Cnaphalocrocis medinalis is a major insect pest of rice in Asia. A few defensive enzymes were reported to show higher activities in a resistant rice line (Qingliu) than in a susceptible rice line (TN1) upon leaffolder infestation. However, the overall molecular regulation of the rice defense response against leaffolder herbivory is unknown. Here, differential proteomic analysis by SWATH-MS was performed to identify differentially expressed proteins between the two rice varieties, Qingliu and TN1, at four time points of leaffolder herbivory, 0, 6, 24, and 72 h. Gene Ontology (GO) enrichment of the differentially expressed proteins indicated overrepresentation of (1) photosynthesis, (2) amino acid and derivative metabolic process, and (3) secondary metabolic process. Phenylalanine ammonia lyase and chalcone synthase, which catalyze flavonoid biosynthesis, and lipoxygenase, which catalyzes jasmonic acid biosynthesis, exhibited higher expression in Qingliu than in TN1 even before insect herbivory. Momentary activation of the light reaction and Calvin cycle was detected in Qingliu at 6 h and 24 h of insect herbivory, respectively. At 72 h of insect herbivory, amino acid biosynthesis and glutathionemediated antioxidation were activated in Qingliu. A defense response involving jasmonic acid signaling, carbon remobilization, and the production of flavonoids and glutathione could underlie the resistance of Qingliu to leaffolder. Rice production in Asia is affected by a harmful insect pest, Cnaphalocrocis medinalis (Guenée) (Lepidoptera: Pyralidae), also known as the rice leaffolder 1. Leaffolder caterpillars feed on mesophyll tissues, interfering with photosynthesis and then reducing yield 2. The affected leaf blades appear white, so heavily infested fields may have 'scorched'-looking patches. This insect herbivore can complete three life cycles during each rice cropping season in Taiwan 3,4. The application of chemical insecticides is the main method used to control pest populations in rice fields 5. However, this method of pest control is rendered less effective because the caterpillars build a feeding chamber by folding a leaf longitudinally with silk, and the chamber indirectly protects the pests from the chemical spray. When plants are infested by insect pests, they can differentiate diverse types of insects based on the insect elicitors or the nature of the damage caused by the insects 6. The plants immediately activate various defensive signaling pathways, including those associated with mitogen-activated protein kinase (MAPK) and phytohormones, such as jasmonic acid (JA), salicylic acid (SA) and ethylene (ET) 7. Chewing insects are generally known to trigger JA signaling pathways, while phloem-feeding insects trigger the SA signaling pathway 8. The JA and SA signaling pathways also participate in antagonistic crosstalk in plant-insect herbivory interactions 8. Because the signaling pathways triggered by different herbivorous insects vary, the production of downstream defensive proteins and secondary metabolites is...