Nutrient status in soil is crucial for the growth and development of plants which indirectly or directly affect the ecophysiological functions of resident soil microorganisms. Soil methanogens and methanotrophs can be affected by soil nutrient availabilities and plant growth, which in turn modulate methane (CH4) emissions. Here, we assessed whether deficits in soil-available phosphorus (P) and potassium (K) modulated the activities of methanogens and methanotrophs in a long-term (20 year) experimental system involving limitation in either one or both nutrients. Results showed that a large amount of CH4 was emitted from paddy soil at rice tillering stage (flooding) while CH4 flux was minimum at ripening stage (drying). Compared to soils amended with NPK fertiliser treatment, the soils without P input significantly reduced methane flux rates, whereas those without K input did not. Under P limitation, methanotroph transcript copy number significantly increased in tandem with a decrease in methanogen transcript abundance, suggesting that P-deficiency-induced changes in soil physio-chemical properties, in tandem with rice plant growth, might constrain the activity of methanogens, whereas the methanotrophs might be adaptive to this soil environment. In contrast, lower transcript abundance of both methanogen and methanotrophs were observed in K-deficient soils. Assessments of community structures based upon transcripts indicated that soils deficient in P induced greater shifts in the active methanotrophic community than K-deficient soils, while similar community structures of active methanogens were observed in both treatments. These results suggested that the population dynamics of methanogens and methanotrophs could vary along with the changes in plant growth states and soil properties induced by nutrient deficiency.