Goddijn O J, de Kam R J, Zanetti A, Schilperoort R A, Hoge J H
Biotechnology Delft-Leiden, Project Group Plant Cell Biotechnology, Leiden University, Netherlands.
Plant Mol Biol. 1992 Apr;18(6):1113-20. doi: 10.1007/BF00047714.
The enzyme tryptophan decarboxylase (TDC) (EC 4.1.1.28) converts tryptophan into tryptamine, and thereby channels primary metabolites into indole alkaloid biosynthesis. The production of these secondary metabolites in suspension cells of Catharanthus roseus depends on medium composition. Of the possible variables, we investigated the effect of hormones on the expression of the tdc gene in cell cultures. Omission of NAA from the growth medium resulted in accumulation of tdc mRNA. The addition of 1-naphthaleneacetic acid (NAA), indoleacetic acid (IAA) or 2,4-dichlorophenoxyacetic acid (2,4-D) rapidly reduced the enhanced tdc transcript level. Cytokinin was unable to suppress the enhanced transcript level. Hairy roots transformed by Agrobacterium rhizogenes also showed a reduction of the tdc mRNA level after NAA addition. Run-off transcription experiments showed that the down-regulation takes place at the transcriptional level within 15 minutes and independent of de novo protein synthesis. Thus one of the mechanisms which control the activity of terpenoid indole alkaloid biosynthesis in C. roseus cell cultures is the negative regulation by auxin of the gene involved in the first committed step.
色氨酸脱羧酶(TDC)(EC 4.1.1.28)可将色氨酸转化为色胺,从而将初级代谢产物导入吲哚生物碱的生物合成过程。长春花悬浮细胞中这些次生代谢产物的产生取决于培养基的成分。在可能的变量中,我们研究了激素对细胞培养物中tdc基因表达的影响。生长培养基中去除萘乙酸(NAA)会导致tdc mRNA积累。添加1-萘乙酸(NAA)、吲哚乙酸(IAA)或2,4-二氯苯氧乙酸(2,4-D)会迅速降低增强的tdc转录水平。细胞分裂素无法抑制增强的转录水平。发根农杆菌转化的毛状根在添加NAA后也显示tdc mRNA水平降低。连续转录实验表明,下调在15分钟内发生在转录水平,且与从头蛋白质合成无关。因此,控制长春花细胞培养物中萜类吲哚生物碱生物合成活性的机制之一是生长素对参与第一步关键反应的基因的负调控。