Abstract: This study aimed to investigate the differences of physical characteristics of flue-cured tobacco leaves (FTLs) across different production areas from Yunnan province. A total of 3,646 FTLs samples from 13 tobacco-planting cities/autonomous prefectures in Yunnan were collected, including upper, middle and lower parts. Tensile force, thickness, and stem content were measured, and then the tensile force-to-thickness ratio was calculated. Multiple comparisons, cluster analysis and partial least squares discriminant analysis (PLS-DA) were employed to reveal the similarities and differences of FTLs in different production areas from Yunnan, and a preliminary identification model was established for specific regions within the province. The results showed that: (1) Tensile force showed positively correlation with thickness in all parts of FTLs, while stem content exhibited negatively correlation with thickness in middle and lower leaves. (2) Physical indices differed significantly among production areas: FTLs from Zhaotong showed the highest tensile force and thickness in upper leaves, that from Lijiang had the highest tensile force in middle leaves. While FTLs from Dehong exhibited the highest tensile force and thickness in lower leaves. Regarding stem content, FTLs from Dehong showed the lowest value in all parts. FTLs from Dali and Lijiang displayed notably higher tensile force-to-thickness ratio. (3) Based on physical indices combined with bootstrap resampling approach, discrimination models including Random Forest and PLS-DA were constructed for FTLs from four production regions in Yunnan Province, achieving accuracies of 93.50% and 87.75%, respectively, on the test set. In both models, the tensile-to-thickness ratio was identified as the most important discriminant variable. (4) Cluster analysis revealed that FTLs from Yunnan could be classified into five categories. FTLs from Dali and Lijiang were always of the same category, exhibiting the characteristics of "high tensile force-to-thickness ratio". That from Dehong have always been classified as an independent category, with the lowest stem content.