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Improved glyphosate resistance of 5-enolpyruvylshikimate-3-phosphate synthase from Vitis vinifera in transgenic Arabidopsis and rice by DNA shuffling

文献类型： 外文期刊

作者： Tian, Yong-Sheng 1 ; Xu, Jing 1 ; Xing, Xiao-Juan 4 ; Zhao, Wei 1 ; Fu, Xiao-Yan 1 ; Peng, Ri-He 1 ; Yao, Quan-Hong 1 ;

作者机构： 1.Shanghai Acad Agr Sci, Biotechnol Res Inst, Shanghai 201106, Peoples R China

2.Shanghai Ruifeng Agr Sci & Technol Co Ltd, Shanghai 201106, Peoples R China

3.Nanjing Agr Univ, Coll Hort, Nanjing 210095, Jiangsu, Peoples R China

4.Shanxi Agr Univ, Coll Hort, Taigu 030801, Peoples R China

关键词： EPSP synthase;Vitis vinifera;DNA shuffling;Glyphosate tolerance;Mutation;Transgenic rice

期刊名称：MOLECULAR BREEDING （ 影响因子：2.589； 五年影响因子：2.75 ）

ISSN：

年卷期：

页码：

收录情况： SCI

摘要： To date, only AroA variant derived from Agrobacterium tumefaciens CP4 has been used to generate the commercial glyphosate-resistant crops currently available in the market. This single source of the EPSPS gene may have caused the decrease in herbicide tolerance, which has become a major concern of those involved in field management programs. Therefore, it is of interest to increase aroA/EPSPS gene diversity and seek new glyphosate-tolerant genes for developing glyphosate-tolerant crops. In the current study, EPSPS gene from Vitis vinifera (VvEPSPS) was cloned using reverse transcription polymerase chain reaction. However, wild type VvEPSPS cannot be directly used for developing transgenic crops because of its extreme glyphosate sensitivity. Recent studies have demonstrated that DNA shuffling is an effective strategy in producing multi-mutated EPSPS resourced from plants (EPSPSplant) with improved glyphosate resistance in bacteria and plants. After performing DNA shuffling on VvEPSPS gene, one highly glyphosate-resistant mutant with seven amino acid variations was isolated after five rounds of shuffling and screening. The mutant showed seven amino acid changes in the EPSPS gene, namely, Q93R, T113A, P117L, G126A, C160Y, N239H, and V343A. The assay of glyphosate resistance further confirmed the potential of the VvEPSPS(mutant) in developing glyphosate-resistant crops.

  • 相关文献

[1]Directed evolution of beta-galactosidase from Escherichia coli into beta-glucuronidase. Xiong, Ai-Sheng,Peng, Ri-He,Zhuang, Jing,Liu, Jin-Ge,Xu, Fang,Cai, Bin,Guo, Zhao-Kui,Qiao, Yu-Shan,Chen, Jian-Min,Zhang, Zhen,Yao, Quan-Hong. 2007

[2]Mutation by DNA shuffling of 5-enolpyruvylshikimate-3-phosphate synthase from Malus domestica for improved glyphosate resistance. Tian, Yong-Sheng,Xu, Jing,Peng, Ri-He,Xiong, Ai-Sheng,Xu, Hu,Zhao, Wei,Fu, Xiao-Yan,Han, Hong-Juan,Yao, Quan-Hong.

[3]Estimating the copy number of transgenes in transformed rice by real-time quantitative PCR. Yang, LT,Ding, JY,Zhang, CM,Jia, JW,Weng, HB,Liu, WX,Zhang, DB. 2005

[4]Dietary safety assessment of genetically modified rice EH rich in beta-carotene. Yao, Quanhong,Peng, Rihe,Tang, Xueming,Zhao, Kai,Yao, Quanhong,Peng, Rihe,Tang, Xueming,Zhao, Kai,Wu, Yangyang,Xu, Yan,Du, Yanan,Hu, Ruili,Fan, Xiaorui,Ren, Fangfang,Zhao, Xiao. 2017

[5]Improving Rice Ability to Degrade Different Polycyclic Aromatic Hydrocarbons Through Multigene Transformation of a Hybrid Dioxygenase System. Peng, Ri-He,Shi, Biao,Fu, Xiao-Yan,Tian, Yong-Sheng,Zhao, Wei,Zhu, Bo,Xu, Jing,Han, Hong-Juan,Yuan, Zhong-Yi,Liu, Kai-Hong,Yao, Quan-Hong.

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