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作物学报 ›› 2009, Vol. 35 ›› Issue (5): 855-860.doi: 10.3724/SP.J.1006.2009.00855

• 作物遗传育种·种质资源·分子遗传学 • 上一篇    下一篇

薤白EPSP合成酶基因转化烟草提高其草甘膦抗性

黄丽华1，蒋向1，李博1，李育强2，张学文1*   

  1. 1湖南农业大学生物科学技术学院，湖南长沙410128；2湖南省棉花科学研究所，湖南常德415101
  • 收稿日期:2008-09-03 修回日期:2008-12-30 出版日期:2009-05-12 网络出版日期:2009-02-16
  • 通讯作者: 张学文
  • 基金资助:

    本研究由湖南省科技计划项目（07JJ5040）资助。

Transformation of EPSP Synthetase Gene from Allium macrostemon Bunge into Tobacco and Improvement of Resistance in Transgenic Phants

HUANG Li-Hua1,JIANG Xiang1,LI Bo1,LI Yu-Qiang2,ZHANG Xue-Wen1*   

  1. 1College of Bioscience and Biotechnology,Hunan Agricultural University, Changsha 410128,China;2Huna Cotton Science Institute, Changde 415101,Hunan,China
  • Received:2008-09-03 Revised:2008-12-30 Published:2009-05-12 Published online:2009-02-16
  • Contact: ZHANG Xue-Wen

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1718

被引次数

8

摘要：

葱属植物薤白具有较强的草甘膦抗性。将薤白中与草甘膦抗性相关的EPSP合成酶基因(EPSPsA) cDNA重组到植物表达载体pWM101中, 构建植物表达薤白EPSPsA重组载体, 采用根癌农杆菌介导法将其转入烟草品种WS38, 获得转薤白EPSPsA基因烟草。PCR分析表明, 目的基因已经整合进烟草基因组中。对转基因烟草愈伤组织及幼苗进行的草甘膦抗性检测表明, 转基因烟草对草甘膦的耐受性明显提高。

关键词: 薤白, EPSP合成酶基因, 烟草转化, 草甘磷抗性

Abstract:

EPSPs (5-enolppyruvylshikimate-3-phosphate synthetase) is an important enzyme involved in the synthesis of the aromatic amino acids in all plants, which catalyzes phosphoenolpyruvate and shikimate 3-phophate to form 5-enolpyruvlshimimate-3-phosphate and phosphate. Glyphosate is a chemical with similar structure of phosphoenolpyruvate, which combines enzyme to inhibit the activity of EPSPs, so it is a broad spectrum herbicide to control weeds. Breeding glyphosate tolerance or resistance crops will be an effective way to control weeds. In the study, a gene cDNA (EPSPsA) isolated from Allium macrostemon Bunge was ligated with vector pWM101 to construct a recombinant vector. The recombinant gene was transformed into WS38 via Agrobacterium-mediated way to obtain transgenic tobacco seedlings. Transgenic seedlings were screened with ordinary antibiotics. PCR analysis showed that EPSPsA was successfully integrated into the tobacco genome in all the transgenic seedlings. The result of RT-PCR also validated that EPSPsA was transcripted at mRNA leve1. At 60 d after spraying 200 mg L-1 glyphosate, the transgenic tobacco plants grew better than the untransformed plants. The average fresh weight of aerial parts in transgenic plants had a significant increase compared with those in untransformed control plants. The average chlorophyll content in transgenic plants was 5.54 mg g-1, while that in the control only 1.16 mg g-1. In the treatment with 400 mg L-1 glyphosate, the untransformed plants died while the transgenic ones grew well in the treatment with 1 000 mg L-1 glyphosate. These results proved that the EPSPsA can  improve the glyphosate tolerance in tobacco .

Key words: Allium macrostemon Bunge, EPSPs cDNA, Tobacco transformation, Glyphosate resistance

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