转2mG2-epsps基因烟草的草甘膦耐受性分析

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。

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

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.