<p>Rodents play a crucial role in seed dispersal through scatter-hoarding behavior. In natural ecosystems, rodents demonstrate selective foraging behavior among co-occurring heterospecific and conspecific seeds. Current studies on rodent-seed interactions primarily employ two contrasting experimental approaches: releasing seeds of each plant species separately and releasing mixed seeds of multiple species, representing fundamentally distinct paradigms that engage different dimensions of optimal foraging theory. Specifically, single-species releases primarily isolate optimal diet selection processes based on energy-to-handling-time ratios, while mixed-species releases incorporate more complex ecological dynamics including patch-use strategies and trait-mediated neighbor effects. This fundamental divergence in experimental design has profound implications for research outcomes, as each approach captures different aspects of rodent foraging decision-making under varying ecological contexts. Surprisingly, most current studies overlook the potential impact of these two experimental methods on research outcomes. Here, we selected three tree species in a subtropical forest in southwest of China and compared the differences in the seed predation and dispersal by rodents of each species between two seed releasing methods. Our study revealed that methodological differences in seed release significantly affect seed predation and dispersal by rodents. These methodological effects are not only manifested in the seed fates of specific species and the effects of seed size, but also in the preferential selection of different species of seeds by rodents. Therefore, we recommend that the choice between mixed releasing of multiple species and individual releasing of each species in actual research should be comprehensively considered based on the research objectives and experimental environmental context.</p>