The sustainable management of agricultural residues is a pivotal element in ensuring the sustainable development of agriculture. This is based on strategies that include the reutilization of residues as a substrate for the cultivation of economically significant mushroom species. The primary aim of this investigation is to assess the viability of utilizing two of the most prevalent agricultural residues in Ecuador as a substrate for the cultivation of hybrids within the Pleurotus genus. This assessment includes an evaluation of the nutritional and productivity parameters exhibited by the resulting mushrooms, employing multivariate statistical methodologies. The hybrid strains were developed by crossing compatible neohaplonts obtained through chemical dedikaryotization. A total of five neohaplonts of Pleurotus ostreatus as parental strain P1 and five monokaryons of Pleurotus djamor as parental strain P2 were randomly crossed in all possible combinations. Two parental hybrid strains, H1 and H2, were produced. These hybrids were cultivated using agricultural waste substrates, specifically, green banana leaves (GBL) and sugarcane bagasse (SB). Two distinct treatments or mixtures were tested: M1 (composed of 80% SB and 20% GBL) and M2 (composed of 20% SB and 80% GBL). It was found that the M1 blend promotes mushroom growth, yielding superior properties attributable to the higher proportion of nutritional content derived from sugarcane bagasse.