Abstract
Winter rapeseed is a high-oil crop that exhibits significant sensitivity to low temperatures, leading to a substantial reduction in production. Hence, it is of great significance to elucidate the genomic genetic mechanism of strong freezing-resistant winter rapeseed to improve their freezing-resistant traits. In this study, global transcriptome expression profiles of the freezing-resistant cultivar NTS57 (NS) under freezing stress were obtained for the years 2015, 2016, and 2017 by RNA sequencing (RNA-seq). Most differentially expressed genes (DEGs) were involved in the plant hormone signal transduction, alpha-linolenic acid metabolism, protein processing, glutathione metabolism, and plant-pathogen interaction pathways. Antioxidant enzyme activities and lipid peroxidation levels were significantly positively and negatively correlated with overwintering rate (OWR), respectively. After freezing treatment, the formation of freezing resistance of NS was attributed to the increase in antioxidant enzyme activities and content of osmotic regulation substances, as well as the decrease in lipid peroxidation level. Furthermore, quantitative reverse transcription polymerase chain reaction (qRT-PCR) and phenotypic verification indicated that heat stress transcription factor A2 (HSFA2) and 17.6 kDa class II heat shock protein (HSP17.6) participated in the response to freezing stress. This study will further refine the regulatory network of plants against freezing stress and help to screen candidate genes for improving plant freezing resistance.