Aspergillus cristatus undergoes sexual and asexual development under conditions of low and high osmotic pressure, respectively. In this study, the expression levels of 107 genes associated with sexual and asexual development were analysed under conditions of low and high osmotic pressure by RNA sequencing. The results showed that 37 genes were up-regulated and other genes were down-regulated under conditions of high osmotic pressure, with most of the up-regulated genes associated with asexual development and most down-regulated genes associated with sexual development. These results suggest that osmotic pressure regulated sexual and asexual development of A. cristatus by controlling the expression levels of key genes. Meanwhile, there were differences in the expression levels of key genes associated with the regulation of sexual and asexual development between A. cristatus and Aspergillus nidulans. Moreover, we verified the reliability of the results by quantitative real-time polymerase chain reaction analysis of some key genes. In this study, the relationship between sporulation-related genes and osmotic pressure at the transcriptome level were analysed, which indicated that A. cristatus was a useful model organism for the study of osmotic pressure regulation on sexual and asexual development.