Liu Feng, Cao Yueping
Department of Plant Science, School of Agriculture and Biology, Shanghai Jiao Tong University , Shanghai , P. R. China.
Turk J Biol. 2018 Apr 27;42(2):187-194. doi: 10.3906/biy-1712-56. eCollection 2018.
Glyphosate is a widely used herbicide that inhibits the 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS)-encoding aroA gene in the shikimate pathway. The discovery and cloning of the aroA gene with high resistance is central to breeding a transgenic glyphosate-resistant plant. A novel aroAPantoea gene from Pantoea G-1 was previously isolated and cloned. The aroA Pantoea enzyme was defined as a new class I EPSPS with glyphosate resistance. The aroA Pantoea gene was introduced into tobacco through Agrobacteriummediated transformation. The transgenic tobacco plants were confirmed by PCR, RT-PCR, and Southern blot. The analysis of glyphosate resistance also showed that the transgenic tobacco plants could survive at 15 mM glyphosate; the glyphosate resistance level of the transgenic plants is higher than the agricultural application level recommended by most manufacturers. Overall, this study shows that aroAPantoea can be used as a candidate gene for the development of genetically modified crops.
草甘膦是一种广泛使用的除草剂，它能抑制莽草酸途径中编码5-烯醇丙酮酰莽草酸-3-磷酸合酶（EPSPS）的aroA基因。发现并克隆具有高抗性的aroA基因是培育抗草甘膦转基因植物的关键。先前已从泛菌属G-1中分离并克隆了一个新的aroA泛菌基因。aroA泛菌酶被定义为一种具有草甘膦抗性的新型I类EPSPS。通过农杆菌介导的转化将aroA泛菌基因导入烟草。通过PCR、RT-PCR和Southern杂交对转基因烟草植株进行了鉴定。草甘膦抗性分析还表明，转基因烟草植株在15 mM草甘膦浓度下能够存活；转基因植株的草甘膦抗性水平高于大多数制造商推荐的农业应用水平。总体而言，本研究表明aroA泛菌可作为转基因作物开发的候选基因。
