Sustainable agriculture urgently requires innovative, pesticide-free strategies to mitigate herbivory and safeguard food security. Ultraviolet-B (UV-B) irradiation, with tunable intensity and cost-effectiveness, has emerged as a promising non-chemical method to enhance plant resistance, yet its underlying mechanisms remain elusive. Here, using tea plant ( Camellia sinensis) and its major pest Ectropis obliqua as a model, we developed a multimodal framework that integrates AI-enhanced electronic nose technology for real-time volatile profiling with in situ hyperspectral stimulated Raman scattering (SRS) microscopy to characterize defense responses under precisely controlled UV-B treatments. This approach identified herbivore-induced volatiles—hexanal, (Z)-3-hexenol, octanal, and (Z)-3-hexenyl acetate—optimally induced at 1.2 kJ·m -2 UV-B and linked to insect deterrence. SRS imaging further revealed elevated jasmonic acid derivatives and L-phenylalanine, coupled with reduced protein levels and altered stomatal dynamics, all correlating with enhanced resistance. Transcriptomic and molecular analyses confirmed transcriptional regulation of these pathways. By bridging volatile detection, metabolic imaging, and molecular validation, this study pioneers a multimodal strategy that provides mechanistic insights into UV-B–mediated plant defense and highlights the potential of multimodal methodologies as powerful tools for developing sustainable, pesticide-free pest management solutions in precision agriculture.