Drought is one of the most widespread stress factors adversely affecting plant growth, crop yield and quality. In Subtropical region, on the face of global warming, temperature extremes aggravate the negative effects of drought. Increased resistance to stress has been achieved in several plants by exogenous application of various organic osmoprotectants. In this study, the role of glycine betaine (GB) as exogenous application, is aimed to investigate for increasing grapevine stress tolerance to drought. The grapevines of Alphonse Lavalléeʼ cultivar, grafted on the rootstock 41 B, were subjected to four different applications; (1) full irrigation (FI) as control (irrigation at field capacity level), (2) deficit irrigation (DI, 50% of FI), (3) DI plus 5000 ppm GB pulverization, and (4) DI plus 10000 ppm GB pulverization in a pot experiment under glasshouse condition. Leaf fresh weight of vines subjected to DI was 31.8% lower than those of FI vines. GB appeared to exert an influence on leaf water statute, slightly alleviating the leaf water loss resulting from water shortage. GB treatments, regardless of the concentration, slightly increased the fresh weight of the leaves (22.2% lower than FI). Investigations on leaf turgid weight and dry weight were also similar to those of fresh weight in that the highest and the lowest values were determined in FI and DI applications, respectively. Reduction in shoot water content in response to water deficit was closely related to the decrease in leaf water content.