Abstract: To breed new tobacco varieties with resistance to tobacco mosaic virus (TMV), a new flue-cured tobacco cv. Ekao No. 2 was bred with a male sterile line of Zhongyan 103 (a quality flue-cured female variety) and K8 (a male breeding intermediate with high resistance to TMV) as the parents, respectively. The results showed that Ekao No. 2 possessed a moderate resistance to black shank and root knot nematode, immunity to TMV, and high resistance to climate spot disease. With improved adaptability, the comprehensive disease resistance of Ekao No. 2 was better than that of the control varieties (K326 and Yunyan 87). The flue-cured leaves of Ekao No. 2 displayed well-distributed golden color and were loose in structure, moderate in body and rich in oil. Its single leaf weight was relatively high, the contents of main chemical components were harmonious, the sensory quality was above moderate. Ekao No. 2 was better in comprehensive smoking quality comparing with K326 and Yunyan 87. Therefore, Ekao No. 2 could be considered as a new variety with balanced potentials for high quality, good productivity, economic performance, wide adaptability and strong resistance to diseases.