| 孙柏林,马骊,曾秀存,牛早霞,王万鹏,呼芳娣,路晓明,祁伟亮,武军艳,刘丽君,李学才,孙万仓,缪纯庆,王学芳,李孝泽,方彦.低温胁迫下白菜型冬油菜脂肪酸组分及FAD3的差异表达分析[J].干旱地区农业研究,2021,39(1):65~74 |
| 低温胁迫下白菜型冬油菜脂肪酸组分及FAD3的差异表达分析 |
| Fatty acid components and differential expression of FAD3 in winter Brassica rapa L. under low temperature stress |
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| DOI：10.7606/j.issn.1000-7601.2021.01.09 |
| 中文关键词: FAD3 白菜型冬油菜；低温胁迫；脂肪酸；脂肪酸不饱和指数；差异表达 |
| 英文关键词:FAD3 winter Brassica rapa L. cold stress fatty acid IUFA differential expression |
| 基金项目:国家自然基金项目(31860388)；科技部“油菜杂交优势利用技术与强优势杂交种创制”（2016YFD0101300）；国家现代农业产业技术体系项目(CARS-12)；甘肃省现代农业产业技术体系项目(GARS-TSZ-1)；国家自然基金项目(31960435) |
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| 中文摘要: |
| 为探究白菜型冬油菜(Brassica rapa L.)脂肪酸组分对低温胁迫的响应及脂肪酸去饱和酶FAD3(ω-3-fatty acid desaturase）基因与抗寒性的关系，采用气相色谱-质谱法测定了3个不同抗寒性白菜型冬油菜品种在低温胁迫下根和叶的脂肪酸组分变化；利用同源克隆法获得白菜型油菜陇油7号FAD3的CDS序列，利用生物信息学方法对其进行分析。结果显示:低温胁迫下，强抗寒品种陇油7号根部亚油酸和亚麻酸的含量较不抗寒品种Lenox分别高157%和89%，陇油7号根部脂肪酸不饱和指数（IUFA）较Lenox高91%,即低温胁迫下，强抗寒品种的亚油酸、亚麻酸和IUFA均显著高于不抗寒品种；陇油7号FAD3序列CDS区全长1 290 bp，编码429 aa，分子质量约49.16 kD，等电点为8.60，是一种稳定的亲水蛋白。多序列比对分析发现其编码蛋白与同一科属中大白菜相似性最高，为100%，进化树分析结果也表明FAD3蛋白与大白菜亲缘关系较近。荧光定量表达分析发现低温胁迫下，FAD3基因在陇油7号、天油2号和Lenox根中的表达量较对照（22℃）分别增加了690%、590%和710%，说明FAD3基因通过上调表达来适应低温，且其表达量与IUFA呈正相关关系。 |
| 英文摘要: |
| In order to explore the response of fatty acid composition to low temperature stress and the cold resistance mechanism of FAD3 (ω-3-Fatty acid desaturase) in winter Brassica rapa L., The changes of fatty acid composition in roots and leaves of three different cold resistant types of winter rapeseed (Brassica campestris L.) under low temperature stress were determined by gas chromatography Mass Spectrometry (GC-MS). The CDS sequence of omega-3 fatty acid desaturase (FAD3) of Longyou 7 was obtained by PCR technique. The FAD3 protein sequence of Longyou 7 was analyzed by bioinformatics. The results showed that under low temperature stress, the contents of linoleic acid and linolenic acid in roots of strong cold\|resistant variety Longyou 7 increased by 157% and 89%, respectively, compared with non\|cold\|resistant variety Lenox, and the value of IUFA in root of Longyou 7 increased by 91% compared with Lenox. That is, under low temperature stress, linoleic acid, linolenic acid, and IUFA of cold\|resistant varieties were significantly higher than those of non\|cold\|resistant varieties. The results showed that the CDS region of Longyou 7 FAD3 sequence encodes 429 amino acids with a molecular weight of 49.16 kD and an isoelectric point of 8.60. It is a stable hydrophilic protein. Multiple sequence alignment analysis showed that its encoded protein had the highest similarity with Brassica rapa in the same family, reaching 100%. The result of evolutionary tree analysis also showed that the relationship between FAD3 protein and Brassica rapa was closely correlated. Fluorescence quantitative expression analysis showed that under low temperature stress, the expression of FAD3 in Longyou 7, Tianyou 2, and Lenox roots increased by 690%, 590%, and 710%, respectively, compared with the control (22℃). The results showed that FAD3 was up\|regulated to adapt to low temperature, and its expression was positively correlated with fatty acid unsaturation index (IUFA). The above studies provided ideas for understanding the function of [STBX]FAD3 protein and the composition of fatty acids in winter Brassica rapa L., exploring its role in stress resistance and cultivating stress\|resistant germplasm resources. |
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