Abstract: To explore the potential of azoxystrobin in controlling target sport diseases of tobacco caused by Rhizoctonia solani, 30 strains isolated from different regions in Guizhou Province were used as test materials. The effects of azoxystrobin on sclerotial formation were analyzed using the plate incorporation method and Biolog FF microplate technology, while its effects on pathogenicity were analyzed using the detached leaf assay. The results showed that all 30 strains were able to form sclerotia on PDA medium. The LK4 strain produced the most sclerotia, demonstrating the strongest sclerotial formation ability. Compared to the PDA medium without azoxystrobin, 8 strains exhibited delayed sclerotial formation, reduced maturity and lower sclerotia yield, while 22 strains failed to form sclerotia on the PDA medium containing azoxystrobin(5 μg/mL). Without azoxystrobin stress, the LK4 strain could metabolize all substrates on the FF microplate expect sebacic acid, and could form sclerotia when induced by 38 substrates including D-cellobiose, β-cyclodextrin, and dextrin. Azoxystrobin stress(5 μg/mL)increased the metabolic capacity of the LK4 strain towards salicin and arbutin in the FF microplate. However, the metabolic capacity towards 90 substrates decreased, excluding L-fucose, L-sorbose, γ-aminobutyric acid and D-lactic acid methyl ester. Furthermore, none of the substrates induced sclerotial formation in the LK4 strain under azoxystrobin stress. Results from the detached leaf assay experiment showed that lesions formed on the leaves within 7 days following inoculation with test strains without azoxystrobin stress. After culturing the strains on a PDA medium containing azoxystrobin(5 μg/mL)for 4 days and inoculating them onto detached leaves, 25 of the tested strains failed to form lesions on the leaves after 7 days. The 5 strains that did form lesions had significantly smaller lesion diameters compared to those without azoxystrobin stress