The substrate is one of the most important factors in the formation of the seedling and must present conditions suitable for germination and development of the seedling root system. The objective of this work was to evaluate the effect of different substrates on the initial growth of Physalis peruviana L. seedlings. The experiment was conducted at the Experimental Farm, of the Federal University of Espírito Santo– São Mateus (Brazil). The experimental design used was completely randomized, with three treatments (Soil, Provaso® and Bioplant®), seven replications and seven plants per plot, using seeds with 427 days of storage. At 56 days after sowing, the 10 most vigorous seedlings were selected for evaluation. The following characteristics were evaluated: percentage of emergence, emergence speed index (ESI), mean time of emergence (MET), number of leaves (NL), stem diameter (SD), aerial part length (APL), aerial part fresh matter (APFM), aerial part dry matter (APDM), root fresh matter (RFM), root dry matter (RDM) and Dickson quality index (DQI). The Provaso® substrate showed the best performance in all studied variables, being recommended for the production of Physalis seedlings.