反义RNA介导GmFAD2-1B基因沉默增强大豆种子中油酸的高效累积

发布时间：2018-03-06 19:44

本文选题：大豆　切入点：油酸　出处：《作物学报》2017年11期 　论文类型：期刊论文

【摘要】：油酸含量是评价大豆油食用品质和稳定性的重要指标。本研究采用农杆菌介导转化法,将反义GmFAD2-1B基因导入栽培大豆品种,获得油酸含量显著提高的转基因大豆新品系。Southern杂交检测表明,外源GmFAD2-1B基因片段已导入大豆基因组,其插入拷贝数为1~5个。q RT-PCR检测表明,外源GmFAD2-1B主要在大豆种子中表达,并导致种子中内源GmFAD2-1 m RNA表达水平显著降低,而根、茎、叶、花组织中内源GmFAD2-1 m RNA表达水平无显著变化。脂肪酸组分分析表明,12份转基因大豆种子油酸含量为27.38%~80.42%,其中,L40和L72油酸含量分别为68.91%~80.42%和65.98%~80.22%,较对照品种Williams 82(17.8%~22.0%)提高2.65倍以上,亚油酸降低至4.84%~14.55%,饱和脂肪酸降低至10.34%~11.16%,但总脂肪和总蛋白含量与对照品种相比没有显著变化。农艺性状分析表明,转基因大豆在熟期、株高、叶形、花色、结荚高度、百粒重等方面与对照品种也没有显著差异。
[Abstract]:The content of oleic acid is an important index to evaluate the edible quality and stability of soybean oil. In this study, antisense GmFAD2-1B gene was introduced into soybean varieties by Agrobacterium tumefaciens mediated transformation. Southern blot analysis of transgenic soybean lines with significant increase in oleic acid content showed that exogenous GmFAD2-1B gene fragment had been introduced into soybean genome, and the inserted copy number was 1 ~ 5. Q RT-PCR analysis showed that exogenous GmFAD2-1B was mainly expressed in soybean seeds. The expression level of endogenous GmFAD2-1 m RNA in seeds was significantly decreased, while the expression of endogenous GmFAD2-1 m RNA in root, stem and leaf was significantly decreased. There was no significant change in the expression level of endogenous GmFAD2-1 m RNA in floral tissues, and fatty acid composition analysis showed that the oleic acid content of 12 transgenic soybean seeds was 27.38 and 80.42, respectively. The oleic acid contents of L40 and L72 were 68.911and 65.980.22%, respectively, which were more than 2.65 times higher than those of the control variety Williams 8217.88.80.22. Linoleic acid decreased to 4.84 ~ 14.55 and saturated fatty acid decreased to 10.34 and 11.16, but the contents of total fat and total protein were not significantly different from those of the control. Agronomic analysis showed that transgenic soybean had high plant height, leaf shape, flower color and pod height at maturity. There was also no significant difference between 100 seed weight and control variety.
【作者单位】：吉林省农业科学院农业生物技术研究所/吉林省农业生物技术重点实验室;
【基金】：国家转基因生物新品种培育科技重大专项(2016ZX08004-003) 国家自然科学基金项目(31671764)资助~~
【分类号】：S565.1

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