Studies On Construction Of HMGR Gene Plant Expression Vector And Agrobacterium Tumefaciens Mediated Genetic Transformation Of Salvia Miltiorrhiza Bunge

Salvia miltiorrhiza Bunge. is a well-known Chinese herbal plant, and its roots, known as Danshen in Chinese, are widely used clinically in the treatment of menstrual disorders and blood circulation diseases etc.. The chief bioactive ingredients of Salvia miltiorrhiza , particularly the phenanthrofurane quinone derivatives generally was known as tanshinones, which have different pharmacological effects such as potent antioxidative effect, the prevention of inflammation and so on.Tanshinones, the chief bioactive ingredients of Salvia miltiorrhiza, which are the diterpenoid pigments, are generally believed to be synthesized in the non-MVA pathway. However, there has been growing evidence for the involvement of the cytosolic MVA pathway in diterpene biosynthesis and the existence of crosstalk between the two pathways for isoprenoid biosynthesis in some plants. 3-hydroxy-3-methylglutaryl coenzyme A reductase (HMGR), catalyses the conversion of 3-hydroxy- 3-methylglutaryl coenzyme A (HMG-CoA) to mevalonate (MVA),the first key enzyme in MVA pathway. HMGR could be involved in separate subcellular pathways to produce specific isoprenoid end-products through metabolic channels. A number of investigators have reported a correlation between the induction of isoprenoid biosynthesis and HMGR enzyme activity. In the article, HMGR were transferred into Salvia miltiorrhiza by Agrobacterium mediated method and the regenerated putative transgenic plants had been obtained in order to enhance the products of tanshinone, investigate effects of MVA pathway in diterpene biosynthesis in S. miltiorrhiza and realize the industrial production of tanshinone in future. The result is following:(1) In the article,3-hydroxy-3-methylglutaryl-coen-zyme A reductase (HMGR) from Aspergillus nidulans. was cloned into a plant expression vector pCAMBIA2301 confused with the Ubiquetin promoter and nos terminator. And leaf discs of S. miltiorrhiza were used to Agrobacterium-mediated gene transfer in order to get the overexpression of HMGR in Salvia miltiorrhiza.And the regenerated putative transgenic plants had been obtained based on kanamycin resistance.(2) A series of factors influencing Agrobacterium-mediated S. miltiorrhiza...