The choice of a substrate that meets the nutritional and physical demands of papaya seedlings with the use of organic wastes is directly linked to increases in crop yield and is an important environmental and economic alternative. Aiming to produce good-quality papaya seedlings while considering the composition and availability of substrates, this study examined the influence of applying increasing rates of poultry litter biochar as a substrate component in the production of papaya seedlings. The study was developed in a protected environment belonging to the Federal University of Campina Grande (UFCG). The experiment was laid out in a completely randomized design with a 6 × 2 factorial arrangement consisting of six rates of biochar (4, 8, 12, 16 and 20 t ha-1 and the control) and two papaya cultivars (Formosa and Ouro), with four replicates. Plant height, stem diameter, number of leaves and leaf area were evaluated 90 days after sowing. The fresh and dry biomass of shoots and root system and the Dickson Quality Index were also evaluated. In conclusion, the rates that provided good development of papaya seedlings were 8 and 16 t ha-1, and papaya cultivar Ouro showed better growth results.