Sugarcane plays a crucial role in global sugar and ethanol production. Conventionally, sugar-cane propagation involves planting billets. However, Brazilian researchers have introduced the innovative pre-sprouted seedlings (PSS) method, widely used in the MEIOSI (Simultaneously Occurring Interrotational Method) system. Although MPB has several advantages over the con-ventional method, its sensitivity to water scarcity is a challenge. The study aimed to evaluate the survival and growth of PSS inoculated with Bacillus licheniformis and Bacillus subtilis under differ-ent water regimes. The experiment was conducted in the field in a randomized block in strips (split-block) using a 2×4 factorial scheme consisting of two inoculation conditions (with and without bacteria) and four water regimes (0%, 33%, 66%, and 100% of the ideal irrigation). The bacteria increased the PSS's survival and water use efficiency in the field. In addition, inocula-tion increased root and shoot growth, as well as the nutrient uptake such as N, P, Mn, Zn, and Cu, which resulted in a higher number of stalks per meter and, therefore, a higher multiplica-tion rate in the MEIOSI system. Inoculation proved to be a promising alternative for establishing PSS under water restriction.