The invention discloses a primer pair for constructing a molecular marker map of a cured tobacco variety of Hunan main tobacco varieties, and the sequence of the primer pair is as shown by SEQ ID No.1-46. The primer pair can be used for identifying cured tobacco varieties of Hunan main tobacco varieties, such as Xiangyan No.3, Xiangyan No.5, Xiangyan No.6, Xiangyan No.7, Yunyan 87, Yunyan 97, Yunyan 99, Yunyan 100, Yunyan 203, Zhongyan 100 yuan, Zhongyan 100, Guangdong tobacco 9, Gui tobacco No.1, Honghua Dajinyuan, Longjiang 911, Nanjiang No.3, NC729, G80 or K326. The primer pair is of great significance in helping tobacco enterprises to quickly, accurately and efficiently identify and grade the cured tobacco.