The seed provenance, the type of substrate and its properties, as well as the watering regime in the first years after sowing are among the most important characteristics affecting the production of containerized seedlings. The objective of this study was to analyse the growth of black locust (Robinia pseudacacia L.) seedlings on different types of substrate (six mineral and two organic) with different textures and pHs under three different regimes of available water content (no-limiting, medium drought, severe drought) over a period of five months (May–October 2023), using seeds from three sources located in the southeast, south, and west of Romania. The highest seedling emergence rate (73.7%) was obtained for the medium seeds with 18.492 g weight (1000 seed weighs) (Bucharest provenance). Direct sowing in the field and containerized seedlings both showed a trend of growing in height during unlimited water and of growing in diameter during low watering. The lowest survival rate of seedlings (86.7%) occurred in the mineral substrate in the case of rendzina with additional sand (pH 8.70), and for the organic substrate, the lowest survival rate (87.0%) occurred for the peat MKS 3 substrate (pH 4.54).