转2mG2-epsps基因烟草的草甘膦耐受性分析

›› 2009, Vol. 11 ›› Issue (4): 100-106.

转2mG2-epsps基因烟草的草甘膦耐受性分析

孙鹤,郎志宏,陆伟,林敏,黄大昉

中国农业科学院生物技术研究所, 北京 100081

收稿日期:2009-06-10 修回日期:2009-07-10 出版日期:2009-08-15 发布日期:2009-07-24
通讯作者: 郎志宏，副研究员，主要从事植物基因工程研究。Tel:010-82109857; E-mail:langzh@caas.net.cn
作者简介:孙鹤,硕士研究生,研究方向为生物化学与分子生物学。
基金资助:
国家自然科学基金项目（30771383）;国家转基因生物新品种培育科技重大专项（2008ZX08003-001）资助。

Analysis of Glyphosate Tolerance in Transgenic Tobacco with 2mG2-epsps Gene

SUN He, LANG Zhi-hong, LU Wei, LIN Min, HUANG Da-fang

Biotechnology Research Institute, Chinese Academy of Agricultural Sciences, Beijing 100081, China

Received:2009-06-10 Revised:2009-07-10 Online:2009-08-15 Published:2009-07-24

摘要/Abstract

摘要：

利用农杆菌介导法将耐草甘膦基因2mG2-epsps转入烟草，获得87株PCR阳性植株，阳性率为43%。对PCR阳性植株进行Southern杂交、RTPCR、Western 杂交和ELISA分析，结果显示外源基因已经整合到烟草基因组中并高效表达，转基因植株的叶片中2mG2-EPSPS蛋白的最高表达量可达26.03 μg/g 鲜重。转基因烟草草甘膦耐受性分析显示，其中有15个转化事件表现出较好的草甘膦耐受性，对草甘膦的耐受性可达到1%。转基因植株的种子可以在10 mmol/L草甘膦异丙铵盐浓度下萌发，T1代转基因烟草幼苗可以耐受浓度为0.8%的草甘膦。研究结果表明转2mG2-epsps基因烟草具有较高的草甘膦耐受性，2mG2-epsps基因可以用于耐除草剂转基因作物的培育。

关键词: 2mG2-epsps基因；转基因烟草；草甘膦耐受性

Abstract:

The glyphosate tolerance 2mG2-epsps gene was transferred into tobacco via Agrobacterium-mediated transformation. The results of PCR detection showed that 87 transformed tobacco plants were positive and the transfer rate was 43%. Southern blot, RT-PCR, Western blot and ELISA results showed that the foreign gene had been integrated into the tobacco genome and expressed effectively. The highest expression level of 2mG2-EPSPS protein reached 26.03 μg/g fresh weight. The glyphosate-tolerance assay indicated that there were 15 events showed good glyphosate tolerance. The glyphosate tolerance level of the transgenic plants reached 1%. Their seeds could germinate at the medium containing 10 mmol/L glyphosate and the transgenic seedlings could survive after spaying with 0.8% glyphosate. The research results indicated that the transgenic tobacco had good tolerance to glyphosate, and the 2mG2-epsps gene could be used for cultivation of glyphosate tolerant crops.

Key words: 2mG2-epsps gene, transgenic tobacco(Nicotinana tabacum); glyphosate tolerance

中图分类号:

孙鹤,郎志宏,陆伟,林敏,黄大昉. 转2mG2-epsps基因烟草的草甘膦耐受性分析[J]. , 2009, 11(4): 100-106.

SUN He, LANG Zhi-hong, LU Wei, LIN Min, HUANG Da-fang. Analysis of Glyphosate Tolerance in Transgenic Tobacco with 2mG2-epsps Gene[J]. , 2009, 11(4): 100-106.

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