| 本生烟草组培再生及遗传转化的研究 |
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| 引用本文： | 苏上,张旋,冯思思,李凤兰.本生烟草组培再生及遗传转化的研究[J].安徽农业科学,2010,38(20):10530-10531,10607. |
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| 作者姓名： | 苏上 张旋 冯思思 李凤兰 |
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| 作者单位： | 北京林业大学生物科学与技术学院,北京,100083;北京林业大学生物科学与技术学院,北京,100083;北京林业大学生物科学与技术学院,北京,100083;北京林业大学生物科学与技术学院,北京,100083 |
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| 基金项目： | 北京林业大学理科基地基金 |
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| 摘 要： | 目的]获得本生烟草转胰高血糖素样肽-1基因植株。方法]以本生烟草无菌苗叶片为外植体材料,建立本生烟草再生系统,通过农杆菌叶盘法浸染进行遗传转化,获得本生烟草转胰高血糖素样肽-1基因植株。结果]通过多重比对和重复,确定本生烟草叶片最适分化培养基为MS＋1.5mg/L6-BA＋0.1mg/LNAA,生根培养基为MS＋0.1mg/LIBA;经由卡那霉素浓度梯度试验,确定选择压力为5mg/L卡那霉素;对经卡那霉素筛选获得的抗性芽进行GUS组织化学检测,获得阳性信号。结论]初步证实外源基因胰高血糖素样肽-1已整合到本生烟草基因组中。
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| 关 键 词： | 本生烟草 组培 转基因 GUS检测 |
Construction of Tissue Culture and Genetic Transformation Systems of Nicotiana benthamiana |
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| Institution: | SU Shang et al(College of Biological Science and Biotechnology,Beijing Forestry University,Beijing 100083) |
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| Abstract: | Objective]The research aimed to obtain the transfer of glucagon likepeptide-1 gene Nicotiana benthamiana plants.Method] With sterile tobacco leaf explants as the experimental material,the regeneration system of tobacco was established and transgenic Nicotiana benthamiana was obtained through Agrobacterium infection.Result]By multiple comparison and repeated experiments,the optimal Nicotiana benthamiana differentiation medium of MS+1.5 mg/L 6-BA +0.1 mg/L NAA was determined,so did the rooting medium MS +0.1 mg/L IBA.By kanamycin selection pressure gradient experiments,the selection concentration was identified as 5 mg/L kanamycin,later disseminated by Agrobacterium leaf disc method for genetic transformation.GUS acquired resistance to staining the buds and the positive signal was obtained.Conclusion]Gene GLP-1 was integrated into the genome of tobacco Nicotiana benthamiana. |
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| Keywords: | Nicotiana benthamiana Tissue culture Transgenic GUS detection |
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