Comparative Transcriptome Analysis of the Response of Two Lines of Rapeseed (Brassica napus L.) to Cold Stress

J. Lee, Kwang-Soo Kim, Young-Lok Cha, Dahee An, Jongwon Byun, Yong-Ku Kang
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Abstract

Rapeseed is a typical winter crop, and its freezing stress tolerance is a major feature for winter survival. Therefore, it is important to comprehend clearly the physical and molecular mechanisms of rapeseed under freezing stress conditions. This study investigates the physical and transcriptome changes of two rapeseed lines, ‘J8634-B-30’ and ‘EMS26’, under cold acclimation and freezing temperature treatments. The proline content of ‘J8634-B-30’ at 5 °C increased 8.7-fold compared to that before treatment, and there was no significant change in that of ‘EMS26’ RNA-sequencing analysis revealed 5,083 differentially expressed genes (DEGs) of ‘J8634-B-30’ under cold acclimation condition. Among the genes, 2,784 (54.8%) were up-regulated and 2,299 (45.2%) were down-regulated. The DEGs of ‘EMS26’ under cold acclimation condition were 5,831 genes, and contained 2,199 up-regulated genes (37.7%) and 3,632 down-regulated genes (62.3%). Among them, only DEGs annotated in the cold response-related signaling pathways were selected, and their expression in the two rapeseed lines was compared. Comparative DEGs analysis indicated that cold response related signaling pathways are proline metabolism and ABA (Abscisic acid) signaling. And ICE (Inducer of CBF expression) CBF (C-repeat-binding factor) COR (Cold-regulated) signaling were the significantly differentially expressed transcripts in the two rapeseed lines. The major induced transcripts of ‘J8634-B-30’ induced P5CS (Δ‘-pyrroline-5-carboxylate synthetase), which is related to proline biosynthesis, PYL (pyrabactin resistance-like protein, ABA receptor) and COR413 (cold-regulated 413 plasma membrane 1). In conclusion, these result provide a foundation for understanding the mechanisms of freezing stress tolerance in rapeseeds. Further functional studies should be performed on the freezing stress-related genes identified in this study, which can contribute to the transgenic and molecular breeding for freezing stress tolerance in rapeseed.
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2个油菜品系对冷胁迫响应的比较转录组分析
油菜籽是典型的冬季作物,其抗冻性是其冬季生存的重要特征。因此,弄清油菜籽在冰冻胁迫条件下的生理和分子机制具有重要意义。研究了两个油菜籽品系J8634-B-30和EMS26在低温驯化和低温处理下的生理和转录组变化。5℃时,‘J8634-B-30’的脯氨酸含量比处理前增加了8.7倍,而‘EMS26’的含量变化不显著。测序结果显示,‘J8634-B-30’在冷驯化条件下存在5083个差异表达基因(DEGs)。其中上调2784个(54.8%),下调2299个(45.2%)。‘EMS26’在冷驯化条件下的基因数量为5831个,其中上调基因2199个(37.7%),下调基因3632个(62.3%)。其中,只选择了在冷响应相关信号通路中注释的deg,并比较了它们在两种油菜籽系中的表达。比较DEGs分析表明,与冷响应相关的信号通路主要是脯氨酸代谢和ABA(脱落酸)信号通路。两个油菜品系的诱导因子ICE (Inducer of CBF expression)、CBF (c -repeat binding factor)和COR (Cold-regulated)信号表达均存在显著差异。J8634-B-30的主要诱导转录本诱导了脯氨酸生物合成相关的P5CS (Δ’-吡roline -5-羧酸合成酶)、PYL (pyrabactin抗性样蛋白,ABA受体)和COR413(冷调节413质膜1)。这些结果为了解油菜抗冻机制提供了基础。本研究鉴定的冷冻胁迫相关基因的功能有待进一步研究,为油菜籽耐冷冻胁迫的转基因和分子育种奠定基础。
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