In order to improve soil, yield and quality of flue-cured tobacco, the effects of different application times of green manure in Vicia villosa var. glabresens-tobacco rotation were studied under the condition of field experiments in Enshi, Hubei Province. The results from 2 years’ experiments showed green manuring in 15-25 days earlier than tobacco transplantation was appropriate when chemical fertilizer amount was 85% of the conventional dose and green manure application rate was 15000 kg/hm2. In this case, some agronomic traits like plant height and largest leaf area, dry matter weight of root, stem, leaves and whole plant, yield and production value of flue-cured tobacco were higher than or similar to the conventional technique of fertilization. When green manure was applied in 35 days earlier than tobacco transplantation, the tobacco could not grow well, its yield and production value were significantly decreased. Compared with conventional technique of fertilization, the upper leaves of tobacco grew better, its dry matter weight was increased by 4.5%-18.5%, the proportion of high and middle grad tobacco was improved by 2.7%-8.6%, when conventional chemical fertilizer rate was reduced by 15% and 15000 kg/hm2 of green manure was applied in 15-35 days ahead of tobacco transplantation.