ABSTRACT
There are many multi-original medicinal materials in Chinese Pharmacopoeia, and the mixed use of medicinal materials from different sources is common, which has certain influence on the stability of clinical medication. In this study, pyrosequencing technology was used to screen species-specific single nucleotide polymorphisms (SNP) from commonly used DNA barcode sequences, and a rapid and accurate molecular identification method for original species in mixed medicinal powder of Epimedii Folium was established. Multiple sequence alignment analysis showed that the 176th (C/T) mutation and the 196th (A/G) mutation of ITS, the 123rd (C/G) mutation of matK and the 892nd (A/C) mutation of rbcL could be used as the unique SNPs of E. sagittatum, E. koreanum, E. brevicornu and E. pubescens, respectively. In this study, the applicability of pyrosequencing and Sanger sequencing methods in the sequencing of mixture samples was investigated from the perspective of sensitivity and stability. Pyrosequencing method has higher detection sensitivity than Sanger sequencing method for low content samples in the mixed samples. Stability analysis showed that pyrosequencing technology could still obtain effective sequencing results for the amplified products of template DNA after 45 min of 95 ℃ high temperature water bath, while the critical point of Sanger sequencing method was 30 min. In this study, a new identification technology of Epimedii Folium mixed powder primordial species based on pyrosequencing and specific SNP was developed, which can quickly and accurately identify the mixed use of Epimedii Folium with high sensitivity and stability, and can also support the identification of different primordial species and mixed powder primordial herbs, which is conducive to ensuring the consistency and stability of clinical medication.
ABSTRACT
Sichuan province is very famous for its abundant resources of traditional Chinese medicine(TCM).However, within the scope of administrative division of Sichuan province, the origin records of Dao-di herbs in different historical periods show a dynamic distribution process. On the basis of carefully sorting out the geographical scope of Sichuan province in different historical periods, this article focuses on the textual research of the Dao-di herbs in Sichuan province recorded in the seven mainstream ancient works of materia medica.The results showed that, according to the records of Mingyi bielu and Bencaojing Jizhu, the main distribution areas of Dao-di herbs were mainly in the central and eastern regions of Sichuan province, mainly including Moschus, Coptidis Rhizoma, Zingiberis Rhizoma, Aconiti Lateralis Radix Praeparata and most of the rest materia medica had become unused in the historical process. Qianjin Yifang records that the distribution areas of Dao-di herbs were mainly in the middle and eastern part of Sichuan province.Aconiti Radix, Lateralis Radix Praeparata, Zingiberis Rhizoma, Notopterygii Rhizoma et Radix are still the Dao-di herbs of Sichuan province. According to the book of Bencao Tujing,the main distribution areas of Dao-di herbs are Chengdu Plain, Yibin and Santai, While Toosendan Fructus, Chuanxiong Rhizoma, Zanthoxyli Pericarpium, Aconiti Radix are still the Dao-di herbs of Sichuan province. Ben Cao Gang Mu records the place of origin as Sichuan.Coptidis Rhizoma, Toosendan Fructus, Cyathulae Radix are still the Dao-di herbs of Sichuan pro-vince. Yaowu Chuchanbian and Zengding Weiyao Tiaobian records the place of origin as Sichuan, as well as Kangding, Songpan, Dujiang-yan, Jiangyou, Nanchong, Ya'an, etc. Moschus, Coptidis Rhizoma, Eucommiae Cortex, Phellodendri Chinensis Cortex are still the Dao-di herbs of Sichuan province. The results of this article provide a new understanding of the history and distribution changes of Dao-di herbs in Sichuan province, and can help to further understand the formation connotation of Sichuan Dao-di herbs.