Oxidation of methane by soil microorganisms is an important mechanism controlling the content of this potent greenhouse gas in the atmosphere. Agricultural soils operate under stressful conditions, and ammonium (N-fertilization) and drying (global warming) may have a significant impact on methane oxidation. In order to investigate how soil methanotrophs respond to drying–rewetting (DW), ammonium addition (100 mg/g) (A), and their combined action (MS), agricultural soil microcosms were incubated over the three months and methane oxidation was measured before and after perturbations, while community composition was monitoring using 16S rRNA gene sequencing. A significant decline in the methane-oxidation activity after perturbations was found, with subsequent restoration, and the combined treatment was more effective than the sum of individual treatments, indicating a synergistic effect. After rewetting, the structure of the bacterial community returned to pre-dry-down levels, but the application of ammonia and combined action lead to irreversible changes in the structure of soil methanotrophic communities. Methanotroph Methylomicrobium were significantly reduced under disturbances, while there was a significant increase in the representation of Methylobacter accompanied by the facultative methylotroph Methylovorus. We concluded that methanotrophic communities in agricultural soil demonstrated flexibility, and even when the abundance of dominant populations drops, ecosystem functions can recover.