Study on DNA molecular identification of mix samples of five species of Baitouweng medicinal materials based on high-throughput sequencing technology
10.16438/j.0513-4870.2018-0696
- VernacularTitle:基于高通量测序技术对五种白头翁药材混合样品的DNA分子鉴定研究
- Author:
Ting-ting ZHANG
1
;
Yong-man LIANG
1
;
Liang XU
1
;
Yan-yun YANG
1
;
Yan-ping XING
1
;
Tao LIU
2
;
Ting-guo KANG
1
Author Information
1. Liaoning University of Traditional Chinese Medicine, Dalian 116000, China
2. China Medical University, Shenyang 110122, China
- Publication Type:ORIGINAL ARTICLES
- Keywords:
Illumina MiSeq;
Pulsatilla chinensis;
mix power;
ITS2
- From:
Acta Pharmaceutica Sinica
2018;53(11):1918-1923
- CountryChina
- Language:Chinese
-
Abstract:
Traditional Chinese medicine Baitouweng have a long history of application. The pharmacopoeia included dry roots of Pulsatilla chinensis (Bge.) Regel of Ranunculaceae. There are easily confused species in the market circulation, such as P. cernua (Thunb.) Bercht. et Opiz., P. dahurica (Fisch.) Spreng., P. turczaninovii Kryl. et Serg., and P. chinensis (Bge.) Regel var. kissii (Mandl) S. H. Li et Y. H. Huang, etc. In this study, using the method of metagenomics, based on high-throughput sequencing technology, the ITS2 sequence of mixed samples of five species of Baitouweng medicinal materials was sequenced. First, the total DNA extraction of medicinal materials mixing powder, and the ITS2 fragment of total DNA was amplified by PCR. Second, the Illumina MiSeq platform was used to carry out Paired-end sequencing for DNA fragments. Last, using FLASH, QⅡME and GraPhlAn software to arrange and analyze, and clustering analysis with the sequences of uploaded to GenBank by our group in the early stage. The results showed that a total of 53 024 sequences of ITS2 were obtained from the mixed samples, there are 52 295 effective sequences, there are a total of 49 079 of five species of medicinal materials of P. Miller. After the representative sequences and the sequence of uploaded to GenBank by our group in the early stage were clustering analysis, 5 species of Baitouweng medicinal materials were clustered into one branch separately, presenting monophyletic. The results showed that using the high-throughput sequencing technology, using ITS2 sequence as DNA barcode, the mix powder of 5 species of Baitouweng medicinal materials could be effectively identified. It provides a new method and thought for the origin identification of mixed Chinese medicinal materials.
