The two disease-resistance genes chitinase and glucanase, which were respectively directed by commelina yellow mottle virus promoter (CoYMV, vascular specific) and CaMV35S promoter, were introduced into cotton genome via Agrobacterium tumefaciens. Transgenic plants were obtained from two popularly cultivated varieties Jihe321 and CRIC35. After screening by spraying kanamycin over unfolding leaves, the kanamycin resistance (KmR) plants were tested by PCR and Southern blot. The results showed that there were one or two inserts of transgenes in cotton genome. Performance test of resistance of T3 families in field and greenhouse showed that seven lines were resistant or tolerant to Verticillium dahliea. Meanwhile, the resistance at seedling stage in greenhouse was in accordance with that at the boll-setting stage in field. Among the seven lines, D9910, D9915 and D9919 had a disease resistance index of 6.5, 9.4 and 9.5, respectively in field, which showed a high resistance level. Genetics analysis of the three lines showed a classical Mendelian pattern of one pair of genes, which meant that each of the three lines contains one copy of transgene. Southern blotting also confirmed the copy number of inserts.