ABSTRACT Nitric oxide (NO) plays an important role in fungal response to abiotic stress, but little is known about whether or not NO is involved in regulating cytochrome P450 (CYP) gene family and mitochondrial related genes (MRG) of Pleurotus eryngii in response to cadmium (Cd) stress. In this study, 125 CYP members and 127 differentially expressed MRG were identified. The analysis of gene length and exon number revealed the complexity of CYP gene family. After adding exogenous NO, the expression patterns showed that PleCYP2 and 4-hydroxybenzoate polyprenyltransferase were significantly up-regulated, the functions of membrane and membrane components were enhanced, and the mitochondrial splicing apparatus component gene was activated, which might be the potential mechanism of NO regulating P. eryngii resistance to Cd stress. This study lays a foundation for further revealing the mechanism of NO in regulating P. eryngii response to abiotic stress and promotes the potential application in fungal cultivation.