The large number of spores produced by edible mushrooms cause many problems, including causing lung disease, depleting natural genetic diversity, and reduced quality of fruiting bodies. Obtaining spore-deficient strains and understanding the underlying molecular mechanisms of such strains are important for breeding work. In this study, we crossed monokaryotic strains isolated from the edible fungi Agrocybe salicacola to obtain three spore-deficient strains with losses of the sterigmata on the surface of the lamella. A mating test revealed that recessive alleles distributed in some strains might control sterigmata development during the mitotic or meiotic phases. Transcriptome analysis revealed that the majority of the genes involved in DNA mismatch repair, base excision repair, and homologous recombination exhibited down-regulated expression patterns in the mutant fruiting bodies. Five genetic fragments, which were highly similar to the GTP-cyclohydrolase encoding gene, the DNA repair gene rad 8, and cell wall integrity and stress response component-encoding genes, were all expressed exclusively in the wild-type strains; these findings provide important information for the study of the spore development of edible fungi.