Optimization of Agrobacterium-mediated genetic transformation of wheat and transformation of HMW-GS 1Bx14 gene

The genetic transformation of wheat Mianyang 19 and Luoyang 8716 immature embryo callus mediated by Agrobacterium tumefaciens was studied.The factors that influence the efficiency of transformation were investigated by using the transient expression of gus gene.The result showed that for both varieties,the best transformation procedure of EHA105 was Agrobacterium cell density of OD600≈0.8,incubation for 30 min.When using Agrobacterium LBA4404,the transformation efficiency of Mianyang 19 and Luoyang 8716 immature embryo callus was highest when cell density was OD600≈1.0,incubation for 30 min and cell density of OD600≈0.8,incubation for 1h,respectively.Using the optimized conditions,the high molecular weight glutenin subunit 1Bx14 gene was transformed.After selected with hygromycin,tested by PCR and PCR-Southern,the positive progenies were obtained,yielding an average transformation frequency of 0.61%.