Abstract: By using layering harvest method, a comparative study was conducted on the biomass allocation and its spatial distribution of 20-year-old-Eucalyptus camaldulensis plantation, Leucaena leucocephala plantation, and E. camaldulensis-L. leucocephala plantation in Yuanmou dry-hot valley of Yunnan Province, Southwest China. The stand biomass in the mixed E. camaldulensis-L. leucocephala plantation (82.99 t·hm^-2) was between that of monoculture E. camaldulensis plantation (60.64 t·hm^-2) and L. leucocephala plantation (127.79 t·hm^-2). The individual tree biomass of E. camaldulensis in the mixed plantation (44.32 kg) was 49.8% higher than that in monoculture plantation (29.58 kg). The branch and leaf biomass of L. leucocephala (25.4%) in monoculture plantation was larger than that of E. camaldulensis (8.9%) in monoculture plantation, and the aboveground biomass distribution ratio (78.0%) of L. leucocephala (25.4%) was also higher than that of E. camaldulensis (73.4%). The roots of L. leucocephala in both monoculture and mixed plantations were mainly distributed in 0-40 cm soil layer, while those of E. camaldulensis in monoculture and mixed plantations were mainly found in 0-80 cm and 0-60 cm, respectively. The proportion of biomass allocated to roots including medium roots, small roots, and fine roots of L. leucocephala in mixed plantation was higher than that in monoculture plantation, but it was contrary for E. camaldulensis. It was suggested that introducing L. leucocephala in E. camaldulensis plantation promoted the growth of E. camaldulensis, especially for its aboveground biomass, and increased the amount of lateral roots in 0-20 cm soil layer, which had significance in soil and water conservation in the study area.