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Acta Agron Sin ›› 2017, Vol. 43 ›› Issue (11): 1588-1595.doi: 10.3724/SP.J.1006.2017.01588

• CROP GENETICS & BREEDING·GERMPLASM RESOURCES·MOLECULAR GENETICS • Previous Articles     Next Articles

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 Online:2017-11-12 Published:2017-08-01
  • Contact: 杨向东, E-mail: xdyang020918@126.com, Tel: +86-431-87063044 E-mail:jinggyang@126.com
  • 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).

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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