[Objectivc] This study aimed to investigate the chilling tolerance of seedlings of different cotton genotypes and screen appropriate indicators for assess- ing chilling tolerance, to establish reliable mathematical evaluation model for chilling tolerance of cotton, thus providing theoretical basis for breeding and promoting new chilling-tolerant cotton germplasms and large-scale evaluation of chilling tolerance of cotton varieties. [Method] Fifteen cotton varieties (lines) were used as experimental materials. The photosynthetic gas exchange parameters, chlorophyll fluorescence ki- netic parameters, chlorophyll content, relative soluble sugar content, malonaldehyde content, relative proiine content, relative conductivity and other 12 physiological indi- cators of seedling leaves under low temperature treatment (5 ℃, 12 h) and recovery treatment (25 ℃. 24 h) were determined; based on the chilling tolerance coefficient (CTC) of various individual indicators, the comprehensive evaluation of chilling toler- ance was conducled by using principal component analysis, hierarchical cluster anal- ysis and stepwise regression analysis. [Result] The results showed that the 12 indi- vidual physiological indicators could be classified into 7 independent comprehensive components by principal component analysis; 15 cotton varieties (lines) were clus- tered into three categories by using membership function method and hierarchical cluster analysis; the mathematical model for evaluating chilling tolerance of cotton seedlings was established: D =0.275 -0.244Fo1 +0.206Fv/Fm1+0.326g,%-0.056SS + 0.225MDA+O.O38REC (FF=0.995), and the evaluation accuracy of the equation was higher than 94.25%,0. Six identification indicators closely related to chilling tolerance were screened, including Fo,, Fv/Fm1, Seedling leaves of cotton varieties (lines) gs2, SS, MDA, and REC. [Conclusion] with high chilling tolerance are less dam- aged under low temperature stress, and are able to maintain relatively high p