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Powdery mildew is the most serious fungal disease of Rosa roxburghii in Guizhou Province, China. In this study, the control role of chitosan-assisted pyraclostrobin against powdery mildew of R. roxburghii and its influences on the resistance, photosynthesis, yield, quality and amino acids of R. roxburghii were evaluated. The results indicate that the foliar application of 30% pyraclostrobin suspension concentrate (SC) 100 mg L−1 + chitosan 500 mg L−1 displayed a superior control potential against powdery mildew, with a control efficacy of 89.30% and 94.58% after 7 d and 14 d of spraying, respectively, which significantly (p < 0.01) exceeded those of 30% pyraclostrobin SC 150 mg L−1, 30% pyraclostrobin SC 100 mg L−1, and chitosan 500 mg L−1. Simultaneously, their co-application could effectively enhance their effect on the resistance and photosynthesis of R. roxburghii leaves compared to their application alone. Meanwhile, their co-application could also more effectively enhance the yield, quality, and amino acids of R. roxburghii fruits compared to their application alone. This work highlights that chitosan can be applied as an effective adjuvant to promote the efficacy of low-dosage pyraclostrobin against powdery mildew in R. roxburghii and improve its resistance, photosynthesis, yield, quality, and amino acids. [ABSTRACT FROM AUTHOR]