The Study On The Tissue Culture Technology Of Dictamnus Dasyarpus Turcz. And The Endophytic Fungi's Separation And Effect

Dictamnus dasycarp Turcz is a plant of Rutaceae and perennial berb. The part used for medical purpose is root and bark.It is a significant dermatology with the medical functions of resisting bacteria and eliminating inflammation,broad-spectrum,antifungal,anticancer, hemostasis,improving the body immunity and widely used in skin and health protection.It is mainly located in northeast,North China, southwest and Yangtze River region.In recent years,the reasons of constantly spreading of application,continuous developing of new products,feeblish feature of fertility of the seeds of plant itself,have caused resources in short supply day by day,which makes an important effect on the widely development and application of Dictamnus dasycarps Turcz.To protect and make use of Dictamnus dasycarps Turcz.effectively, and solve the problem of shortage of Dictamnus dasycarps Turcz.,this article studies the tissue cultivation process of Dictamnus dasycarps Turcz.and obtains endophytic fungi through separating and has application in suspended cell generating inducement of dictamnine.The results are as follows:1.The study receives callus of Dictamnus dasycarps Turcz.by stem successfully and optimizes its inductive medium.Finally,culture media callus MS+2.4-D_0.5+KT_1.5+6-BA_1.5(M42) and MS+2.4-D_0.5+KT_0.5 +6-BA_0.5(M52) are eugenic,loose,of high biomass and light yellow-green.However,first,culture media M52 starts slowly,later grows rapidly and gains ideal callus.2.The study has obtained callus of acceptable suspended cell culture through further optimizing of secondary culture.Then,on the basis of regeneration subculture medium.The study gets suspended cell culture media which is made of MS+2,4-D_0.5+KT_1.5+BA_1.5(M4-16),MS+2, 4-D_0.5+KT_1.5+BA_1.0(M4-17) and constructs the system of suspended cell culture of Dictamnus dasycarps Turcz.3.The study has separated and acquired fifty endophytic fungus monocolonies from ten objects when inducing the callus.They are M 1-1toM 1-5,M 2-1toM 2-7,M 3-1toM 3-8,M 4-1toM 4-6,M 6-1toM 6-6,M 7-1,M 7-2,M 8-1toM 8-6,M 9-1toM 9-4,M10-1to M10-6.4.The suspended cells of Dictamnus dasycarps Turcz.produce dictamnine under the condition of seven epiphytes M 1-3,M 3-5,M 1-2,M 1-4,M 6-5,M 1-5,M 6-1,M 1-6 as elicitors,which optimizes the inducing prerequisite.The epiphytic elicitors have most influence on suspended cell generating dictamnine when they are added at the telekinesis of cell culturing(12d).Each elicitor has obvious promoter action on producing dictamnine.During the induction period,the best induction is the inducer M1-5,M1-2 and M6-1 second,M1-4 worst, M6-5 M1-6 and M1-3 have indistinctively promoter action on suspended cell generating dictamnine. 5.The study has built a simple test method of dictamnine.That is to use ultraviolet spectrophotometer at 237nm to detect the contents of dictamnine and use the thin layer chromatography.The difference of verification results is between 0.1%and 0.2%.And the inaccuracy is in the permitted experiment.Therefore,the study can use ultraviolet spectrophotometer directly to detect the contents of dictamnine at 237nm.