This study aimed to explore transcriptomic distinctions between resistant (CS-319) and susceptible (JAL-42) groundnut (Arachis hypogaea) genotypes exposed to Sclerotium rolfsii infection across different developmental stages. Employing a de novo assembly-based approach, we analysed the transcriptomic response in these groundnut plants under control and infected conditions at 24, 72 and 120 hours post-inoculation (hpi). Our RNA-Seq data yielded a total of 133,900,261 reads, revealing 7796 differentially expressed genes (DEGs). We constructed a gene regulatory network with 59 hub genes, identified 6783 transcription factors and uncovered 88,424 putative markers, including 17,236 simple-sequence repeats (SSRs), 10,099 single-nucleotide polymorphisms (SNPs) and 78,332 indels. Notably, the majority of DEGs were upregulated at 24 hpi in the resistant genotype, encompassing diverse functional categories such as pathogenesis-related genes, defence-related (R) genes, genes involved in plant-fungus interactions, oxidation-reduction-related genes, transport, metabolism and proteolysis genes, along with transcription factors (FAR1, B3, GATA, NAC, WRKY, MYBC1 and bHLH), secondary metabolic pathway-related genes and photosynthesis-related genes. The up-regulation of WRKY transcripts, associated with the activation of the jasmonic acid defence signalling pathway, potentially induced systemic acquired resistance (SAR). Conversely, these DEGs exhibited down-regulation in the susceptible genotype. Furthermore, a total of 17,236 expressed sequence tag (EST)-SSRs were identified from the unigenes, holding significant potential for advancing plant breeding through marker-assisted methods, facilitating quantitative trait locus (QTL) mapping and evaluating genetic diversity among genotypes. This study's approach contributes to a more profound understanding of the molecular-level defence mechanisms involved in the interaction between groundnuts and S. rolfsii.