基于玉米自交系E28的农杆菌介导与基因枪转化体系的建立及比较

Comparison and Established the Transformation Systems of Agrobacterium-mediated and Gene-gun in Maize

以玉米自交系E28的胚性愈伤组织为受体,分别利用农杆菌介导法和基因枪轰击法对携带有GUS基因和Bar基因的质粒进行转化,借助GUS基因的瞬时表达率和抗性愈伤组织比例建立体系,并对两种方法进行比较。农杆菌介导法以OD6000.6、浸染30 min、25℃共培养3 d、加入0.01%的silwet L-77为最佳参数;基因枪轰击法为4.5 MPa、11 cm轰击距离最佳参数。农杆菌介导法与基因枪轰击法的GUS瞬时表达率没有显著差异,基因枪法的抗性愈伤组织比例显著高于农杆菌介导法,基因枪轰击法较农杆菌介导法更有利于获得转基因后代。

By respectively using agrobacterium-mediated transformation and gene-gun bombardment,GUS and Bar gene were transformed to the embryogenic callus of line E28.With the transient GUS expression and the proportion of PPT-resistant callus establish the transformation system,two methods were compared.In the agrobacterium-mediated transformation,OD600 0.6,30 min,25℃,co-cultivation 3 days and adding 0.01% silwet L-77 were thought the appropriate parameters.Four point five MPa bombardment pressure combine with 11cm bombardment distance were most appropriate in the gene-gun transformation.Compared the agrobacterium-mediated with gene-gun,the transient GUS expression was no significant difference between two methods,but the proportion of PPT-resistant callus of the gene-gun method were extremely significant higher than agrobacterium-mediated transformation.So,the gene-gun method maybe more conducive to require the transgenic offspring,facilitation of the late management.