Study On The Artificial Propagation And Extractive Of Huperzia Serrata (Thunb.ex Murray) Trev.

The Huperzia serrata,also called "qiancengta" in Chinese,is perennial herbaceous plant,which is mainly used to treat the gynecological diseases and relieve swelling and detoxification and so on in folk and the alzheimer's disease,myasthenia gravis as well as memory deterioration in clinical application currently.With the coming of aging population and as people's life pressures increase,people's demand for the Huperzia serrata gradually increases.As a result,the wild resources are destroyed seriously,and in some areas even on the brink of extinction.At present,there are research scholars have explored tissue culture to expand breeding of Huperzia serrata.But there are also some difficulties with the culture medium preparation and sterilization of endophytic.What's more,high cost of production facilities is another problem.Besides,there is still some way between synthetic Huperzia serrata and market application.Therefore,this research discusses about the effects of matrices of physical and chemical properties,treatment processes and management on cutting propagation,alkaloid extraction and detection of Huperzia serrata.Intending to offer theoretical supports for normalized and industrial artificial cultivation and breeding.The main results are as follows:(1)Wild plants of Huperzia serrata were domesticated after their transplantation that used three mixed matrix.The main statistical methods were PCA,analysis of variance and multiple comparison.The comparison results show that the volumetric weight is 0.29 g/cm~3,the size of the bulk porosity is 77.77%,the void ratio is 0.45 of mixed matrix of Z1.The rate of survival after acclimatization is 95%after 90d,adventitious roots grow well,it is the most large of cumulative growth(40.75 mm).Compared with the other two mixed matrix the rises of 46.17%and 14.49%in Z1 and Z2.The total amount of dry matter is 35.53g,root-shoot ratio is 0.49,other physiological indexes are better than the rest of the mixed matrixes.The results,which proves that the mixed matrix of Z1(perlite:fine ceramsite:peat = 2:1:1)is suitable for transplanting of Huperzia serrata.(2)This study conducts a cuttage test in a sealed bottle and analyzes if mixed matrix,biological hormone and sterilization would make effect on physiology and growth targets of cutting of Huperzia serrata.The test result shows that the mixed matrix of JZ3 mixed matrix of perlite:peat:sand =1:1:1,the concentration of IBA for 100 mg/L,soaking time for 30 min give better cuttage effects than that of JZ1 and ZJ2.Indexes investigated were rooting rate,root number,length and cumulative growth,which is 91.67%,3.70 per plant,13.52mm,28.63mm respectively,which are optimal.(3)In the process of alkaloid's extracting of Huperzia Serrata,it was soaked by 1.0%(V/V)hydrochloric acid,at room temperature for 24 hours(solid-liquid ratio was 1:10),use ultrasonic techniques for 45 minutes,regulate the pH value to 9 by ammonia,and extracted using chloroform for three times and finally retrieved it.Through integrated comparative analysis demonstrate that the extraction rate and purity were higher in the above condition.(4)Compared the Huperzia serrata planed in Kunming University of Science and Technology with the wild plants grow in other three provinces.The thesis established a LC-MS method for rapid determination of Hup A content in Huperzia serrata(Thunb.ex Murray)Trev.And compare the content of Hup A from different parts of the plants and from different places.The mass spectrometer was operated in the positive ionization electrospray(ESI)mode using selection ion monitoring(SIM).The transition m/z:50-2000.Agilent ZORBAX SB-C18(4.6x250 mm)was used the mobile phase was methanol:water(90:10),the detection wave length was 300 nm,with flow rate at 0.5 ml/min,the column temperature was 28?,sample size was 10 ?l.Results there is a satisfactory linear relationship with in the range of 50-400 ?g/ml for Hup A(r=0.9994).It was found that the quantity of Hup A in leaf was Yunnan(0.0559%)>Jiangxi(0.0433%)>Hunan(0.0462%)>Fujian(0.0274%).And the quantity of Hup A in stem was Yunnan(0.0444%)>Jiangxi(0.0426%)>Hunan(0.0326%)>Fujian(0.0217%).The quantity of Hup A in leaf was higher than in stem in different places of Huperzia serrata.