摘要： 大白菜小孢子培养获得的子叶期胚状体，经粉碎的玻璃碴摩擦后，与农杆菌共培养，在加筛选剂Basta（含phosphinothricin 5 mg/L）的培养基上，再生出数株绿苗。自交留种后，对其后代苗期进行的Basta抗性鉴定显示，有4株抗此除草剂。Southern杂交结果显示，抗性植株的基因组中各有一个bar基因插入位点。对转化株的小孢子进行再培养，后代小孢子植株对Basta抗性的分离比显示，此转基因株为杂合体。

关键词: 大白菜, 小孢子胚胎, 根癌农杆菌, bar基因, 除草剂抗性, Southern杂交

Abstract: Microspore derived embryos of Chinese cabbage (Brassica campestris ssp.pekinensis)were bruised with crushed glass while coculturing with Agrobacterium,strain C58 The strain harbours vector pDHB321 with CaMV 35S driving bar gene that confers resistance to Basta,a phophinothricin (PPT) based herbicide.There are six green plants regenerated on the selective medium with 5 mg/L PPT out of nearly one thousand treated embryos.Having sprayed a 1% solution of the commercial preparation Basta on one leaf of each plant,we found that all of the treated leaves withered then died.When doing the leaf disc assay on MS medium containing 2 mg/L BA,1 mg/L NAA,10 mg/L PPT,it was found that one of these plants remained clear green,while the control turned completely yellow The former was selfed and its seeds were collected Four of its progenies showed distinctly resistance to the spraying of 1% Basta Results of Southern blottings realved that the bar gene had been integrated into the genome of the plants Microspores of these transformants were cultured further,and plants induced The Basta resistance segregation ratio showed that the transformants were heterogametic.

Key words: Brassica campestris ssp pekinensis, Microspore derived embryo, Agrobacterium tumefaciens, Bar gene, Herbicide resistance, Southern blot

中图分类号: 

• S634.103