Low temperature germination (LTG) is one of crucial agronomic traits for field-grown rapeseed in the Yangtze River basin, where delayed direct sowing frequently exposes seeds to cold stress during germination. Despite its importance, the genetic basis underlying rapeseed germination under different temperatures remains largely unexplored. Here we screened out two cultivars with different LTG performance (JY1621 and JY1605) and conducted transcriptome sequencing at five key timepoints under different temperatures. Comparative analysis among multiple groups of differentially expressed genes (DEGs) revealed sets of early and late temperature response germination (ETRG and LTRG) genes, as well as cold-tolerant (CDT) and temperature-insensitive (TPI) candidate regulators at different germination stages. Furthermore, we performed a genome-wide association study (GWAS) using germination index (GI) of 273 rapeseed accessions and identified 24 significant loci associated with the germination trait under the normal temperature. Through integrated analysis of transcriptome sequencing and GWAS, we identified a series of candidate genes involved in temperature-dependent germination. To further verify the results, we analyzed expression patterns and haplotype distributions of BnaA3.CYP77A4and BnaA3.NAC078 among the genetic population. This study performed a comprehensive multi-omics analysis on temperature-dependent germination and provides insights for low-temperature germination breeding of rapeseed.