| 叶片衰老诱导FLP重组酶删除转基因烟草外源基因的研究 |
| |
| 引用本文： | 秦利军,李启龙,赵德刚.叶片衰老诱导FLP重组酶删除转基因烟草外源基因的研究[J].中国烟草学报,2019,25(6):90-97. |
| |
| 作者姓名： | 秦利军 李启龙 赵德刚 |
| |
| 作者单位： | 1.贵州大学农业生物工程研究院/山地植物资源保护与种质创新省部共建教育部重点实验室，贵州贵阳甲秀南路 550025 |
| |
| 基金项目： | 贵州省科技计划项目“烟草钾离子通道蛋白及BR对植株抗非生物胁迫研究”黔科合LH[2016]7449号贵大人基合字“利用基因编辑技术创制抗除草剂烟草新种质研究”[2015]24号 |
| |
| 摘 要： | 目的 “Gene-deletor”系统可实现转基因植物中外源基因的清除。为了研究该系统在转基因烟草中对清除外源基因的作用并最终创制不含外源基因的烟草新种质。 方法 以含有“Gene-deletor”系统的转基因烟株为材料，分析外源筛合报告选融基因Bar::GUS、重组酶基因FLP在不同叶龄叶片中的表达水平，同时通过观察转基因烟草花粉中GUS蛋白的表达活性，分析外源基因GUS在花粉中的删除情况。 结果 （1）相同叶龄不同转基因烟株叶片均表现出GUS活性和对除草剂草铵膦抗性，且3个转基因烟草株系中GUS活性和对除草剂抗性都表现出随着叶龄增大而降低的特点。（2）Real-time PCR和RT-PCR结果进一步证明外源Bar::GUS基因表达水平随叶片发育成熟持续下降，而外源FLP基因的表达水平则出现先增后降的变化趋势，在测定后期两者的表达量均达到最低，（3）花粉GUS组织染色结果表明，“明，色结果降的变化趋势，特系统诱导各转基因植株中花粉外源基因清除效率不同，平均清除率分别78.3%，54.7%和75.2%。 结论 在转基因烟草中，叶片衰老特异表达基因SAG12基因启动子驱动“动启动子驱动达基因，平均清系统（LoxP/FRT）的表达不仅可引起转基因植株叶片中外源基因的特异性清除，还能诱导花粉中外源基因的删除，该技术可为进一步创制培育不含外源基因的烟草新种质提供参考。
|
| 关 键 词： | 烟草 外源基因清除 筛选报告基因Bar::GUS 重组酶基因FLP |
| 收稿时间： | 2019-07-24 |
Study on exogenous gene deletion from transgenic tobacco by leaf senescence-induced FLP recombinase |
| |
| Affiliation: | 1.Institute of Agro-Bioengineering / Key Laboratory of Plant Resources Conservation and Germplasm Innovation in Mountainous Region (Ministry of Education), Guizhou University, Guiyang 550025, China2.Guizhou Academy of Agricultural Sciences, Guiyang 550006, China |
| |
| Abstract: | The "gene-deletor" system is able to achieve the excision of exogenous genes in transgenic plants.In order to study the effect of this system on the excision of exogenous genes in transgenic tobacco and create new tobacco germplasm without exogenous genes, transgenic tobacco lines containing "gene-deletor" system were used as materials to analyze the expression levels of the reporter gene Bar::GUS and recombinase gene FLP in leaves at different leaf ages after exogenous screening.Meanwhile, GUS deletion in pollen was also analyzed by observing the expression activity of GUS in transgenic tobacco pollen.The results showed 1) at the same leaf age, leaves from different transgenic tobacco plants displayed GUS activity and resistance to herbicides phosphinothricin, and the GUS activity and herbicides-resistance in three transgenic tobacco lines showed a decreasing trend of GUS activity and herbicides-resistance with the increase of leaf age (2) The results of Real-time PCR and RT-PCR further proved that the expression level of Bar::GUS gene declined with mature leaf development, while the expression level of FLP showed a tendency of first increasing and then decreasing (3) GUS tissue staining results showed that the Gene-deletor" system led to the removal of exogenous gene in pollen of transgenic plants, with an average excision rate of 78.3%, 54.7 and 75.2%, respectively.In transgenic tobaccos, the the expression of "Gene-deletor" system (LoxP/FRT)driven by leaf senescence gene SAG12 not only could cause the specific excision of the foreign genes in transgenic tobacco leaves, but also could induce the excision of the exogenous gene in pollen.Moreover, the system can be used as a reference for further fostering new tobacco germplasm without exogenous genes. |
| |
| Keywords: | |
|
| 点击此处可从《中国烟草学报》浏览原始摘要信息 |
|
点击此处可从《中国烟草学报》下载全文 |
