Jilin Provincial Key Laboratory of Agricultural Biotechnology, Agro-Biotechnology Institute, Jilin Academy of Agricultural Sciences, Changchun, 130033, China.
Transgenic Res. 2018 Apr;27(2):155-166. doi: 10.1007/s11248-018-0063-4. Epub 2018 Feb 23.
Soybean oil contains approximately 20% oleic acid and 63% polyunsaturated fatty acids, which limits its uses in food products and industrial applications because of its poor oxidative stability. Increasing the oleic acid content in soybean seeds provides improved oxidative stability and is also beneficial to human health. Endoplasmic reticulum-associated delta-12 fatty acid desaturase 2 (FAD2) is the key enzyme responsible for converting oleic acid (18:1) precursors to linoleic acid (18:2) in the lipid biosynthetic pathway. In this study, a 390-bp conserved sequence of GmFAD2-1B was used to trigger a fragment of RNAi-mediated gene knockdown, and a seed-specific promoter of the β-conglycinin alpha subunit gene was employed to downregulate the expression of this gene in soybean seeds to increase the oleic acid content. PCR and Southern blot analysis showed that the T-DNA had inserted into the soybean genome and was stably inherited by the progeny. In addition, the expression analysis indicated that GmFAD2-1B was significantly downregulated in the seeds by RNAi-mediated post-transcription gene knockdown driven by the seed-specific promoter. The oleic acid content significantly increased from 20 to ~ 80% in the transgenic seeds, and the linoleic and linolenic acid content decreased concomitantly in the transgenic lines compared with that in the wild types. The fatty acid profiles also exhibited steady changes in three consecutive generations. However, the total protein and oil contents and agronomic traits of the transgenic lines did not show a significant difference compared with the wild types.
大豆油含有约 20%的油酸和 63%的多不饱和脂肪酸,由于其氧化稳定性差,限制了其在食品和工业应用中的使用。增加大豆种子中的油酸含量可以提高氧化稳定性,也有利于人类健康。内质网相关的 delta-12 脂肪酸去饱和酶 2(FAD2)是负责将油酸(18:1)前体转化为油脂生物合成途径中亚油酸(18:2)的关键酶。在这项研究中,使用 390bp 的 GmFAD2-1B 保守序列触发 RNAi 介导的基因敲低片段,并用β-伴大豆球蛋白α亚基基因的种子特异性启动子下调该基因在大豆种子中的表达,以增加油酸含量。PCR 和 Southern blot 分析表明,T-DNA 已插入大豆基因组,并通过种子特异性启动子稳定遗传给后代。此外,表达分析表明,GmFAD2-1B 在种子中的表达通过种子特异性启动子驱动的 RNAi 介导的转录后基因敲低显著下调。与野生型相比,转基因种子中的油酸含量从 20%显著增加到约 80%,亚油酸和亚麻酸含量同时减少。脂肪酸图谱在三个连续的世代中也表现出稳定的变化。然而,与野生型相比,转基因系的总蛋白和油含量以及农艺性状没有显著差异。