ORCA3转录因子对红豆杉细胞中紫杉醇生物合成的影响  

Effects of Transcriptional Factor ORCA3 on Taxol Yield in Suspension-cultured Taxus

作　　者：于小青[1] 杨海燕[2] 张蒙[1] 栗茂腾[1,3] 付春华[1,3] 余龙江[1,3] 

机构地区：[1]华中科技大学生命科学与技术学院,资源生物学与生物技术研究所,湖北武汉430074 [2]华中科技大学生命科学与技术学院 [3]华中科技大学分子生物物理教育部重点实验室,湖北武汉430074

出　　处：《山地农业生物学报》2012年第3期189-193,共5页Journal of Mountain Agriculture and Biology

基　　金：国家自然科学基金青年基金项目(200906036);国家自然科学基金面上项目(20776058)

摘　　要：ORCA3是一个受MeJA诱导的调控植物基础和次生代谢的转录调控因子。本文拟探讨ORCA3转录因子对红豆杉细胞中紫杉醇生物合成的影响,旨在为创制紫杉醇稳定高产的红豆杉细胞系,解决细胞大规模培养中紫杉醇含量低的瓶颈问题提供参考。从长春花中克隆ORCA3基因后,采用农杆菌介导法转入红豆杉细胞中,通过PCR、组织化学活性染色等检测法,筛选得到20余株转基因细胞系。HPLC检测转基因细胞系内紫杉醇含量表明:与阴性对照相比,转基因细胞系的紫杉醇含量增加了0.5～2.5倍,最高可达159.315μg/g。因此,ORCA3转录因子可以调控红豆杉细胞中紫杉醇合成。ORCA3, induced by MeJA, is a transcriptional factor regulating primary and secondary metabolism of plants. In the present research, effects of ORCA3 on taxol yield in suspension-cultured Taxus were investi- gated via transformation. Firstly, ORCA3 gene was cloned from Catharanthua roseus, then it was transferred into Taxus cell via Agrobacterium-mediated method. Totally, 20 transgenie cell lines were obtained, which were confirmed by PCR and [3-Glucuronidase assay. Taxol contents of five positive transgenic cell lines quan- tified by HPLC, were 0.5 - 2.5 times higher than that of the negative control, and the highest one was 159. 315 μg/g. Therefore, the over-expression of ORCA3 in transgenic Taxua cells might lead to an elevation of taxol yield, which lay a foundation for the creation of the high and stable taxol yield cell lines, and over- come the bottleneck for taxol production via the large-scale culture of Taxus cell.

关 键 词：红豆杉 遗传转化 ORCA3 紫杉醇 

分 类 号：Q789[生物学—分子生物学]