Abstract: In order to provide the data of nicotine content in tobacco from Yunnan for cigarette manufacturers,13912 flue-cured tobacco samples of 4 major cultivars,Yunyan 85,K326,Honghuadajinyuan(Hongda) and Yunyan 87 from 12 growing areas in Yunnan from 2004 to 2007 were analyzed.The results showed that:1) The nicotine contents in upper,middle and lower leaf samples averaged 3.31%,2.56% and 1.77%,respectively,the difference between stalk positions was significant.The nicotine level in flue-cured tobacco from Yunnan was appropriate in general;2) The difference of nicotine content in the samples from the same stalk position between prefectures(cities) was significant;3) The varieties in the order of nicotine content in lower and upper leaves were Hongda K326 Yunyan85 Yunyan87,the difference between varieties was significant.The varieties in the order of nicotine content in middle leaves were Yunyan85 K326 Hongda Yunyan87.The nicotine content in Yunyan87 was significantly lower than that in Hongda,K326 and Yunyan85,while there was no significant difference between K326,Hongda and Yunyan85.