To demonstrate the phylogenetic evolution, the molecular characteristics of the motif of HA protein cleavage site and the varieties at the receptor binding sites of the hemagglutinin gene of the duck-origin H9N2 subtype avian influenza viruses, sequence alignment and phylogenetic analysis were performed by MEGA 4.1 Neighbor-Joining method.. The results revealed that the duck-origin H9N2 AIV viruses originated from CK/BJ/1/94-like and North-Ame-like, all the duck-origin H9N2 AIV viruses from mainland China belonged to CK/BJ/1/94-like and formed multiple genotypes through complicated re-assortment, while other duck-origin H9N2 AIV, isolated from other countries in Aisa, American and European such as Korea, Japan, Alberta, Austria, Switzerland, Iran, belonged to the North-Ame-like phylogenetic lineage. The amino acids at positions 183, 190, and 226 of the receptor binding sites of North-Ame-like group isolates had highly conserved H, E and Q respectively. In contrast with duck-origin H9N2 AIV viruses isolates from mainland China, the amino acids had N at positions 183, A, T, or V at 190, L or Q at 226, which was the same as the chicken-origin H9N2 AIV from mainland China. Most newly isolated chicken-origin H9N2 AIV in Fujian Province in Southern China had L at position 226 emphasized the higher risk of cross-infection between the chicken-origin and duck-origin H9N2 AIV in China.