Production of useful substances by factory of elite plant cells

植物可以在自己的微细胞工厂里从简单的无机化合物生产复杂的有机化合物。利用这一复杂的功能，在离体植物组织培养系统中进行了用于化学工程合成的有用工业材料的生产。通过基因导入代谢工程、胁迫诱导抑制代谢途径和/或选择优质克隆，建立表达目标化合物高产的优质克隆。利用根生农杆菌将阿魏酸（C_6-C_3）转化为香兰素（C_6-C_1）的细菌HCHL基因引入到甜菜毛状根中。所选毛状根无性系的pHBA（C_1）葡萄糖酯含量在干重基础上可达14% 2。2 .将能将山莨菪碱转化为东莨菪碱的植物H6H引入莱茵杜氏菌（Duboisia leichhardtii），与前文相同，建立了产总莨菪碱95%以上东莨菪碱的优良无性系。1)酵母提取物在叶培养物中诱导呋喃香豆素，而在根培养物中不诱导。2)抗坏血酸在根培养和叶培养中高诱导呋喃香豆素。这是根培养系统中呋喃香豆素诱导的第一例。白芍毛状根产生核黄素（维生素B_2）在缺铁条件下，白芍毛状根将核黄素分泌到培养基中。产量与根尖大小和根尖数量有关。选择大部分高产优良无性系作进一步研究。

Plants can produce complex organic compounds from simple inorganic compounds in their own micro cell factory. Using this sophisticated function, productions of useful industrial materials, which are used to be synthesized by chemical engineering, were performed with in vitro plant tissue culture system. To establish elite clones expressing high productivity of aimed compounds, metabolic engineering by gene introduction, induction of suppressed metabolic pathway with stress treatments, and/or selection of superior clones were carried out as follows:1.p-Hydroxy benzoic acid(pHBA)production by Beta vulgaris hairy rootsThe gene of bacterial HCHL, which converts ferulic acid(C_6-C_3) to vanillin(C_6-C_1), was introduced to Beta vulgaris using Agrobacterium rhizogenes. A selected elite hairy root clone produced glucose ester of pHBA(C_1)up to 14% on the basis of dry weight2. Scopolamine production by Duboisia leichhardtii hairy rootsPlant H6H, which converts hyoscyamine to scopolamine, was introduced into Duboisia leichhardtii, same as above, establishing an elite clone that produced more than 95% of scopolamine in total tropane alkaloids.3.Coumarin production by Glehnia littoralis organ cultures1)Yeast extract induced furanocoumarin in leaf cultures, but not in root cultures.2)Ascorbic acid highly induced furanocoumarin in root culture as well as leaf culture. This is the first case of furanocoumarin induction in a root culture system.4.Riboflavin(vitamin B_2) production by Hyoscyamus albus hairy rootsRiboflavin was excreted by Hyoscyamus albus hairy roots into the medium cultured under iron-deficient condition. The productivity was dependent to size and number of root-tip. Most producing elite clone was chosen for further study.