Gossypium barbadense/G. hirsutum grafted cotton seedlings (S/R) were obtained by using G. hirsutum cotton Pima90 (R) resistant to Verticillium wilt as the rootstock and G. barbadense cotton variety Emian 22 (S) susceptible to Verticillium wilt in Hubei Province as scion. The disease resistance after incubation of Pima90 (R),Emian 22(S) and grafted cotton seedlings (S/R) was studied. H2O2 content in leaves was determined. The differences in the levels of POD/PAL enzyme activity and the expression patterns of PAL/GST genes were compared. Results showed that wilting degree of the G. barbadense/G. hirsutum grafted cotton (S/R) after the incubation of damaged cotton root with pathogen suspensions was between that of the rootstock Pima90（R）and scion Emian 22 (S),indicating that grafting improved the resistance of cotton seedlings to Verticillium wilt. H2O2 contents in leaves and the enzymatic activities of POD and PAL in Emian 22 (S),Pima90(R) and the grafted cotton (S/R) were obviously increased after incubation with Verticillium dahliae Kleb. However,H2O2 content in leaves,POD and PAL activities increased slowly in Emian 22 (S),rapidly in Pima90 (R),and moderately in the S/R grafted cotton. Pima90(R) and the grafted cotton (S/R) showed similar tendency of changing. Results of real-time PCR showed that the mRNA expression level of GST gene in three cottons increased firstly and then decreased. The duration of GST gene expression for Emian 22 (S),Pima90 (R) and grafted cotton (S/R) was 8-16 h and 8-48 h,respectively. The patterns of PAL gene expression in the three cottons were similar. Expression level of PAL gene was increased firstly,then decreased,and increased subsequently,with twice peak at 8 h and 48 h. The peak value in Emian 22 (S),Pima90(R) and grafted cotton (S/R) was low,high,and moderate.