| Summary: | <i>Fusarium solani</i> is the most significant pathogen that causes root rot in wolfberry, which has led to serious economic losses in terms of production. As an important enzyme in organisms, NADPH oxidase produces ROS. However, the mechanism of ROS mediated by NADPH oxidase in the growth of <i>F. solani</i> has not been studied. In this study, <i>F. solani</i> colonies were treated with 40 µmol/L DPI and 0.0012% H<sub>2</sub>O<sub>2</sub>. The growth rate in terms of colonies, number of spores, key gene expression levels, activity of key enzymes and the content of key products of ROS metabolic pathways were determined. The results showed that the growth rate of colonies treated by DPI decreased by 19.43%, the number of macroconidia increased by 231.03%, the IOD/area values of O<sub>2</sub><sup>−</sup> and H<sub>2</sub>O<sub>2</sub> decreased by 34.88% and 16.97%, respectively, the expression levels in terms of <i>NCF2</i>, <i>SOD1</i>, <i>CTA1</i> and <i>PXMP4</i> significantly decreased and the activities of SOD, CAT and POD decreased significantly, while the MDA content increased significantly. Additionally, in the case of the colonies treated with exogenous H<sub>2</sub>O<sub>2</sub>, the MDA content decreased significantly while the other indicators increased. Taken together, the <i>NCF2</i> gene is involved in regulating the activity of NADPH oxidase and regulates the products of O<sub>2</sub><sup>−</sup> and ROS metabolism enzyme genes and their activities to affect colony growth in the <i>F. solani</i> growth process.
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