A Study On Production Of Isoflavones By Embryos Of Sophora Japonica L.Cell

Flavonoids has functions such as scavenging free radicals, antioxidation, antimutagenicity, antitumor, antibacterial, antiviral, regulating immunity, preventing hemal sclerosis, and reducing blood glucose, ect. It is also called "plant estrogen" by scientists. Highly pure isoflavone, flavonols and flavonoid can be obtained from fruits of S. japonica L., and cell culture is technological base of producting isoflavone by fruits of S. japonica L.. In this paper, three different growth period of embryos of Sophora japonica L. were taken as the explant for studying the technique of callus induction and cell suspension culture at first. And setting up callus cultivating system about different growth of embryos of S. japonica L.. In the second place, callus that had trained in the past were taken as the explants for adding to all kinds of inducting factors in the cell-culturing process. Establish system that elicitor promote isoflavones content about callus of embryos of S. japonica L.. At last, used ethanol extraction to optimize extraction temperature, extraction times, ethanol concentration and extracting liquid and extraction agent ratio separately. First of all, the single factor experiments were conducted, then the orthogonal experiment of L9(34)(four factors three levels) by colorimetric estimation was designed. Establish the best system that ethanol extraction of isoflavone from callus of embryos of S. japonica L.The result about three different growth period of callus culture of S. japonica L. embryos showed that adding plant growth regulator of different kinds and concentration could induce callus inⅠ,ⅡandⅢ. But there was different from induction time and induction frequency. Induction time of callus culture of S. japonica L. embryos showed that:Ⅲ>Ⅱ>Ⅰ, induction frequency of callus culture of S. japonica L. embryos showed that:Ⅱ>Ⅲ>Ⅰ, and the induction frequency ofⅡwas significantly different fromⅠandⅢ. In solid medium, The total content of isoflavones showed that:Ⅲ>Ⅰ>Ⅱ, and the isoflavones ofⅢwas significantly different fromⅠandⅡ. The suspension cultured cells, its growth showed that:Ⅱ>Ⅰ>Ⅲin each growth stage. and theⅡwas significantly different fromⅠandⅢ. But its total isoflavones-content showed that:Ⅱ>Ⅲ>Ⅰ, and the isoflavones-content ofⅢwas significantly different fromⅠandⅡ.According to induction time, induction frequenc, total isoflavones-content of solid medium and suspension cultured cells from embryos of S. a japonica L., we selectedⅡof S. japonica L. embryos to cultivate.The results of the study on elicitor promote isoflavones-content about callus of S. japonica L. embryos showed that:MeJA and SA all can increase the isoflavones-content about suspension cell of S. japonica L. embryos, but its effect was not evident, and the induction effect of SA was better than MeJA, it was contrary to results of previous. So in promoting isoflavones-content about callus of S. japonica L. embryos, we should do further study.The studying showed that the optimum process of ethanol extraction of isoflavone from S. japonica L. embryos were extraction temperature at 75℃, extraction time reaching 9h, ethanol concentration being 60% and extracting liquid and extraction agent ratio being 70:1. The maximum quantity of extracted of isoflavone of S. japonica L. was 12.045mg/g.