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This research investigates the cold tolerance mechanisms and physiological responses of winter rapeseed to freezing stress.

April 7, 2026Open Access

Revealing the response mechanism of winter rapeseed (Brassica rapa L.) to freezing stress based on transcriptome analysis

Key Points

This research investigates the cold tolerance mechanisms and physiological responses of winter rapeseed to freezing stress.
Utilized strong and moderate cold-tolerant rapeseed varieties for field planting.
Analyzed growth, physiological characteristics, hormone levels, and transcriptomic responses.
Implemented RNA sequencing to explore molecular regulatory mechanisms.
Conducted weighted gene co-expression network analysis to identify candidate genes.
Higher levels of proline, soluble sugars, soluble proteins, and salicylic acid were observed in the cold-tolerant variety.
Lower levels of malondialdehyde and gibberellin were found in the strong cold-tolerant variety.
Differentially expressed genes were notably engaged in metabolism and signaling pathways.
Nine candidate genes related to signal transduction and plant hormones were identified.

Abstract

Winter rapeseed (Brassica rapa L. subsp.oleifera) is an important oilseed crop in northern China, where freezing stress imposes severe constraints on its yield. Its leaves dry up and wither during the overwintering period, but its roots stay deeply embedded in the soil. Following March of the next year, new leaves grow from the growing point and it enters reproductive growth. Therefore, this study aims to investigate the cold tolerance regulatory mechanisms and physiological-biochemical responses of field-grown roots in rapeseed cultivars with significant differences in cold tolerance during cold stress.This study used a strong cold-tolerant variety “Longyou 7 (L7)” and a moderate cold-tolerant variety “Longyou 99 (L99)” as experimental materials. The materials were planted in field to analyze the effects of natural freezing stress on rapeseed growth, physiological characteristics, hormone levels, and transcriptional levels before wintering. Additionally, transcriptome sequencing (RNA-seq) was combined to decipher the molecular regulatory mechanisms. Results showed that as temperature decrease, contents of proline (Pro), soluble sugar (SS), soluble protein (SP), and salicylic acid (SA) in roots of the strong cold-tolerant variety L7 were all significantly higher than those in L99, while contents of malondialdehyde (MDA) and gibberellin (GA₃) in L7 were significantly lower than those in L99. RNA-seq results revealed that differentially expressed genes (DEGs) were significantly involved in phenylpropane biosynthesis, carbon metabolism, starch and sucrose metabolism, MAPK signaling pathway, ribosome, proteasome, and protein processing. Among these, the differences in the expression of genes related to cell signal transduction (MAPK signaling pathway) and metabolism were particularly prominent. Through weighted gene co-expression network analysis (WGCNA), 9 candidate genes related to protein kinases, plant hormones, transcription factors, and signal transduction were identified in the MAPK and 2 other modules.

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