烟草科技2016，Vol.49Issue(4)：8-15,8.DOI:10.16135/j.issn1002-0861.20160402
烟草牻牛儿基牻牛儿基焦磷酸合成酶基因NtGGPPS1的克隆和功能分析
Cloning and functional analysis of geranylgeranyl pyrophosphate synthase gene NtGGPPS1 from Nicotiana tabacum
摘要
Abstract
To further study the function of geranylgeranyl pyrophosphate synthase (GGPPS) gene from Nicotiana tabacum, NtGGPPS1 was successfully cloned from tobacco cultivar K326 using gene-specific primers which were designed according to the coding DNA sequence of GGPPS1 (GenBank: GQ911583.1) from Nicotiana tabacum. Fluorescent expression vector of NtGGPPS1 (PFF19-NtGGPPS1-GFP) was constructed for subcellular localization in tobacco. After treated with different phytohormones, the expression level of NtGGPPS1 in K326 was studied by real-time PCR. RNAi vector pHellsgate2-NtGGPPS1 was introduced into K326 via Agrobacterium-mediated transformation and the contents of plastid pigments in NtGGPPS1 down-regulated tobacco plants were detected. The results showed that green fluorescence protein NtGGPPS1-GFP fusion was distributed in the plastids of transgenic BY-2 cells, suggesting that NtGGPPS1 located in plastids. The expression level of NtGGPPS1 in K326 was significantly increased after MeJA treatment, while it was contrary to IAA treatment. Comparing with the control, the contents of plastid pigments significantly decreased in NtGGPPS1 down-regulated plants, which indicated that NtGGPPS1 was involved in the biosynthesis ofβ-carotene in Nicotiana tabacum.关键词
普通烟草/牻牛儿基牻牛儿基焦磷酸合成酶/亚细胞定位/激素处理/质体色素Key words
Nicotiana tabacum/Geranylgeranyl pyrophosphate synthase/Subcellular localization/Phytohormone treatment/Plastid pigment分类
轻工纺织引用本文复制引用
魏攀,杨军,林福呈,李锋,孟利军,陈千思,刘萍萍,谢小东,王燃,武明珠,张剑锋,魏春阳..烟草牻牛儿基牻牛儿基焦磷酸合成酶基因NtGGPPS1的克隆和功能分析[J].烟草科技,2016,49(4):8-15,8.基金项目
中国烟草总公司郑州烟草研究院科技项目“烟草萜类化合物合成关键基因Ntggpps功能分析和调控机制研究”（902012CZ340）；中国烟草总公司郑州烟草研究院院长科技发展基金项目“DNA条形码技术在烟叶害虫检测中的应用”（902014CA0420）。 （）
