博落回愈伤抗除草剂Bar基因的转化以及抗性功能的检测

Transformation of Bar Gene for Callus of Macleaya and Detection of Resistance Function

本研究成功建立了一个全新的以博落回愈伤为外植体的遗传转化体系。该遗传体系将抗草丁膦的Bar基因整合到pcambia2301质粒,构建植物载体,通过农杆菌介导转染博落回愈伤,选用Bar基因为筛选标记,采用PPT进行筛选,对转染的愈伤和组培苗用PCR、RT-PCR、GUS染色等方法验证为阳性,获得了抗除草剂的博落回植株。该遗传体系相比以叶片为外植体、抗生素为筛选标记的遗传体系,更能加快转化时间,缩短转化周期,而且假阳性更低,对于博落回生物学研究具有一定帮助。

A new genetic transformation system of Macleaya callus which used callus as explants was established in the research. The resistance PPT gene Bar was integrated into the pcambia2301 plasmid to construct plant carrier and it was transfected into the callus by Agrobacterium mediated in this integretion system. The Bar gene was used as the screening marker, and the transgenic callus and plantlets were positive by PCR, RT-PCR and GUS staining methods. Then, the herbicide resistant plants were obtained. Compared with the system using leaves as explants, antibiotics as sceening marker, the genetic system could speed up the conversion time, improve efficiency, shorten the cycle of transformation and had lower false positives. It was very helpful to the biological study of Macleaya callus.