Seedling production is one of the most important stages of plant cultivation, with quality mainly depending on the substrate and plant material used. This study aimed to evaluate the production of Pereskia aculeata seedlings combining substrates and cutting diameters. The experimental design was completely randomized, in a 5 × 3 factorial arrangement, corresponding to five substrate combinations (commercial, kapok, palm, commercial + kapok and commercial + palm) and three cutting diameter ranges (0.5-3.0, 3.1-5.5 and 5.6-8.0 mm). The following parameters were evaluated: plant height, leaf length and width, number of shoots and leaves, root length, shoot and root fresh mass, shoot and root dry mass, total fresh and dry mass and Dickson quality index. There was a significant interaction effect between substrates and cutting diameters on the seedlings formed, except for number of shoots, leaf length and root dry mass. Seedlings of higher quality can be achieved with larger cutting diameters combined with commercial substrate. The palm and kapok substrates can be used alternatively when combined with larger cutting diameters.