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

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.