Callus induction and regeneration of wheat is highly genotype dependent. To establish an optimal system for callus induction, regeneration and highly efficient genetic transformation of the wheat variety Alondra’s, we studied the effects of culture medium, hormones on the callus induction and plant regeneration of immature embryos. Use of the N₆ medium, 3 mg·L^–1 2,4-D, together with 1 000 mg·L^–1 CH, achieved the best callus induction, and use of 4 mg·L^–1 ZT without indoleacetic acid had the highest regeneration frequency. The vector pCAMBIA1301-220.6 was constructed and transformed into Alondra’s by particle bombardment. Thirty hygromycin-resistant plants were selected by use of 100 mg·L^–1 hygromycin and regenerated. Among them, 5 plants were identified to be positive by PCR, with transformation frequency of 0.5%. This system has further enriched the wheat variety Alondra’s for transformation and will be helpful for both genetic improvement and gene function analysis in various backgrounds by genetic transformation.