Abstract: Using stable isotope techniques, water sources and water use efficiencies of Eucalyptus urophylla ×E. grandis (exotic tree species) and Liquidambar formosana (native tree species, as a reference) were studied in a typical karst artificial forest, and the risk of drought stress of the plantation was discussed. The results showed that the isotope value of shallow soil water (0-50 cm) had obvious gradient features and was similar to the recent rain, while that of deep water was more stable and different from the shallow soil water. The soil water content in wet season (May and September) was higher than in dry season (October), and was higher in upslope than in downslope segment. The main water source of L. formosana was shallow soil water, which corresponded to high water use efficiency in different seasons. The main water sources for E. urophylla ×E. grandis on the upper slope and downslope segments were both shallow soil water in wet season, however, the sum of average water using the proportion of deep soil water increased for the downslope segment in the end of wet season. In dry season, E. urophylla ×E. grandis on the downslope segment mainly used shallow soil water, while it used deeper soil water on the upslope segment when soil water was relatively low. Meanwhile, the water use efficiency of E. urophylla ×E. grandis was lower compared with L. formosana, which revealed E. urophylla ×E. grandis did not get enough water supply under drought stress. The above results indicated that E. urophylla ×E. grandis would encounter high risk of water stress if extreme drought events happened in karst regions in the coming future.