Freshwater lakes, especially eutrophic ones, have become a hotspot of methanogenesis. However, the effects of eutrophication and seasonality on methanogenesis activity and methanogenic microbial community remain unclear. In the current study, for two adjacent lakes at different trophic status, their methanogenesis potential in different seasons was evaluated using incubation experiments. The density, diversity, and community structure of methanogens were analyzed based on the mcrA gene. Correlation analysis and redundancy analysis were carried out to identify the environmental factors driving the variations of methanogenesis potential and methanogen community. The results showed that eutrophication could result in active methanogenesis with relatively high seasonal variance. The methanogenesis variation could be well explained by carbon input in association with algal growth, as well as the change of methanogen population density. With the dominance of Methanomicrobiales in both lakes, the hydrogenotrophic pathway had a major contribution to total methane production. The considerable proportion of Methanomassiliicocales in eutrophic lake implied that methylotrophic methanogenesis might be previously underestimated. These results added new insights towards methanogenesis process in eutrophic freshwater lakes.