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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。
Using the embryogenic callus of maize inbred line E28 as the receptor, the plasmid carrying GUS gene and Bar gene was transformed by Agrobacterium-mediated method and gene gun bombardment method respectively. With the help of the transient expression rate and anti-GUS gene The establishment of sexual callus ratio system, and compare the two methods. Agrobacterium-mediated method with OD6000.6, dipping for 30 min, co-cultured at 25 ℃ for 3 d, adding 0.01% silwet L-77 as the best parameters; particle bombardment method 4.5 MPa, 11 cm bombardment distance best parameters. Agrobacterium-mediated method and gene gun bombardment GUS transient expression rate was not significantly different, the gene gun method of callus resistance was significantly higher than Agrobacterium-mediated method, gene gun bombardment method is more conducive to Agrobacterium-mediated method Get transgenic offspring.