The root distribution and water uptake of irrigated vines (Vitis vinifera L.) in arid oasis areas remain poorly understood, particularly in terms of the variations in water uptake resulting from plant type and growing period. In this study, excavation and soil coring were employed to investigate the root distribution of vines. Meanwhile, the water uptake dynamics of irrigated vines in an arid oasis area were developed using stable isotopes of hydrogen and oxygen in water bodies (δD and δ18O) and in crops and soil water, coupled with a direct inference approach and a linear mixed model (IsoSource). The soil layers were divided into nine layers via hierarchical cluster analysis. The results indicated that the vertical distributions of the total and fine roots of grapevines were mainly in the range of 40–160 cm, accounting for 93.1% (91.2%) and 92.5% (90.0%) of the total root (200 cm) distribution during May and October, respectively. In the horizontal direction, both the total root and fine root systems were mainly distributed within 0–100 cm from the trunk and contributed 81.2% and 79.8% of the total root distribution, respectively. Meanwhile, both the total root weight (length) density and fine root weight (length) density decreased gradually with increasing radial distance from the trunk in the 0–80 cm range. The main water uptake of vines was at 0–40 cm in June, 20–80 cm in July, and 40–100 cm in August. These findings suggest that the main depth of root water uptake increases during the whole growth stage for grapevines in arid oasis ecosystems. This information will help growers to improve irrigation efficiency and provide a data analysis of water conservation in an arid oasis area during water stress.