Sesame (Sesamum indicum L., 2n = 2× = 26) from the Pedaliaceae family is primarily grown for its high oil content, rich in unsaturated fatty acids like linoleic acid (LA) and alpha-linolenic acid (ALA). However, the molecular mechanisms of sesame oil accumulation remain poorly understood. This study analyzed transcriptomes at two seed development stages: Young Stage (YS, pods 1.5-2.5 cm) and Mature Stage (MS, brown pods >
2.5 cm), to explore regulatory mechanisms and identify key genes involved in lipid biosynthesis. From 25,173 genes, 18,820 with expression values >
10 CPM in at least 70 % of replicates were included in differential expression (DE) analysis. Active expression (LFC >
0) was observed in 9372 and 9448 genes at YS and MS, respectively. DEGs were annotated, revealing roles in various biological processes, (e.g., mRNA metabolic process, reproduction-related developmental processes, seed development), molecular functions (e.g., aminoacyltransferase activity, ubiquitin-like protein and ubiquitin-protein transferase activities), and cellular components (e.g., peroxisome, microbody, lipid droplet). KEGG analysis highlighted genes involved in fatty acid synthesis (e.g., fabG, fabZ), TAG biosynthesis (DGAT1, GPAT), and alpha-linolenic acid metabolism (AOS, LCAT3). Key genes upregulated at MS included SIN_1025205 (protein transport), SIN_1006853 (acetylajmalan esterase), and SIN_1003267 (gamma-cadinene synthase). The study generated a valuable transcriptome dataset and gene list for seed development and lipid biosynthesis, which will be validated through functional studies. An interactive webpage is provided for data exploration.