Clubroot disease, caused by the soil-borne pathogen Plasmodiophora brassicae, poses a severe threat to the global production of Brassicaceae crops, including radish (Raphanus sativus L.). Although resistance breeding is an important method for sustainable disease management, the molecular mechanism underlying clubroot resistance remains elusive in radish compared to other Brassicaceae species. In this study, 52 radish inbred lines were screened for disease responses following P. brassicae inoculation, with the resistant line T6 and the susceptible line T14 selected for transcriptome analysis. RNA-Seq was performed at 10, 20, and 30 days post inoculation (DPI) to elucidate transcriptional responses. The susceptible line T14 exhibited a higher number of differentially expressed genes (DEGs) and persistent upregulation across all time points, indicating ineffective defense responses and metabolic hijacking by the pathogen. In contrast, the resistant line T6 displayed temporally coordinated defense activation marked by rapid induction of core immune mechanisms: enhanced plant–pathogen interaction recognition, MAPK cascade signaling, and phytohormone transduction pathways, consistent with effector-triggered immunity priming and multilayered defense orchestration. These findings indicate that resistance in T6 could be mediated by the rapid activation of multilayered defense mechanisms, including R gene-mediated recognition, MAPK-Ca²⁺-ROS signaling, and jasmonic acid (JA) pathway modulation. The outcomes of this study would not only facilitate clarifying the molecular mechanism underlying clubroot resistance, but also provide valuable resources for genetic improvement of clubroot resistance in radish.