Abstract: Wheat crown rot caused by Fusarium pseudograminearum is an important disease of wheat and leads to a severe impact on the yield and quality of wheat. At present, there are no chemical fungicides registered for the control of this disease. In this study, samples were collected from 17 cities in Henan Province and 82 strains of wheat crown rot were isolated and purified, including 76 strains of F. pseudograminearum and 6 strains of Fusarium graminearum. The EC₅₀ values of azoxystrobin on spore germination of F. pseudograminearum 76 strains are 0.02-1.54 μg/mL and the average EC₅₀ value is (0.33 ± 0.29) μg/mL. According to the test of abnormal data, LHWY-6 and SQYC-6 were the abnormal strains. The average EC₅₀ values of azoxystrobin to 74 isolates (97.37%) was (0.30 ± 0.24) μg/mL after discarding the two outliers, which could be used as the sensitivity baseline of F. pseudograminearum to azoxystrobin. The sensitivity of strain from different cities varies. The sensitivity to azoxystrobin is also fluctuant for the strains from the same city. The fold changes of strain sensitivity from Xuchang, Jiaozuo, Luoyang and Shangqiu are all above 20.00, that from Nanyang, Sanmenxia, and Kaifeng are all below 2.00, and from the other cities are between 2.70-12.00. When the sensitivity of F. pseudograminearum spore germination to azoxystrobin was compared with that to prothioconazole, carbendazim, fludioxonil, phenamacril, pyraclostrobin, and tebuconazole, the results showed that the inhibitory effects of 5 fungicides on spore germination are lower than those of azoxystrobin, except pyraclostrobin. The EC₅₀ values of azoxystrobin against 6 strains of F. graminearum are 0.10-0.42 μg/mL and the average EC₅₀ value is (0.19 ± 0.12) μg/mL. The inhibitory activity of azoxystrobin on the mycelial growth of the two species of Fusarium is faint with EC₅₀ of above 7 μg/mL. The 15% azoxystrobin FS agent has a 63.64% control effect against wheat crown rot with a dose of 260 g/(100 kg seeds) in vitro and a control effect of 55.24% with a dose of 220 g/(100 kg seeds) in the greenhouse. Azoxystrobin shows a notable inhibitory effect on spore germination of wheat crown rot fungi, has a good effect on disease control in vitro and greenhouse conditions and can be used as a high-quality alternative fungicide in the field.