Enhancing terpenoid indole alkaloid production by inducible expression of mammalian Bax in Catharanthus roseus cells.
Bax, a mammalian pro-apoptotic member of the Bcl-2 family, triggers hypersensitive reactions when expressed in plants. To investigate the effects of Bax on the biosynthesis of clinically important natural products in plant cells, we generate transgenic Catharanthus roseus cells overexpressing a mouse Bax protein under the beta-estradiol-inducible promoter. The expression of Bax in transgenic Catharanthus roseus cells is highly dependent on beta-estradiol concentrations applied. Contents of catharanthine and total terpenoid indole alkaloid of the transgenic cells treated with 30 micromol/L beta-estradiol are 5.0-and 5.5-fold of the control cells. Northern and Western blotting results show that expression of mammalian Bax induces transcriptional activation of Tdc and Str, two key genes in terpenoid indole alkaloid biosynthetic pathway of Catharanthus roseus cells, and stimulates the accumulation of defense-related protein PR1 in the cells, showing that the mouse Bax triggers the defense responses of Catharanthus roseus cells and activates the terpenoid indole alkaloid biosynthetic pathway. Thus, our data suggest that the mammalian Bax might be a potential regulatory factor for secondary metabolite biosynthesis in plant cells and imply a new secondary metabolic engineering strategy for enhancing the metabolic flux to natural products by activating the whole biosynthetic pathway rather than by engineering the single structural genes within the pathways.
Xu
MaoJun
M
Department of Biotechnology, Zhejiang Gongshang University, Hangzhou 310035, China. maojunxu@163.com.
Dong
JuFang
J
D000818:Animals; D002460:Cell Line; D004958:Estradiol; D018014:Gene Transfer Techniques; D031313:Gentianella / Q000235:genetics*; D051379:Mice; D016366:Open Reading Frames; D010940:Plant Proteins / Q000235:genetics*; D030821:Plants, Genetically Modified; D010957:Plasmids; D011401:Promoter Regions, Genetic; D014162:Transfection; D051028:bcl-2-Associated X Protein / Q000235:genetics*
D010940:Plant Proteins; D051028:bcl-2-Associated X Protein; D004958:Estradiol