16S rRNA and pmoA functional genes were used as biomarker genes to investigate the temporal and spatial distribution of community diversity of Candidatus Methylomirabilis oxyfera (M. oxyfera) in the sediments of the Hunhe River by clone library technology. The dependence relationship between the physicochemical property of water and sediment samples and the diversity characteristics of the M. oxyfera community were analyzed by multivariate direct gradient analysis. Among the examples collected in September 2014, the OTU number of the 16S rRNA gene of M. oxyfera was 2-5, the Shannon-Wiener diversity index was 0.21-1.4, and the distribution characteristics in the middle reaches > upstream > downstream were presented. The OTU number and Shannon-Wiener diversity index of the pmoA functional gene in upstream samples are significantly higher than those in the middle and lower reach samples, and the Shannon-Wiener diversity indices in the upstream samples are 3.5 times and 2.3 times higher than that of the middle and the downstream samples, respectively. The community diversity of M. oxyfera showed a distinct regional distribution. Samples were collected at 3 sampling points in March 2015. The OTU values of 16S rRNA and pmoA for M. oxyfera are 6 and 5 respectively, which were obviously higher than those in September 2014. The Shannon-Wiener index is also higher than that in September 2014 (1.4>0.68; 57>0.00). The community diversity of M. oxyfera showed obvious seasonal distribution characteristics. Multivariate direct gradient analysis results showed that the concentration of DOC in water, sediment conductivity, TOC concentration, and nitrite concentration in the sediment are the main environmental factors affecting the community diversity of M. oxyfera.