Bioaerosols and humic-like substances (HULIS) are important components of atmospheric aerosols, which can affect regional climate by acting as cloud condensation nuclei and some of which can damage human health. Up to date, release of bioaerosols and HULIS initiated by precipitation is still poorly understood. Here we present different release processes for bioaerosols, non-bioaerosols and HULIS during a precipitation event in Beijing, China. Large fungal-spore-like aerosols were emitted at the onset and later weak stage of precipitation, the number concentration of which increased by more than two folds, while the number concentration of bacteria-like particles doubled when the precipitation strengthened. Besides, a good correlation between protein-like substances that were measured simultaneously by on-line and off-line fluorescence techniques consolidated their applications to measure bioaerosols. Furthermore, our EEM results suggest that the relative contribution of water-soluble HULIS to microbial materials was enhanced gradually by the rain event.