Research On The Selection Of Ginkgo Biloba Cell Lines Highly Producing Flavone Glycoside And Terpene As Well As The Regulation Of Flavone Glycoside Synthesis

The ginkgo biloba extract (Egb) which contains flavonoids and terpenes (ginkgolide and bilobalides) has important pharmacologic functions : The ginkgolide as a potent plate-let activating factor (PAF) antagonist has a inhibition of anaphylaxis induced by antigen and neuroprotective properties under conditions hypoxia/ ischemia, and peripheral nerve damage; Flavonoids as oxygen-free radical scavengers could attenuate lipid peroxide. Egb is in great demand in pharmacentical market In order to meet the need of pharmaceutical market for Egb, the large scale suspension culture of ginkgo biloba cells is a means could be adopted. In this article, we studied that the selecting of cell producing flavonoids and terpenes, the control of browning and the estabolishment of self-immobilization and the regulation of flavonoids synthesis in cell suspension culture. The results as follow.1. The best condition inducing ginkgo biloba callus is MS medium, adding 3.0 mg/L 2,4-D, 0.5 mg/L KT, inducing under black, the best medium to subculture callus is also MS , which could make callus loose and friable. The nutritional component and the kind of plant hormones and their combination obviously influence the growths speed and texture characteristic of callus.2. Firstly adopted small cell groups by hypoxia stress to select cell line of higher productivity , which obviously enhance the capability of flavone glycoside synthesis. The flavone glycoside content in cell line G2-1 is 0.458%, that is 3.98 times more than in callus before selecting (0.115%). The total terpenes in G2-1 is 0.105%, Which is nearly as same as in ginkgo biloba leaf.3. Through the optimization of nutritional component and kind of plant homones and their combination in media, adding antioxidants in media, that can inhibit the browning in suspension culture of ginkgo biloba cell. The optimal conditions is that: SH medium, the kind and combination of plant homones being NAA (mg/L):6-BA (mg/L) (3:1, 3:3), NAA:KT (3:0.5), 2,4-D:6-BA (3:1), 2,4-D:KT (3:0.5), and adding antioxdant PVP 500 mg/L at day 10 in suspension culture.4. Firstly estabolished ginkgo biloba cell self-immobilization suspension culture...