An experiment was carried out to investigate the effect of different substrates on growth and yield of oyster mushroom (Pleurotus florida). Four different agricultural substrates viz., wheat straw, wheat straw + water hyacinth (1:1), maize straw, maize straw + bajra straw (1:1) were used to evaluate growth period, growth parameters and yield potential of three strains of oyster mushroom (PL-20-204, PL-20-205 and PL-20-206). Maximum average weight of fruiting bodies were recorded in bags where wheat straw+ water hyacinth was used as substrate and it was 19.30 g in case of PL-20-206 followed by 18.03 g and 16.09 g in PL-20-204 and PL-20-205 strains, respectively. Wheat straw + water hyacinth substrate showed best results among all substrates with a total yield of 900.1 kg bag−1 for PL20-206, 795.1 kg bag−1 for PL-20-204 and 731.9 kg bag−1 for PL-20-205 in three harvests. The results would be useful to mushroom growers in selecting suitable substrate for higher yield.