Identification of a phosphinothricin-resistant mutant of rice glutamine synthetase using DNA shuffling
文献类型： 外文期刊
作者： Tian, Yong-Sheng 1 ; Xu, Jing 1 ; Zhao, Wei 1 ; Xing, Xiao-Juan 4 ; 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
期刊名称：SCIENTIFIC REPORTS （ 影响因子：4.379； 五年影响因子：5.133 ）
ISSN： 2045-2322
年卷期： 2015 年 5 卷
页码：
收录情况： SCI
摘要： To date, only bar/pat gene derived from Streptomyces has been used to generate the commercial PPT-resistant crops currently available in the market. The limited source of bar/pat gene is probably what has caused the decrease in PPT-tolerance, which has become the main concern of those involved in field management programs. Although glutamine synthetase (GS) is the target enzyme of PPT, little study has been reported about engineering PPT-resisitant plants with GS gene. Then, the plant-optimized GS gene from Oryza sativa (OsGS1S) was chemically synthesized in the present study by PTDS to identify a GS gene for developing PPT-tolerant plants. However, OsGS1S cannot be directly used for developing PPT-tolerant plants because of its poor PPT-resistance. Thus, we performed DNA shuffling on OsGS1S, and one highly PPT-resistant mutant with mutations in four amino acids (A63E, V193A, T293A and R295K) was isolated after three rounds of DNA shuffling and screening. Among the four amino acids substitutions, only R295K was identified as essential in altering PPT resistance. The R295K mutation has also never been previously reported as an important residue for PPT resistance. Furthermore, the mutant gene has been transformed into Saccharomyces cerevisiae and Arabidopsis to confirm its potential in developing PPT-resistant crops.
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