Mechanism Analysis of Increased Erucic Acid Content in Brassica napus L. Seeds Resulting Low Nighttime Temperature

IF 2.6 3区 生物学 Q2 GENETICS & HEREDITY Gene Pub Date : 2024-11-21 DOI:10.1016/j.gene.2024.149119
Chao Mi , Yanning Zhao , Liangbin Lin , Jinxiong Wang
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Abstract

Brassica napus L. (B. napus L.), a crop of Brassica in the family Cruciferae, is a major sources of edible vegetable oil and one of the four most widely grown. Oil accumulation is determined by genetic and environmental factors, including temperature, light, humidity, edatope, latitude, and altitude. Temperature also plays a crucial part in the maturation stage. A highly temperature-sensitive line (STSL, DH0729DH0815) and weakly temperature-sensitive line (WTSL, DH0729) were used to express LOC106368911 and its effect on the erucic acid content of seeds under low nighttime temperatures. Condition of 20/18 °C (±0.5 °C, daytime/nighttime temperature, CK) and 20/13 °C (low nighttime temperature, LNT) were used in tests. Under LNT, the erucic acid content in STSL seeds increased significantly, whereas the WTSL change was not significant. The relative expression of LOC106368911 was significantly increased in STSL at 27, 35 and 43 days after flowering (DAF), whereas the change in WTSL was not significant. The CDS sequence of the LOC106368911 was cloned. Compared to the protein sequence encoded by the reference gene, STSL changed the 96th amino acid sequence from I (leucine) to L (isoleucine). However, there was no difference in the secondary and tertiary structures. Based on this, we cloned the LOC106368911 promoter sequence and found that the mutation of C to T at position −790 in WTSL caused the loss of LTR (cis-acting element involved in low-temperature responsiveness, −791 to −786) element function in response to low temperature and increased erucic acid content. The LOC106368911 promoter had 2 LTR elements in STSL (−791 to −786 and −588 to −583). A GUS reporter vector was constructed to study the transient expression in tobacco leaf transformations. GUS gene expression at 13 ℃ after 2 h was significantly higher than that at 18 ℃. Base and number differences in the LTR element were found in the LOC106368911 promoter sequence of STSL and WTSL. Base mutations occurred in the LTR element in WTSL, which resulted in decrease or loss of erucic acid content in response to low nighttime temperatures.
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夜间低温增加油菜籽芥酸含量的机理分析
油菜(Brassica napus L.,B. napus L.)是十字花科芸薹属作物,是食用植物油的主要来源,也是种植面积最大的四种植物之一。油脂积累由遗传和环境因素决定,包括温度、光照、湿度、纬度和海拔。温度在成熟阶段也起着至关重要的作用。利用高温度敏感品系(STSL,DH0729-DH0815)和弱温度敏感品系(WTSL,DH0729)表达 LOC106368911 及其对夜间低温条件下种子芥酸含量的影响。试验条件为 20/18 °C(±0.5 °C,昼夜温差,CK)和 20/13 °C(夜间低温,LNT)。在 LNT 条件下,STSL 种子中芥酸含量显著增加,而 WTSL 变化不明显。在开花后 27 天、35 天和 43 天(DAF),STSL 中 LOC106368911 的相对表达量明显增加,而 WTSL 的变化不明显。克隆了 LOC106368911 的 CDS 序列。与参考基因编码的蛋白质序列相比,STSL 的第 96 个氨基酸序列从 I(亮氨酸)变为 L(异亮氨酸)。然而,二级和三级结构并无差异。在此基础上,我们克隆了 LOC106368911 启动子序列,发现 WTSL 中 -790 位的 C 突变为 T,导致 LTR(参与低温响应的顺式作用元件,-791 至 -786)元件对低温和芥酸含量增加的响应功能丧失。LOC106368911 启动子在 STSL 中有两个 LTR 元件(-791 至 -786 和 -588 至 -583)。为了研究烟草叶片转化中的瞬时表达,我们构建了一个 GUS 报告载体。2 h后,13 ℃时的GUS基因表达量明显高于18 ℃时的表达量。在STSL和WTSL的LOC106368911启动子序列中发现了LTR元件的碱基和数目差异。WTSL的LTR元件发生碱基突变,导致芥酸含量对夜间低温的反应降低或丧失。
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来源期刊
Gene
Gene 生物-遗传学
CiteScore
6.10
自引率
2.90%
发文量
718
审稿时长
42 days
期刊介绍: Gene publishes papers that focus on the regulation, expression, function and evolution of genes in all biological contexts, including all prokaryotic and eukaryotic organisms, as well as viruses.
期刊最新文献
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