Annual branches with a diameter of 0.5-0.7 cm from grape rootstocks and varieties of ‘Zuoshan No. 1’, 10114, 3309C, SO4, 140R, ‘Pinot noir’, etc. were used as the materials. After the treatment of low temperature gradient (0 ℃, -14 ℃, -19 ℃, -24 ℃, -29 ℃ and -34 ℃), the temperature and time of TTC staining were optimized, and the survival of different tissues from grape branches was observed and counted. The relative electrical conductivity of the branches, the contents of soluble protein, free proline, soluble sugar and starch in phloem and xylem, and the ratio of bound water to free water (bundle to body) were measured. The semilethal temperature (LT₅₀) of branches was calculated by Logistic equation fitting the dyeing area of longitudinal section and relative electrical conductivity to evaluate the cold resistance of branches. Meanwhile, the cold resistance of the phloem and xylem of the branches was comprehensively evaluated by the physiological index membership function method. The evaluation results of the three methods were compared to establish an intuitive method for identifying the cold resistance of grape varieties. The results showed that: (1) the optimal conditions for TTC viability staining of grape shoots were pH=7.0, 0.5% TTC0.1 mol/L phosphate buffer at 35 ℃ for 36 h in the dark. With the decrease of stress temperature, the dyeing area of the longitudinal section of each variety decreased gradually. According to the optimized TTC method, the LT₅₀ of branches form ‘Zuoshan No 1’, 10114, 3309C, SO4, 140R, ‘Pinot noir’ were -31.38 ℃, -26.51 ℃, -26.10 ℃, -23.60 ℃, -23.33adually increased, and the relative conductivity of ‘Pinot noir’ basically maintained the highest and the largest increase (51.93%), while the relative electrical conductivity of ‘Zuoshan No.1’ always had the lowest and the smallest increase (44.07%). According to the relative conductivity, LT₅₀ of ‘Zuoshan No.1’, 10114, 3309C, SO4, 140R and ‘Pinot noir’ were -30.02 ℃, -26.40 ℃, -25.75 ℃, -23.16 ℃, -21.13 ℃ and -17.72 ℃, respectively. (3) The comprehensive evaluation results of the membership function of branch cold resistance through five physiological indicators showed that the cold resistance of the phloem in the same variety was stronger than that of the xylem. The cold resistance of different varieties in the same part was as follows: ‘Zuoshan No. 1’ > 10114 > 3309C > SO4 > 140R > ‘Pinot noir’. This study found that the TTC staining method was consistent with conductance method and comprehensive membership function method in evaluating the cold resistance of grape branches. However, compared with the other two methods, TTC staining was more intuitive and effective in evaluating the cold resistance of grape branches.