一种根癌农杆菌介导的简单、快捷转化雪柑的新方法

A Simple and Rapid Method for Transformation of "Xuegan" Using Agrobacterium tumefaciens

以雪柑种子为外植体,以pC1301-PMI-LFY重组载体为转化载体,将蘸有农杆菌侵染液的接种针在种子胚的顶端分生组织部位刺2-3次,深度1 mm左右,当实生苗成活后移栽。通过PCR检测和PCR-Southern杂交检测得到了1株转基因植株,转化率为4.35%。针刺法简便快速,在雪柑上能够成功应用,为木本植物的遗传转化提供了新的思路。

To inoculate A.tumefaciens into the embryonic apical meristerm of the soaked seeds,a region on the seed suiface where a shoot would later emerge was pierced twice up to a depth of about 1 mm with a needle dipped in the A.tumefaciens inoculum.Germinated seedings were transplanted.One transgenic plantlet was obtained,which was confirmed by PCR assay and PCR-Southern blot.The transformation rate reached to 4.35%,and the feasibility of this method was proved,which could provid reference for the transgene of woody trees.