The pot experiment was carried out by the low-temperature stress under controlled conditions. Five species (strains) of Euonymus were selected as experimental materials, aiming to study the properties of cell membrane permeability, contents of osmotic regulators, protective enzymes activities and other physiological indexes in different E. species when suffered to various temperature conditions. Otherwise, leaf anatomical structure and subordinate function, as well the cold resistance capability of five species (strains) were also measured by lab analysis. The analysis method of subordinated function was applied to describe evaluation index. The results showed that: (1) E. japonicus ‘CuZhi’ NO.6 had three close layers of palisade cell, while E. japonicus "CuZhi", E. kiautschovicus, E. japonicus had two close and one loose ones. In contrast E. fortunei had only two close layers of palisade cell. Low-temperature stress led to varieties in leaf tissue tightness of the five E. species, and the order indicated that E. japonicus ‘CuZhi’ NO.6 > E. japonicus "Cu Zhi" > E. kiautschovicus > E. japonicus > E. fortunei. (2) The logistic equation was applied to analyze the relative conductivity of five E. species and the lethal temperature of 50% (LT50), and the arrangement was that E. japonicus ‘CuZhi’ NO.6 (-19.69℃) < E. japonicus "CuZhi" (-17.41℃) < E. kiautschovicus (-16.03℃) < E. japonicus (-13.33℃) < E. fortunei (-10.45℃). (3) According to the influences of each index by cold-stress, the resistance ability of five species was obtained by using membership function. The sequences was that E. japonicus ‘CuZhi’ NO.6 > E. japonicus "CuZhi" > E. kiautschovicus > E. japonicus > E. fortunei. In conclusion, cold resistance of five E. species was evaluated correctly by studying leaf anatomical structure, lethal temperature of 50% and subordinate function. It provides a theoretical basis for cold resistant screening and applying of E. germplasm resource.