Edible chrysanthemum is widely cultivated and used as an important ingredient of medicine, tea and multifunctional food. During the planting of chrysanthemum, pesticides are extensively used for preventing plant diseases and insect pests. To ensure the food safety of edible chrysanthemum, rapid detection methods are urgently needed for on-site inspection. In this study, a graphene oxide/Au nanoparticle (GO/Au NP) cellulose substrate was prepared through layer-by-layer assembly of GO and Au NPs on a mixed cellulose ester membrane. Surface-enhanced Raman spectroscopy (SERS) detection of four types of organophosphorus and organosulfur pesticides was achieved by filtering the extracting solution through the substrate and analysing SERS spectra. Qualitative and semi-quantitative detection of fenthion, phoxim, isocarbophos and thiram was accomplished with the detection limits of 38.01, 8.13, 48.97 and 8.74 ng mL-1, respectively. A spiking experiment further demonstrated the feasibility of this method for rapid and on-site detection of mixed pesticides in chrysanthemum. This study provides a new approach for rapid detection of multiple hazardous substances in flowering and herbal plants.