Saito K
Faculty of Pharmaceutical Sciences, Chiba University, Japan.
Yakugaku Zasshi. 1994 Jan;114(1):1-20. doi: 10.1248/yakushi1947.114.1_1.
The advances in molecular genetics and biotechnology in the field of medicinal plant research are discussed with focusing on the works using transgenic plants. Differentiated organ cultures and transgenic teratomas, incited by the infection with mutants of Agrobacterium Ti and Ri plasmids, were established in quinolizidine-alkaloid producing plants and Solanaceae plants. These cultured cells were used for the production and bioconversion of specific alkaloids produced in these plants. The methods of integration of foreign genes into medicinal plants were developed using an Ri binary vector. The mode of gene expression driven by TR1'-2' promoters was elucidated in transgenic medicinal plants, e.g., Nicotiana tabacum, Glycyrrhiza uralensis, Digitalis purpurea and Atropa belladonna. The genes for herbicide resistance, mammalian cytochrome P450 and bacterial beta-hydroxydecanoylthioester dehydrase were transferred and expressed in plants either to confer herbicide-resistant trait or to change the pattern of metabolites. The cDNA clones encoding cysteine synthase responsible for sulfur assimilation and biosynthesis of non-protein amino acids were isolated and characterized from Spinacea oleracea and Citrullus vulgaris. The functional lysine residue was identified by site-directed mutagenesis experiments. An over-expression system in Escherichia coli was constructed for the bacterial production of the plant specific non-protein amino acids. We made transgenic N. tabacum integrated with sense- and antisense-constructs of cysteine synthase cDNA driven by cauliflower mosaic virus 35S promoter for the purpose of genetic manipulation of biosynthetic flow of cysteine in plants. The future prospects of medicinal plant research are also discussed in the context of modern plant molecular biology.
本文讨论了药用植物研究领域分子遗传学和生物技术的进展,重点关注利用转基因植物开展的研究工作。通过感染根癌农杆菌Ti质粒和发根农杆菌Ri质粒的突变体,在喹诺里西啶生物碱生产植物和茄科植物中建立了分化器官培养物和转基因畸胎瘤。这些培养细胞被用于这些植物中特定生物碱的生产和生物转化。利用Ri二元载体开发了将外源基因整合到药用植物中的方法。在转基因药用植物,如烟草、甘草、毛地黄和颠茄中,阐明了由TR1'-2'启动子驱动的基因表达模式。将抗除草剂基因、哺乳动物细胞色素P450基因和细菌β-羟基癸酰硫酯脱水酶基因导入植物并使其表达,以赋予植物抗除草剂特性或改变代谢物模式。从菠菜和西瓜中分离并鉴定了编码负责硫同化和非蛋白质氨基酸生物合成的半胱氨酸合酶的cDNA克隆。通过定点诱变实验鉴定了功能性赖氨酸残基。构建了大肠杆菌中的过表达系统,用于细菌生产植物特异性非蛋白质氨基酸。为了对植物中半胱氨酸的生物合成途径进行遗传操作,我们构建了由花椰菜花叶病毒35S启动子驱动的半胱氨酸合酶cDNA正义和反义构建体的转基因烟草。本文还在现代植物分子生物学的背景下讨论了药用植物研究的未来前景。