280 

分 类 号：Q789[生物学—分子生物学]