Summary1. Disturbances play a central role in determining the spatial and temporal dynamics of many plant communities. In our study of macrophyte assemblages at 150 sites in five arms of a large subtropical reservoir (Itaipu Reservoir, Brazil–Paraguay border), we used co‐occurrence null models and spatiotemporal analyses to describe the patterns in the assemblages during a historically large water drawdown in 2000, in comparison with the previous year (1999) and subsequent years (2001–07). A C‐Score co‐occurrence index tested the null hypothesis of random structure during the drawdown period. A detrended correspondence analysis and multiresponse permutation procedure were used to verify whether species composition differed before, during and after the disturbance.2. In contrast to our expectations, the null models showed that the macrophyte assemblages were spatially structured during the drawdown (2000), although species composition was significantly different from the previous year (1999) and also changed in the following years (2001–07). Significant species co‐occurrence patterns were generated by the drawdown disturbance, with species extinctions and colonisation by new species from propagules and seed bank germination.3. The randomness we expected in 2000 actually occurred in 2001, probably because the reestablishment of normal water level enabled both submersed and free‐floating species to recolonise the shore that emergent species had inhabited since the drawdown. Biotic interactions appeared to increase during the years after the disturbance and the habitat preferences of the aquatic macrophytes were re‐established, resulting in higher similarities in aquatic macrophyte species composition in the years after the drawdown.