To improve the wheat crown rot (CR), an important disease in wheat area of the middle and lower reaches of Yangtze River, we have transferred three genes, TaPIMP1, AtNPR1, Gastrodianin, to Yangmai12 using particle bombardment method. Forty-three of the homozygous transgenic lines with three alien genes were obtained, and the mRNA expression levels were analyzed. Meanwhile, we evaluated the transgenic plants resistance for the diseases of wheat crown rot, sharp eyespot (SE) and Fusarium head blight (FHB). The results have showed that the alien genes could express normally in the transgenic lines. However, the resistance levels differ among 43 transgenic lines. Twenty-four transgenic lines exhibit significant elevated resistance to wheat CR comparing to the recipient variety Yangmai12. The notable correlation was also observed between CR and SE but not CR and FHB. In the study, we have obtained a series of resistant materials, including two lines with the resistance to CR, SE and FHB and one line both resistant to CR and FHB, which will play a significant role in disease resistance breeding in the future.