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作物学报 ›› 2017, Vol. 43 ›› Issue (11): 1588-1595.doi: 10.3724/SP.J.1006.2017.01588

• 作物遗传育种·种质资源·分子遗传学 • 上一篇    下一篇

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*   

  1. 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?
  • 收稿日期:2017-03-20 修回日期:2017-07-19 出版日期:2017-11-12 网络出版日期:2017-08-01
  • 通讯作者: 杨向东, E-mail: xdyang020918@126.com, Tel: +86-431-87063044
  • 基金资助:

    本研究由国家转基因生物新品种培育科技重大专项(2016ZX08004-003)和国家自然科学基金项目(31671764)资助。

Antisense RNA-Mediated GmFAD2-1B Gene Silencing Enhances Accumulation of Oleic Acid in Transgenic Soybean Seeds

YANG Jing,XING Guo-Jie,NIU Lu,HE Hong-Li,DU Qian,GUO Dong-Quan,YUAN Ying*,YANG Xiang-Dong*   

  1. Agro-Biotechnology Institute, Jilin Academy of Agricultural Sciences / Jilin Provincial Key Laboratory of Agricultural Biotechnology, Changchun 130033, China
  • Received:2017-03-20 Revised:2017-07-19 Published:2017-11-12 Published online:2017-08-01
  • Contact: 杨向东, E-mail: xdyang020918@126.com, Tel: +86-431-87063044
  • Supported by:

    This study was supported by the National Major Project for Developing New GM Crops (2016ZX08004-003) and the National Natural Science foundation of China (31671764).

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。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。

关键词: 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.

Key words: Soybean, Oleic acid, GmFAD2-1B, Agrobacterium-mediated transformation

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