Soil water deficit is the main obstacle factor of plant reconstruction in rocky desertification area of Southwest China. Mycorrhizal mulberry (Morus alba L.) can cope with the main ecological barriers during the seasonal drought in the rocky desertification area. However, it is unclear the water resource of mulberry. In this study, we studied the water use strategy of mulberry root in different seasons by hydrogen and oxygen stable isotope technique. The mulberry was grown in typical rocky desertification area in Qingshuipu town, Qixingguan district, Bijie city (105°35′02′′E, 27°41′19′′N) in Guizhou Province. The precipitation was mainly concentrated in June-September. The mulberry was not irrigated. The annual growth of mulberry was in 2 seasons: from April to June and from September to November. The samples of rainfall water, soil and plant branch were collected from April to November of 2014. Soil was sampled at 0-60 cm depth with 10 cm as a sampling layer. The abundance of deuterium and oxygen stable isotope were measured. The correction analysis method was used to preliminarily evaluate water sources of mulberry. The contribution rate of water at each soil layer was also calculated. The results showed that in rainfall water, the abundance of deuterium from April to November averaged -25.9‰ and the oxygen stable isotope abundance averaged -4.82‰. In spring from April to June, the abundance of deuterium and oxygen stable isotope in rainfall water averaged -0.24‰ and -1.68‰, respectively. In autumn from September to November, the abundance of deuterium and oxygen stable isotope in rainfall water averaged -60.0‰ and -8.17‰, respectively. In spring, the abundance of deuterium and oxygen stable isotope in soil water averaged -29.4‰ and -4.77‰, respectively while the abundance of deuterium and oxygen stable isotope in xylem water averaged -33.2‰ and -5.39‰, respectively. In autumn, the abundance of deuterium and oxygen stable isotope in soil water averaged -70.1‰ and -9.27‰, respectively while the abundance of deuterium and oxygen stable isotope in xylem water averaged -55.7‰ and -7.92‰, respectively. The abundance of both stable isotopes in soil water and xylem water were below that in the rainfall water and that of xylem water were between soil water and rainfall water. It indicated that the mulberry mainly used soil water. Both isotopes in rainwater were more enriched in the early stage of mulberry growth (spring, from April to June) than in the late growth stages (autumn, from September to November). With the seasonal variation of rainfall, the both isotopes of soil water showed a downward trend. According to rainfall water equations, the mulberry absorbed mainly soil water. Both isotopes had clear seasonal trend. In spring, the isotopes in soil water decreased with the increase of soil depth. In autumn, isotopes in soil water decreased then increased with soil depth. Based on the correlation method, the mulberry absorbed water from different soil depth. In spring, it absorbed water from soil below 20 cm while it absorbed water from soil 0-20 cm. In September, it also absorbed water from 50-60 cm soil layer. Based on results from the IsoSource software, in spring, mulberry mainly used soil water from soil layers of 10-30 to 50-60 cm. In autumn, it mainly used soil water from soil layers of 50-60 to 0-10 cm. In spring, the contribution rate was 69.1% by 10-30 cm in soil, 31.8% and 66% by 50-60 cm layer in May and June, 70.2% by 50-60 cm in September and 94.3% and 71.9% by 0-10 cm in October and November, respectively. The study will be helpful to mulberry manage and application in rocky desertification area. [ABSTRACT FROM AUTHOR]