Phytoplankton and bacteria play key roles in material cycling and their consequent eco-functions in lakes, which are threatened by anthropogenic pressures, especially agricultural activities, which, in the watershed, are effective in changing the material composition and hydrodynamic conditions of the lake through material input and water withdrawal. This process theoretically changes the interaction and assembly pattern of microorganisms, which are important factors driving the structural and functional evolution of ecological communities in lakes. In this research, the community structure, interactions, and assembly of phytoplankton and bacteria were investigated during agro-irrigation seasons in a typical agricultural drainage receiving lake, Wuliangsuhai. The results showed that the seasonal variations in the community were driven by nitrogen and phosphorus. In particular, Cyanobacteria increased significantly during the seasons with the regulation of TP (λ = 0.56, p < 0.01, n = 30). The TN positively drove Chlorophyta and Bacillariophyta (λ = 0.42 and 0.65, p < 0.05, n = 30). Furthermore, MENA showed that planktonic algae and bacterial community interactions were enhanced, and interspecific competition increased at high trophic levels. The community assembly is primarily a stochastic process that is mostly related to hydrodynamic conditions. The second related factor, nitrogen and phosphorus inputs, had obvious effects on community assembly, which responded to its effects on species diversity, niche width, and interactions, and they jointly controlled community assembly. This study reveals that the assembly processes of bacteria and planktonic algae were driven by different environmental factors in specific ways, which provides a new view for understanding agriculture’s impacts on microecology and helps in developing lake protection strategies.

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