SDAU18, an amphiploid of Ae.ventricosa with Ae.cylindrica, was identified by cytological analysis, seed storage protein electrophoresis, genomic in situ hybridization (GISH) and inoculation assessment. The results are as follows: The chromosome number of root tip cells (RTCs) of SDAU18 plants varied from 52 to 56. 28 bivalents were observed in most PMCs MI of SDAU18 with 56 chromosomes, meanwhile, a few univalents, multivalents also existed in some PMCs MI, and the average chromosome configuration was 2n = 56 = 3.21 I +19.78 II, (Ring)+6.50 II (Rod)+0.01 III +0.04 IV (Ring)R+0.01 IV (Rod). There were both Ae. ventricosa-specific bands and Ae. cylindrica-specific bands in the seed storage protein electrophoretogram of SDAU18, furthermore, SDAU18 had one novel HMW-GS not found in the parents and two novel ones not found in common wheats. By labeling the total genomic DNA of Ae. ventricosa and Ae. cylindrica as probes respectively, and using that of another parent as block, GISH of RTCs spread of SDAU18 was carried out. The green hybridization signal was observed in 14 chromosomes respectively, within 56 ones in RTCs of SDAU18. SDAU18 was immune to powdery mildew and stripe rusts. SDAU18 was an amphiploid of Ae. ventricosa with Ae. cylindrica, and had very important significance in wheat breeding and genetic improvement.