烟草NtGGPPS 3基因的亚细胞定位和组织表达分析

Subcellular Localization and Tissue Expression Analysis of NtGGPPS 3 Gene in Tobacco

为研究烟草牻牛儿基牻牛儿基焦磷酸合成酶基因NtGGPPS 3在烟草萜类物质合成中的功能,构建NtGGPPS 3与GFP融合蛋白的植物表达载体,并通过基因枪法瞬时转化烟草悬浮细胞BY2,研究该基因的亚细胞定位情况,并对该基因在烟草不同生育期不同器官中的表达量进行分析。结果表明:NtGGPPS 3定位在烟草的叶绿体和质膜上;NtGGPPS 3基因在普通烟草主要生育期各组织中均表达,在叶片中表达量较高,在打顶期的茎中表达量明显上升。表明,该基因可能参与叶绿素、类胡萝卜素和质体醌等与光合作用相关萜类化合物的合成,也可能参与激素和防御相关的萜类物质合成。

The subcellular location of NtGGPPS gene was determined by building the plant expression vector of NtGGPPS3 and GFP fusion protein, and transiently transforming tobacco suspension cultured cell BY2 by particle bombardment and then expression quantity of GGPPS gene in different organs of tobacco at different growth stages was analyzed to study the function of geranylgeranyl pyrophosphate synthase gene （NtGGPPS） in terpenoid synthesis of tobacco. The results show that NtGGPPS gene is located in chloroplast and plasmalemma and generally expresses in different tissues of common tobacco at main growth stages. The expression quantity of NtGGPPS3 in leaves is the highest and expression quantity of NtC-GPPS3 gene in stems of tobacco plants at topping stage increases obviously, which indicates that NtGGPPS3 gene may involve in synthesis of terpenoid including chlorophyll, carotenoid and plastoquinone related to photosynthesis and may involve in synthesis of terpenoid related to hormone and defense also.