Based On The Pcr - Sscp And Miniature Clematis Root Molecular Identification Of Bar Code

The Clematis, whose source is from roots parts of the plants, Clematis chinensis Osbeck.、Clematis hexapetala Pall. and Clematis manshurica Rupr., family Ranunculaceae Clematis, is a kind of traditional Chinese medicine. According to research reports and market varieties survey, the Clematis has a complicated varieties in our medicine market. Adulterants’ origin and medicine property are different from the authentic medicine. These phenomena will undoubtedly disrupt the normal medicine market and clinical applications. Therefore, it is very critical to identify and sort out the variety of Clematis accurately, in order to control its quality.The identification of herbs is a important issue in Traditional Chinese Medicine research.The methods of traditional identification include character identification, microscopic identification and physical and chemical identification. With the continuous development of molecular techniques, DNA barcoding identification methods plays an important role in the identification of medicine. The steps of this method is simple, and it can complete the work stably, highly efficiently, and accurately The mini-DNA barcoding which is a shorter fragment, based on the stable nucleotide differences between spieces, is suitable for identification of DNA serious degradation of the experimental material. In this study,we also use PCR-SSCP-PAGE method to identify Clematis herbs and their origin plantIn This study, we selected three origin plants and commercially available herbs. Based on mini-DNA barcoding technology and PCR-SSCP-PAGE method, we sort out Clematis varieties medicinal herbs and provide quality control for Clematis.The results are as follows:1According to the Clematis three specific sites of origin, we establish a mini-barcoding identification method by PCR amplification technology.The results shows that the primer designed to target fragment can amplify short fragment about200bp length, from DNA poor quality samples, which sequencing success rate is 100%.Identification of herbs can be completed accurately and quickly.2On the basis of the traditional identification, using PCR-SSCP-PAGE, we establish the system of identification of Clematis herbs. In this study, the group of three original plants can be authenticaty by PCR-SSCP-PAGE.But for Clematis herbs, its identification is ineffective. PCR-SSCP technique has the advantages of high-throughput, sensitive, and economic, but the technique is applied in the field of medicine identification less, it still has great prospects for development.3, After character identification and microscopic identification of Clematis herbs, we learn Clematis herbs mainly derived from Clematis chinensis Osbeck. and Clematis manshurica Rupr..Combined with the content determination of herbs, we find Clematis chinensis Osbeck., Clematis manshurica Rupr. and Clematis hexapetala Pall, oleanolic acid and Hederagenin content are quite different. Therefore, it is recommended that Clematis chinensis Osbeck. as the main cultivars in medicinal herbs cultivation.4We suggest that Clematis chinensis Osbeck., Clematis manshurica Rupr. and Clematis hexapetala Pall, oleanolic acid and Hederagenin content, separate regulation, in order to control and supervision of the quality of the herbs.