Using the PMS pressure chamber and isotope mass spectrometer (MAT-252), the leaf juice of Acacia mangium was obtained, and the carbon isotope discrimination (D) representing the most recently fixed carbon in the juice was determined. At the same time, the water-use efficiency of A. mangium was estimated. The results indicated that the carbon isotope ratio in the air of forest canopy (δa), 10 m high a bove ground averaged - 7.57 ± 1.41 % in cloudy days, and - 8.54±0.67% in sunny days, respectively. The diurnal change of the carbon isotope ratio in the photosynthetic products of the leaf juice (δp) was of saddle type in cloudy days, but dropped down from morning to later afternoon in sunny days. A strong negative correlation between δp and leaf-to-air vapor pressure deficit (D) was observed in sunny days, but a slight change in δp was found in cloudy days. The δp also decreased with decreasing leaf water potential (ω), reflecting that water stress could cause the decrease of δp. The carbon isotope discrimination of the leaf juice was positively correlated with the ratio between intercellular (Pi) and atmospheric (Pa) partial pressure of CO2. For A. mangium, the isotope effect on diffusion of atmospheric CO2 via stomata was denoted by a = 4.6%, and that in net C3 diffusion with respect to Pi was indicated by b = 28.2%. The results were in reasonable accord with the theoretically diffusive and biochemical fractionation of carbon isotope. It was defined that carbon isotope discrimination of photosynthetic products in A. mangium leaf juice was in proportion to that from photosynthetic products in dry material. The water-use efficiency estimated by the carbon isotope discrimination in leaf juice, fit well with that measured by gas exchange system (R2 = 0.86, p < 0.0001). The application of leaf juice in measuring the stable carbon isotope discrimination would reduce the effects of fluctuating environmental factors during the synthesis of dry matter, and improve the ecophysiological studies on carbon and water balance when scaling from the plant to canopy in the fields.