Study Of Provenance Identified Of Genuine Medicinal Materials (Liriope Muscari And Tripterygium Wilfordii) Base On RAPD Molecular Markers

The identification and quality evaluation of chinese medicinal materials is an indispensable prerequisite for clinical application,preparation of patent medicine and traditional Chinese medicine research. Because of the region differethe and the wide variety, what have happened are the confusion of specific name and the uncertain of the efficacy.Liriope muscari of Liliaceae, which is distributed mainly in guangxi and FuJian province, has positive effects on the nourishment of Yin, Lung, stomach and fluid.Tripterygium Wilfordii is the Tripterygii plant of Celastraceae, which mainly distributes in the southern mountain area of the drainage area of the Changjiang River and the Changbai Mountain in the northeast area. Men named it as duanchangcao, shanpishuang and nansheteng too. Because of the toxicity in the body, it is used to kill bug and mouse in folk. From the 1960's, Tripterygium Preparations has been used on the treatmen of inflammation , cancer , chronic nephritis , leprosy , Systemic Lupus Erythematosus , some dermatosisand so on.In this paper, the identification's study by RAPD on Liriope muscari population in Fujian province and 28 populations of Tripterygium Wilfordii in the south china is concluded as follows:(1) The genomic DNA samples of high purity was prepared by the method of CTAB from Liriope muscari and by the method of improved CTAB from Tripterygium Wilfordii.And then we have study the rapid extraction method of them.(2) The reliable RAPD analysis system of the both was established. 15 primers (Liriope muscari),14 primers (Tripterygium Wilfordii) which were produced distinct bands, showing unmistakable polymorphism were selected from screening of 96 RAPD primer .(3) Analysis of the experimental data with the aid of POPGEN32 and SPSS13.0 revealed that the 14 plants can be classified into three genus at the genetic distance of 0.60 as was done in the classical classification (Ophiopogon japonicus , Liriope spicata Lout (Thunb).var. prolifera.Y.T.Ma , Liriope muscari (Decne) Bailey ).The Liriope muscari from quanzhou provenance can be clustered together, while the genetic distance between planted provenances and wild provenances from xianyou city was bigger . Base on the ending of the two cluster softwares (POPGEN32,SPSS13.0),we can find that the provenance of Liriope muscari was conducted by using RAPD .(4) The distance between planted provenances and wild provenances from XianYou city is the biggest in the all provenances of Liriope muscari. So, we can consider the distance of 0.4532 as the dividing crest between Liriope muscari and the others.(5) The polymorphism of 15 primers analysis for Liriope muscari showed a total of 118 bands , among which 113 bands were polymorphic , classifing the 14 plants into three genus .The 118 bands was marked as"1"if there is a band ,"0"if there is no band to make a finger print of the 14 Liriope muscari provenances .(6) Analysis of the experimental data of Tripterygium Wilfordii with the aid of POPGEN32 and SPSS13.0 revealed that the YunNan, GuiZhou and GuangXi provenances can be clustered together. The HuNan provenances were clustered together, while JiangXi provenances were mixed with FuJian and HuNan provences. The ZheJiang provenance was mixed with FuJian provenances too.(7) The genetic distance of 28 populations of Tripterygium Wilfordii range from 0.0466 to 0.5028. The the range of variation is large, most of them range from 0.3000 to 0.4500 , less than 5% of them range from 0.1000 to 0.2000, only the distance between QuJing provenance in YunNan province and XiangTan provenance in HuNan province over 0.5.(8) The distance of the accurately geo-herbal research identification relates with genetic diversity of each population, most of them range from 0.2000 to 0.3000, referring to the distance of all populations.(9) The polymorphism of 14 primers analysis for Tripterygium Wilfordii showed a total of 164 bands, among which 151 bands were polymorphic, identifying the 85 provenance.The 164 bands was marked as"1"if there is a band ,"0"if there is no band to make a finger print of the 85 provenances .(10) Random amplified polymorphic DNA (RAPD) marker can be used to identify the provenance of Liriope muscari and Tripterygium Wilfordii.