杨 静 邢国杰 牛 陆 贺红利 杜 茜 郭东全 袁 英 杨向东

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, 吉林长春130033

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

杨 静 邢国杰 牛 陆 贺红利 杜 茜 郭东全 袁 英*杨向东*

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, 吉林长春130033

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。qRT-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 mRNA表达水平显著降低, 而根、茎、叶、花组织中内源GmFAD2-1 mRNA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。

大豆; 油酸; GmFAD2-1B; 农杆菌介导转化法

Abstract:The content of oleic acid is usually taken as a crucial indicator of cooking quality and stability for soybean oil. Breeding soybean cultivars with high oleic acid has always been considered as one of the principal aims for soybean improvement. In this study, the anti-sense GmFAD2-1B gene from soybean was transformed into the cultivated soybean by Agrobacterium-mediated method and several soybean transgenic lines with dramatically increased oleic acid content were obtained. Southern blot analysis confirmed integration of the anti-sense GmFAD2-1B fragment into the soybean genome with one to five copies of T-DNA insertions in the selected independent transgenic plants. The results of qRT-PCR showed that the foreign anti-sense GmFAD2-1B was mainly expressed in the transgenic seeds and simultaneously resulted in a significant decrease of the endogenous GmFAD2-1 mRNA, however, there were no significant differences observed for GmFAD2-1 mRNA contents in the root, leaf,stem and flower tissues of the transgenic plants compared to the non-transgenic ones. Analysis of the fatty acid compositions indicated that the content of oleic acid increased to 27.38%–80.42% in twelve transgenic lines. Specifically, in transgenic lines L40 and L72, oleic acid content was raised to 68.91%–80.42% and 65.98%–80.22% respectively, with over 2.65-fold increase compared with the wild type Williams 82 (containing 17.8%–22.0% of oleic oil). Concomitantly, linoleic acid and saturated fatty acid were reduced to 4.84%–14.55% and 10.34%–11.16% respectively in the seeds of transgenic lines L40 and L72. However, there was no significant difference in total oil and protein contents, as well as agronomic traits such as maturity period, plant height, leaf shape, flower color, pod height and weight per 100 seeds between the transgenic lines and non-transformed controls.

Keywords:Soybean; Oleic acid; GmFAD2-1B; Agrobacterium-mediated transformation

大豆(Glycine max L. Merrill)是我国最重要的油料作物和饲料作物之一, 也是人类植物性蛋白和油脂的重要来源。大豆主要加工产品-大豆油约占我国食用植物油总量的 40%左右。大豆油中富含多种人体必需的脂肪酸, 包括硬脂酸(stearic acid, SA,18:0)、棕榈……