Optimization of Regeneration System of Tissue Culture and Transformation of 1Dx5 Gene without Markers in Wheat

摘要

To improve bread making quality of flour and produce transgenic plants free of selectable markers, Immature embryo scutella from Xindong No. 26 were used as explants for establishing an efficient and stable wheat regeneration system, then he linear expressing box of 1Dx5 without selectable markers was transformed into the immature embryo of Xin Dong No. 26 via particle bombardment. The result showed that MS medium containing 2, 4-D 1.5 mg/L and Dicamba 0.5 mg/L for callus induction, R medium containing ZT 1 mg/L and 2,4-D 0.01 mg/L for callus differentiation were the best efficiency, the differentiation frequency of callus was 90%; Transformed plants were screened by PCR, three transgenic plants were detected among 1000 transformed plants, only yielding the transformation rate of 0.3%. The compositions of HWM-GS were analyzed by SDS-PAGE. The HMW-GS gene 1Dx5 was expressed in some seeds of transgenic plants. Our studies lay the foundations for obtaining marker-free transformants using the linear gene via particle bombardment.