The agronomic traits, chemical components, and diseases are very important factors affecting the yield and leaf quality of tobacco. QTLs linked to the three aspects of traits can be used in marker-assisted selection for the yield and leaf quality improvement in tobacco. The objective of this study was to identify QTLs controlling important traits in tobacco, which could promote the basic research of molecular marker-assisted selection breeding. A population with 127 F2 and F2∶3 individuals from the cross between Taiyan 7 and Burley 21 was used to establish a genetic linkage map of tobacco, 190 marker loci were mapped into 26 linkage groups which spanned a total map length of 3 483 cM. With a randomized complete block design, two location field tests with three replicates one year were conducted to charactererize six important traits, including nicotine, total chlorine, total kalium, leaf length, leaf angle, and powdery mildew. Utilizing data of field trials and MCIM method, QTLs for six important characters were mapped and their genetic interactions were analyzed. As the results, two QTLs for nicotine, two QTLs for total chlorine, one QTL for total kalium, four QTLs for leaf length, one QTL for leaf angle, and one QTL for powdery mildew were detected, of which six QTLs had additive effect, and the others were epistatic. The results indicated that epistatic QTLs, the additive QTLs also played important roles in the genetic basis of the six important traits with eleven gene loci in tobacco.